Effect of Azotobacter vinelandii and compatible solutes on germination wheat seeds and root concentrations of sodium and potassium under salt stress.

PubMed

Silini, A; Silini-Chérif, H; Ghoul, M

2012-02-01

The effect of plant growth-promoting Rhizobacteria (PGPR) and exogenous application of compatible solutes on seed germination and root concentrations of sodium and potassium of two wheat varieties (Triticum durum L.) were evaluated under saline stress. In this experiment, Azotobacter vinelandii strain DSM85, glycine betaine and proline were used. Inoculated seeds for each variety were placed on Whatman paper in 9 cm Petri dishes containing 15 mL of distilled water or NaCl solutions at various concentrations (control, 100, 200, 300 mM) supplemented with or without glycine betaine (GB) or proline at 5 mM. The results indicated that addition of proline (5 mM) stimulated the production of indol acetic acid and the growth of A. vinelandii at 200 and 300 mM NaCl, respectively. The germination rate index and the germination final percentage decreased significantly (p < 0.05) with increasing salinity level. The germination was significantly diminished at 300 mM with significant variation among varieties and Waha variety had higher germination percentage than Bousselam variety. Inoculation of seeds by A. vinelandii and exogenous application of proline had significantly positive effect on the germination at this concentration of NaCl. The rate of accumulation of Na+ in roots was important at 100 mM and increased at 200 mM. The concentration of K+ decreased when salinity increased. The effect of inoculation or inoculation with proline decreased the accumulation of Na' and reduced the loss of K+ under salt stress. From the present study we can conclude that the use of A. vinelandii strain DSM85 and external application of low concentrations of proline on seeds might be considered as a strategy for the protection of plants under saline stress.

Alleviation of heavy metals toxicity by the application of plant growth promoting rhizobacteria and effects on wheat grown in saline sodic field.

PubMed

Hassan, Tamoor Ul; Bano, Asghari; Naz, Irum

2017-06-03

The aim of the study was to determine tolerance of plant growth promoting rhizobacteria (PGPR) in different concentrations of Cu, Cr, Co, Cd, Ni, Mn, and Pb and to evaluate the PGPR-modulated bioavailability of different heavy metals in the rhizosphere soil and wheat tissues, grown in saline sodic soil. Bacillus cereus and Pseudomonas moraviensis were isolated from Cenchrus ciliaris L. growing in the Khewra salt range. Seven-day-old cultures of PGPR were applied on wheat as single inoculum, co-inoculation and carrier-based biofertilizer (using maize straw and sugarcane husk as carrier). At 100 ppm of Cr and Cu, the survival rates of rhizobacteria were decreased by 40%. Single inoculation of PGPR decreased 50% of Co, Ni, Cr and Mn concentrations in the rhizosphere soil. Co-inoculation of PGPR and biofertilizer treatment further augmented the decreases by 15% in Co, Ni, Cr and Mn over single inoculation except Pb and Co where decreases were 40% and 77%, respectively. The maximum decrease in biological concentration factor (BCF) was observed for Cd, Co, Cr, and Mn. P. moraviensis inoculation decreases the biological accumulation coefficient (BAC) as well as translocation factor (TF) for Cd, Cr, Cu Mn, and Ni. The PGPR inoculation minimized the deleterious effects of heavy metals, and the addition of carriers further assisted the PGPR.

Power Doppler evaluation of joint effusions: investigation in a rabbit model.

PubMed

Strouse, P J; DiPietro, M A; Teo, E L; Doi, K; Chrisp, C E

1999-08-01

To study the power Doppler findings of septic arthritis and noninfectious synovitis in an animal model. The right knees of 10 rabbits were inoculated with an aqueous suspension of Staphylococcus aureus. The right knees of 5 rabbits were injected with talc suspension. The right knees of 5 rabbits were injected with saline. All 20 left knees were injected with saline. Serial power Doppler images were obtained using constant-imaging parameters. Images were reviewed by blinded observers who assessed for increased power Doppler signal. All 10 knees inoculated with S. aureus developed septic arthritis. Each infected rabbit knee demonstrated increased signal on power Doppler on at least one examination, ranging from 1-6 days after inoculation. Only 23 of 45 examinations of infected knees were unequivocally positive by power Doppler on examinations performed 1 to 6 days after inoculation. No knee with talc synovitis demonstrated increased power Doppler signal. No control knee demonstrated increased power Doppler signal. Increased power Doppler signal may be seen with septic arthritis; however, its intensity and timing may vary from subject to subject. A normal power Doppler examination does not exclude septic arthritis.

Can Tomato Inoculation with Trichoderma Compensate Yield and Soil Health Deficiency due to Soil Salinity?

NASA Astrophysics Data System (ADS)

Wagner, Karl; Apostolakis, Antonios; Daliakopoulos, Ioannis; Tsanis, Ioannis

2016-04-01

Soil salinity is a major soil degradation threat, especially for arid coastal environments where it hinders agricultural production and soil health. Protected horticultural crops in the Mediterranean region, typically under deficit irrigation and intensive cultivation practices, have to cope with increasing irrigation water and soil salinization. This study quantifies the beneficial effects of the Trichoderma harzianum (TH) on the sustainable production of Solanum lycopersicum (tomato), a major greenhouse crop of the RECARE project Case Study in Greece, the semi-arid coastal Timpaki basin in south-central Crete. 20 vigorous 20-day-old Solanum lycopersicum L. cv Elpida seedlings are treated with TH fungi (T) or without (N) and transplanted into 35 L pots under greenhouse conditions. Use of local planting soil with initial Electrical Conductivity (ECe) 1.8 dS m-1 and local cultivation practices aim to simulate the prevailing conditions at the Case Study. In order to simulate seawater intrusion affected irrigation, plants are drip irrigated with two NaCl treatments: slightly (S) saline (ECw = 1.1 dS m-1) and moderately (M) saline water (ECw = 3.5 dS m-1), resulting to very high and excessively high ECe, respectively. Preliminary analysis of below and aboveground biomass, soil quality, salinity, and biodiversity indicators, suggest that TH pre-inoculation of tomato plants at both S and M treatments improve yield, soil biodiversity and overall soil health.

Hypothalamic-Pituitary-Adrenal Axis Physiology and Cognitive Control of Behavior in Stress Inoculated Monkeys

ERIC Educational Resources Information Center

Parker, Karen J.; Buckmaster, Christine L.; Lindley, Steven E.; Schatzberg, Alan F.; Lyons, David M.

2012-01-01

Monkeys exposed to stress inoculation protocols early in life subsequently exhibit diminished neurobiological responses to moderate psychological stressors and enhanced cognitive control of behavior during juvenile development compared to non-inoculated monkeys. The present experiments extended these findings and revealed that stress inoculated…

Effects of plant growth promoting bacteria and mycorrhizal on Capsicum annuum L. var. aviculare ([Dierbach] D'Arcy and Eshbaugh) germination under stressing abiotic conditions.

PubMed

Rueda-Puente, Edgar Omar; Murillo-Amador, Bernardo; Castellanos-Cervantes, T; García-Hernández, José Luís; Tarazòn-Herrera, Mario Antonio; Moreno Medina, Salomòn; Gerlach Barrera, Luis Ernesto

2010-08-01

Capsicum annuum var. aviculare to Tarahumara and Papago Indians and farmers of Sonora desert is a promising biological and commercial value as a natural resource from arid and semiarid coastal zones. Traditionally, apply synthetic fertilizers to compensate for soil nitrogen deficiency. However, indiscriminate use of these fertilizers might increase salinity. The inoculation by plant growth promoting bacteria (PGPB) and arbuscular mycorrhizal fungi (AMF) represents an alternative as potential bio fertilizer resources for salty areas. Seeds ecotypes from four areas of Sonora desert (Mazocahui, Baviacora, Arizpe, La Tortuga), in order to inoculate them with one species of PGPB and AMF. Two germination tests were carried out to study the effect of salinity, temperature regime (night/day) and inoculation with PGPB and AMF growth factors measured on germination (percentage and rate), plant height, root length, and produced biomass (fresh and dry matter). The results indicated that from four studied ecotypes, Mazocahui was the most outstanding of all, showing the highest germination under saline and non-saline conditions. However, the PGPB and AMF influenced the others variables evaluated. This study is the first step to obtain an ideal ecotype of C. a. var. aviculare, which grows in the northwest of México and promoting this type of microorganisms as an efficient and reliable biological product. Studies of the association of PGPB and AMF with the C. a. var. aviculare-Mazocahui ecotype are recommended to determine the extent to which these observations can be reproduced under field conditions. Copyright 2010 Elsevier Masson SAS. All rights reserved.

Influence of irrigation protocols on the bond strength of fiber posts cemented with a self-adhesive luting agent 24 hours after endodontic treatment.

PubMed

Lima, Jessica Ferraz Carvalho; Lima, Adriano Fonseca; Humel, Maria Malerba Colombi; Paulillo, Luis Alexandre Maffei Sartini; Marchi, Giselle Maria; Ferraz, Caio Cezar Randi

2015-01-01

The aim of this in vitro study was to evaluate the influence of different irrigation protocols on the bond strength, at different root depths, of fiber posts cemented with a self-adhesive cement 24 hours after endodontic treatment. Fifty-six bovine incisor roots were endodontically prepared and separated into 7 groups (n = 8) according to irrigation protocols: group 1, sterile saline (control); group 2, chlorhexidine (CHX) gel 2% and saline; group 3, sodium hypochlorite (NaOCl) 5.25% and saline; group 4, CHX and saline (final irrigation with ethylenediaminetetraacetic acid [EDTA] 17%); group 5, NaOCl and saline (final irrigation with EDTA); group 6, CHX and saline (final irrigation with NaOCl and EDTA); and group 7, NaOCl (final irrigation with CHX and EDTA). No statistically significant difference was found among the groups. Within the limitations of this study, it can be concluded that the different irrigation protocols did not influence the bond strength of self-adhesive resin cement, which presented similar behaviors at the 3 root depths studied.

The ACC deaminase expressing endophyte Pseudomonas spp. Enhances NaCl stress tolerance by reducing stress-related ethylene production, resulting in improved growth, photosynthetic performance, and ionic balance in tomato plants.

PubMed

Win, Khin Thuzar; Fukuyo, Tanaka; Keiki, Okazaki; Ohwaki, Yoshinari

2018-06-01

Plant growth promoting bacteria (PGPB) endophytes that express 1-aminocyclopropane-1-carboxylate (ACC) deaminase reportedly confer plant tolerance to abiotic stresses such as salinity by lowering stress-related ethylene levels. Two preselected ACC deaminase expressing endophytic Pseudomonas spp. strains, OFT2 and OFT5, were compared in terms of their potential to promote plant growth, leaf water contents, photosynthetic performance, and ionic balance of tomato plants under conditions of moderate NaCl stress (75 mM). Salinity stress strongly affected growth, leaf water contents, and photosynthetic performance of tomato seedlings, and inoculation with either OFT2 or OFT5 ameliorated these adverse effects. Decreases in plant biomass due to salinity stress were significant in both uninoculated control plants and in plants inoculated with OFT2 compared with plants without NaCl stress. However, no reductions in total biomass were observed in plants that were inoculated with the OFT5 strain. Strain OFT5 influenced growth, physiological status, and ionic balance of tomato plants more efficiently than strain OFT2 under NaCl stress. In particular, inoculated OFT5 reduced salt-induced ethylene production by tomato seedlings, and although it did not reduce shoot uptake of Na, it promoted shoot uptake of other macronutrients (P, K, and Mg) and micronutrients (Mn, Fe, Cu, and Zn). These nutrients may activate processes that alleviate the effects of salt, suggesting that OFT5 can be used to improve nutrient uptake and plant growth under moderate salt-affected conditions by reducing stress-related ethylene levels. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

A phytobeneficial strain Planomicrobium sp. MSSA-10 triggered oxidative stress responsive mechanisms and regulated the growth of pea plants under induced saline environment.

PubMed

Shahid, M; Akram, M S; Khan, M A; Zubair, M; Shah, S M; Ismail, M; Shabir, G; Basheer, S; Aslam, K; Tariq, M

2018-06-01

The study was planned to characterize Planomicrobium sp. MSSA-10 for plant-beneficial traits and to evaluate its inoculation impact on physiology of pea plants under different salinity levels. Strain MSSA-10 was isolated from pea rhizosphere and identified by the analysis of 16S rRNA gene sequence. The strain demonstrated phosphate solubilization and auxin production up to 2 mol l -1 NaCl and exhibited 1-aminocyclopropane-1-carboxylic acid deaminase activity up to 1·5 mol l -1 salt. In an inoculation experiment under different salinity regimes, a significant increase in growth was observed associated with decreased levels of reactive oxygen species and enhanced antioxidative enzyme activities. The strain also promoted the translocation of nutrients in plants with subsequent increase in chlorophyll and protein contents as compared to noninoculated plants. It has been observed that rifampicin-resistant derivatives of MSSA-10 were able to survive for 30 days at optimum cell density with pea rhizosphere. Growth-stimulating effect of MSSA-10 on pea plants may be attributed to its rhizosphere competence, nutrient mobilization and modulation of plant oxidative damage repair mechanisms under saline environment. Planomicrobium sp. MSSA-10 might be used as potent bioinoculant to relieve pea plants from deleterious effects of salinity. © 2018 The Society for Applied Microbiology.

Comparative study of protective activities of Neospora caninum bradyzoite antigens, NcBAG1, NcBSR4, NcMAG1, and NcSAG4, in a mouse model of acute parasitic infection.

PubMed

Uchida, Masaki; Nagashima, Kotomi; Akatsuka, Yui; Murakami, Takashi; Ito, Akira; Imai, Soichi; Ike, Kazunori

2013-02-01

Neospora caninum is an obligate intracellular protozoan parasite that causes severe neuromuscular diseases, repeated abortion, stillbirth, and congenital infection in livestock and companion animals. The development of an effective vaccine against neosporosis in cattle is an important issue due to the significant worldwide economic impact of this disease. We evaluated the immunogenicity of four bradyzoite antigens, NcBAG1 (first described in this study), NcBSR4, NcMAG1, and NcSAG4, using an acute infection mouse model to determine synergistic effects with the tachyzoite antigen as a candidate for vaccine production. Mice were inoculated with the recombinant vaccines (r-)NcBAG1, rNcBSR4, rNcMAG1, rNcSAG4, or phosphate-buffered saline (PBS) (adjuvant control group) in an oil-in-water emulsion with bitter gourd extract, a Th1 immune stimulator, or PBS alone as the infection control group. Mice inoculated with each vaccine developed antigen-specific IgG1 and IgG2a antibodies and isolated splenocytes from mice produced high levels of interferon-γ when infected with the N. caninum tachyzoite. The mice inoculated with rNcBAG1, rNcMAG1, or rNcSAG4 developed slight to moderate clinical symptoms but did not succumb to infection. In contrast, rNcBSR4 and both control groups developed severe disease and some mice required euthanasia. The parasitic burden in the brain tissues of vaccinated mice was assessed by N. caninum-specific real-time PCR at 5 weeks after infection. The parasite load in rNcBAG1-, rNcMAG1-, and rNcSAG4-inoculated mice was significantly lower than that in adjuvant and infection control mice. Therefore, these antigens may be useful for the production of a N. caninum-specific vaccination protocol.

Relationship between clinical signs and postmortem test status in cattle experimentally infected with the bovine spongiform encephalopathy agent

PubMed Central

2010-01-01

Background Various clinical protocols have been developed to aid in the clinical diagnosis of classical bovine spongiform encephalopathy (BSE), which is confirmed by postmortem examinations based on vacuolation and accumulation of disease-associated prion protein (PrPd) in the brain. The present study investigated the occurrence and progression of sixty selected clinical signs and behaviour combinations in 513 experimentally exposed cattle subsequently categorised postmortem as confirmed or unconfirmed BSE cases. Appropriate undosed or saline inoculated controls were examined similarly and the data analysed to explore the possible occurrence of BSE-specific clinical expression in animals unconfirmed by postmortem examinations. Results Based on the display of selected behavioural, sensory and locomotor changes, 20 (67%) orally dosed and 17 (77%) intracerebrally inoculated pathologically confirmed BSE cases and 21 (13%) orally dosed and 18 (6%) intracerebrally inoculated but unconfirmed cases were considered clinical BSE suspects. None of 103 controls showed significant signs and were all negative on diagnostic postmortem examinations. Signs indicative of BSE suspects, particularly over-reactivity and ataxia, were more frequently displayed in confirmed cases with vacuolar changes in the brain. The display of several BSE-associated signs over time, including repeated startle responses and nervousness, was significantly more frequent in confirmed BSE cases compared to controls, but these two signs were also significantly more frequent in orally dosed cattle unconfirmed by postmortem examinations. Conclusions The findings confirm that in experimentally infected cattle clinical abnormalities indicative of BSE are accompanied by vacuolar changes and PrPd accumulation in the brainstem. The presence of more frequently expressed signs in cases with vacuolar changes is consistent with this pathology representing a more advanced stage of disease. That BSE-like signs or sign combinations occur in inoculated animals that were not confirmed as BSE cases by postmortem examinations requires further study to investigate the potential causal relationship with prion disease. PMID:21143919

Correlation of cytotoxicity with elimination of iodine-125 from nude mice inoculated with prelabeled human melanoma cells

DOE Office of Scientific and Technical Information (OSTI.GOV)

Lockshin, A.; Giovanella, B.C.; Quian, C.

1984-08-01

BRO human melanoma cells were prelabeled in vitro with (125I)5-iodo-2'-deoxyuridine ((125I)IdUrd) and inoculated into NIH-II nude mice ip, im, sc, or iv. Saline or diphtheria toxin (DT), which is selectively toxic to human cells compared to those of mice, was injected, and the loss of 125I from the animals was monitored daily with a whole-body gamma scintillation detector. For most of the inoculation sites DT accelerated the rate of 125I excretion and in all cases was cytotoxic for the inoculated cells as determined by host survival or measurement of visible tumor growth. Differences between the rates of 125I loss formore » DT-treated mice compared to untreated mice were most evident for cells inoculated ip or im. These results indicate that (125I)IdUrd prelabeling of human tumor cells inoculated in nude mice offers a rapid method for determination of cytotoxicity in vivo.« less

Alleviation of Salt Stress in Pepper (Capsicum annum L.) Plants by Plant Growth-Promoting Rhizobacteria.

PubMed

Hahm, Mi-Seon; Son, Jin-Soo; Hwang, Ye-Ji; Kwon, Duk-Kee; Ghim, Sa-Youl

2017-10-28

In the present study, we demonstrate that the growth of salt-stressed pepper plants is improved by inoculation with plant growth-promoting rhizobacteria (PGPR). Three PGPR strains ( Microbacterium oleivorans KNUC7074, Brevibacterium iodinum KNUC7183, and Rhizobium massiliae KNUC7586) were isolated from the rhizosphere of pepper plants growing in saline soil, and pepper plants inoculated with these PGPR strains exhibited significantly greater plant height, fresh weight, dry weight, and total chlorophyll content than non-inoculated plants. In addition, salt-stressed pepper plants that were inoculated with B. iodinum KNUC7183 and R. massiliae KNUC7586 possessed significantly different total soluble sugar and proline contents from non-inoculated controls, and the activity of several antioxidant enzymes (ascorbate peroxidase, guaiacol peroxidase, and catalase) was also elevated in PGPR-treated plants under salt stress. Overall, these results suggest that the inoculation of pepper plants with M. oleivorans KNUC7074, B. iodinum KNUC7183, and R. massiliae KNUC7586 can alleviate the harmful effects of salt stress on plant growth.

High-quality forage production under salinity by using a salt-tolerant AtNXH1-expressing transgenic alfalfa combined with a natural stress-resistant nitrogen-fixing bacterium.

PubMed

Stritzler, Margarita; Elba, Pagano; Berini, Carolina; Gomez, Cristina; Ayub, Nicolás; Soto, Gabriela

2018-06-20

Alfalfa, usually known as the "Queen of Forages", is the main source of vegetable protein to meat and milk production systems worldwide. This legume is extremely rich in proteins due to its highly efficient symbiotic association with nitrogen-fixing strains. In the last years, alfalfa culture has been displaced to saline environments by other important crops, including major cereals, a fact that has reduced its biomass production and symbiotic nitrogen fixation. In this short communication, we report the high forage production and nutrient quality of alfalfa under saline conditions by alfalfa transformation with the AtNHX1 Na + /H + antiporter and inoculation with the stress-resistant nitrogen-fixing strain Sinorhizobium meliloti B401. Therefore, the incorporation of transgenic traits into salt-sensitive legumes in association with the inoculation with natural stress-resistant isolates could be a robust approach to improve the productivity and quality of these important nitrogen-fixing crops. Copyright © 2018. Published by Elsevier B.V.

Effect of salt-tolerant plant growth-promoting rhizobacteria on wheat plants and soil health in a saline environment.

PubMed

Upadhyay, S K; Singh, D P

2015-01-01

Salt-tolerant plant growth-promoting rhizobacteria (ST-PGPR) significantly influence the growth and yield of wheat crops in saline soil. Wheat growth improved in pots with inoculation of all nine ST-PGPR (ECe = 4.3 dS·m(-1) ; greenhouse experiment), while maximum growth and dry biomass was observed in isolate SU18 Arthrobacter sp.; simultaneously, all ST-PGPR improved soil health in treated pot soil over controls. In the field experiment, maximum wheat root dry weight and shoot biomass was observed after inoculation with SU44 B. aquimaris, and SU8 B. aquimaris, respectively, after 60 and 90 days. Isolate SU8 B. aquimaris, induced significantly higher proline and total soluble sugar accumulation in wheat, while isolate SU44 B. aquimaris, resulted in higher accumulation of reducing sugars after 60 days. Percentage nitrogen (N), potassium (K) and phosphorus (P) in leaves of wheat increased significantly after inoculation with ST-PGPR, as compared to un-inoculated plants. Isolate SU47 B. subtilis showed maximum reduction of sodium (Na) content in wheat leaves of about 23% at both 60 and 90 days after sowing, and produced the best yield of around 17.8% more than the control. © 2014 German Botanical Society and The Royal Botanical Society of the Netherlands.

A simple method to evaluate the number of bradyrhizobia on soybean seeds and its implication on inoculant quality control

PubMed Central

2011-01-01

Soybean seeds are non-sterile and their bacterial population interferes with the enumeration of beneficial bacteria, making it difficult to assess survival under different conditions. Within this context, the principal aims of this work were: (1) to improve a selective media for the enumeration of B. japonicum recovered from inoculated soybean seeds; (2) to establish the most representative mathematical function for B. japonicum mortality on soybean seeds after inoculation; (3) to evaluate if environmental or physiological conditions modify B. japonicum mortality on soybean seeds; and (4) to create a new protocol for quality control of soybean inoculants. We successfully evaluated the combination of pentachloronitrobenzene and vancomycin added to the yeast-mannitol medium to inhibit most fungi and Gram-positive soybean microbiota, thus producing reliable counts of B. japonicum from inoculated soybean seeds. Percentages of recovery and survival factors were obtained and used to construct a two-phase exponential decay non-linear regression function. High temperature and desiccation decreased these parameters, while the optimization of temperature and the use of osmoprotective compounds with inoculants increased them. The use of this protocol minimized heterogeneity between experiments and may be considered more reliable than the simple expression of direct colony count of bacteria recovered from seeds. PMID:21906377

The effect of four-phasic versus three-phasic contrast media injection protocols on extravasation rate in coronary CT angiography: a randomized controlled trial.

PubMed

Karády, Júlia; Panajotu, Alexisz; Kolossváry, Márton; Szilveszter, Bálint; Jermendy, Ádám L; Bartykowszki, Andrea; Károlyi, Mihály; Celeng, Csilla; Merkely, Béla; Maurovich-Horvat, Pál

2017-11-01

Contrast media (CM) extravasation is a well-known complication of CT angiography (CTA). Our prospective randomized control study aimed to assess whether a four-phasic CM administration protocol reduces the risk of extravasation compared to the routinely used three-phasic protocol in coronary CTA. Patients referred to coronary CTA due to suspected coronary artery disease were included in the study. All patients received 400 mg/ml iomeprol CM injected with dual-syringe automated injector. Patients were randomized into a three-phasic injection-protocol group, with a CM bolus of 85 ml followed by 40 ml of 75%:25% saline/CM mixture and 30 ml saline chaser bolus; and a four-phasic injection-protocol group, with a saline pacer bolus of 10 ml injected at a lower flow rate before the three-phasic protocol. 2,445 consecutive patients were enrolled (mean age 60.6 ± 12.1 years; females 43.6%). Overall rate of extravasation was 0.9% (23/2,445): 1.4% (17/1,229) in the three-phasic group and 0.5% (6/1,216) in the four-phasic group (p = 0.034). Four-phasic CM administration protocol is easy to implement in the clinical routine at no extra cost. The extravasation rate is reduced by 65% with the application of the four-phasic protocol compared to the three-phasic protocol in coronary CTA. • Four-phasic CM injection-protocol reduces extravasation rate by 65% compared to three-phasic. • The saline pacer bolus substantially reduces the risk of CM extravasation. • The implementation of four-phasic injection-protocol is at no cost.

Comparison of irrigation solutions and devices in a contaminated musculoskeletal wound survival model.

PubMed

Owens, Brett D; White, Daniel W; Wenke, Joseph C

2009-01-01

There is much to learn about the effectiveness of different methods currently used for the irrigation of open wounds. The purpose of this study was to compare various approaches in a survival animal model. We used an established goat model involving the creation of a reproducible complex musculoskeletal wound followed by inoculation with Pseudomonas aeruginosa (lux) bacteria. This genetically altered luminescent bacterium provides the ability for quantitative analysis with a photon-counting camera system. For Study 1, wound irrigation was performed six hours after the injury and inoculation; the goats were assigned to four treatment groups: normal saline solution, bacitracin solution, castile soap, and benzalkonium chloride. All wounds received sharp débridement and irrigation with use of a pulsatile lavage device (19 psi). Images and photon counts were obtained prior to irrigation, after irrigation, and forty-eight hours after injury and inoculation. For Study 2, we used the same animal model and compared bulb syringe and pulsatile lavage irrigation with saline solution. In Study 1, the irrigation treatment lowered the bacterial counts in all treatment groups. The greatest reduction was seen with castile soap, which lowered the photon count to 13% of the pretreatment level. This was followed by benzalkonium chloride, bacitracin, and saline solution at 18%, 22%, and 29%, respectively. At forty-eight hours, imaging showed a rebound in bacterial counts in every group. The highest rebound was measured in the castile soap group, which rebounded to 120% of the pretreatment level. The benzalkonium chloride group experienced a rebound to 94% of the pretreatment level. These were followed by bacitracin solution (89%) and normal saline solution (68%). In Study 2, both treatment methods were effective in removing 75% of the bacteria initially. At forty-eight hours, the bacterial levels in the pulsed lavage group rebounded to 94% of the original levels (compared with 48% in the bulb syringe group). The difference in the mean photon count ratios at forty-eight hours was significant (p = 0.048). Approaches used to remove bacteria from wounds, such as irrigants other than saline solution or high-pressure devices, may not have the best clinical outcome.

Validation of Single and Pooled Manure Drag Swabs for the Detection of Salmonella Serovar Enteritidis in Commercial Poultry Houses.

PubMed

Kinde, Hailu; Goodluck, Helen A; Pitesky, Maurice; Friend, Tom D; Campbell, James A; Hill, Ashley E

2015-12-01

Single swabs (cultured individually) are currently used in the Food and Drug Administration (FDA) official method for sampling the environment of commercial laying hens for the detection of Salmonella enterica ssp. serovar Enteritidis (Salmonella Enteritidis). The FDA has also granted provisional acceptance of the National Poultry Improvement Plan's (NPIP) Salmonella isolation and identification methodology for samples taken from table-egg layer flock environments. The NPIP method, as with the FDA method, requires single-swab culturing for the environmental sampling of laying houses for Salmonella Enteritidis. The FDA culture protocol requires a multistep culture enrichment broth, and it is more labor intensive than the NPIP culture protocol, which requires a single enrichment broth. The main objective of this study was to compare the FDA single-swab culturing protocol with that of the NPIP culturing protocol but using a four-swab pool scheme. Single and multi-laboratory testing of replicate manure drag swab sets (n  =  525 and 672, respectively) collected from a Salmonella Enteritidis-free commercial poultry flock was performed by artificially contaminating swabs with either Salmonella Enteritidis phage type 4, 8, or 13a at one of two inoculation levels: low, x¯  = 2.5 CFU (range 2.5-2.7), or medium, x¯  = 10.0 CFU (range 7.5-12). For each replicate, a single swab (inoculated), sets of two swabs (one inoculated and one uninoculated), and sets of four swabs (one inoculated and three uninoculated), testing was conducted using the FDA or NPIP culture method. For swabs inoculated with phage type 8, the NPIP method was more efficient (P < 0.05) for all swab sets at both inoculation levels than the reference method. The single swabs in the NPIP method were significantly (P < 0.05) better than four-pool swabs in detecting Salmonella Enteritidis at the lower inoculation level. In the collaborative study (n  =  13 labs) using Salmonella Enteritidis phage type 13a inoculated swabs, there was no significant difference (P > 0.05) between the FDA method (single swabs) and the pooled NPIP method (four-pool swabs). The study concludes that the pooled NPIP method is not significantly different from the FDA method for the detection of Salmonella Enteritidis in drag swabs in commercial poultry laying houses. Consequently based on the FDA's Salmonella Enteritidis rule for equivalency of different methods, the pooled NPIP method should be considered equivalent. Furthermore, the pooled NPIP method was more efficient and cost effective.

Allium sativum (garlic) treatment for murine transitional cell carcinoma.

PubMed

Riggs, D R; DeHaven, J I; Lamm, D L

1997-05-15

Currently, immunotherapy with Bacillus Calmette-Guerin (BCG) is the most effective treatment for superficial bladder carcinoma, but treatment-related toxicity may limit its use in some patients. Alternative treatments are needed for patients who fail to respond to BCG immunotherapy. Allium sativum (AS), or garlic, is known to have a broad range of biologic activities, including immune stimulation and reported antitumor activity. For these reasons, the authors conducted a series of experiments designed to explore the possible therapeutic effects of AS in the MBT2 murine bladder carcinoma model. C3H/HeN mice were randomized prior to initiation of each experimental protocol. Mice received 1 x 10(3) MBT2 cells in 0.1 mL RPMI-1640, administered subcutaneously into the right thigh, on Day 0 of the experiment. AS was injected at the site of tumor transplantation on Day 1 and at 2- to 7-day intervals up to Day 28. To evaluate the effects of oral AS in this model, treatment was initiated 30 days prior to tumor inoculation and continued for 30 days after tumor inoculation. Animals in all experiments were followed for tumor incidence, tumor growth, and survival. In the initial experiments, subcutaneous AS significantly reduced tumor volume compared with the saline control (P < 0.05). Unfortunately, treatment-related death was also observed, requiring reduction in the total dose of AS. Animals that received 5 weekly immunizations of AS (5 mg, 5 mg, 1 mg, 1 mg, and 1 mg; cumulative dose = 13 mg) had significantly reduced tumor incidence, tumor growth, and increased survival when compared with animals that received the saline control. No treatment-related deaths were observed with this treatment schedule. To determine whether systemic AS administration might be effective, orally administered AS was tested at doses of 5 mg, 50 mg, and 500 mg per 100 mL of drinking water. Mice that received 50 mg oral AS had significant reductions in tumor volume (P < 0.05) when compared with animals that received the saline control, and mice that received 500 mg oral AS had significant reductions in both tumor volume and mortality (P < 0.05). The significant antitumor efficacy of subcutaneous and oral AS warrants further investigation and suggests that AS may provide a new and effective form of therapy for transitional cell carcinoma of the bladder.

Purification of Encephalitozoon Cultures Contaminated by Mycoplasmas by Murine Intraperitoneal Inoculation

PubMed Central

Ridoux, Olivier; Foucault, Cédric; Drancourt, Michel

1998-01-01

Encephalitozoon species are strict intracellular microsporidia. Cocultures with eukaryotic cell lines can become accidently contaminated by mycoplasmas. We propose a decontamination protocol based on differential cell targeting after intraperitoneal inoculation in mice. Mycoplasma-free microsporidia were isolated from the brains and spleens of inoculated mice 24 h postinoculation by using the centrifugation shell vial system. Identification was confirmed by direct sequencing of PCR-amplified 16S rRNA. PMID:9666031

Bone Marrow–Derived Mesenchymal Stem Cells Enhance Bacterial Clearance and Preserve Bioprosthetic Integrity in a Model of Mesh Infection

PubMed Central

Criman, Erik T.; Kurata, Wendy E.; Matsumoto, Karen W.; Aubin, Harry T.; Campbell, Carmen E.

2016-01-01

Background: The reported incidence of mesh infection in contaminated operative fields is as high as 30% regardless of the material used. Recently, mesenchymal stem cells (MSCs) have been shown to possess favorable immunomodulatory properties and improve tissue incorporation when seeded onto bioprosthetics. The aim of this study was to evaluate whether seeding noncrosslinked bovine pericardium (Veritas Collagen Matrix) with allogeneic bone marrow–derived MSCs improves infection resistance in vivo after inoculation with Escherichia coli (E. coli). Methods: Rat bone marrow–derived MSCs at passage 3 were seeded onto bovine pericardium and cultured for 7 days before implantation. Additional rats (n = 24) were implanted subcutaneously with MSC-seeded or unseeded mesh and inoculated with 7 × 105 colony-forming units of E. coli or saline before wound closure (group 1, unseeded mesh/saline; group 2, unseeded mesh/E. coli; group 3, MSC-seeded mesh/E. coli; 8 rats per group). Meshes were explanted at 4 weeks and underwent microbiologic and histologic analyses. Results: MSC-seeded meshes inoculated with E. coli demonstrated superior bacterial clearance and preservation of mesh integrity compared with E. coli–inoculated unseeded meshes (87.5% versus 0% clearance; p = 0.001). Complete mesh degradation concurrent with abscess formation was observed in 100% of rats in the unseeded/E. coli group, which is in contrast to 12.5% of rats in the MSC-seeded/E. coli group. Histologic evaluation determined that remodeling characteristics of E. coli–inoculated MSC-seeded meshes were similar to those of uninfected meshes 4 weeks after implantation. Conclusions: Augmenting a bioprosthetic material with stem cells seems to markedly enhance resistance to bacterial infection in vivo and preserve mesh integrity. PMID:27482490

Nitric oxide contributes to the augmented vasodilatation during hypoxic exercise

PubMed Central

Casey, Darren P; Madery, Brandon D; Curry, Timothy B; Eisenach, John H; Wilkins, Brad W; Joyner, Michael J

2010-01-01

We tested the hypotheses that (1) nitric oxide (NO) contributes to augmented skeletal muscle vasodilatation during hypoxic exercise and (2) the combined inhibition of NO production and adenosine receptor activation would attenuate the augmented vasodilatation during hypoxic exercise more than NO inhibition alone. In separate protocols subjects performed forearm exercise (10% and 20% of maximum) during normoxia and normocapnic hypoxia (80% arterial O2 saturation). In protocol 1 (n= 12), subjects received intra-arterial administration of saline (control) and the NO synthase inhibitor NG-monomethyl-l-arginine (l-NMMA). In protocol 2 (n= 10), subjects received intra-arterial saline (control) and combined l-NMMA–aminophylline (adenosine receptor antagonist) administration. Forearm vascular conductance (FVC; ml min−1 (100 mmHg)−1) was calculated from forearm blood flow (ml min−1) and blood pressure (mmHg). In protocol 1, the change in FVC (Δ from normoxic baseline) due to hypoxia under resting conditions and during hypoxic exercise was substantially lower with l-NMMA administration compared to saline (control; P < 0.01). In protocol 2, administration of combined l-NMMA–aminophylline reduced the ΔFVC due to hypoxic exercise compared to saline (control; P < 0.01). However, the relative reduction in ΔFVC compared to the respective control (saline) conditions was similar between l-NMMA only (protocol 1) and combined l-NMMA–aminophylline (protocol 2) at 10% (−17.5 ± 3.7 vs.−21.4 ± 5.2%; P= 0.28) and 20% (−13.4 ± 3.5 vs.−18.8 ± 4.5%; P= 0.18) hypoxic exercise. These findings suggest that NO contributes to the augmented vasodilatation observed during hypoxic exercise independent of adenosine. PMID:19948661

Essential oil composition of sweet basil (Ocimum basilicum L.) in symbiotic relationship with Piriformospora indica and paclobutrazol application under salt stress.

PubMed

Keramati, Sara; Pirdashti, Hemmatollah; Babaeizad, Valliollah; Dehestani, Ali

2016-12-01

Essential oil content and oil composition of paclobutrazol treated sweet basil (Ocimum basilicum L.) plant inoculated with Piriformospora indica under salt stress were investigated by GC-MS. The results show a slight increase in essential oil content when basil plants subjected to moderate salinity stress (3 dS m -1 of NaCl). It decreased signifiicantly with increasing salinity level to 9 dS m -1 . The findings revealed that leaf area, above ground and leaf dry weights, essential oil content and yield were significantly affected by P. indica inoculation, however paclobutrazol application significantly influenced essential oil yield but not content. Fungal symbiosis as well as paclobutrazol application ameliorated the negative effects of salinity on dry matter and essential oil yield. The main constituents found in the volatile oil of O. basilicum in control treatment were Geranial (26.03%), Neral (24.88%) and Estragole (24.78%). The compounds concentrations showed some differences in P. indica and paclobutrazol treatments. The results demonstrate that micorrhiza-like fungi concomitantly increase essential oil production and biomass in sweet basil, a medicinal herb rich in commercially valuable essential oils.

Volatile-Mediated Effects Predominate in Paraburkholderia phytofirmans Growth Promotion and Salt Stress Tolerance of Arabidopsis thaliana

PubMed Central

Ledger, Thomas; Rojas, Sandy; Timmermann, Tania; Pinedo, Ignacio; Poupin, María J.; Garrido, Tatiana; Richter, Pablo; Tamayo, Javier

2016-01-01

Abiotic stress has a growing impact on plant growth and agricultural activity worldwide. Specific plant growth promoting rhizobacteria have been reported to stimulate growth and tolerance to abiotic stress in plants, and molecular mechanisms like phytohormone synthesis and 1-aminocyclopropane-1-carboxylate deamination are usual candidates proposed to mediate these bacterial effects. Paraburkholderia phytofirmans PsJN is able to promote growth of several plant hosts, and improve their tolerance to chilling, drought and salinity. This work investigated bacterial determinants involved in PsJN stimulation of growth and salinity tolerance in Arabidopsis thaliana, showing bacteria enable plants to survive long-term salinity treatment, accumulating less sodium within leaf tissues relative to non-inoculated controls. Inactivation of specific bacterial genes encoding ACC deaminase, auxin catabolism, N-acyl-homoserine-lactone production, and flagellin synthesis showed these functions have little influence on bacterial induction of salinity tolerance. Volatile organic compound emission from strain PsJN was shown to reproduce the effects of direct bacterial inoculation of roots, increasing plant growth rate and tolerance to salinity evaluated both in vitro and in soil. Furthermore, early exposure to VOCs from P. phytofirmans was sufficient to stimulate long-term effects observed in Arabidopsis growth in the presence and absence of salinity. Organic compounds were analyzed in the headspace of PsJN cultures, showing production of 2-undecanone, 7-hexanol, 3-methylbutanol and dimethyl disulfide. Exposure of A. thaliana to different quantities of these molecules showed that they are able to influence growth in a wide range of added amounts. Exposure to a blend of the first three compounds was found to mimic the effects of PsJN on both general growth promotion and salinity tolerance. To our knowledge, this is the first report on volatile compound-mediated induction of plant abiotic stress tolerance by a Paraburkholderia species. PMID:27909432

Volatile profiles and chromatic characteristics of red wines produced with Starmerella bacillaris and Saccharomyces cerevisiae.

PubMed

Englezos, Vasileios; Rantsiou, Kalliopi; Cravero, Francesco; Torchio, Fabrizio; Giacosa, Simone; Ortiz-Julien, Anne; Gerbi, Vincenzo; Rolle, Luca; Cocolin, Luca

2018-07-01

The use of mixed fermentations with Starmerella bacillaris and Saccharomyces cerevisiae is gaining attention in recent years due to their ability to modulate the metabolites production of enological interest. In the present study, four of the most popular planted red grape varieties (Cabernet sauvignon, Merlot, Pinot noir and Shiraz) were fermented using the aforementioned species and two different inoculation protocols (inoculation of S. cerevisiae after 24 and 48 h from the Starm. bacillaris inoculation), in order to evaluate their impact on the volatile composition and chromatic characteristics of wines. Analysis from chemical composition showed that titratable acidity and glycerol content exhibited marked differences among wines after fermentation. For volatile compounds, mixed fermented wines using an inoculation delay of 48 h led to reduction of volatile compounds (mainly esters). A shorter 24 h delay produced wines with higher values of color intensity than pure fermented wines. The differences observed between the inoculation protocols can be explained by the growth dynamics of both species during fermentation. These findings suggest that mixed fermentations posed a great potential in reducing metabolites which are considered negative for wine quality (mainly ethyl acetate and volatile fatty acids) and with an improvement of the chromatic profile of the wines. Copyright © 2018 Elsevier Ltd. All rights reserved.

The Effect of Growth Environment and Salinity on Lipid Production and Composition of Salicornia virginica

NASA Technical Reports Server (NTRS)

Bomani, Bilal Mark McDowell; Link, Dirk; Kail, Brian; Morreale, Bryan; Lee, Eric S.; Gigante, Bethany M.; Hendricks, Robert C.

2014-01-01

Finding a viable and sustainable source of renewable energy is a global task. Biofuels as a renewable energy source can potentially be a viable option for sustaining long-term energy needs. Biodiesel from halophytes shows great promise due to their ability to serve not only as a fuel source, but a food source as well. Halophytes are one of the few biomass plant species that can tolerate a wide range of saline conditions. We investigate the feasibility of using the halophyte, Salicornia virginica as a biofuel source by conducting a series of experiments utilizing various growth and salinity conditions. The goal is to determine if the saline content of Salicornia virginica in our indoor growth vs outdoor growth conditions has an influence on lipid recovery and total biomass composition. We focused on using standard lipid extraction protocols and characterization methods to evaluate twelve Salicornia virginica samples under six saline values ranging from freshwater to seawater and two growth conditions. The overall goal is to develop an optimal lipid extraction protocol for Salicornia virginica and potentially apply this protocol to halophytes in general.

Comparative effects of wild type Stenotrophomonas maltophilia and its indole acetic acid-deficient mutants on wheat.

PubMed

Hassan, T U; Bano, A

2016-09-01

The present investigation evaluated the role of Stenotrophomonas maltophilia and its IAA-deficient mutant on soil health and plant growth under salinity stress in the presence of tryptophan. In the first phase, S. maltophilia isolated from roots of the halo- phytic herb, Cenchrus ciliaris was used as bio-inoculant on wheat grown in saline sodic soil. A field experiment was conducted at Soil Salinity Research Institute during 2010-2011. Treatments included seed inoculation with S. maltophilia with or without tryptophan; uninoculated untreated plants were taken as control. An aqueous solution of tryptophan was added to rhizosphere soil at 1 μg l(_1) after seed germination. Inoculation with S. maltophilia significantly increased soil organic matter, enhanced (20-30%) availability of P, K, Ca and NO3 -N and decreased Na content and electrical conductivity of rhizosphere soil. Plant height, fresh weight, proline and phytohormone content of leaves were increased 30-40% over the control. Activities of superoxide dismutase (SOD) and peroxidase (POD) were 40-50% higher than control. Addition of tryptophan further augmented (10-15%) growth parameters, whereas NO3 -N, P, K and Ca content, proline content and SOD and POD increased 20-30%. In a second phase, indoleacetic acid (IAA)-deficient mutants of S. maltophilia were constructed and evaluated for conversion of tryptophan to IAA at the University of Calgary, Canada, during 2013-2014. About 1800 trans-conjugants were constructed that were unable to produce IAA in the presence of tryptophan. The results suggest that tryptophan assisted S. maltophilia in the amelioration of salt stress, and that IAA played positive role in induction of salt tolerance. © 2016 German Botanical Society and The Royal Botanical Society of the Netherlands.

Hydrocarbon biodegradation in hypersaline environments.

PubMed

Ward, D M; Brock, T D

1978-02-01

When mineral oil, hexadecane, and glutamate were added to natural samples of varying salinity (3.3 to 28.4%) from salt evaporation ponds and Great Salt Lake, Utah, rates of metabolism of these compounds decreased as salinity increased. Rate limitations did not appear to relate to low oxygen levels or to the availability of organic nutrients. Some oxidation of l-[U-C]glutamic acid occurred even at extreme salinities, whereas oxidation of [1-C]hexadecane was too low to be detected. Gas chromatographic examination of hexane-soluble components of tar samples from natural seeps at Rozel Point in Great Salt Lake demonstrated no evidence of biological oxidation of isoprenoid alkanes subject to degradation in normal environments. Some hexane-soluble components of the same tar were altered by incubation in a low-salinity enrichment culture inoculated with garden soil. Attempts to enrich for microorganisms in saline waters able to use mineral oil as a sole source of carbon and energy were successful below, but not above, about 20% salinity. This study strongly suggests a general reduction of metabolic rate at extreme salinities and raises doubt about the biodegradation of hydrocarbons in hypersaline environments.

[Survival capacity of Corynebacterium pseudotuberculosis biovar ovis in different soil types from Chubut, Argentine Patagonia].

PubMed

Alvarez, Laura; William, Aillin; Castro, Isabel; Valenzuela, Fernanda; Estevao Belchior, Silvia

Corynebacterium pseudotuberculosis is transmitted among sheep in Argentine Patagonia causing pseudotuberculosis. The bacterium penetrates the skin or mucous membrane wounds, infecting the superficial lymph nodes and viscera. When surface abscesses are cut during shearing, they drain their purulent contents and contaminate tools and the soil. The objective of this work was to evaluate the survival capacity of C. pseudotuberculosis over time, in soils from the extra-Andean Patagonia region. Five types of superficial soils were collected from different areas in Chubut province (extra-Andean Patagonia), having distinctive physicochemical properties including organic matter content (very high to nonexistent), pH (neutral to strongly alkaline), electrical conductivity (saline to non-saline) and texture (sandy, clayey, silty loam). Different aliquots of each type of soil were inoculated with C. pseudotuberculosis PAT10 strain isolated from a Patagonian sheep, and were stored at room temperature. The number of surviving bacteria was determined at various times. Sixty percent (60%) of the inoculated C. pseudotuberculosis population survived for 80 to 210 days in soils with moderate to high organic matter content respectively. Silty soils favored bacterial survival, whereas the variables pH and salinity had no effect on survival. Copyright © 2016 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

Endophytic Bacteria Improve Plant Growth, Symbiotic Performance of Chickpea (Cicer arietinum L.) and Induce Suppression of Root Rot Caused by Fusarium solani under Salt Stress.

PubMed

Egamberdieva, Dilfuza; Wirth, Stephan J; Shurigin, Vyacheslav V; Hashem, Abeer; Abd Allah, Elsayed F

2017-01-01

Salinity causes disturbance in symbiotic performance of plants, and increases susceptibility of plants to soil-borne pathogens. Endophytic bacteria are an essential determinant of cross-tolerance to biotic and abiotic stresses in plants. The aim of this study was to isolate non-rhizobial endophytic bacteria from the root nodules of chickpea ( Cicer arietinum L.), and to assess their ability to improve plant growth and symbiotic performance, and to control root rot in chickpea under saline soil conditions. A total of 40 bacterial isolates from internal root tissues of chickpea grown in salinated soil were isolated. Four bacterial isolates, namely Bacillus cereus NUU1 , Achromobacter xylosoxidans NUU2, Bacillus thuringiensis NUU3, and Bacillus subtilis NUU4 colonizing root tissue demonstrated plant beneficial traits and/or antagonistic activity against F. solani and thus were characterized in more detail. The strain B. subtilis NUU4 proved significant plant growth promotion capabilities, improved symbiotic performance of host plant with rhizobia, and promoted yield under saline soil as compared to untreated control plants under field conditions. A combined inoculation of chickpea with M. ciceri IC53 and B. subtilis NUU4 decreased H 2 O 2 concentrations and increased proline contents compared to the un-inoculated plants indicating an alleviation of adverse effects of salt stress. Furthermore, the bacterial isolate was capable to reduce the infection rate of root rot in chickpea caused by F. solani . This is the first report of F. solani causing root rot of chickpea in a salinated soil of Uzbekistan. Our findings demonstrated that the endophytic B. subtilis strain NUU4 provides high potentials as a stimulator for plant growth and as biological control agent of chickpea root rot under saline soil conditions. These multiple relationships could provide promising practical approaches to increase the productivity of legumes under salt stress.

The Multifarious PGPR Serratia marcescens CDP-13 Augments Induced Systemic Resistance and Enhanced Salinity Tolerance of Wheat (Triticum aestivum L.)

PubMed Central

Singh, Rajnish Prakash; Jha, Prabhat Nath

2016-01-01

The present study demonstrates the plant growth promoting (PGP) potential of a bacterial isolate CDP-13 isolated from ‘Capparis decidua’ plant, and its ability to protect plants from the deleterious effect of biotic and abiotic stressors. Based on 16S rRNA gene sequence analysis, the isolate was identified as Serratia marcescens. Among the PGP traits, the isolate was found to be positive for ACC deaminase activity, phosphate solubilization, production of siderophore, indole acetic acid production, nitrogen fixation, and ammonia production. CDP-13 showed growth at an increased salt (NaCl) concentration of up to 6%, indicating its potential to survive and associate with plants growing in saline soil. The inoculation of S. marcescens enhanced the growth of wheat plant under salinity stress (150–200 mM). It significantly reduced inhibition of plant growth (15 to 85%) caused by salt stressors. Application of CDP-13 also modulated concentration (20 to 75%) of different osmoprotectants (proline, malondialdehyde, total soluble sugar, total protein content, and indole acetic acid) in plants suggesting its role in enabling plants to tolerate salt stressors. In addition, bacterial inoculation also reduced the disease severity caused by fungal infection, which illustrated its ability to confer induced systemic resistance (ISR) in host plants. Treatment of wheat plants with the test organism caused alteration in anti-oxidative enzymes activities (Superoxide dismutase, Catalase, and Peroxidase) under various salinity levels, and therefore minimizes the salinity-induced oxidative damages to the plants. Colonization efficiency of strain CDP-13 was confirmed by CFU count, epi-fluorescence microscopy, and ERIC-PCR-based DNA fingerprinting approach. Hence, the study indicates that bacterium CDP-13 enhances plant growth, and has potential for the amelioration of salinity stress in wheat plants. Likewise, the results also provide insights into biotechnological approaches to using PGPR as an alternative to chemicals and pesticides. PMID:27322827

The Multifarious PGPR Serratia marcescens CDP-13 Augments Induced Systemic Resistance and Enhanced Salinity Tolerance of Wheat (Triticum aestivum L.).

PubMed

Singh, Rajnish Prakash; Jha, Prabhat Nath

2016-01-01

The present study demonstrates the plant growth promoting (PGP) potential of a bacterial isolate CDP-13 isolated from 'Capparis decidua' plant, and its ability to protect plants from the deleterious effect of biotic and abiotic stressors. Based on 16S rRNA gene sequence analysis, the isolate was identified as Serratia marcescens. Among the PGP traits, the isolate was found to be positive for ACC deaminase activity, phosphate solubilization, production of siderophore, indole acetic acid production, nitrogen fixation, and ammonia production. CDP-13 showed growth at an increased salt (NaCl) concentration of up to 6%, indicating its potential to survive and associate with plants growing in saline soil. The inoculation of S. marcescens enhanced the growth of wheat plant under salinity stress (150-200 mM). It significantly reduced inhibition of plant growth (15 to 85%) caused by salt stressors. Application of CDP-13 also modulated concentration (20 to 75%) of different osmoprotectants (proline, malondialdehyde, total soluble sugar, total protein content, and indole acetic acid) in plants suggesting its role in enabling plants to tolerate salt stressors. In addition, bacterial inoculation also reduced the disease severity caused by fungal infection, which illustrated its ability to confer induced systemic resistance (ISR) in host plants. Treatment of wheat plants with the test organism caused alteration in anti-oxidative enzymes activities (Superoxide dismutase, Catalase, and Peroxidase) under various salinity levels, and therefore minimizes the salinity-induced oxidative damages to the plants. Colonization efficiency of strain CDP-13 was confirmed by CFU count, epi-fluorescence microscopy, and ERIC-PCR-based DNA fingerprinting approach. Hence, the study indicates that bacterium CDP-13 enhances plant growth, and has potential for the amelioration of salinity stress in wheat plants. Likewise, the results also provide insights into biotechnological approaches to using PGPR as an alternative to chemicals and pesticides.

Endophytic Bacteria Improve Plant Growth, Symbiotic Performance of Chickpea (Cicer arietinum L.) and Induce Suppression of Root Rot Caused by Fusarium solani under Salt Stress

PubMed Central

Egamberdieva, Dilfuza; Wirth, Stephan J.; Shurigin, Vyacheslav V.; Hashem, Abeer; Abd_Allah, Elsayed F.

2017-01-01

Salinity causes disturbance in symbiotic performance of plants, and increases susceptibility of plants to soil-borne pathogens. Endophytic bacteria are an essential determinant of cross-tolerance to biotic and abiotic stresses in plants. The aim of this study was to isolate non–rhizobial endophytic bacteria from the root nodules of chickpea (Cicer arietinum L.), and to assess their ability to improve plant growth and symbiotic performance, and to control root rot in chickpea under saline soil conditions. A total of 40 bacterial isolates from internal root tissues of chickpea grown in salinated soil were isolated. Four bacterial isolates, namely Bacillus cereus NUU1, Achromobacter xylosoxidans NUU2, Bacillus thuringiensis NUU3, and Bacillus subtilis NUU4 colonizing root tissue demonstrated plant beneficial traits and/or antagonistic activity against F. solani and thus were characterized in more detail. The strain B. subtilis NUU4 proved significant plant growth promotion capabilities, improved symbiotic performance of host plant with rhizobia, and promoted yield under saline soil as compared to untreated control plants under field conditions. A combined inoculation of chickpea with M. ciceri IC53 and B. subtilis NUU4 decreased H2O2 concentrations and increased proline contents compared to the un-inoculated plants indicating an alleviation of adverse effects of salt stress. Furthermore, the bacterial isolate was capable to reduce the infection rate of root rot in chickpea caused by F. solani. This is the first report of F. solani causing root rot of chickpea in a salinated soil of Uzbekistan. Our findings demonstrated that the endophytic B. subtilis strain NUU4 provides high potentials as a stimulator for plant growth and as biological control agent of chickpea root rot under saline soil conditions. These multiple relationships could provide promising practical approaches to increase the productivity of legumes under salt stress. PMID:29033922

Salinity tolerance and mycorrhizal responsiveness of native xeroriparian plants in semi-arid western USA

USGS Publications Warehouse

Beauchamp, Vanessa B.; Walz, C.; Shafroth, P.B.

2009-01-01

Restoration of salt-affected soils is a global concern. In the western United States, restoration of salinized land, particularly in river valleys, often involves control of Tamarix, an introduced species with high salinity tolerance. Revegetation of hydrologically disconnected floodplains and terraces after Tamarix removal is often difficult because of limited knowledge regarding the salinity tolerance of candidate native species for revegetation. Additionally, Tamarix appears to be non-mycorrhizal. Extended occupation of Tamarix may deplete arbuscular mycorrhizal fungi in the soil, further decreasing the success of revegetation efforts. To address these issues, we screened 42 species, races, or ecotypes native to southwestern U.S. for salinity tolerance and mycorrhizal responsiveness. As expected, the taxa tested showed a wide range of responses to salinity and mycorrhizal fungi. This variation also occurred between ecotypes or races of the same species, indicating that seed collected from high-salinity reference systems is likely better adapted to harsh conditions than seed originating from less saline environments. All species tested had a positive or neutral response to mycorrhizal inoculation. We found no clear evidence that mycorrhizae increased salinity tolerance, but some species were so dependent on mycorrhizal fungi that they grew poorly at all salinity levels in pasteurized soil. ?? 2009 Elsevier B.V.

In vitro culture of coconut (Cocos nucifera L.) zygotic embryos.

PubMed

Engelmann, Florent; Malaurie, Bernard; N'Nan, Oulo

2011-01-01

Coconut is a very important crop for millions of people in tropical countries. With coconut, in vitro culture protocols have been developed with two main objectives, viz. the large scale production of particular types of coconuts and the international exchange and conservation of coconut germplasm. The methods described in this chapter have been developed in the framework of collaborative activities between research institutes in Côte d'Ivoire and France. Two coconut embryo in vitro collecting protocols have been established, one consisting of storing the disinfected embryos in a KCl solution until they are brought back to the laboratory, where they are re-disinfected and inoculated in vitro under sterile conditions, and the other including in vitro inoculation of the embryos in the field. For international germplasm exchange, zygotic embryos inoculated in vitro in plastic test tubes or endosperm cylinders containing embryos in plastic bags are used. For in vitro culture, embryos are inoculated on semi-solid medium supplemented with sucrose and activated charcoal and placed in the dark, and then transferred to light conditions with the same (solid or liquid) medium once the first true leaf is visible and the root system has started developing.

Effect of plant growth-promoting bacteria (PGPR) and arbuscular mycorrhizal fungi (AMF) inoculation on oats in saline-alkali soil contaminated by petroleum to enhance phytoremediation.

PubMed

Xun, Feifei; Xie, Baoming; Liu, Shasha; Guo, Changhong

2015-01-01

To investigate the effect of plant growth-promoting bacteria (PGPR) and arbuscular mycorrhizal fungi (AMF) on phytoremediation in saline-alkali soil contaminated by petroleum, saline-alkali soil samples were artificially mixed with different amount of oil, 5 and 10 g/kg, respectively. Pot experiments with oat plants (Avena sativa) were conducted under greenhouse condition for 60 days. Plant biomass, physiological parameters in leaves, soil enzymes, and degradation rate of total petroleum hydrocarbon were measured. The result demonstrated that petroleum inhibited the growth of the plant; however, inoculation with PGPR in combination with AMF resulted in an increase in dry weight and stem height compared with noninoculated controls. Petroleum stress increased the accumulation of malondialdehyde (MDA) and free proline and the activities of the antioxidant enzyme such as superoxide dismutase, catalase, and peroxidase. Application of PGPR and AMF augmented the activities of three enzymes compared to their respective uninoculated controls, but decreased the MDA and free proline contents, indicating that PGPR and AMF could make the plants more tolerant to harmful hydrocarbon contaminants. It also improved the soil quality by increasing the activities of soil enzyme such as urease, sucrase, and dehydrogenase. In addition, the degradation rate of total petroleum hydrocarbon during treatment with PGPR and AMF in moderately contaminated soil reached a maximum of 49.73%. Therefore, we concluded the plants treated with a combination of PGPR and AMF had a high potential to contribute to remediation of saline-alkali soil contaminated with petroleum.

Effect of Plant Growth Promoting Bacteria Associated with Halophytic Weed (Psoralea corylifolia L) on Germination and Seedling Growth of Wheat Under Saline Conditions.

PubMed

Sorty, Ajay M; Meena, Kamlesh K; Choudhary, Khushboo; Bitla, Utkarsh M; Minhas, P S; Krishnani, K K

2016-11-01

Halotolerant bacteria associated with Psoralea corylifolia L., a luxuriantly growing annual weed in salinity-affected semi-arid regions of western Maharashtra, India were evaluated for their plant growth-promoting activity in wheat. A total of 79 bacteria associated with different parts viz., root, shoot and nodule endophytes, rhizosphere, rhizoplane, and leaf epiphytes, were isolated and grouped based on their habitat. Twelve bacteria isolated for their potential in plant growth promotion were further selected for in vitro studies. Molecular identification showed the presence of the genera Bacillus, Pantoea, Marinobacterium, Acinetobacter, Enterobacter, Pseudomonas, Rhizobium, and Sinorhizobium (LC027447-53; LC027455; LC027457, LC027459, and LC128410). The phylogenetic studies along with carbon source utilization profiles using the Biolog® indicated the presence of novel species and the in planta studies revealed promising results under salinity stress. Whereas the nodule endophytes had minute plant growth-promoting (PGP) activity, the cell free culture filtrates of these strains enhanced seed germination of wheat (Triticum aestivum L). The maximum vigor index was monitored in isolate Y7 (Enterobacter sp strain NIASMVII). Indole acetic acid (IAA) production by the isolates ranged between 0.22 and 25.58 μg mL -1 . This signifies the need of exploration of their individual metabolites for developing next-generation bio-inoculants through co-inoculation with other compatible microbes. This study has potential in utilization of the weed-associated microbiome in terms of alleviation of salinity stress in crop plants.

A novel hapten and monoclonal-based enzyme-linked immunosorbent assay for sulfonamides in edible animal tissues.

PubMed

Zhou, Qi; Peng, Dapeng; Wang, Yulian; Pan, Yuanhu; Wan, Dan; Zhang, Xiya; Yuan, Zonghui

2014-07-01

For high-throughput monitoring of the residues of sulfonamides (SAs) in edible animal tissues, a novel hapten and monoclonal-based indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) was developed. The novel hapten was synthesized and conjugated to carrier protein as immunogen. The spleen cells of the inoculated mice expressing group-specificity against SAs were fused. The obtained monoclonal antibody 4E5 showed the cross-reactivity (CR) to 16 structurally different SAs. Based on this antibody, an optimised icELISA protocol was carried out with only phosphate-buffered saline for the fast extraction of SAs in the tissues. The limits of detection of SAs in chicken ranged from 1.5 to 22.3μgkg(-1). The recoveries were 70.6-121% with less than 24.1% relative standard deviation. The developed ic-ELISA showed a good correlation with high performance liquid chromatography. It would be a useful tool for the screening of residues of SAs in edible animal tissues. Copyright © 2014 Elsevier Ltd. All rights reserved.

Mining Halophytes for Plant Growth-Promoting Halotolerant Bacteria to Enhance the Salinity Tolerance of Non-halophytic Crops

PubMed Central

Etesami, Hassan; Beattie, Gwyn A.

2018-01-01

Salinity stress is one of the major abiotic stresses limiting crop production in arid and semi-arid regions. Interest is increasing in the application of PGPRs (plant growth promoting rhizobacteria) to ameliorate stresses such as salinity stress in crop production. The identification of salt-tolerant, or halophilic, PGPRs has the potential to promote saline soil-based agriculture. Halophytes are a useful reservoir of halotolerant bacteria with plant growth-promoting capabilities. Here, we review recent studies on the use of halophilic PGPRs to stimulate plant growth and increase the tolerance of non-halophytic crops to salinity. These studies illustrate that halophilic PGPRs from the rhizosphere of halophytic species can be effective bio-inoculants for promoting the production of non-halophytic species in saline soils. These studies support the viability of bioinoculation with halophilic PGPRs as a strategy for the sustainable enhancement of non-halophytic crop growth. The potential of this strategy is discussed within the context of ensuring sustainable food production for a world with an increasing population and continuing climate change. We also explore future research needs for using halotolerant PGPRs under salinity stress. PMID:29472908

Hydrocarbon Biodegradation in Hypersaline Environments

PubMed Central

Ward, David M.; Brock, T. D.

1978-01-01

When mineral oil, hexadecane, and glutamate were added to natural samples of varying salinity (3.3 to 28.4%) from salt evaporation ponds and Great Salt Lake, Utah, rates of metabolism of these compounds decreased as salinity increased. Rate limitations did not appear to relate to low oxygen levels or to the availability of organic nutrients. Some oxidation of l-[U-14C]glutamic acid occurred even at extreme salinities, whereas oxidation of [1-14C]hexadecane was too low to be detected. Gas chromatographic examination of hexane-soluble components of tar samples from natural seeps at Rozel Point in Great Salt Lake demonstrated no evidence of biological oxidation of isoprenoid alkanes subject to degradation in normal environments. Some hexane-soluble components of the same tar were altered by incubation in a low-salinity enrichment culture inoculated with garden soil. Attempts to enrich for microorganisms in saline waters able to use mineral oil as a sole source of carbon and energy were successful below, but not above, about 20% salinity. This study strongly suggests a general reduction of metabolic rate at extreme salinities and raises doubt about the biodegradation of hydrocarbons in hypersaline environments. PMID:16345276

Combining essential oils and olive extract for control of multi-drug resistant Salmonella enterica on organic leafy greens

USDA-ARS?s Scientific Manuscript database

We investigated the combined antimicrobial effects of plant essential oils and olive extract. Organic baby spinach, mature spinach, romaine lettuce, and iceberg lettuce were inoculated with the pathogen and then dip-treated in phosphate buffered saline (PBS) control, 3.0% hydrogen peroxide, a 0.1% ...

Development of a method for detection and quantification of B. brongniartii and B. bassiana in soil

NASA Astrophysics Data System (ADS)

Canfora, L.; Malusà, E.; Tkaczuk, C.; Tartanus, M.; Łabanowska, B. H.; Pinzari, F.

2016-03-01

A culture independent method based on qPCR was developed for the detection and quantification of two fungal inoculants in soil. The aim was to adapt a genotyping approach based on SSR (Simple Sequence Repeat) marker to a discriminating tracing of two different species of bioinoculants in soil, after their in-field release. Two entomopathogenic fungi, Beauveria bassiana and B. brongniartii, were traced and quantified in soil samples obtained from field trials. These two fungal species were used as biological agents in Poland to control Melolontha melolontha (European cockchafer), whose larvae live in soil menacing horticultural crops. Specificity of SSR markers was verified using controls consisting of: i) soil samples containing fungal spores of B. bassiana and B. brongniartii in known dilutions; ii) the DNA of the fungal microorganisms; iii) soil samples singly inoculated with each fungus species. An initial evaluation of the protocol was performed with analyses of soil DNA and mycelial DNA. Further, the simultaneous detection and quantification of B. bassiana and B. brongniartii in soil was achieved in field samples after application of the bio-inoculants. The protocol can be considered as a relatively low cost solution for the detection, identification and traceability of fungal bio-inoculants in soil.

Development of a method for detection and quantification of B. brongniartii and B. bassiana in soil

PubMed Central

Canfora, L.; Malusà, E.; Tkaczuk, C.; Tartanus, M.; Łabanowska, B.H.; Pinzari, F.

2016-01-01

A culture independent method based on qPCR was developed for the detection and quantification of two fungal inoculants in soil. The aim was to adapt a genotyping approach based on SSR (Simple Sequence Repeat) marker to a discriminating tracing of two different species of bioinoculants in soil, after their in-field release. Two entomopathogenic fungi, Beauveria bassiana and B. brongniartii, were traced and quantified in soil samples obtained from field trials. These two fungal species were used as biological agents in Poland to control Melolontha melolontha (European cockchafer), whose larvae live in soil menacing horticultural crops. Specificity of SSR markers was verified using controls consisting of: i) soil samples containing fungal spores of B. bassiana and B. brongniartii in known dilutions; ii) the DNA of the fungal microorganisms; iii) soil samples singly inoculated with each fungus species. An initial evaluation of the protocol was performed with analyses of soil DNA and mycelial DNA. Further, the simultaneous detection and quantification of B. bassiana and B. brongniartii in soil was achieved in field samples after application of the bio-inoculants. The protocol can be considered as a relatively low cost solution for the detection, identification and traceability of fungal bio-inoculants in soil. PMID:26975931

Effects of inhaled fine dust on lung tissue changes and antibody response induced by spores of opportunistic fungi in goats.

PubMed

Purdy, Charles W; Layton, Robert C; Straus, David C; Ayers, J R

2008-04-01

To investigate the effects of sterile fine dust aerosol inhalation on antibody responses and lung tissue changes induced by Mucor ramosissimus or Trichoderma viride spores following intratracheal inoculation in goats. 36 weanling Boer-Spanish goats. 6 goats were allocated to each of 2 M ramosissimus-inoculated groups, 2 T viride-inoculated groups, and 2 control (tent or pen) groups. One of each pair of sporetreated groups and the tent control group were exposed 7 times to sterilized fine feedyard dust (mean+/-SD particle diameter, <7.72+/-0.69 microm) for 4 hours in a specially constructed tent. Goats in the 4 fungal treatment groups were inoculated intratracheally 5 times with a fungal spore preparation (30 mL), whereas tent control goats were intratracheally inoculated with physiologic saline (0.9% NaCl) solution (30 mL). Pen control goats were not inoculated or exposed to dust. Goats received an IV challenge with equine RBCs to assess antibody responses to foreign antigens. Postmortem examinations were performed at study completion (day 68) to evaluate lung tissue lesions. 5 of 7 deaths occurred between days 18 and 45 and were attributed to fine dust exposures prior to fungal treatments. Fine dust inhalation induced similar lung lesions and precipitating antibodies among spore-treated goats. Following spore inoculations, dust-exposed goats had significantly more spores per gram of consolidated lung tissue than did their nonexposed counterparts. Fine dust inhalation appeared to decrease the ability of goats to successfully clear fungal spores from the lungs following intratracheal inoculation.

Different prion disease phenotypes result from inoculation of cattle with two temporally separated sources of sheep scrapie from Great Britain

PubMed Central

Konold, Timm; Lee, Yoon Hee; Stack, Michael J; Horrocks, Claire; Green, Robert B; Chaplin, Melanie; Simmons, Marion M; Hawkins, Steve AC; Lockey, Richard; Spiropoulos, John; Wilesmith, John W; Wells, Gerald AH

2006-01-01

Background Given the theoretical proposal that bovine spongiform encephalopathy (BSE) could have originated from sheep scrapie, this study investigated the pathogenicity for cattle, by intracerebral (i.c.) inoculation, of two pools of scrapie agents sourced in Great Britain before and during the BSE epidemic. Two groups of ten cattle were each inoculated with pools of brain material from sheep scrapie cases collected prior to 1975 and after 1990. Control groups comprised five cattle inoculated with sheep brain free from scrapie, five cattle inoculated with saline, and for comparison with BSE, naturally infected cattle and cattle i.c. inoculated with BSE brainstem homogenate from a parallel study. Phenotypic characterisation of the disease forms transmitted to cattle was conducted by morphological, immunohistochemical, biochemical and biological methods. Results Disease occurred in 16 cattle, nine inoculated with the pre-1975 inoculum and seven inoculated with the post-1990 inoculum, with four cattle still alive at 83 months post challenge (as at June 2006). The different inocula produced predominantly two different disease phenotypes as determined by histopathological, immunohistochemical and Western immunoblotting methods and biological characterisation on transmission to mice, neither of which was identical to BSE. Whilst the disease presentation was uniform in all scrapie-affected cattle of the pre-1975 group, the post-1990 inoculum produced a more variable disease, with two animals sharing immunohistochemical and molecular profile characteristics with animals in the pre-1975 group. Conclusion The study has demonstrated that cattle inoculated with different pooled scrapie sources can develop different prion disease phenotypes, which were not consistent with the phenotype of BSE of cattle and whose isolates did not have the strain typing characteristics of the BSE agent on transmission to mice. PMID:17044917

Attempts to Produce Experimental Edema Disease in Swine by Parenterally Injecting Escherichia Coli Serotype 0139:K82:H1*

PubMed Central

Pickrell, J. A.; Link, R. P.; Simon, J.; Rhoades, H. E.; Gossling, J.

1969-01-01

Twenty-two pigs were inoculated parenterally with various E. coli 0139:K82:H1 preparations. Clinical signs of disease in pigs injected with freeze-thaw extract consisted of early listlessness, diarrhea and, later, hyperirritability of varying intensity in some animals. Hemorrhagic gastroenteritis involving the duodenum, spiral colon and the fundic portion of the stomach, and ulceration of the fundic stomach were observed at post-mortem examination of pigs inoculated parenterallly with living culture or freeze-thaw extract. No significant lesions were observed in pigs inoculated with ultrasonic or hypotonic acid-saline extract. In pigs injected with living culture or freeze-thaw extract, the histological alterations consisted of moderate perivascular edema of the brain, marked hepatic parenchymal cell degeneration, hepatic subserosal edema and “toxic” lymph nodes, when compared to the control group. ImagesFig. 1.Fig. 2.Fig. 3.Fig. 4. PMID:4237302

Assessing the role of endophytic bacteria in the halophyte Arthrocnemum macrostachyum salt tolerance.

PubMed

Navarro-Torre, S; Barcia-Piedras, J M; Mateos-Naranjo, E; Redondo-Gómez, S; Camacho, M; Caviedes, M A; Pajuelo, E; Rodríguez-Llorente, I D

2017-03-01

There is an increasing interest to use halophytes for revegetation of salt affected ecosystems, as well as in understanding their mechanisms of salt tolerance. We hypothesized that bacteria from the phyllosphere of these plants might play a key role in its high tolerance to excessive salinity. Eight endophytic bacteria belonging to Bacillus and closely related genera were isolated from phyllosphere of the halophyte Arthrocnemum macrostachyum growing in salty agricultural soils. The presence of plant-growth promoting (PGP) properties, enzymatic activities and tolerance towards NaCl was determined. Effects of inoculation on seeds germination and adult plant growth under experimental NaCl treatments (0, 510 and 1030 mM NaCl) were studied. Inoculation with a consortium including the best performing bacteria improved considerably the kinetics of germination and the final germination percentage of A. macrostachyum seeds. At high NaCl concentrations (1030 mM), inoculation of plants mitigated the effects of high salinity on plant growth and physiological performance and, in addition, this consortium appears to have increased the potential of A. macrostachyum to accumulate Na + in its shoots, thus improving sodium phytoextraction capacity. Bacteria isolated from A. macrostachyum phyllosphere seem to play an important role in plant salt tolerance under stressing salt concentrations. The combined use of A. macrostachyum and its microbiome can be an adequate tool to enhance plant adaptation and sodium phytoextraction during restoration of salt degraded soils. © 2016 German Botanical Society and The Royal Botanical Society of the Netherlands.

A meta-analysis of arbuscular mycorrhizal effects on plants grown under salt stress.

PubMed

Chandrasekaran, Murugesan; Boughattas, Sonia; Hu, Shuijin; Oh, Sang-Hyon; Sa, Tongmin

2014-11-01

Salt stress limits crop yield and sustainable agriculture in most arid and semiarid regions of the world. Arbuscular mycorrhizal fungi (AMF) are considered bio-ameliorators of soil salinity tolerance in plants. In evaluating AMF as significant predictors of mycorrhizal ecology, precise quantifiable changes in plant biomass and nutrient uptake under salt stress are crucial factors. Therefore, the objective of the present study was to analyze the magnitude of the effects of AMF inoculation on growth and nutrient uptake of plants under salt stress through meta-analyses. For this, data were compared in the context of mycorrhizal host plant species, plant family and functional group, herbaceous vs. woody plants, annual vs. perennial plants, and the level of salinity across 43 studies. Results indicate that, under saline conditions, AMF inoculation significantly increased total, shoot, and root biomass as well as phosphorous (P), nitrogen (N), and potassium (K) uptake. Activities of the antioxidant enzymes superoxide dismutase, catalase, peroxidase, and ascorbate peroxidase also increased significantly in mycorrhizal compared to nonmycorrhizal plants growing under salt stress. In addition, sodium (Na) uptake decreased significantly in mycorrhizal plants, while changes in proline accumulation were not significant. Across most subsets of the data analysis, identities of AMF (Glomus fasciculatum) and host plants (Acacia nilotica, herbs, woody and perennial) were found to be essential in understanding plant responses to salinity stress. For the analyzed dataset, it is concluded that under salt stress, mycorrhizal plants have extensive root traits and mycorrhizal morphological traits which help the uptake of more P and K, together with the enhanced production of antioxidant enzymes resulting in salt stress alleviation and increased plant biomass.

Semi-automated method to measure pneumonia severity in mice through computed tomography (CT) scan analysis

NASA Astrophysics Data System (ADS)

Johri, Ansh; Schimel, Daniel; Noguchi, Audrey; Hsu, Lewis L.

2010-03-01

Imaging is a crucial clinical tool for diagnosis and assessment of pneumonia, but quantitative methods are lacking. Micro-computed tomography (micro CT), designed for lab animals, provides opportunities for non-invasive radiographic endpoints for pneumonia studies. HYPOTHESIS: In vivo micro CT scans of mice with early bacterial pneumonia can be scored quantitatively by semiautomated imaging methods, with good reproducibility and correlation with bacterial dose inoculated, pneumonia survival outcome, and radiologists' scores. METHODS: Healthy mice had intratracheal inoculation of E. coli bacteria (n=24) or saline control (n=11). In vivo micro CT scans were performed 24 hours later with microCAT II (Siemens). Two independent radiologists scored the extent of airspace abnormality, on a scale of 0 (normal) to 24 (completely abnormal). Using the Amira 5.2 software (Mercury Computer Systems), a histogram distribution of voxel counts between the Hounsfield range of -510 to 0 was created and analyzed, and a segmentation procedure was devised. RESULTS: A t-test was performed to determine whether there was a significant difference in the mean voxel value of each mouse in the three experimental groups: Saline Survivors, Pneumonia Survivors, and Pneumonia Non-survivors. It was found that the voxel count method was able to statistically tell apart the Saline Survivors from the Pneumonia Survivors, the Saline Survivors from the Pneumonia Non-survivors, but not the Pneumonia Survivors vs. Pneumonia Non-survivors. The segmentation method, however, was successfully able to distinguish the two Pneumonia groups. CONCLUSION: We have pilot-tested an evaluation of early pneumonia in mice using micro CT and a semi-automated method for lung segmentation and scoring system. Statistical analysis indicates that the system is reliable and merits further evaluation.

Increased expression of aquaporin-1 on the pleura of rats with a tuberculous pleural effusion.

PubMed

Du, Hongchun; Xie, Canmao; He, Qiao; Deng, Xiaohua

2007-12-01

The purpose of this study was to investigate whether the expression of AQP-1 on the pleura is altered in a rat model with a tuberculous pleural effusion (TPE) and to study its function. A TPE model was established by intrapleural inoculation with 0.03 mg (2 ml) standard tuberculosis bacillus (H(37)Rv). The rats with TPE were sacrificed at different time points (day 1, 3, or 5) after inoculation. The control group received a 2-ml intrapleural injection of saline. The visceral and parietal pleural tissues were harvested and processed for real-time RT-PCR, Western blot, immunohistochemistry, and determination of tissue AQP-1 levels. Recombinant adenovirus Ad-rAQP-1 containing full-length cDNA of AQP-1 was constructed. Six groups of seven Wistar rats were assigned to receive the following treatments: group 1: intrapleural administration of normal saline; group 2: intrapleural administration of tuberculosis bacilli (TB); group 3: intrapleural inoculation with TB at day 7 following intrapleural administration of Ad-rAQP-1 vector; group 4: intrapleural inoculation with 0.03 mg TB at day 7 following intrapleural administration of control Ad-GFP vector; group 5: intrapleural administration of Ad-rAQP-1; group 6: intrapleural administration of control Ad-GFP vector. The expression of AQP-l on the pleural tissue was detected by immunohistochemistry and Western blot analysis. Histopathologic changes of the pleura and the volume of pleural fluid were examined on day 7 following gene intervention or on day 3 following TB inoculation. Bilateral pleural effusions appeared within 5 days in all rats who received an intrapleural inoculation with TB. The peak amount of pleural fluid occurred on day 3. The AQP-1 expression at protein and mRNA was increased in the early phase of TPE. The expression of AQP-1 was increased in the Ad-rAQP-1 gene transfer group, indicating successful adenovirus gene transfer. The volume of pleural fluid in group 3 (6.1 +/- 0.7 ml) was significantly increased compared with that in group 2 (3.8 +/- 1.0 ml) and group 4 (4.0 +/- 1.1 ml). These findings suggested that AQP-1 was increased in TPE and it may be involved in the formation of TPE.

Endophytic fungal association via gibberellins and indole acetic acid can improve plant growth under abiotic stress: an example of Paecilomyces formosus LHL10

PubMed Central

2012-01-01

Background Endophytic fungi are little known for exogenous secretion of phytohormones and mitigation of salinity stress, which is a major limiting factor for agriculture production worldwide. Current study was designed to isolate phytohormone producing endophytic fungus from the roots of cucumber plant and identify its role in plant growth and stress tolerance under saline conditions. Results We isolated nine endophytic fungi from the roots of cucumber plant and screened their culture filtrates (CF) on gibberellins (GAs) deficient mutant rice cultivar Waito-C and normal GAs biosynthesis rice cultivar Dongjin-byeo. The CF of a fungal isolate CSH-6H significantly increased the growth of Waito-C and Dongjin-byeo seedlings as compared to control. Analysis of the CF showed presence of GAs (GA1, GA3, GA4, GA8, GA9, GA12, GA20 and GA24) and indole acetic acid. The endophyte CSH-6H was identified as a strain of Paecilomyces formosus LHL10 on the basis of phylogenetic analysis of ITS sequence similarity. Under salinity stress, P. formosus inoculation significantly enhanced cucumber shoot length and allied growth characteristics as compared to non-inoculated control plants. The hypha of P. formosus was also observed in the cortical and pericycle regions of the host-plant roots and was successfully re-isolated using PCR techniques. P. formosus association counteracted the adverse effects of salinity by accumulating proline and antioxidants and maintaining plant water potential. Thus the electrolytic leakage and membrane damage to the cucumber plants was reduced in the association of endophyte. Reduced content of stress responsive abscisic acid suggest lesser stress convened to endophyte-associated plants. On contrary, elevated endogenous GAs (GA3, GA4, GA12 and GA20) contents in endophyte-associated cucumber plants evidenced salinity stress modulation. Conclusion The results reveal that mutualistic interactions of phytohormones secreting endophytic fungi can ameliorate host plant growth and alleviate adverse effects of salt stress. Such fungal strain could be used for further field trials to improve agricultural productivity under saline conditions. PMID:22235902

Bactec™ blood culture bottles allied to MALDI-TOF mass spectrometry: rapid etiologic diagnosis of bacterial endophthalmitis.

PubMed

Tanaka, Tatiana; Oliveira, Luiza Manhezi de Freitas; Ferreira, Bruno Fortaleza de Aquino; Kato, Juliana Mika; Rossi, Flavia; Correa, Karoline de Lemes Giuntini; Pimentel, Sergio Luis Gianotti; Yamamoto, Joyce Hisae; Almeida Junior, João Nóbrega

2017-07-01

Matrix-assisted laser desorption ionization-time of flight (MALDI-TOF) mass spectrometry (MS) has been used for direct identification of pathogens from blood-inoculated blood culture bottles (BCBs). We showed that MALDI-TOF MS is an useful technique for rapid identification of the causative agents of endophthalmitis from vitreous humor-inoculated BCBs with a simple protocol. Copyright © 2017 Elsevier Inc. All rights reserved.

Immune Responses and Protection against Experimental Brucella suis biovar 1 Challenge in Non-vaccinated or RB51-Vaccinated Cattle

USDA-ARS?s Scientific Manuscript database

Twenty Hereford heifers, approximately 9 months of age, were vaccinated with saline (control) or 2 x 10**10 CFU of Brucella abortus strain RB51 (RB51) vaccine. Immunologic responses after inoculation demonstrated significantly greater (P<0.05) antibody and proliferative responses to RB51 antigens i...

The impact of arbuscular mycorrhizal fungi in mitigating salt-induced adverse effects in sweet basil (Ocimum basilicum L.).

PubMed

Elhindi, Khalid M; El-Din, Ahmed Sharaf; Elgorban, Abdallah M

2017-01-01

Salinity is one of the serious abiotic stresses adversely affecting the majority of arable lands worldwide, limiting the crop productivity of most of the economically important crops. Sweet basil ( Osmium basilicum ) plants were grown in a non-saline soil (EC = 0.64 dS m -1 ), in low saline soil (EC = 5 dS m -1 ), and in a high saline soil (EC = 10 dS m -1 ). There were differences between arbuscular mycorrhizal ( Glomus deserticola ) colonized plants (+AMF) and non-colonized plants (-AMF). Mycorrhiza mitigated the reduction of K, P and Ca uptake due to salinity. The balance between K/Na and between Ca/Na was improved in +AMF plants. Growth enhancement by mycorrhiza was independent from plant phosphorus content under high salinity levels. Different growth parameters, salt stress tolerance and accumulation of proline content were investigated, these results showed that the use of mycorrhizal inoculum (AMF) was able to enhance the productivity of sweet basil plants under salinity conditions. Mycorrhizal inoculation significantly increased chlorophyll content and water use efficiency under salinity stress. The sweet basil plants appeared to have high dependency on AMF which improved plant growth, photosynthetic efficiency, gas exchange and water use efficiency under salinity stress. In this study, there was evidence that colonization with AMF can alleviate the detrimental salinity stress influence on the growth and productivity of sweet basil plants.

Effects of arbuscular mycorrhizal fungi on growth and nitrogen uptake of Chrysanthemum morifolium under salt stress

PubMed Central

Wang, Minqiang; Li, Yan; Wu, Aiping; Huang, Juying

2018-01-01

Soil salinity is a common and serious environmental problem worldwide. Arbuscular mycorrhizal fungi (AMF) are considered as bio-ameliorators of soil salinity tolerance in plants. However, few studies have addressed the possible benefits of AMF inoculation for medicinal plants under saline conditions. In this study, we examined the effects of colonization with two AMF, Funneliformis mosseae and Diversispora versiformis, alone and in combination, on the growth and nutrient uptake of the medicinal plant Chrysanthemum morifolium (Hangbaiju) in a greenhouse salt stress experiment. After 6 weeks of a non-saline pretreatment, Hangbaiju plants with and without AMF were grown for five months under salinity levels that were achieved using 0, 50 and 200 mM NaCl. Root length, shoot and root dry weight, total dry weight, and root N concentration were higher in the mycorrhizal plants than in the non-mycorrhizal plants under conditions of moderate salinity, especially with D. versiformis colonization. As salinity increased, mycorrhizal colonization and mycorrhizal dependence decreased. The enhancement of root N uptake is probably the main mechanism underlying salt tolerance in mycorrhizal plants. These results suggest that the symbiotic associations between the fungus D. versiformis and C. morifolium plants may be useful in biotechnological practice. PMID:29698448

Effects of arbuscular mycorrhizal fungi on growth and nitrogen uptake of Chrysanthemum morifolium under salt stress.

PubMed

Wang, Yanhong; Wang, Minqiang; Li, Yan; Wu, Aiping; Huang, Juying

2018-01-01

Soil salinity is a common and serious environmental problem worldwide. Arbuscular mycorrhizal fungi (AMF) are considered as bio-ameliorators of soil salinity tolerance in plants. However, few studies have addressed the possible benefits of AMF inoculation for medicinal plants under saline conditions. In this study, we examined the effects of colonization with two AMF, Funneliformis mosseae and Diversispora versiformis, alone and in combination, on the growth and nutrient uptake of the medicinal plant Chrysanthemum morifolium (Hangbaiju) in a greenhouse salt stress experiment. After 6 weeks of a non-saline pretreatment, Hangbaiju plants with and without AMF were grown for five months under salinity levels that were achieved using 0, 50 and 200 mM NaCl. Root length, shoot and root dry weight, total dry weight, and root N concentration were higher in the mycorrhizal plants than in the non-mycorrhizal plants under conditions of moderate salinity, especially with D. versiformis colonization. As salinity increased, mycorrhizal colonization and mycorrhizal dependence decreased. The enhancement of root N uptake is probably the main mechanism underlying salt tolerance in mycorrhizal plants. These results suggest that the symbiotic associations between the fungus D. versiformis and C. morifolium plants may be useful in biotechnological practice.

Evaluation of two types of swabs for sampling allograft musculoskeletal tissue.

PubMed

Varettas, Kerry

2015-01-01

Allograft musculoskeletal tissue is commonly sampled by a swab for bioburden screening. To determine if bioburden recovery could be improved at the pre-analytical stage, two swab systems were evaluated: the Amies gel swab and the ESwab. In vitro studies were performed to determine the recovery of each swab system with <100 colony-forming unit of challenge organisms using inoculated swabs and by sampling inoculated femoral heads. The standard culture protocol used in this laboratory was also evaluated after sampling of inoculated femoral heads. A prospective study was performed with both swab systems used in parallel to sample cadaveric allograft musculoskeletal tissue. The challenge organisms could be recovered from the in vitro inoculated studies. The standard culture protocol in this laboratory recovered all challenge organisms from both swab systems. One hundred and six paired Amies and ESwabs were collected from eight cadaveric donors with skin commensals the predominant isolates. The sampling of an inoculated femoral head was included to reflect routine swab sampling practice as was the inclusion of the standard method used in this laboratory. This appears to be the first study to compare Amies gel swabs with ESwabs to sample allograft femoral heads and in a prospective study with cadaveric allograft musculoskeletal tissue. Other comparative studies of swab systems have used a much higher inoculum to mimic an infection; however, sepsis is an exclusion criterion for allograft donors. It was found that the Amies gel swab and ESwab are both suitable sampling devices for bioburden testing of allograft musculoskeletal tissue. © 2014 Royal Australasian College of Surgeons.

Quantifying the degradation of TNT and RDX in a saline environment with and without UV-exposure.

PubMed

Sisco, Edward; Najarro, Marcela; Bridge, Candice; Aranda, Roman

2015-06-01

Terrorist attacks in a maritime setting, such as the bombing of the USS Cole in 2000, or the detection of underwater mines, require the development of proper protocols to collect and analyse explosive material from a marine environment. In addition to proper analysis of the explosive material, protocols must also consider the exposure of the material to potentially deleterious elements, such as UV light and salinity, time spent in the environment, and time between storage and analysis. To understand how traditional explosives would be affected by such conditions, saline solutions of explosives were exposed to natural and artificial sunlight. Degradation of the explosives over time was then quantified using negative chemical ionization gas chromatography mass spectrometry (GC/NCI-MS). Two explosives, trinitrotoluene (TNT) and cyclotrimethylenetrinitramine (RDX), were exposed to different aqueous environments and light exposures with salinities ranging from freshwater to twice the salinity of ocean water. Solutions were then aged for up to 6 months to simulate different conditions the explosives may be recovered from. Salinity was found to have a negligible impact on the degradation of both RDX and TNT. RDX was stable in solutions of all salinities while TNT solutions degraded regardless of salinity. Solutions of varying salinities were also exposed to UV light, where accelerated degradation was seen for both explosives. Potential degradation products of TNT were identified using electrospray ionization mass spectrometry (ESI-MS), and correspond to proposed degradation products discussed in previously published works [1]. Published by Elsevier Ireland Ltd.

Epizootic rabbit enteropathy inoculum (TEC4): antibiograms and antibiotic fractionation.

PubMed

Huybens, Nathalie; Houeix, Julien; Licois, Dominique; Mainil, Jacques; Marlier, Didier

2011-01-01

Epizootic rabbit enteropathy (ERE) emerged and spread in Europe within the last 13 years causing major economical loss. The aims of the study was to evaluate antibiograms of TEC4, an inoculum composed of an extract of intestinal content of affected rabbits, and to test the potential of different antibiotic-based TEC4 fractions to reproduce the disease. Twenty nine different antibiotic discs were incubated for determining bacteria resistance. In a complementary study, nine tubes of liquid medium were inoculated with TEC4, incubated and added individually with amoxicillin/clavulanic acid, bacitracin, ceftiofur, doxycycline, novobiocin, streptomycyin, tylosin, vancomycin and 0.9% saline solution as control. The content of each tube was washed by centrifugation and suspended in saline. The three most effective antibiotics are florfenicol, amoxycillin/clavulanic acid and tylosin. A high concentration of Clostridium sordelli and Bacillus firmus were isolated in all fractions. Species never cultured from TEC4 were identified as Fusobacterium necrogenes (in vancomycin fraction), Cellulomonas sp (in novobiocin fraction) and Bacteroides distasonis (in doxycycline fraction). The ERE was reproduced when bacitracin, doxycycline and 0.9% fractions were inoculated. Rabbits showed ERE clinical signs with the specific drop in daily weight gain.

DNA vaccines: protective immunizations by parenteral, mucosal, and gene-gun inoculations.

PubMed Central

Fynan, E F; Webster, R G; Fuller, D H; Haynes, J R; Santoro, J C; Robinson, H L

1993-01-01

Plasmid DNAs expressing influenza virus hemagglutinin glycoproteins have been tested for their ability to raise protective immunity against lethal influenza challenges of the same subtype. In trials using two inoculations of from 50 to 300 micrograms of purified DNA in saline, 67-95% of test mice and 25-63% of test chickens have been protected against a lethal influenza challenge. Parenteral routes of inoculation that achieved good protection included intramuscular and intravenous injections. Successful mucosal routes of vaccination included DNA drops administered to the nares or trachea. By far the most efficient DNA immunizations were achieved by using a gene gun to deliver DNA-coated gold beads to the epidermis. In mice, 95% protection was achieved by two immunizations with beads loaded with as little as 0.4 micrograms of DNA. The breadth of routes supporting successful DNA immunizations, coupled with the very small amounts of DNA required for gene-gun immunizations, highlight the potential of this remarkably simple technique for the development of subunit vaccines. Images Fig. 1 PMID:8265577

Intermediate-level disinfection with accelerated hydrogen peroxide prevents accumulation of bacteria in Versajet™ tubing during repeated daily debridement using simulated-use testing with an inoculated pork hock.

PubMed

Gawaziuk, J P; Alfa, M J; Olson, N; Logsetty, S

2014-05-01

This study assesses the feasibility of using the Versajet™ system (VJS) on an inoculated pork hock (PH) skin surface sequentially for 8 days with daily cleaning and intermediate-level disinfection (ILD). Daily, PHs were inoculated with bacteria suspended in artificial test soil (ATS). An ILD protocol with accelerated hydrogen peroxide (AHP, OxivirTB(®)) was employed to clean and disinfect the VJS between debridements. PH skin contains 6.1-6.8×10(6)cfu/cm(2) bacteria. Bacterial counts in the handpiece and discharge hoses immediately after debridement of the PHs, and before cleaning, increased throughout the study period (5.19-6.43log10cfu/mL). Cleaning with the ILD protocol was reduced bacterial counts on the VJS by 6-log. Protein, a surrogate marker of organic contamination, was also reduced post-cleaning and ILD. Compared to a maximum post-debridement level of protein (57.9 μg/mL) obtained before ILD, VJS protein levels dropped to 9.8 (handpiece) and 13.8 μg/mL (discharge hose). Disinfection of the handpiece and discharge hose after debridement with AHP resulted in a 6-log reduction in bacterial count and 4.2 fold reduction in protein. An ILD protocol with an AHP may be a feasible method for serial skin surface debridements with the VJS for up to eight days. Copyright © 2013 Elsevier Ltd and ISBI. All rights reserved.

Evaluation of two immunomagnetic separation techniques for the detection and recovery of E. coli O157:H7 from finished composts

USDA-ARS?s Scientific Manuscript database

Two rapid immunomagnetic separation (IMS) protocols were evaluated to recover 1-2 log CFU/g inoculated E. coli O157:H7 from 30 different commercial, finished compost samples. Both protocols detected E. coli O157:H7 in compost samples; PCR techniques required the removal of inhibitors to reduce poss...

Facial paralysis induced by ear inoculation of herpes simplex virus in rat.

PubMed

Fujiwara, Takashi; Matsuda, Seiji; Tanaka, Junya; Hato, Naohito

2017-02-01

Bell's palsy is caused by the reactivation of herpes simplex virus type 1 (HSV-1). Using Balb/c mice inoculated with the KOS strain of HSV-1, we previously developed an animal disease model that simulated mild Bell's palsy. The current study developed an animal disease model of more severe facial palsy than that seen in the mouse model. Three-week-old female Wister rats weighing 60-80g were inoculated on the auricle with HSV-1 and acyclovir was administered intraperitoneally to deactivate the infected HSV-1. Instead of HSV-1, phosphate-buffered saline was used for inoculation as a negative control. Quantitative polymerase chain reaction (PCR), behavior testing (blink reflex), electroneuronography, histopathology of the peripheral nerve, and immunohistochemistry of the facial nerve nucleus were evaluated. Facial palsy occurred 3-5 days after virus inoculation, and the severity of the facial palsy progressed for up to 7 days. Quantitative PCR showed an increase in HSV-1 DNA copies in the facial nerve from 24 to 72h, suggesting that HSV-1 infection occurred in the nerve. Electroneuronography values were 33.0±15.3% and 110.0±18.0% in HSV-1-inoculated and control rats, respectively. The histopathology of the peripheral nerve showed demyelination and loss of the facial nerve, and the facial nerve nucleus showed degeneration. Facial palsy developed in Wister rats following inoculation of the KOS strain of HSV-1 onto the auricles. The behavioral, histopathological, and electroneuronography data suggested that the severity of facial palsy was greater in our rats than in animals in the previous mouse disease model. Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.

Establishment of an efficient plant regeneration culture protocol and achievement of successful genetic transformation in Jatropha curcas L.

PubMed

Liu, Ying; Liu, Guoxuan; Yang, Yali; Niu, Sufang; Yang, Fuguang; Yang, Shaoxia; Tang, Jianian; Chen, Jianping

2017-12-01

An efficient and reproducible protocol is described for shoot-bud regeneration and Agrobacterium tumefaciens-mediated genetic transformation of J. curcas. Treating the explants with high concentrations (5-120 mg/L) of TDZ for short durations (5-80 min) before inoculation culture increased significantly the regeneration frequency and improved the quality of the regenerated buds. The highest shoot-buds induction rate (87.35%) was achieved when petiole explants were treated with 20 mg/L TDZ solution for 20 min and inoculated on hormone-free MS medium for 30 days. Regenerated shoots of 0.5 cm or a little longer were isolated and grafted to seedling stocks of the same species, and then the grafted plantlets were planted on half-strength MS medium containing 0.1 mg/L IBA and 2 mg/L sodium nitroprusside (SNP). This grafting strategy was found to be very effective, to obtain that healthy grafted plantlets ready for acclimatization within 20 days. By the above mentioned protocol and with general Agrobacterium - mediated genetic transformation methods only 65 days were needed to obtain intact transgenic plants.

An evaluation of the use of chlorine dioxide (Tristel One-Shot) in an automated washer/disinfector (Medivator) fitted with a chlorine dioxide generator for decontamination of flexible endoscopes.

PubMed

Coates, D

2001-05-01

Microbiological tests were carried out to evaluate a new chlorine dioxide sterilant: Tristel OneShot. Preliminary in vitro suspension tests showed that solutions containing around 140 ppm chlorine dioxide achieved a reduction factor exceeding 10(6) of Staphylococcus aureus in 1min and of Bacillus subtilis spores in 2.5 min in the presence of 3g/L bovine albumin. Subsequent tests evaluated the effectiveness of Tristel One-Shot in a Medivator washer/disinfector fitted with a Tristel Generator for processing flexible endoscopes. Each test run involved three stages. In the first, the instrument and air-water channels of a gastroscope were inoculated with a suspension of Pseudomonas aeruginosa (10(8)cfu/ml) in 10% sodium glutamate and serum (0, 5 or 10%) and then drained, partially dried, and saline flushed through for total viable counts (TVCs). In the second stage, the channels were re-inoculated with test organisms; detergent was flushed through the channels which were then brushed; and saline was flushed through for TVCs. In the third stage, the channels were re-inoculated; detergent was flushed through the channels which were then brushed; the endoscope was processed in the Medivator; and saline was flushed through for TVCs. Carrying out all three stages enabled determination of (1) the contribution played by manual cleaning of channels prior to processing in the Medivator, and (2) the combined effect of manual cleaning followed by processing. Two series of test runs were done. In the first, the Tristel Generator was set to generate 230ppm chlorine dioxide, and in the second 150ppm. In the first, cleaning followed by processing in the Medivator consistently achieved a >/= 10(6)-fold reduction of test organisms, and in the second a >/= 10(5)-fold reduction. Pre-cleaning of channels was very important-when done the initial concentration of serum in the inoculum (0-10%) had no affect on the results obtained after processing. Copyright 2001 The Hospital infection Society.

Strategies for merging microbial fuel cell technologies in water desalination processes: Start-up protocol and desalination efficiency assessment

NASA Astrophysics Data System (ADS)

Borjas, Zulema; Esteve-Núñez, Abraham; Ortiz, Juan Manuel

2017-07-01

Microbial Desalination Cells constitute an innovative technology where microbial fuel cell and electrodialysis merge in the same device for obtaining fresh water from saline water with no energy-associated cost for the user. In this work, an anodic biofilm of the electroactive bacteria Geobacter sulfurreducens was able to efficiently convert the acetate present in synthetic waste water into electric current (j = 0.32 mA cm-2) able to desalinate water. .Moreover, we implemented an efficient start-up protocol where desalination up to 90% occurred in a desalination cycle (water production:0.308 L m-2 h-1, initial salinity: 9 mS cm-1, final salinity: <1 mS cm-1) using a filter press-based MDC prototype without any energy supply (excluding peristaltic pump energy). This start-up protocol is not only optimized for time but also simplifies operational procedures making it a more feasible strategy for future scaling-up of MDCs either as a single process or as a pre-treatment method combined with other well established desalination technologies such as reverse osmosis (RO) or reverse electrodialysis.

Differences in time until dispersal between cryptic species of a marine nematode species complex.

PubMed

De Meester, Nele; Derycke, Sofie; Moens, Tom

2012-01-01

Co-occurrence of closely related species may be achieved in environments with fluctuating dynamics, where competitively inferior species can avoid competition through dispersal. Here we present an experiment in which we compared active dispersal abilities (time until first dispersal, number and gender of dispersive adults, and nematode densities at time of dispersal) in Litoditis marina, a common bacterivorous nematode species complex comprising four often co-occurring cryptic species, Pm I, II, III, and IV, as a function of salinity and food distribution. The experiment was conducted in microcosms consisting of an inoculation plate, connection tube, and dispersal plate. Results show species-specific dispersal abilities with Pm I dispersing almost one week later than Pm III. The number of dispersive adults at time of first dispersal was species-specific, with one dispersive female in Pm I and Pm III and a higher, gender-balanced, number in Pm II and Pm IV. Food distribution affected dispersal: in absence of food in the inoculation plate, all species dispersed after ca four days. When food was available Pm I dispersed later, and at the same time and densities irrespective of food conditions in the dispersal plate (food vs no food), suggesting density-dependent dispersal. Pm III dispersed faster and at a lower population density. Salinity affected dispersal, with slower dispersal at higher salinity. These results suggest that active dispersal in Litoditis marina is common, density-dependent, and with species, gender- and environment-specific dispersal abilities. These differences can lead to differential responses under suboptimal conditions and may help to explain temporary coexistence at local scales.

Multi-scale soil salinity mapping and monitoring with proximal and remote sensing

USDA-ARS?s Scientific Manuscript database

This talk is part of a technical short course on “Soil mapping and process modelling at diverse scales”. In the talk, guidelines, special considerations, protocols, and strengths and limitations are presented for characterizing spatial and temporal variation in soil salinity at several spatial scale...

Bacillus licheniformis SA03 Confers Increased Saline–Alkaline Tolerance in Chrysanthemum Plants by Induction of Abscisic Acid Accumulation

PubMed Central

Zhou, Cheng; Zhu, Lin; Xie, Yue; Li, Feiyue; Xiao, Xin; Ma, Zhongyou; Wang, Jianfei

2017-01-01

Soil saline-alkalization is a major abiotic stress that leads to low iron (Fe) availability and high toxicity of sodium ions (Na+) for plants. It has recently been shown that plant growth promoting rhizobacteria (PGPR) can enhance the ability of plants to tolerate multiple abiotic stresses such as drought, salinity, and nutrient deficiency. However, the possible involvement of PGPR in improving saline–alkaline tolerance of plants and the underlying mechanisms remain largely unknown. In this study, we investigated the effects of Bacillus licheniformis (strain SA03) on the growth of Chrysanthemum plants under saline–alkaline conditions. Our results revealed that inoculation with SA03 alleviated saline–alkaline stress in plants with increased survival rates, photosynthesis and biomass. The inoculated plants accumulated more Fe and lower Na+ concentrations under saline–alkaline stress compared with the non-inoculated plants. RNA-Sequencing analyses further revealed that SA03 significantly activated abiotic stress- and Fe acquisition-related pathways in the stress-treated plants. However, SA03 failed to increase saline–alkaline tolerance in plants when cellular abscisic acid (ABA) and nitric oxide (NO) synthesis were inhibited by treatment with fluridone (FLU) and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO), respectively. Importantly, we also found that NO acted downstream of SA03-induced ABA to activate a series of adaptive responses in host plants under saline–alkaline stress. These findings demonstrated the potential roles of B. licheniformis SA03 in enhancing saline–alkaline tolerance of plants and highlighted the intricate integration of microbial signaling in regulating cellular Fe and Na+ accumulation. PMID:28706529

Can combined use of low-level lasers and hyaluronic acid injections prolong the longevity of degenerative knee joints?

PubMed Central

Ip, David; Fu, Nga Yue

2015-01-01

Background This study evaluated whether half-yearly hyaluronic acid injection together with low-level laser therapy in addition to standard conventional physical therapy can successfully postpone the need for joint replacement surgery in elderly patients with bilateral symptomatic tricompartmental knee arthritis. Methods In this prospective, double-blind, placebo-controlled study, 70 consecutive unselected elderly patients with bilateral tricompartmental knee arthritis were assigned at random to either one of two conservative treatment protocols to either one of the painful knees. Protocol A consisted of conventional physical therapy plus a sham light source plus saline injection, and protocol B consisted of protocol A with addition of half-yearly hyaluronic acid injection as well as low-level laser treatment instead of using saline and a sham light source. Treatment failure was defined as breakthrough pain necessitating joint replacement. Results Among the 140 painful knees treated with either protocol A or protocol B, only one of the 70 painful knees treated by protocol B required joint replacement, whereas 15 of the 70 painful knees treated by protocol A needed joint replacement surgery (P<0.05). Conclusion We conclude that half-yearly hyaluronic acid injections together with low-level laser therapy should be incorporated into the standard conservative treatment protocol for symptomatic knee arthritis, because it may prolong the longevity of the knee joint without the need for joint replacement. PMID:26346122

The Activity of Antimicrobial Surfaces Varies by Testing Protocol Utilized

PubMed Central

Campos, Matias D.; Zucchi, Paola C.; Phung, Ann; Leonard, Steven N.; Hirsch, Elizabeth B.

2016-01-01

Background Contaminated hospital surfaces are an important source of nosocomial infections. A major obstacle in marketing antimicrobial surfaces is a lack of efficacy data based on standardized testing protocols. Aim We compared the efficacy of multiple testing protocols against several “antimicrobial” film surfaces. Methods Four clinical isolates were used: one Escherichia coli, one Klebsiella pneumoniae, and two Staphylococcus aureus strains. Two industry methods (modified ISO 22196 and ASTM E2149), a “dried droplet”, and a “transfer” method were tested against two commercially available antimicrobial films, one film in development, an untreated control, and a positive (silver) control film. At 2 (only ISO) and 24 hours following inoculation, bacteria were collected from film surfaces and enumerated. Results Compared to untreated films in all protocols, there were no significant differences in recovery on either commercial brand at 2 or 24 hours after inoculation. The silver surface demonstrated significant microbicidal activity (mean loss 4.9 Log10 CFU/ml) in all methods and time points with the exception of 2 hours in the ISO protocol and the transfer method. Using our novel droplet method, no differences between placebo and active surfaces were detected. The surface in development demonstrated variable activity depending on method, organism, and time point. The ISO demonstrated minimal activity at 2 hours but significant activity at 24 hours (mean 4.5 Log10 CFU/ml difference versus placebo). The ASTEM protocol exhibited significant differences in recovery of staphylococci (mean 5 Log10 CFU/ml) but not Gram-negative isolates (10 fold decrease). Minimal activity was observed with this film in the transfer method. Conclusions Varying results between protocols suggested that efficacy of antimicrobial surfaces cannot be easily and reproducibly compared. Clinical use should be considered and further development of representative methods is needed. PMID:27494336

Leakage of Intracellular UV Materials of High Hydrostatic Pressure-Injured Escherichia Coli O157:H7 Strains in Tomato Juice

USDA-ARS?s Scientific Manuscript database

The effect of high hydrostatic pressure (HHP) treatment on inactivation, injury and recovery of Salmonella Enteritidis and Escherichia coli O157:H7 cocktail inoculated in tomato juice (pH 4.1) and phosphate buffer saline (PBS. pH 7.2) at 8.0 log CFU/ml and treated at 350, 400, 450 MPa for 20 min at ...

The Risk of Microbial Contamination in Multiple-Dose Preservative-Free Ophthalmic Preparations.

PubMed

Saisyo, Atsuyuki; Shimono, Rima; Oie, Shigeharu; Kimura, Kazuhiro; Furukawa, Hiroyuki

2017-01-01

Multiple-dose ophthalmic preparations that do not contain preservatives carry high risks of microbial contamination. However, there are various types of hospital preparations, with different physicochemical properties. In the present study, we evaluated the association between physicochemical properties and microbial contamination in ophthalmic preparations. The investigated hospital preparations included ophthalmic preparations of physiological saline, 0.2% fluconazole, 0.5% vancomycin hydrochloride, and 2% cyclosporine. We investigated the microbial dynamics of each ophthalmic preparation and microbial contamination in ophthalmic preparations used by patients. Remarkable growth of Pseudomonas aeruginosa, Burkholderia cepacia, and Serratia marcescens was observed in ophthalmic preparations of physiological saline and 0.2% fluconazole. All tested microorganisms displayed decreased counts after inoculation in 0.5% vancomycin hydrochloride. In 2% cyclosporine, all investigated microorganisms were below the limit of detection after inoculation for 6 h. The microbial contamination rates of ophthalmic preparations used by patients were 16.7% (3/18 samples) for 0.5% vancomycin hydrochloride and 0% (0/30 samples) for 2% cyclosporine. All detected contaminants in 0.5% vancomycin hydrochloride were Candida spp., one of which was present at a level of 1×10 4 colony-forming units/mL. The storage method for in-use ophthalmic preparations should be considered on the basis of their physicochemical properties.

Bovine Colostral Antibody Against Verotoxin 2 Derived from Escherichia coli O157:H7: Resistance to Proteases and Effects in Beagle Dogs

PubMed Central

Kuribayashi, Takashi; Seita, Tetsuro; Matsumoto, Mariko; Furuhata, Katsunori; Tagata, Kazutoshi; Yamamoto, Shizuo

2009-01-01

A bovine colostral antibody against verotoxin (VT) 2 of Escherichia coli O157:H7 was administered orally to beagle dogs. The antibody remained in the dogs’ small intestine for at least 2 h, whereas little serum antibody remained 1.5 h after administration. Furthermore, the antibody activity of secretory IgA did not change until 2 h after administration; however, the activity of IgG and IgM antibodies decreased by approximately 60% and 40% at 2 h after administration, respectively. Seven beagle dogs inoculated with Escherichia coli O157:H7 producing VT2 were administered bovine colostral antibody or bovine colostral whey without antibody. With administration of bovine colostral whey without antibody, the amount of VT2 in feces decreased gradually after administration and increased again at 5 d after inoculation, whereas bovine colostral antibody significantly reduced the amount of VT2 in feces on the day after administration. In addition, 9 beagle dogs were given bovine colostral antibody, bovine plasma antibody, or saline. The amount of VT2 in feces again decreased significantly more rapidly after administration of bovine colostral antibody than after administration of bovine plasma antibody or saline. PMID:19389308

Use of an agar-gel technique for large scale application to recover Ascaris suum larvae from intestinal contents of pigs.

PubMed

Slotved, H C; Barnes, E H; Eriksen, L; Roepstorff, A; Nansen, P; Bjørn, H

1997-01-01

Four groups each of 3 pigs were inoculated with Ascaris suum eggs. Pigs in groups 1 and 3 were inoculated with 1000 eggs, and pigs in groups 2 and 4 with 10,000 eggs. On day 10 and 21 post-inoculation (p.i.), respectively, groups 1 + 2 and 3 + 4 were slaughtered, and the contents from the small intestines collected. The contents were mixed with agar to a final concentration of 1% agar and allowed to sediment. The larvae were allowed to migrate from the agar-gel into 38 degrees C 0.9% saline overnight, and were then collected on a sieve (20 microns mesh) and counted. The larvae retained in the agar-gel were counted after pouring the melted agar through a sieve (20 microns mesh). The results showed that more than 97% of the larvae migrated out of the agar-gel and were available for counting in an almost clean suspension. The inoculation dose level did not significantly affect the recovery percentage, neither did the larval stage (10 or 21 days old larvae). The variation in the time interval from slaughtering to start of incubation (interval 57-155 min) did not significantly affect the recovery percentage.

Development of porcine circovirus 2 (PCV2) open reading frame 2 DNA vaccine with different adjuvants and comparison with commercial PCV2 subunit vaccine in an experimental challenge.

PubMed

Park, Changhoon; Jeong, Jiwoon; Choi, Kyuhyung; Park, Su-Jin; Kang, Ikjae; Chae, Chanhee

2017-07-01

The objective of this study was to compare the protection against challenge with porcine circovirus 2 (PCV2) induced by an experimental vaccine based on open reading frame (ORF) 2 of PCV2 DNA plus an adjuvant (aluminum hydroxide, cobalt oxide, or liposome) and a commercial PCV2 subunit vaccine. A total of 35 colostrum-fed, cross-bred, conventional piglets were randomly divided into 7 groups. The commercial vaccine was more efficacious against PCV2 challenge than the 4 experimental vaccines according to immunologic, virologic, and pathological outcomes. The pigs inoculated with the experimental vaccine containing the liposome adjuvant had significantly higher levels ( P < 0.05) of neutralizing antibodies and interferon-γ-secreting cells, and significantly lower levels ( P < 0.05) of PCV2 viremia than the pigs inoculated with the other experimental vaccines. The pigs inoculated with the experimental vaccines containing either the liposome adjuvant or the cobalt oxide adjuvant had significantly lower lymphoid lesion scores ( P < 0.05) than the pigs in the group inoculated with the PCV2 DNA vaccine dissolved in phosphate-buffered saline. Liposome proved to be a potent adjuvant that efficiently enhanced both humoral and cellular immune responses induced by the PCV2 DNA vaccine.

Development of porcine circovirus 2 (PCV2) open reading frame 2 DNA vaccine with different adjuvants and comparison with commercial PCV2 subunit vaccine in an experimental challenge

PubMed Central

Park, Changhoon; Jeong, Jiwoon; Choi, Kyuhyung; Park, Su-Jin; Kang, Ikjae; Chae, Chanhee

2017-01-01

The objective of this study was to compare the protection against challenge with porcine circovirus 2 (PCV2) induced by an experimental vaccine based on open reading frame (ORF) 2 of PCV2 DNA plus an adjuvant (aluminum hydroxide, cobalt oxide, or liposome) and a commercial PCV2 subunit vaccine. A total of 35 colostrum-fed, cross-bred, conventional piglets were randomly divided into 7 groups. The commercial vaccine was more efficacious against PCV2 challenge than the 4 experimental vaccines according to immunologic, virologic, and pathological outcomes. The pigs inoculated with the experimental vaccine containing the liposome adjuvant had significantly higher levels (P < 0.05) of neutralizing antibodies and interferon-γ-secreting cells, and significantly lower levels (P < 0.05) of PCV2 viremia than the pigs inoculated with the other experimental vaccines. The pigs inoculated with the experimental vaccines containing either the liposome adjuvant or the cobalt oxide adjuvant had significantly lower lymphoid lesion scores (P < 0.05) than the pigs in the group inoculated with the PCV2 DNA vaccine dissolved in phosphate-buffered saline. Liposome proved to be a potent adjuvant that efficiently enhanced both humoral and cellular immune responses induced by the PCV2 DNA vaccine. PMID:28725106

The Plasma-Lyte 148 v Saline (PLUS) study protocol: a multicentre, randomised controlled trial of the effect of intensive care fluid therapy on mortality.

PubMed

Hammond, Naomi E; Bellomo, Rinaldo; Gallagher, Martin; Gattas, David; Glass, Parisa; Mackle, Diane; Micallef, Sharon; Myburgh, John; Saxena, Manoj; Taylor, Colman; Young, Paul; Finfer, Simon

2017-09-01

0.9% sodium chloride (saline) is the most commonly administered resuscitation fluid on a global basis but emerging evidence suggests that its high chloride content may have important adverse effects. To describe the study protocol for the Plasma- Lyte 148 v Saline study, which will test the hypothesis that in critically ill adult patients the use of Plasma-Lyte 148 (a buffered crystalloid solution) for fluid therapy results in different 90-day all-cause mortality when compared with saline. We will conduct this multicentre, blinded, randomised controlled trial in approximately 50 intensive care units in Australia and New Zealand. We will randomly assign 8800 patients to either Plasma-Lyte 148 or saline for all resuscitation fluid, maintenance fluid and compatible drug dilution therapy while in the ICU for up to 90 days after randomisation. The primary outcome is 90-day all-cause mortality; secondary outcomes include mean and peak creatinine concentration, incidence of renal replacement therapy, incidence and duration of vasoactive drug treatment, duration of mechanical ventilation, ICU and hospital length of stay, and quality of life and health services use at 6 months. The PLUS study will provide high-quality data on the comparative safety and efficacy of Plasma-Lyte 148 compared with saline for resuscitation and compatible crystalloid fluid therapy in critically ill adult patients.

Comparative photochemistry activity and antioxidant responses in male and female Populus cathayana cuttings inoculated with arbuscular mycorrhizal fungi under salt

PubMed Central

Wu, Na; Li, Zhen; Wu, Fei; Tang, Ming

2016-01-01

We investigated the impact of arbuscular mycorrhizal fungi (AMF) on the morphology and physiology of two genders of the typical dioecious plant Populus cathayana under salt stress. We conducted a pot experiment containing seedlings of the two genders that were subjected to salt or non-salt and filled with soil that was either inoculated with Rhizophagus intraradices or not. The results showed that males had higher mycorrhizal dependency than females. Salt stress decreased growth, the relative water content and chlorophyll fluorescence. Meanwhile, salt increased the superoxide radical (O2−), and hydrogen peroxide (H2O2) contents and antioxidant enzyme activities. Mycorrhizal male seedlings performed better than females in shoot morphological growth under both conditions and in chlorophyll fluorescence parameters, O2− and H2O2 contents, MDA concentration, proline content and antioxidant enzymes activities under salt stress. In females, under saline conditions, a lower MDA concentration and H2O2, O2− and proline contents were observed in the leaves and roots. In addition, inoculated female plants performed better in chlorophyll fluorescence parameters than non-inoculated plants. AMF inoculation had either slight or no effects on the performance of females. These findings suggested that when subjected to stress and AMF, differences in the genders existed, followed by the alleviation of the damage to P. cathayana by AMF via improving growth and photosynthesis and antioxidant systems under salt stress. PMID:27898056

Influence of Bacillus spp. strains on seedling growth and physiological parameters of sorghum under moisture stress conditions.

PubMed

Grover, Minakshi; Madhubala, R; Ali, Sk Z; Yadav, S K; Venkateswarlu, B

2014-09-01

Microorganisms isolated from stressed ecosystem may prove as ideal candidates for development of bio-inoculants for stressed agricultural production systems. In the present study, moisture stress tolerant rhizobacteria were isolated from the rhizosphere of sorghum, pigeonpea, and cowpea grown under semiarid conditions in India. Four isolates KB122, KB129, KB133, and KB142 from sorghum rhizosphere exhibited plant growth promoting traits and tolerance to salinity, high temperature, and moisture stress. These isolates were identified as Bacillus spp. by 16S rDNA sequence analysis. The strains were evaluated for growth promotion of sorghum seedlings under two different moisture stress conditions (set-I, continuous 50% soil water holding capacity (WHC) throughout the experiment and set-II, 75% soil WHC for 27 days followed by no irrigation for 5 days) under greenhouse conditions. Plate count and scanning electron microscope studies indicated successful root surface colonization by inoculated bacteria. Plants inoculated with Bacillus spp. strains showed better growth in terms of shoot length and root biomass with dark greenish leaves due to high chlorophyll content while un-inoculated plants showed rolling of the leaves, stunted appearance, and wilting under both stress conditions. Inoculation also improved leaf relative water content and soil moisture content. However, variation in proline and sugar content in the different treatments under two stress conditions indicated differential effect of microbial treatments on plant physiological parameters under stress conditions. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Physiological and biochemical characterization of Azospirillum brasilense strains commonly used as plant growth-promoting rhizobacteria.

PubMed

Di Salvo, Luciana P; Silva, Esdras; Teixeira, Kátia R S; Cote, Rosalba Esquivel; Pereyra, M Alejandra; García de Salamone, Inés E

2014-12-01

Azospirillum is a plant growth-promoting rhizobacteria (PGPR) genus vastly studied and utilized as agriculture inoculants. Isolation of new strains under different environmental conditions allows the access to the genetic diversity and improves the success of inoculation procedures. Historically, the isolation of this genus has been performed by the use of some traditional culture media. In this work we characterized the physiology and biochemistry of five different A. brasilense strains, commonly used as cereal inoculants. The aim of this work is to contribute to pose into revision some concepts concerning the most used protocols to isolate and characterize this bacterium. We characterized their growth in different traditional and non-traditional culture media, evaluated some PGPR mechanisms and characterized their profiles of fatty acid methyl esters and carbon-source utilization. This work shows, for the first time, differences in both profiles, and ACC deaminase activity of A. brasilense strains. Also, we show unexpected results obtained in some of the evaluated culture media. Results obtained here and an exhaustive knowledge revision revealed that it is not appropriate to conclude about bacterial species without analyzing several strains. Also, it is necessary to continue developing studies and laboratory techniques to improve the isolation and characterization protocols. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Influence of arbuscular mycorrhizal fungi and treated wastewater on water relations and leaf structure alterations of Viburnum tinus L. plants during both saline and recovery periods.

PubMed

Gómez-Bellot, María José; Nortes, Pedro Antonio; Ortuño, María Fernanda; Romero, Cristina; Fernández-García, Nieves; Sánchez-Blanco, María Jesús

2015-09-01

Nowadays, irrigation with low quality water is becoming an alternative to satisfy the needs of crops. However, some plant species have to deal with high salinity of reclaimed water, by adapting their physiological behaviour during both saline and recovery periods and developing morphological changes in their leaves. The application of arbuscular mycorrhizal fungi (AMF) could also be a suitable option to mitigate the negative effects of this kind of water, although the effectiveness of plant-AMF association is influenced by many factors. In this work, during forty weeks, the combined effect of Glomus iranicum var. tenuihypharum and two types of water: control, C, EC<0.9 dS m(-1) and reclaimed water, RW (with EC: 4 dS m(-1) during a first saline period and EC: 6 dS m(-1) during a second saline period) was evaluated for laurustinus plants (Viburnum tinus L.) transplanted in soil. This was followed by a recovery period of eight weeks, when all the plants were irrigated in the control irrigation conditions. Seasonal and daily changes in stem water potential (Ψstem), stomatal conductance (gs), photosynthesis (Pn) and leaf internal CO2 concentration (Ci) of laurustinus plants were evaluated. Leaf structure alterations, nutrient imbalance, height and leaf hydraulic conductivity (Kleaf) were also determined. Due to the high difficulty of absorbing water from the soil, RW plants showed a high volumetric water content (θv) in soil. The stem water potential and the stomatal conductance (gs) values were reduced in RW plants throughout the second saline period. These decreases were also found during the day. Leaf Ca(2+)/Na(+) and K(+)/Na(+) ratios diminished in RW plants respect to the C plants due to the Na(+) accumulation, although height and chlorophyll content values did not show statistical differences. Leaves from RW plants showed a significantly thicker mesophyll than Control leaves as a consequence of high EC. The area of palisade parenchyma (PP) increased while the area of spongy parenchyma (SP) decreased in RW leaves with respect to the C leaves. These structural changes could be considered as a strategy to maximize photosynthesis potential in saline conditions. Mycorrhizal inoculation improved the water status of both C and RW plants by increasing their Ψstem and gs values. As regards leaf structure, AMF showed an opposite effect to salinity for PP and SP. At the end of the recovery period, hardly any statistical differences of physiological parameters were found between treatments, although a tendency to improve them was observed in inoculated plants. In any case, the leaf structural changes and the great reduction in Kleaf observed at Ψleaf below -1.5 MPa would constitute an important mechanism for laurustinus plants to reduce the water loses produced by salinity. Copyright © 2015 Elsevier GmbH. All rights reserved.

Remediation of saline soils contaminated with crude oil using the halophyte Salicornia persica in conjunction with hydrocarbon-degrading bacteria.

PubMed

Ebadi, Ali; Khoshkholgh Sima, Nayer Azam; Olamaee, Mohsen; Hashemi, Maryam; Ghorbani Nasrabadi, Reza

2018-08-01

The negative impact of salinity on plant growth and the survival of rhizosphere biota complicates the application of bioremediation to crude oil-contaminated saline soils. Here, a comparison was made between the remedial effect of treating the soil with Pseudomonas aeruginosa, a salinity tolerant hydrocarbon-degrading consortium in conjunction with either the halophyte Salicornia persica or the non-halophyte Festuca arundinacea. The effect of the various treatments on salinized soils was measured by assessing the extent of total petroleum hydrocarbon (TPH) degradation, the soil's dehydrogenase activity, the abundance of the bacteria and the level of phytotoxicity as measured by a bioassay. When a non-salinized soil was assessed after a treatment period of 120 days, the ranking for effectiveness with respect to TPH removal was F. arundinacea > P. aeruginosa > S. persica > no treatment control, while in the presence of salinity, the ranking changed to S. persica > P. aeruginosa > F. arundinacea > no treatment control. Combining the planting of S. persica or F. arundinacea with P. aeruginosa inoculation ("bioaugmentation") boosted the degradation of TPH up to 5-17%. Analyses of the residual oil contamination revealed that long chain alkanes (above C20) were particularly strongly degraded following the bioaugmentation treatments. The induced increase in dehydrogenase activity and the abundance of the bacteria (3.5 and 10 fold respectively) achieved in the bioaugmentation/S. persica treatment resulted in 46-76% reduction in soil phytotoxicity in a saline soil. The indication was that bioaugmentation of halophyte can help to mitigate the adverse effects on the effectiveness of bioremediation in a crude oil-contaminated saline soil. Copyright © 2018 Elsevier Ltd. All rights reserved.

Marine Bacteria Cause False-Positive Results in the Colilert-18 Rapid Identification Test for Escherichia coli in Florida Waters

PubMed Central

Pisciotta, John M.; Rath, Damon F.; Stanek, Paul A.; Flanery, D. Michael; Harwood, Valerie J.

2002-01-01

The Colilert-18 system for enumeration of total coliforms and Escherichia coli is approved by the U.S. Environmental Protection Agency for use in drinking water analysis and is also used by various agencies and research studies for enumeration of indicator organisms in fresh and saline waters. During monitoring of Pinellas County, Fla., marine waters, estimates of E. coli numbers (by Colilert-18) frequently exceeded fecal coliform counts (by membrane filtration) by 1 to 3 orders of magnitude. Samples from freshwater sites did not display similar discrepancies. Fecal coliforms, including E. coli, could be cultured from 100% of yellow fluorescent wells (denoting E. coli-positive results) inoculated with freshwater samples but could be cultured from only 17.1% of the “positive” wells inoculated with marine samples. Ortho-nitrophenyl-β-d-galactopyranoside (ONPG)-positive or 4-methylumbelliferyl-β-d-glucuronide (MUG)-positive noncoliform bacteria were readily cultured from Colilert-18 test wells inoculated with marine samples. Filtered cell-free seawater did not cause false positives. Coculture preparations of as few as 5 CFU of Vibrio cholerae (ONPG positive) and Providencia sp. (MUG positive) ml−1 inoculated into Colilert-18 caused false-positive E. coli results. Salinity conditions influenced coculture results, as the concentration of coculture inoculum required to cause false positives in most wells increased from about 5 CFU ml−1 in seawater diluted 1:10 with freshwater to ≈5,000 CFU ml−1 in seawater diluted 1:20 with freshwater. Estimated E. coli numbers in various marine water samples processed at the 1:10 dilution ranged from 10 to 7,270 CFU·100 ml−1, while E. coli numbers in the same samples processed at the 1:20 dilution did not exceed 40 CFU·100 ml−1. The lower estimates of E. coli numbers corresponded well with fecal coliform counts by membrane filtration. This study indicates that assessment of E. coli in subtropical marine waters by Colilert-18 is not accurate when the recommended 1:10 sample dilution is used. The results suggest that greater dilution may diminish the false-positive problem, but further study of this possibility is recommended. PMID:11823188

Intravenous versus intramuscular cobinamide compared to intravenous saline (control) in the treatment of acute, survivable, mitochondrial toxins in swine (Sus Scrofa): a pilot study

DTIC Science & Technology

2018-04-10

Type of Research: Animal Research 3. Title: Intravenous versus intramuscular cobinamide compared to intravenous saline ( control ) in the treatment...the hyperkalemia under control and in our upcoming protocol we feel we will finally be able to induce apnea with the toxin and calcium channel...intramuscular cobinamide compared to intravenous saline ( control ) in the treatment of acute, survivable, mitochondrial toxins in swine (Sus Scrofa): a pilot

Inoculation of Soil with Plant Growth Promoting Bacteria Producing 1-Aminocyclopropane-1-Carboxylate Deaminase or Expression of the Corresponding acdS Gene in Transgenic Plants Increases Salinity Tolerance in Camelina sativa

PubMed Central

Heydarian, Zohreh; Yu, Min; Gruber, Margaret; Glick, Bernard R.; Zhou, Rong; Hegedus, Dwayne D.

2016-01-01

Camelina sativa (camelina) is an oilseed crop touted for use on marginal lands; however, it is no more tolerant of soil salinity than traditional crops, such as canola. Plant growth-promoting bacteria (PGPB) that produce 1-aminocyclopropane-1-carboxylate deaminase (ACC deaminase) facilitate plant growth in the presence of abiotic stresses by reducing stress ethylene. Rhizospheric and endophytic PGPB and the corresponding acdS- mutants of the latter were examined for their ability to enhance tolerance to salt in camelina. Stimulation of growth and tolerance to salt was correlated with ACC deaminase production. Inoculation of soil with wild-type PGPB led to increased shoot length in the absence of salt, and increased seed production by approximately 30–50% under moderately saline conditions. The effect of ACC deaminase was further examined in transgenic camelina expressing a bacterial gene encoding ACC deaminase (acdS) under the regulation of the CaMV 35S promoter or the root-specific rolD promoter. Lines expressing acdS, in particular those using the rolD promoter, showed less decline in root length and weight, increased seed production, better seed quality and higher levels of seed oil production under salt stress. This study clearly demonstrates the potential benefit of using either PGPB that produce ACC deaminase or transgenic plants expressing the acdS gene under the control of a root-specific promoter to facilitate plant growth, seed production and seed quality on land that is not normally suitable for the majority of crops due to high salt content. PMID:28018305

Effects of Thrips Density, Mode of Inoculation, and Plant Age on Tomato Spotted Wilt Virus Transmission in Peanut Plants.

PubMed

Shrestha, Anita; Sundaraj, Sivamani; Culbreath, Albert K; Riley, David G; Abney, Mark R; Srinivasan, Rajagopalbabu

2015-02-01

Spotted wilt caused by tomato spotted wilt virus (TSWV; family Bunyaviridae; genus Tospovirus) is a serious disease of peanut (Arachis hypogaea L.) in the southeastern United States. Peanut genotypes with field resistance to TSWV are effective in suppressing spotted wilt. All commercially available genotypes with field resistance to TSWV were developed through conventional breeding. As a part of the breeding process, peanut genotypes are regularly screened under field situations. Despite numerous advantages associated with field screening, it is often limited by inconsistent vector (thrips) and TSWV pressure. A greenhouse transmission protocol would aid in thorough screening of selected genotypes and conserve time. In this study, various parameters associated with TSWV transmission, including tobacco thrips, Frankliniella fusca (Hinds) density, mode of inoculation, and plant age, were evaluated. Greater incidences of TSWV infection were obtained with thrips-mediated inoculation when compared with mechanical inoculation. TSWV inoculation with three, five, and 10 thrips resulted in greater incidences of TSWV infection in plants than inoculation with one thrips. However, incidences of TSWV infection did not vary between plants inoculated with three, five, and 10 viruliferous thrips. With both thrips-mediated and mechanical inoculation methods, incidences of TSWV infection in 1-wk-old plants were greater than in 4-wk-old plants. TSWV copy numbers, as determined by qPCR, also decreased with plant age. Results suggest that using at least three thrips per plant and 1- to 2-wk-old plants would maximize TSWV infection in inoculated plants. © The Author 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Assessing the viability of microorganisms in the ballast water of vessels transiting the North Atlantic Ocean.

PubMed

Steichen, Jamie L; Quigg, Antonietta

2015-12-15

Testing phytoplankton viability within ballast tanks and receiving waters of ballast water discharge remain understudied. Potentially harmful dinoflagellates and diatoms are transported via ballast water to Galveston Bay, Texas (USA), home to three major ports: Houston, Texas City and Galveston. Ballast water from vessels transiting the North Atlantic Ocean was inoculated into treatments representing low and high salinity conditions similar to the Ports of Houston and Galveston respectively. Phytoplankton in ballast water growout experiments were deemed viable and showed growth in low and mid salinities with nutrient enrichment. Molecular methods identified several genera: Dinophysis, Gymnodinium, Gyrodinium, Heterocapsa, Peridinium, Scrippsiella, Chaetoceros and Nitzschia. These phytoplankton genera were previously identified in Galveston Bay except Scrippsiella. Phytoplankton, including those capable of forming harmful algal blooms leading to fish and shellfish kills, are transported to Galveston Bay via ballast water, and are viable when introduced to similar salinity conditions found in Galveston Bay ports. Copyright © 2015 Elsevier Ltd. All rights reserved.

Adjuvant Activity of a Novel Metabolizable Lipid Emulsion with Inactivated Viral Vaccines

DTIC Science & Technology

1980-06-01

glycerol and lecithin . Hamsters and mice inoculated with lipid 0 emulsion plus western or Venezuelan equine encephalitis vaccine were signifi- cantly...antibody titer. soybean lecithin (Centrolex-F, Central Soya, Ft. Immunization and challenge studies. Mice or Wayne, Ind.) was dissolved in 10 parts...vaccine combined a sterile Teflon magnetic bar. The glycerol and lecithin with LE or saline as shown in the tables of individual mixture was transferred

Virological and clinico-pathological features of orf virus infection in experimentally infected rabbits and mice.

PubMed

Cargnelutti, J F; Masuda, E K; Martins, M; Diel, D G; Rock, D L; Weiblen, R; Flores, E F

2011-01-01

Many aspects of the biology of orf virus (ORFV) infection remain poorly understood and attempts to establish animal models have yielded conflicting and non-reproducible results. We herein describe the characterization of ORFV infection and disease in rabbits and mice. A protocol of intradermal inoculation was employed to inoculate 10(8.5)TCID₅₀/mL of ORFV strain IA-82 in the skin of ears, of the back and labial commissures. All inoculated rabbits presented a clinical course characterized by erythema, macules, papules/vesicles or pustules that eventually dried originating scabs. Local signs started around days 3 and 4 post-inoculation (pi) and lasted 3-10 days. Virus was recovered from lesions between days 2 and 14pi. Histological examination of lesions revealed focal proliferative dermatitis with ballooning degeneration and eosinophilic intracytoplasmic inclusion bodies in keratinocytes, histological hallmarks of contagious ecthyma in sheep. A similar, albeit milder clinical course occurred in 5/10 inoculated mice; virus was recovered from lesions from three animals. Inoculated lambs - used as controls - developed severe lesions of contagious ecthyma. VN tests performed at day 28pi failed to detect neutralizing antibodies in all inoculated animals. In contrast, convalescent rabbit sera were positive by ELISA at dilutions from 100 to 400. These results show that rabbits are susceptible to ORFV infection and thus may be used to study selected aspects of ORFV biology. Copyright © 2010 Elsevier Ltd. All rights reserved.

Oxygen Mass Transfer Coefficient (k sub L a’) Consideration for Scale-Up Fermentation Systems at the Biotechnology Laboratory U.S. Army Edgewood Chemical Biological Center

DTIC Science & Technology

2011-02-01

30 °C Stock vials were prepared following standard microbial protocol. A single colony from the overnight nutrient agar plate was inoculated into...500-mL seed cultures were prepared by inoculating with 2 vials of frozen stock and incubating at 30 °C and 200 rpm until the OD reached approximately...and OD profiles vs. elapsed fermentation time ( EFT ) from 20- and 1000-L scales are shown in Figure 9. In both runs, no DO limitation was apparent, and

Effect of High Dietary Manganese on the Immune Responses of Broilers Following Oral Salmonella typhimurium Inoculation.

PubMed

Pan, Shuqin; Zhang, Keying; Ding, Xuemei; Wang, Jianping; Peng, Huanwei; Zeng, Qiufeng; Xuan, Yue; Su, Zuowei; Wu, Bing; Bai, Shiping

2018-02-01

Manganese (Mn) is an essential nutrient for both host and pathogen. Recent studies have demonstrated the nutritional immunity of Mn against Salmonella infection in mammals. To investigate the effect of high dietary Mn on immune responses of broilers following Salmonella challenge, 144 1-day-old male broilers were fed a basal diet (containing 20.04 mg Mn/kg) plus an additional 40 (the control group) or 400 mg Mn/kg (the H-Mn group) for 7 days. The 72 broilers in each group were then orally inoculated with 5 × 10 7 CFUs of Salmonella typhimurium (ATCC#14028) or phosphate-buffered saline. Peripheral blood, spleens, cecal tonsils, and bursa of Fabricius were collected from Salmonella-inoculated and Salmonella-noninoculated broilers (n = 6) at 2 days post inoculation (2 DPI) and 7 days post inoculation (7 DPI). Peripheral blood lymphocyte subpopulations were determined by flow cytometry. The messenger RNA (mRNA) abundance of genes was determined by quantitative real-time polymerase chain reaction. Salmonella counts were higher (P < 0.05) in the H-Mn group than that in the control group at 2 DPI in the cecal contents of Salmonella-inoculated broilers. High dietary Mn increased CD3 + CD4 + and CD3 + CD8 + percentages in the peripheral blood of Salmonella-inoculated broilers at 2 DPI. Salmonella inoculation increased interleukin (IL)-6 mRNA expression in spleens and bursa of Fabricius at 2 DPI and increased IL-1β and IL-6 mRNA expression in cecal tonsils at 7 DPI in the H-Mn group. These changes were not observed in the control group. High dietary Mn increased interferon-γ (IFN-γ) in spleens and decreased IFN-γ and IL-12 mRNA expression in cecal tonsils of Salmonella-inoculated broilers at 2 DPI. High dietary Mn decreased IL-17 mRNA expression in the bursa of Fabricius at 7 DPI, but increased this expression in cecal tonsils at 2 and 7 DPI in Salmonella-inoculated broilers. These results suggested that dietary Mn level affected T helper (Th) 1-cytokine reaction in spleens and cecal tonsils, and Th17-mediated immunity in cecal tonsils and the bursa of Fabricius of broilers when challenged with Salmonella.

Halotolerant plant-growth promoting rhizobacteria modulate gene expression and osmolyte production to improve salinity tolerance and growth in Capsicum annum L.

PubMed

Yasin, Nasim Ahmad; Akram, Waheed; Khan, Waheed Ullah; Ahmad, Sajid Rashid; Ahmad, Aqeel; Ali, Aamir

2018-06-04

Some rhizobacteria have demonstrated a noteworthy role in regulation of plant growth and biomass production under biotic and abiotic stresses. The present study was intended to explicate the ameliorative consequences of halotolerant plant growth-promoting rhizobacteria (HPGPR) on growth of capsicum plants subjected to salt stress. Salt stress was ascertained by supplementing 1 and 2 g NaCl kg -1 soil. The HPGPR positively invigorated growth attributes, chlorophyll, protein contents, and water use efficiency (WUE) of supplemented capsicum plants under salinity stress conditions. Bacillus fortis strain SSB21 caused highest significant increase in shoot length, root length, and fresh and dry biomass production of capsicum plants grown under saline conditions. This multi-trait bacterium also increased biosynthesis of proline and up-regulated the expression profiles of stress related genes including CAPIP2, CaKR1, CaOSM1, and CAChi2. On the other hand, B. fortis strain SSB21 inoculated plants exhibited reduced level of ethylene, lipid peroxidation, and reactive oxygen species (ROS). All these together contribute to activate physiological and biochemical processes involved in the mitigation of the salinity induced stress in capsicum plants.

Effects of different remediation treatments on crude oil contaminated saline soil.

PubMed

Gao, Yong-Chao; Guo, Shu-Hai; Wang, Jia-Ning; Li, Dan; Wang, Hui; Zeng, De-Hui

2014-12-01

Remediation of the petroleum contaminated soil is essential to maintain the sustainable development of soil ecosystem. Bioremediation using microorganisms and plants is a promising method for the degradation of crude oil contaminants. The effects of different remediation treatments, including nitrogen addition, Suaeda salsa planting, and arbuscular mycorrhiza (AM) fungi inoculation individually or combined, on crude oil contaminated saline soil were assessed using a microcosm experiment. The results showed that different remediation treatments significantly affected the physicochemical properties, oil contaminant degradation and bacterial community structure of the oil contaminated saline soil. Nitrogen addition stimulated the degradation of total petroleum hydrocarbon significantly at the initial 30d of remediation. Coupling of different remediation techniques was more effective in degrading crude oil contaminants. Applications of nitrogen, AM fungi and their combination enhanced the phytoremediation efficiency of S. salsa significantly. The main bacterial community composition in the crude oil contaminated saline soil shifted with the remediation processes. γ-Proteobacteria, β-Proteobacteria, and Actinobacteria were the pioneer oil-degraders at the initial stage, and Firmicutes were considered to be able to degrade the recalcitrant components at the later stage. Copyright © 2014 Elsevier Ltd. All rights reserved.

Swabs as a tool for monitoring the presence of norovirus on environmental surfaces in the food industry.

PubMed

Rönnqvist, Maria; Rättö, Marjaana; Tuominen, Pirkko; Salo, Satu; Maunula, Leena

2013-08-01

Human norovirus (HuNoV), which causes gastroenteritis, can be transmitted to food and food contact surfaces via viruscontaminated hands. To investigate this transmission in food processing environments, we developed a swabbing protocol for environmental samples, evaluated the stability of HuNoV in the swabs, and applied the method in the food industry. Swabs made of polyester, flocked nylon, cotton wool, and microfiber were moistened in either phosphate-buffered saline (PBS) or glycine buffer (pH 9.5) and used to swab four surfaces (latex, plastic, stainless steel, and cucumber) inoculated with HuNoV. HuNoV was eluted with either PBS or glycine buffer and detected with quantitative reverse transcription PCR. HuNoV recoveries were generally higher with an inoculation dose of 100 PCR units than 1,000 PCR units. The highest recoveries were obtained when surfaces were swabbed with microfiber cloth moistened in and eluted with glycine buffer after a HuNoV inoculation dose of 100 PCR units: 66% ± 18% on latex, 89% ±2% on plastic, and 79% ±10% on stainless steel. The highest recovery for cucumber, 45% ±5%, was obtained when swabbing the surface with microfiber cloth and PBS. The stability of HuNoV was tested in microfiber cloths moistened in PBS or glycine buffer. HuNoV RNA was detected from swabs after 3 days at 4 and 22°C, although the RNA levels decreased more rapidly in swabs moistened with glycine buffer than in those moistened with PBS at 22°C. In the field study, 172 microfiber and 45 cotton wool swab samples were taken from environmental surfaces at three food processing companies. Five (5.6%) of 90 swabs collected in 2010 and 7 (8.5%) of 82 swabs collected in 2012 were positive for HuNoV genogroup II; all positive samples were collected with microfiber swabs. Three positive results were obtained from the production line and nine were obtained from the food workers' break room and restroom areas. Swabbing is a powerful tool for HuNoV RNA detection from environmental surfaces and enables investigation of virus transmission during food processing.

Establishing fungal entomopathogens as endophytes: towards endophytic biological control

USDA-ARS?s Scientific Manuscript database

Beauveria basssiana is a fungal entomopathogen with the ability to colonize plants endophytically. As an endophyte, B. bassiana may play a role in protecting plants from herbivory and disease. This protocol demonstrates two inoculation methods to establish B. bassiana endophytically in the common be...

Fruit body formation on silkworm by Cordyceps militaris

USDA-ARS?s Scientific Manuscript database

Injection inoculation protocols for fruit body formation of Cordyceps militaris were investigated to improve the incidence of infection in the silkworm species Bombyx mori. Injection, with suspensions of C. militaris hyphal bodies into living silkworm pupae, was used to test for fruit body productio...

Establishing Mouse Models for Zika Virus-induced Neurological Disorders Using Intracerebral Injection Strategies: Embryonic, Neonatal, and Adult.

PubMed

Herrlinger, Stephanie A; Shao, Qiang; Ma, Li; Brindley, Melinda; Chen, Jian-Fu

2018-04-26

The Zika virus (ZIKV) is a flavivirus currently endemic in North, Central, and South America. It is now established that the ZIKV can cause microcephaly and additional brain abnormalities. However, the mechanism underlying the pathogenesis of ZIKV in the developing brain remains unclear. Intracerebral surgical methods are frequently used in neuroscience research to address questions about both normal and abnormal brain development and brain function. This protocol utilizes classical surgical techniques and describes methods that allow one to model ZIKV-associated human neurological disease in the mouse nervous system. While direct brain inoculation does not model the normal mode of virus transmission, the method allows investigators to ask targeted questions concerning the consequence after ZIKV infection of the developing brain. This protocol describes embryonic, neonatal, and adult stages of intraventricular inoculation of ZIKV. Once mastered, this method can become a straightforward and reproducible technique that only takes a few hours to perform.

Application of bacteria from non-cultivated plants to promote growth, alter root architecture and alleviate salt stress of cotton.

PubMed

Irizarry, I; White, J F

2017-04-01

Cotton seeds are frequently treated with acid to remove fibres and reduce seed-transmitted diseases. This process also eliminates beneficial bacteria on the seed surface. The goal of this research was to seek and apply beneficial bacteria to acid delinted cotton seeds to evaluate their growth-promoting and salt stress alleviating effects in seedlings. Bacteria were isolated from non-cultivated plants in the Malvaceae. Seeds were collected from Portia tree (Thespesia populnea) and wild cotton (Gossypium hirsutum) from coastal and arid areas of Puerto Rico. Bacillus amyloliquefaciens, Curtobacterium oceanosedimentum and Pseudomonas oryzihabitans were inoculated onto acid delinted cotton seeds. Bacteria increased cotton seed germination and length of emerging seedling radicles. Cotton seeds were inoculated with B. amyloliquefaciens to evaluate growth and root architecture of non-stressed and salt stressed seedlings. Inoculating cotton seeds with B. amyloliquefaciens led to a greater percentage of seedlings with expanded cotyledons after 8 days, enhanced primary and lateral root growth, and altered root architecture. Similar results were obtained when okra seeds were inoculated with B. amyloliquefaciens. The data supported the hypothesis that non-cultivated plants in the Malvaceae growing in stressful environments possess bacteria that promote growth, alter root architecture and alleviate salt stress of cotton and okra seedlings. This study demonstrated the effects of applying beneficial bacteria on acid delinted cotton seeds. Inoculating seeds with salt stress alleviating bacteria could improve the growth of crop seedlings that are vulnerable to soil salinization. © 2017 The Society for Applied Microbiology.

Induction of tumor necrosis factor by Legionella pneumophila.

PubMed Central

Blanchard, D K; Djeu, J Y; Klein, T W; Friedman, H; Stewart, W E

1987-01-01

Mice were inoculated with Legionella pneumophila via an intratracheal route to establish an experimental model of infection. Lung lavage fluid obtained from infected mice contained a cytolytic factor identified as tumor necrosis factor (TNF). Peak levels of TNF were produced at about 24 h postinfection and rapidly declined thereafter. Treatment of the mice with dextran sulfate before inoculation with the bacteria resulted in lowered amounts of TNF in the lung lavage fluid, suggesting that macrophages were responsible for production of the cytokine. Furthermore, cultures of adherent lung leukocytes and a macrophage cell line, PU 5-1.8, were stimulated to produce TNF by exposure to Legionella antigens. In addition, adherent lung leukocytes from Legionella-infected mice spontaneously released TNF into the culture supernatant. Inoculation of mice with saline or latex particles failed to induce TNF in vivo, indicating that bacterial antigens or products were the stimulating signals. Since there was no detectable TNF activity in sera at any time after intratracheal inoculation, TNF production appeared to be confined to the site of infection. Pretreatment of PU 5-1.8 cultures with gamma interferon, which was detected in the lung lavage fluid before TNF, resulted in augmented TNF production, suggesting cooperativity may exist between the two cytokines, either in the pathogenicity of the bacterium or in a possible immunomodulatory function of TNF and interferon during infection. PMID:2433220

Evaluation of a Biological Pathogen Decontamination Protocol for Animal Feed Mills.

PubMed

Huss, Anne R; Cochrane, Roger A; Deliephan, Aiswariya; Stark, Charles R; Jones, Cassandra K

2015-09-01

Animal feed and ingredients are potential vectors of pathogenic bacteria. Contaminated ingredients can contaminate facility equipment, leading to cross-contamination of other products. This experiment was conducted to evaluate a standardized protocol for decontamination of an animal feed manufacturing facility using Enterococcus faecium (ATCC 31282) as an indicator. A pelleted swine diet inoculated with E. faecium was manufactured, and environmental samples (swabs, replicate organism detection and counting plates, and air samples) were collected (i) before inoculation (baseline data), (ii) after production of inoculated feed, (iii) after physical removal of organic material using pressurized air, (iv) after application of a chemical sanitizer containing a quaternary ammonium-glutaraldehyde blend, (v) after application of a chemical sanitizer containing sodium hypochlorite, (vi) after facility heat-up to 60 8 C for 24 h, (vii) for 48 h, and (viii) for 72 h. Air samples collected outside the facility confirmed pathogen containment; E. faecium levels were equal to or lower than baseline levels at each sample location. The decontamination step and its associated interactions were the only variables that affected E. faecium incidence (P < 0.0001 versus P > 0.22). After production of the inoculated diet, 85.7% of environmental samples were positive for E. faecium. Physical cleaning of equipment had no effect on contamination (P = 0.32). Chemical cleaning with a quaternary ammonium-glutaraldehyde blend and sodium hypochlorite each significantly reduced E. faecium contamination (P < 0.0001) to 28.6 and 2.4% of tested surfaces, respectively. All samples were negative for E. faecium after 48 h of heating. Both wet chemical cleaning and facility heating but not physical cleaning resulted in substantial E. faecium decontamination. These results confirmed both successful containment and decontamination of biological pathogens in the tested pilot-scale feed mill.

A segmental chronic pain syndrome in rats associated with intrathecal infusion of NMDA: evidence for selective action in the dorsal horn.

PubMed

Zochodne, D W; Murray, M; Nag, S; Riopelle, R J

1994-02-01

We explored the effects of chronic lumbar intrathecal NMDA infusion (mini-osmotic pumps) in Sprague-Dawley rats on motor and sensory axon integrity. Several different infusion protocols, each given over a 4 week period were examined: 0.15 M NMDA in phosphate buffered saline; phosphate buffered saline without NMDA; and 0.20 M magnesium sulfate plus 0.15 M NMDA; 0.35 M NMDA. In two additional protocols, 0.15 M NMDA or phosphate buffered saline were infused for a total of 8 weeks. Within 1-2 weeks of the onset of NMDA, but not phosphate buffered saline infusions, the rats exhibited irritability, circling, biting and excessive grooming resulting in loss of hair, and skin ulcerations from autotomy localized to lumbar and sacral innervated dermatomes. Co-infusion of NMDA with magnesium sulfate almost completely prevented these findings. The behavioural changes were not associated with abnormalities of sensory or motor conduction. Intrathecal infusion of NMDA induces a chronic "central" experimental pain disorder in rats, localized to the cord segment with the greatest exposure to the infusion, without involvement of peripheral sensory axons and sparing the axonal integrity of anterior horn cells.

Plant growth promotion properties of bacterial strains isolated from the rhizosphere of the Jerusalem artichoke (Helianthus tuberosus L.) adapted to saline-alkaline soils and their effect on wheat growth.

PubMed

Liu, Xiaolin; Li, Xiangyue; Li, Yan; Li, Runzhi; Xie, Zhihong

2017-03-01

The Jerusalem artichoke (JA; Helianthus tuberosus), known to be tolerant to saline-alkaline soil conditions, has been cultivated for many years in the Yellow River delta, Shandong Province coastal zone, in China. The aim of our study was to isolate nitrogen-fixing bacteria colonizing the rhizosphere of JA and to characterize other plant growth promotion properties. The ultimate goal was to identify isolates that could be used as inoculants benefiting an economic crop, in particular for improving wheat growth production in the Yellow River delta. Bacterial strains were isolated from the rhizosphere soil of JA on the basis of growth on nitrogen-free Ashby medium. Identification and phylogenetic analysis was performed after nucleotide sequencing of 16S rRNA gene. Plant-growth-promoting traits, such as nitrogen fixation activity, phosphate solubilization activity, indole-3-acetic acid production, were determined using conventional methods. Eleven strains were isolated and 6 of them were further examined for their level of salt tolerance and their effect on plant growth promotion. Inoculation of Enterobacter sp. strain N10 on JA and wheat led to significant increases in both root and shoot dry mass and shoot height. Enterobacter sp. strain N10 appeared to be the best plant-growth-promoting rhizobacteria to increase wheat productivity in future field applications.

Optimization of 64-MDCT urography: effect of dual-phase imaging with furosemide on collecting system opacification and radiation dose.

PubMed

Portnoy, Orith; Guranda, Larisa; Apter, Sara; Eiss, David; Amitai, Marianne Michal; Konen, Eli

2011-11-01

The purpose of this study was to compare opacification of the urinary collecting system and radiation dose associated with three-phase 64-MDCT urographic protocols and those associated with a split-bolus dual-phase protocol including furosemide. Images from 150 CT urographic examinations performed with three scanning protocols were retrospectively evaluated. Group A consisted of 50 sequentially registered patients who underwent a three-phase protocol with saline infusion. Group B consisted of 50 sequentially registered patients who underwent a reduced-radiation three-phase protocol with saline. Group C consisted of 50 sequentially registered patients who underwent a dual-phase split-bolus protocol that included a low-dose furosemide injection. Opacification of the urinary collecting system was evaluated with segmental binary scoring. Contrast artifacts were evaluated, and radiation doses were recorded. Results were compared by analysis of variance. A significant reduction in mean effective radiation dose was found between groups A and B (p < 0.001) and between groups B and C (p < 0.001), resulting in 65% reduction between groups A and C (p < 0.001). This reduction did not significantly affect opacification score in any of the 12 urinary segments (p = 0.079). In addition, dense contrast artifacts overlying the renal parenchyma observed with the three-phase protocols (groups A and B) were avoided with the dual-phase protocol (group C) (p < 0.001). A dual-phase protocol with furosemide injection is the preferable technique for CT urography. In comparison with commonly used three-phase protocols, the dual-phase protocol significantly reduces radiation exposure dose without reduction in image quality.

Development of a greenhouse-based inoculation protocol for the fungus Colletotrichum cereale pathogenic to annual bluegrass (Poa annua)

USDA-ARS?s Scientific Manuscript database

The fungus Colletotrichum cereale incites anthracnose disease on Poa annua (annual bluegrass) turfgrass. Anthracnose disease is geographically widespread highly destructive, with infections by C. cereale resulting in extensive turfgrass loss. Comprehensive research aimed at controlling turfgrass a...

Plant Growth Promoting Rhizobacteria in Amelioration of Salinity Stress: A Systems Biology Perspective

PubMed Central

Ilangumaran, Gayathri; Smith, Donald L.

2017-01-01

Salinity affects plant growth and is a major abiotic stress that limits crop productivity. It is well-understood that environmental adaptations and genetic traits regulate salinity tolerance in plants, but imparting the knowledge gained towards crop improvement remain arduous. Harnessing the potential of beneficial microorganisms present in the rhizosphere is an alternative strategy for improving plant stress tolerance. This review intends to elucidate the understanding of salinity tolerance mechanisms attributed by plant growth promoting rhizobacteria (PGPR). Recent advances in molecular studies have yielded insights into the signaling networks of plant–microbe interactions that contribute to salt tolerance. The beneficial effects of PGPR involve boosting key physiological processes, including water and nutrient uptake, photosynthesis, and source-sink relationships that promote growth and development. The regulation of osmotic balance and ion homeostasis by PGPR are conducted through modulation of phytohormone status, gene expression, protein function, and metabolite synthesis in plants. As a result, improved antioxidant activity, osmolyte accumulation, proton transport machinery, salt compartmentalization, and nutrient status reduce osmotic stress and ion toxicity. Furthermore, in addition to indole-3-acetic acid and 1-aminocyclopropane-1-carboxylic acid deaminase biosynthesis, other extracellular secretions of the rhizobacteria function as signaling molecules and elicit stress responsive pathways. Application of PGPR inoculants is a promising measure to combat salinity in agricultural fields, thereby increasing global food production. PMID:29109733

INFLUENCE OF IODOFORM ON ANTIMICROBIAL POTENTIAL OF CALCIUM HYDROXIDE

PubMed Central

Estrela, Carlos; Estrela, Cyntia Rodrigues de Araújo; Hollanda, Augusto César Braz; Decurcio, Daniel de Almeida; Pécora, Jesus Djalma

2006-01-01

The purpose of this research was to verify the influence of Iodoform on antimicrobial potential of calcium hydroxide. S. aureus, E. faecalis, P. aeruginosa, B. subtilis, C. albicans were the biological indicators. The substances tested were: calcium hydroxide + saline; calcium hydroxide + Iodoform + saline; Iodoform + saline. For the agar diffusion test, 18 Petri plates with 20 ml of BHI agar were inoculated with the microbial suspensions. Fifty-four cavities were made and filled with the substances tested. The diameters of microbial inhibition were then measured. In direct exposure test, 162 #50 sterile absorbent paper points were immersed in the experimental suspensions for 5 min, and covered with the pastes. At intervals of 24, 48 and 72 hours, the paper points were immersed in 10 ml of Letheen Broth, followed by incubation at 37°°C for 48h. Microbial growth was evaluated by turbidity of the culture medium. A 0.1 ml inoculum obtained from the Letheen Broth was transferred to 7 ml of BHI, and incubated at 37°°C for 48h. Bacterial growth was again evaluated by turbidity of the culture medium. The calcium hydroxide associated with the saline or the iodoform plus saline showed antimicrobial effectiveness in both experimental methods. The iodoform paste presented antimicrobial ineffectiveness for the agar diffusion test on all biological microorganisms and for the direct exposure test on B. subtilis and on the mixture. PMID:19089027

A protocol for the 0.9% saline versus Plasma-Lyte 148 for intensive care fluid therapy (SPLIT) study.

PubMed

Reddy, Sumeet K; Bailey, Michael J; Beasley, Richard W; Bellomo, Rinaldo; Henderson, Seton J; Mackle, Diane M; McArthur, Colin J; Mehrtens, Jan E; Myburgh, John A; McGuinness, Shay P; Psirides, Alex J; Young, Paul J

2014-12-01

0.9% saline is the most commonly used intravenous (IV) fluid in the world. However, recent data raise the possibility that, compared with buffered crystalloid fluids such as Plasma-Lyte 148, the administration of 0.9% saline to intensive care unit patients might increase their risk of acute kidney injury (AKI). To describe the protocol for the 0.9% Saline v Plasma-Lyte 148 for ICU Fluid Therapy (SPLIT) study. This is a multicentre, cluster-randomised, double crossover feasibility study to be conducted in four New Zealand tertiary ICUs over a 28-week period and will enroll about 2300 participants. All ICU patients who need crystalloid IV fluid therapy (except those with established renal failure needing dialysis and those admitted to the ICU for palliative care) will be enrolled. Participating ICUs will be randomly assigned to 0.9% saline or Plasma-Lyte 148 as the routine crystalloid IV fluid, in a blinded fashion, in four alternating 7-week blocks. The primary outcome will be the proportion of patients who develop AKI in the ICU. Secondary outcomes will include the difference between the most recent serum creatinine level measured before study enrollment and the peak serum creatinine level in the ICU; use of renal replacement therapy; and ICU and in hospital mortality. All analyses will be conducted on an intention-to-treat basis. The SPLIT study started on 1 April 2014 and will provide preliminary data on the comparative effectiveness of using 0.9% saline v Plasma- Lyte 148 as the routine IV fluid therapy in ICU patients.

Deciphering Staphylococcus sciuri SAT-17 Mediated Anti-oxidative Defense Mechanisms and Growth Modulations in Salt Stressed Maize (Zea mays L.)

PubMed Central

Akram, Muhammad S.; Shahid, Muhammad; Tariq, Mohsin; Azeem, Muhammad; Javed, Muhammad T.; Saleem, Seemab; Riaz, Saba

2016-01-01

Soil salinity severely affects plant nutrient use efficiency and is a worldwide constraint for sustainable crop production. Plant growth-promoting rhizobacteria, with inherent salinity tolerance, are able to enhance plant growth and productivity by inducing modulations in various metabolic pathways. In the present study, we reported the isolation and characterization of a salt-tolerant rhizobacterium from Kallar grass [Leptochloa fusca (L.) Kunth]. Sequencing of the 16S rRNA gene revealed its lineage to Staphylococcus sciuri and it was named as SAT-17. The strain exhibited substantial potential of phosphate solubilization as well as indole-3-acetic acid production (up to 2 M NaCl) and 1-aminocyclopropane-1-carboxylic acid deaminase activity (up to 1.5 M NaCl). Inoculation of a rifampicin-resistant derivative of the SAT-17 with maize, in the absence of salt stress, induced a significant increase in plant biomass together with decreased reactive oxygen species and increased activity of cellular antioxidant enzymes. The derivative strain also significantly accumulated nutrients in roots and shoots, and enhanced chlorophyll and protein contents in comparison with non-inoculated plants. Similar positive effects were observed in the presence of salt stress, although the effect was more prominent at 75 mM in comparison to higher NaCl level (150 mM). The strain survived in the rhizosphere up to 30 days at an optimal population density (ca. 1 × 106 CFU mL-1). It was concluded that S. sciuri strain SAT-17 alleviated maize plants from salt-induced cellular oxidative damage and enhanced growth. Further field experiments should be conducted, considering SAT-17 as a potential bio-fertilizer, to draw parallels between PGPR inoculation, elemental mobility patterns, crop growth and productivity in salt-stressed semi-arid and arid regions. PMID:27375588

Welfare Assessment following Heterotopic or Orthotopic Inoculation of Bladder Cancer in C57BL/6 Mice.

PubMed

Miller, Amy; Burson, Hannah; Söling, Ariane; Roughan, Johnny

2016-01-01

Few studies have assessed whether mice used as cancer models experience pain. Despite this possibility, the usual practice is to withhold analgesics as these are generally viewed as confounding. However, pain also alters cancer progression, so preventing it might not only be beneficial to welfare but also to study validity. Establishing the extent to which different cancer models result in pain is an important first step towards their refinement. We used conditioned place preference (CPP) testing and body-weight and behaviour analyses to evaluate the assumption that heterotopically implanted tumours result in less pain and fewer welfare concerns than those implanted orthotopically. C57Bl/6 mice received MB49Luc luciferase expressing bladder cancer cells or saline implanted subcutaneously or into the bladder. These tumour-bearing or control groups underwent 2 daily 45 minute conditioning trials to saline or morphine (2mg/kg) and then a 15 minute drug-free preference test on day 3 of a 3 day cycle, continuing until the study ended. Tumours were imaged and behaviour data obtained following preference tests. Development of preference for the morphine-paired chamber (morphine-seeking) was determined over time. Heterotopic tumour development had no effect on morphine-seeking, and although the restraint used for heterotopic inoculation caused greater initial weight losses than anaesthesia, these mice steadily gained weight and behaved comparatively normally throughout the study. Orthotopic tumour inoculation caused no initial weight losses, but over the final 7 days these mice became less active and lost more body weight than cancer-free controls. This indicated orthotopic implantation probably caused a more negative impact on welfare or conceivably pain; but only according to the current test methods. Pain could not be confirmed because morphine-seeking in the tumour-bearing groups was similar to that seen in controls. Imaging was not found to be an effective method of monitoring tumour development surpassing manual tumour inspection.

Comparison of efficacies of bovine immune colostral antibody and each immunoglobulin class against verotoxin 2, flagellum and somatic cells of Escherichia coli O157:H7 in mice.

PubMed

Seita, Tetsurou; Kuribayashi, Takashi; Honjo, Toshio; Yamamoto, Shizuo

2013-04-01

The efficacy of bovine immune colostral (colostral) antibodies against verotoxin (VT) 2, flagellum and somatic cells of Escherichia coli (E. coli) O157:H7 in mice was determined. Three major immunoglobulin (Ig) classes were isolated from the colostral antibody against VT2 by affinity chromatography and were used for estimation. Mice inoculated with VT2 were administered each Ig class from the colostral antibody, colostral antibody (colostral whey containing antibody) or serum antibody against VT2 at 1 hour after VT2 inoculation. All control mice (20/20) died after administration of sterilized saline instead of the colostral antibody. The survival rate was 93.3% (14/15) after administration of S-IgA or IgM antibody, or colostral antibody. Survival rates for IgG antibody and serum antibody administration were 80% (12/15) and 60% (9/15), respectively. Serum concentrations of VT2, which was absorbed from the small intestine in mice after administration of VT2 and colostral antibody, were measured by fluorescence enzyme immunoassay (FEIA). Serum concentrations of VT2 after administration of colostral antibody were lower than those after administration of sterilized saline. Mice inoculated with VT2-producing E. coli 157:H7 were administered anti-flagellum or anti-somatic colostral antibodies. Survival rates for E. coli O157:H7-infected mice administered the anti-flagellum and anti-somatic colostral antibodies were 52.4% (11/21) and 22.2% (4/18), respectively. Furthermore, survival rates increased to 89.5% (17/19) with combined administration of anti-flagellum and anti-VT2 colostral antibodies. These results suggest that colostral antibodies against VT2, flagellum and somatic cells are effective against E. coli O157:H7 infection. Copyright © 2012. Published by Elsevier B.V.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Hendrick, G.K.; Frizzell, R.T.; Cherrington, A.D.

In the 7-days fasted conscious dog, unlike the postabsorptive conscious dog, somatostatin infusion results in decreased levels of nonesterified fatty acids (NEFA) and increased glucose utilization (R{sub d}) even when insulin and glucagon levels are held constant. The aim of this study was to determine whether NEFA replacement in such animals would prevent the increase in R{sub d}. In each of three protocols there was an 80-min tracer equilibration period, a 40-min basal period, and a 3-h test period. During the test period in the first protocol saline was infused, in the second protocol somatostatin was infused along with intraportalmore » replacement amounts of insulin and glucagon (hormone replacement), while in the third protocol somatostatin plus the pancreatic hormones were infused with concurrent heparin plus Intralipid infusion. Glucose turnover was assessed using (3-{sup 3}H)glucose. The peripheral levels of insulin, glucagon, and glucose were similar and constant in all three protocols; however, during somatostatin infusion, exogenous glucose infusion was necessary to maintain euglycemia. The NEFA level was constant during saline infusion and decreased in the hormone replacement protocol. In the hormone replacement plus NEFA protocol, the NEFA level did not change during the first 90-min period and then increased during the second 90-min period. After a prolonged fast in the dog, (1) somatostatin directly or indirectly inhibits adipose tissue NEFA release and causes a decrease in the plasma NEFA level, and (2) this decrease in the NEFA level causes an increase in R{sub d}.« less

Effects of imposed salinity gradients on dissimilatory arsenate reduction, sulfate reduction, and other microbial processes in sediments from two California soda lakes

USGS Publications Warehouse

Kulp, T.R.; Han, S.; Saltikov, C.W.; Lanoil, B.D.; Zargar, K.; Oremland, R.S.

2007-01-01

Salinity effects on microbial community structure and on potential rates of arsenate reduction, arsenite oxidation, sulfate reduction, denitrification, and methanogenesis were examined in sediment slurries from two California soda lakes. We conducted experiments with Mono Lake and Searles Lake sediments over a wide range of salt concentrations (25 to 346 g liter-1). With the exception of sulfate reduction, rates of all processes demonstrated an inverse relationship to total salinity. However, each of these processes persisted at low but detectable rates at salt saturation. Denaturing gradient gel electrophoresis analysis of partial 16S rRNA genes amplified from As(V) reduction slurries revealed that distinct microbial populations grew at low (25 to 50 g liter-1), intermediate (100 to 200 g liter-1), and high (>300 g liter-1) salinity. At intermediate and high salinities, a close relative of a cultivated As-respiring halophile was present. These results suggest that organisms adapted to more dilute conditions can remain viable at high salinity and rapidly repopulate the lake during periods of rising lake level. In contrast to As reduction, sulfate reduction in Mono Lake slurries was undetectable at salt saturation. Furthermore, sulfate reduction was excluded from Searles Lake sediments at any salinity despite the presence of abundant sulfate. Sulfate reduction occurred in Searles Lake sediment slurries only following inoculation with Mono Lake sediment, indicating the absence of sulfate-reducing flora. Experiments with borate-amended Mono Lake slurries suggest that the notably high (0.46 molal) concentration of borate in the Searles Lake brine was responsible for the exclusion of sulfate reducers from that ecosystem. Copyright ?? 2007, American Society for Microbiology. All Rights Reserved.

Effects of Platelet-Rich Plasma and Indomethacin on Biomechanics of Rotator Cuff Repair.

PubMed

Meadows, Molly C; Levy, David M; Ferry, Christopher M; Gardner, Thomas R; Teratani, Takeshi; Ahmad, Christopher S

We conducted a study to determine if platelet-rich plasma (PRP) enhances the strength of rotator cuff repair (RCR) and if concomitant use of nonsteroidal anti-inflammatory drugs (NSAIDs) affects PRP efficacy. We also wanted to determine the optimal centrifugation protocol for making PRP from rats. This study used 48 rats, 14 in a centrifugation protocol and 34 in an operative protocol. Six syngeneic rats from the operative group were used as PRP blood donors; the other 28 operative rats underwent bilateral RCRs. The Autologous Conditioned Plasma system (Arthrex) was used to prepare leukocyte-poor PRP. One shoulder was randomized to an intratendinous PRP injection, and the other received normal saline. Each rat was also randomly placed on a postoperative diet, either a regular diet or an indomethacin-enhanced diet. After rats were euthanized at 3 weeks, specimens were dissected to isolate the supraspinatus tendon at its humeral attachment, which was subjected to biomechanical testing. PRP prepared with a protocol of 5 minutes × 1300 revolutions per minute had the highest platelet index. Mean (SD) energy to failure was significantly higher (P = .03) in tendons treated with PRP, 11.7 (7.3) N-mm, than in tendons treated with saline, 8.7 (4.6) N-mm. Both groups (PRP, saline) showed no significant differences between tendons treated with NSAIDs and those not treated with NSAIDs. Intraoperative application of PRP enhances energy to failure after RCR in rats. There were no differences in biomechanical strength with NSAID use and no interactions between PRP and NSAID use.

The role of the cardiac nerves in regulation of sodium excretion in conscious dogs.

PubMed

Kaczmarczyk, G; Drake, A; Eisele, R; Mohnhaupt, R; Noble, M I; Simgen, B; Stubbs, J; Reinhardt, H W

1981-05-01

Conscious, chronically instrumented dogs, maintained on a high sodium intake, were used to investigate whether surgical cardiac denervation impairs the natriuresis associated with left atrial pressure increase produced in three ways: during an increase in left atrial pressure by means of a reversible mitral stenosis (protocol 1); after an i.v. saline load (1.0 ml 0.9% saline min-1 . kg-1 over 60 min) (protocol 2); after an oral saline load (14.5 mmol Na . kg-1 given with the food as isotonic solution) (protocol 3). During a reversible mitral stenosis, in intact dogs, urine volume and sodium excretion increased markedly (from 34--145 microliters . min-1 . kg-1 and from 3--12 mumol . min-1 . kg-1); mean arterial pressure increased by an average of 2 kPa (15 mm Hg) and heart rate by 53 b/min; plasma renin activity fell from 0.37--0.21 ng AI . ml-1 . h-1 . Cardiac denervation eliminated these effects of left atrial distension except for a small increase in heart rate (12 b/min). This indicates that the natriuresis and diuresis during left atrial distension resulted from stimulation of receptors located in the left atrium. In contrast, during protocol 2 and 3, the same amounts of sodium and water were excreted in the cardiac denervated dogs as compared to the intact dogs. A comparable decrease in plasma renin activity also was observed. -- Apparently the presence of the cardiac nerves is not a prerequisite for maintenance of sodium and water homeostasis.

Characterization of a halophilic heterotrophic nitrification-aerobic denitrification bacterium and its application on treatment of saline wastewater.

PubMed

Duan, Jinming; Fang, Hongda; Su, Bing; Chen, Jinfang; Lin, Jinmei

2015-03-01

A novel halophilic bacterium capable of heterotrophic nitrification-aerobic denitrification was isolated from marine sediments and identified as Vibrio diabolicus SF16. It had ability to remove 91.82% of NH4(+)-N (119.77 mg/L) and 99.71% of NO3(-)-N (136.43 mg/L). The nitrogen balance showed that 35.83% of initial NH4(+)-N (119.77 mg/L) was changed to intracellular nitrogen, and 53.98% of the initial NH4(+)-N was converted to gaseous denitrification products. The existence of napA gene further proved the aerobic denitrification ability of strain SF16. The optimum culture conditions were salinity 1-5%, sodium acetate as carbon source, C/N 10, and pH 7.5-9.5. When an aerated biological filter system inoculated with strain SF16 was employed to treat saline wastewater, the average removal efficiency of NH4(+)-N and TN reached 97.14% and 73.92%, respectively, indicating great potential of strain SF16 for future full-scale applications. Copyright © 2014 Elsevier Ltd. All rights reserved.

Characterization and crop production efficiency of diazotrophic bacterial isolates from coastal saline soils.

PubMed

Barua, Shilajit; Tripathi, Sudipta; Chakraborty, Ashis; Ghosh, Sagarmoy; Chakrabarti, Kalyan

2012-01-20

Use of eco-friendly area specific salt tolerant bioinoculants is better alternatives to chemical fertilizer for sustainable agriculture in coastal saline soils. We isolated diverse groups of diazotrophic bacteria from coastal saline soils of different forest and agricultural lands in the Sundarbans, West Bengal, India, to study their effect on crop productivity in saline soils. Phenotypic, biochemical and molecular identifications of the isolates were performed. The isolates produced indole acetic acid, phosphatase, and solubilized insoluble phosphates. Sequence analysis of 16S rDNA identified the SUND_BDU1 strain as Agrobacterium and the strains SUND_LM2, Can4 and Can6 belonging to the genus Bacillus. The ARA activity, dinitrogen fixation and presence of nifH genes indicated they were diazotrophs. Field trials with these strains as bioinoculants were carried out during 2007-2009, with rice during August-December followed by Lady's finger during April-June. Microplots, amended with FYM inoculated with four bioinoculants individually were compared against sole FYM (5 t ha(-1)) and a sole chemical fertilizer (60:30:30 kg ha(-1) NPK) treated plot. The strain Can6 was by far the best performer in respect of yield attributes and productivity of studied crops. Copyright © 2011 Elsevier GmbH. All rights reserved.

Sm14 of Schistosoma mansoni in Fusion with Tetanus Toxin Fragment C Induces Immunoprotection against Tetanus and Schistosomiasis in Mice

PubMed Central

Abreu, Patrícia A. E.; Miyasato, Patrícia A.; Vilar, Mônica M.; Dias, Waldely O.; Ho, Paulo L.; Tendler, Míriam; Nascimento, Ana L. T. O.

2004-01-01

We have constructed vectors that permit the expression in Escherichia coli of Schistosoma mansoni fatty acid-binding protein 14 (Sm14) in fusion with the nontoxic, but highly immunogenic, tetanus toxin fragment C (TTFC). The recombinant six-His-tagged proteins were purified by nickel affinity chromatography and used in immunization and challenge assays. Animals inoculated with TTFC in fusion with or coadministered with Sm14 showed high levels of tetanus toxin antibodies, while animals inoculated with Sm14 in fusion with or coadministered with TTFC showed high levels of Sm14 antibodies. In both cases, there were no changes in the type of immune response (Th2) obtained with the fusion proteins compared to those obtained with the nonfused proteins. Mice immunized with the recombinant proteins (TTFC in fusion with or coadministered with Sm14) survived the challenge with tetanus toxin and did not show any symptoms of the disease. Control animals inoculated with either phosphate-buffered saline (PBS) or Sm14 died with severe symptoms of tetanus after 24 h. Mice immunized with the recombinant proteins (Sm14 in fusion with or coadministered with TTFC) showed a 50% reduction in worm burden when they were challenged with S. mansoni cercariae, while control animals inoculated with either PBS or TTFC were not protected. The results show that the expression of other antigens in fusion at the carboxy terminus of TTFC is feasible for the development of a multivalent recombinant vaccine. PMID:15385496

In vitro propagation of fraser photinia using Azospirillum-mediated root development.

PubMed

Llorente, Berta E; Larraburu, Ezequiel E

2013-01-01

Fraser photinia (Photinia × fraseri Dress.) is a woody plant of high ornamental value. The traditional propagation system for photinia is by rooting apical cuttings using highly concentrated auxin treatments. However, photinia micropropagation is an effective alternative to traditional in vivo propagation which is affected by the seasonal supply of cuttings, the long time required to obtain new plants, and the difficulties in rooting some clones.A protocol for in vitro propagation of fraser photinia using the plant growth-promoting ability of some rhizobacteria is described here. Bacterial inoculation is a new tool in micropropagation protocols that improves plant development in in vitro culture. Shoots culture on a medium containing MS macro- and microelements, Gamborg's vitamins (BM), N (6)-benzyladenine (BA, 11.1 μM), and gibberellic acid (1.3 μM) produce well-established explants. Proliferation on BM medium supplemented with 4.4 μM BA results in four times the number of shoots per initial shoot that develops monthly. Consequently, there is a continuous supply of plant material since shoot production is independent of season. Azospirillum brasilense inoculation, after 49.2 μM indole-3-butyric acid pulse treatment, stimulates early rooting of photinia shoots and produces significant increase in root fresh and dry weights, root surface area, and shoot fresh and dry weights in comparison with controls. Furthermore, inoculated in vitro photinia plants show anatomical and morphological changes that might lead to better adaptation in ex vitro conditions after transplanting, compared with the control plants.

The development of a new three-step protocol to determine the efficacy of disinfectant wipes on surfaces contaminated with Staphylococcus aureus.

PubMed

Williams, G J; Denyer, S P; Hosein, I K; Hill, D W; Maillard, J-Y

2007-12-01

We developed a three-step protocol to quantify the efficacy of disinfectant wipes, their ability to remove and prevent microbial transfer from surfaces and their overall antimicrobial activity. Meticillin-resistant (MRSA) or -susceptible (MSSA) Staphylococcus aureus (6-7 log(10)cfu) were inoculated onto stainless steel discs with or without organic load and dried. Grapefruit extract-containing test wipes and unmedicated control wipes were used. In step 1, wipes were mechanically rotated against surfaces for 10s at 60rpm, exerting a weight of 100+/-5g. Bacterial removal was assessed by transferring the steel discs to neutraliser, resuspending and counting remaining bacteria. In step 2, bacterial transfer from wipes was assessed by eight consecutive mechanical adpression transfers to agar/neutraliser plates. Step 3 was the measurement of antimicrobial activity by direct inoculation of the wipes for 10s followed by neutralisation and enumeration. Test wipes achieved a significantly higher bacterial cell removal than control wipes on all surfaces (P<0.05). The low bactericidal activity of the wipes (<1 log(10) reduction when directly inoculated) and the subsequent survival of bacteria on the wipes, however, led to repeated microbial transfer when initially high contamination levels were present. There were no differences between MRSA and MSSA in removal, transfer or antimicrobial activity. The three-step method is a useful tool for developing future guidelines to assess the ability of wipes to disinfect surfaces.

Establishment of an efficient virus-induced gene silencing (VIGS) assay in Arabidopsis by Agrobacterium-mediated rubbing infection.

PubMed

Manhães, Ana Marcia E de A; de Oliveira, Marcos V V; Shan, Libo

2015-01-01

Several VIGS protocols have been established for high-throughput functional genomic screens as it bypasses the time-consuming and laborious process of generation of transgenic plants. The silencing efficiency in this approach is largely hindered by a technically demanding step in which the first pair of newly emerged true leaves at the 2-week-old stage are infiltrated with a needleless syringe. To further optimize VIGS efficiency and achieve rapid inoculation for a large-scale functional genomic study, here we describe a protocol of an efficient VIGS assay in Arabidopsis using Agrobacterium-mediated rubbing infection. The Agrobacterium inoculation is performed by simply rubbing the leaves with Filter Agent Celite(®) 545. The highly efficient and uniform silencing effect was indicated by the development of a visibly albino phenotype due to silencing of the Cloroplastos alterados 1 (CLA1) gene in the newly emerged leaves. In addition, the albino phenotype could be observed in stems and flowers, indicating its potential application for gene functional studies in the late vegetative development and flowering stages.

Lactobacillus crustorum KH: novel prospective probiotic strain isolated from Iranian traditional dairy products.

PubMed

Sharafi, Hakimeh; Derakhshan, Venos; Paknejad, Mojgan; Alidoust, Leila; Tohidi, Azadeh; Pornour, Majid; Hajfarajollah, Hamidreza; Zahiri, Hossein Shahbani; Noghabi, Kambiz Akbari

2015-02-01

In recent years, exploring novel probiotic strains for therapeutic intervention has been raised due to the significant increase in market demand. This study aimed to investigate the certain probiotic properties of 15 Lactobacillus isolates from Iranian traditional dairy products. Among them, a novel potential probiotic strain was isolated and identified as Lactobacillus crustorum. The characteristics of potential probiotics were examined in terms of resistance to acidity, bile, and salinity as well as antibiotic tolerance and antibacterial activity. L. crustorum KH has shown tolerance property to bile (0.3 % w), acidity (pH 2-9), and salinity (1-5 % NaCl) and strong antibacterial activity against tested enteropathogens by well-diffusion assay. Furthermore, in vivo study and histological assays were performed to study whether live and heat-killed cells of L. crustorum KH are able to protect against the challenge of Escherichia coli O157:H7 in the gastrointestinal tract of mice used as an experimental model. Therefore, heat-killed and live cells of L. crustorum KH were inoculated by gavage to different groups of 4-6-week-old female BALB/c mice in doses of 10(8) colony-forming unit (CFU)/dose. Thereafter, these mice were challenged with E. coli O157:H7 also inoculated in the gastrointestinal tract (GIT) of the animals. The results showed that heat-killed cells of L. crustorum KH exert a protective effect against E. coli O157:H7 colonization at different degrees, being lower than that produced by viable cells.

Disinfection of rigid nasal endoscopes following in vitro contamination with Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa, and Haemophilus influenzae.

PubMed

Bradford, Benjamin D; Seiberling, Kristin A; Park, Francine E; Hiebert, Jared C; Chang, Dennis F

2013-06-01

If not adequately cleaned, rigid nasal endoscopes (RNEs) have the potential to cause iatrogenic cross-contamination. To test the efficacy of various disinfection methods in reducing bacterial load on RNEs in vitro. In vitro model. Staphylococcus aureus, Streptococcus pneumoniae, Pseudomonas aeruginosa, and Haemophilus influenzae contamination was separately induced on RNEs in vitro. Two experimental sets were completed. The RNEs were disinfected using the following protocols: 30-second scrub with antimicrobial soap (ABS) and water, 30-second scrub with 70% isopropyl alcohol (IA), 30-second scrub with ABS followed by 30-second scrub with IA, 30-second scrub with germicidal cloth, isolated 5-minute soak in an enzymatic soap solution, 5- and 10-minute soaks in ortho-phthalaldehyde, 0.55%, solution (Cidex OPA), and isolated 30-second rinse with tap water, all with 30-second precleaning and postcleaning rinses with tap water. Two sets of experiments (experiment sets A and B) were carried out with a 30-second tap water rinse after inoculation of each RNE. This was followed by immediate cleaning in set A and a 1-hour air-dry delay in set B. Otherwise there were no differences in the disinfection protocols between sets for each method noted. Effectiveness of various disinfection protocols in cleaning rigid nasal endoscopes experimentally inoculated with bacteria commonly found in the upper aerodigestive tract. Positive cultures following disinfection indicated ineffective or incomplete disinfection. Most cleaning methods were effective in eliminating S aureus, S pneumoniae, and H influenzae from the scopes following experimental contamination. Continued growth of P aeruginosa was found after all of the disinfection trials in experiment set A with the exception of a 10-minute immersion in Cidex OPA, and in set B except for the 10-minute Cidex OPA immersion and ABS plus IA trials. Most cleaning methods used in our trials appear to properly disinfect RNEs after in vitro inoculation with S aureus, S pneumoniae, and H influenzae. However, it appears that disinfectants may be less effective in cleaning rigid scopes experimentally inoculated with P aeruginosa. There is a paucity of published data regarding cross-contamination during rigid nasal endoscopy, and these results should guide future studies and to some extent practice to avoid iatrogenic spread of contamination.

Efficacy of Detergent and Water Versus Bleach for the Disinfection of Direct Contact Ophthalmic Lenses

PubMed Central

Abbey, Ashkan M.; Gregori, Ninel Z.; Surapaneni, Krishna; Miller, Darlene

2014-01-01

Purpose While manufacturers recommend cleaning ophthalmic lenses with detergent and water and then a specific disinfectant, disinfectants are rarely used in ophthalmic practices. The aim of this pilot study was to evaluate the efficacy of detergent and water versus bleach, a recommended disinfectant, to eliminate common ocular bacteria and viruses from ophthalmic lenses. Methods Three bacterial strains (Staphylococcus epidermidis, Corynebacterium straitum, and methicillin-resistant Staphylococcus aureus (MRSA) and two viral strains (adenovirus and herpes simplex virus (HSV) type-1) were individually inoculated to 20 gonioscopy and laser lenses. Lenses were washed with detergent and water and then disinfected with 10% bleach. All lenses were cultured after inoculation, after detergent and water, and after the bleach. Bacterial cultures in thioglycollate broth were observed for 3 weeks and viral cultures for 2 weeks. The presence of viruses was also detected by multiplex polymerase chain reaction (PCR). Results All 20 lenses inoculated with Staphylococcus epidermidis, Corynebacterium straitum, adenovirus, and HSV-1 showed growth after inoculation, but no growth after detergent/water and after the bleach. All lenses showed positive HSV and adenovirus PCR after inoculation and negative PCR after detergent/water and after bleach. All MRSA contaminated lenses showed growth after inoculation and no growth after detergent and water. However, one lens showed positive growth after bleach. Conclusions Cleaning with detergent and water appeared to effectively eliminate bacteria and viruses from the surface of contaminated ophthalmic lenses. Further studies are warranted to design practical disinfection protocols that minimize lens damage. PMID:24747806

Baroreceptor unloading does not limit forearm sweat rate during severe passive heat stress.

PubMed

Schlader, Zachary J; Gagnon, Daniel; Lucas, Rebekah A I; Pearson, James; Crandall, Craig G

2015-02-15

This study tested the hypothesis that sweat rate during passive heat stress is limited by baroreceptor unloading associated with heat stress. Two protocols were performed in which healthy subjects underwent passive heat stress that elicited an increase in intestinal temperature of ∼1.8°C. Upon attaining this level of hyperthermia, in protocol 1 (n = 10, 3 females) a bolus (19 ml/kg) of warm (∼38°C) isotonic saline was rapidly (5-10 min) infused intravenously to elevate central venous pressure (CVP), while in protocol 2 (n = 11, 5 females) phenylephrine was infused intravenously (60-120 μg/min) to return mean arterial pressure (MAP) to normothermic levels. In protocol 1, heat stress reduced CVP from 3.9 ± 1.9 mmHg (normothermia) to -0.6 ± 1.4 mmHg (P < 0.001), while saline infusion returned CVP to normothermic levels (5.1 ± 1.7 mmHg; P > 0.999). Sweat rate was elevated by heat stress (1.21 ± 0.44 mg·cm(-2)·min(-1)) but remained unchanged during rapid saline infusion (1.26 ± 0.47 mg·cm(-2)·min(-1), P = 0.5), whereas cutaneous vascular conductance increased from 77 ± 10 to 101 ± 20% of local heating max (P = 0.029). In protocol 2, MAP was reduced with heat stress from 85 ± 7 mmHg to 76 ± 8 mmHg (P = 0.048). Although phenylephrine infusion returned MAP to normothermic levels (88 ± 7 mmHg; P > 0.999), sweat rate remained unchanged during phenylephrine infusion (1.39 ± 0.22 vs. 1.41 ± 0.24 mg·cm(-2)·min(-1); P > 0.999). These data indicate that both cardiopulmonary and arterial baroreceptor unloading do not limit increases in sweat rate during passive heat stress. Copyright © 2015 the American Physiological Society.

The effect of nebulized salbutamol or isotonic saline on exercise-induced bronchoconstriction in elite skaters following a 1,500-meter race: study protocol for a randomized controlled trial.

PubMed

Driessen, Jean M M; Gerritsma, Margryt; Westbroek, Jaap; ten Hacken, Nick H T; de Jongh, Frans H C

2013-07-09

Prevalence of exercise-induced bronchoconstriction (EIB) is high in elite athletes, especially after many years training in cold and dry air conditions. The primary treatment of EIB is inhaling a short-acting beta-2-agonist such as salbutamol. However, professional speed skaters also inhale nebulized isotonic saline or tap water before and after a race or intense training. The use of nebulized isotonic saline or tap water to prevent EIB has not been studied before, raising questions about safety and efficacy. The aim of this study is to analyze the acute effect of nebulized isotonic saline or salbutamol on EIB in elite speed skaters following a 1,500-meter race. This randomized controlled trial compares single dose treatment of 1 mg nebulized salbutamol in 4 mL of isotonic saline, or with 5 mL of isotonic saline. A minimum of 13 participants will be allocated in each treatment group. Participants should be between 18 and 35 years of age and able to skate 1,500 m in less than 2 min 10 s (women) or 2 min 05 s (men). Repeated measurements of spirometry, forced oscillation technique, and electromyography will be performed before and after an official 1,500-m race. Primary outcome of the study is the difference in fall in FEV1 after exercise in the different treatment groups. The trial is currently enrolling participants. Elite athletes run the risk of pulmonary inflammation and remodeling as a consequence of their frequent exercise, and thus increased ventilation in cold and dry environments. Although inhalation of nebulized isotonic saline is commonplace, no study has ever investigated the safety or efficacy of this treatment. This trial protocol was registered with the Dutch trial registration for clinical trials under number NTR3550.

INHALED HYPERTONIC SALINE IN INFANTS AND CHILDREN LESS THAN SIX YEARS OF AGE WITH CYSTIC FIBROSIS: THE ISIS RANDOMIZED TRIAL

PubMed Central

Rosenfeld, Margaret; Ratjen, Felix; Brumback, Lyndia; Daniel, Stephen; Rowbotham, Ron; McNamara, Sharon; Johnson, Robin; Kronmal, Richard; Davis, Stephanie D

2013-01-01

Context Inhaled hypertonic saline is recommended as therapy for cystic fibrosis (CF) patients 6 years of age and older, but its efficacy has never been evaluated in CF patients <6 years of age. Objective To determine if hypertonic saline reduces the rate of protocol-defined pulmonary exacerbations in CF patients <6 years of age. Design and Setting A multicenter, randomized, double-blind placebo-controlled trial was conducted from April 2009 to October 2011 at 30 CF care centers in the United States and Canada. Participants Participants had an established diagnosis of CF and were 4 to 60 months of age. A total of 344 patients were assessed for eligibility; 321 participants were randomized; 29 (9%) withdrew prematurely. Intervention The active group (n=158) received 7% hypertonic saline and the control group (n=163) received 0.9% isotonic saline nebulized twice daily for 48 weeks. Both groups received albuterol or levalbuterol prior to each study drug dose. Main Outcome Measures the rate of protocol-defined pulmonary exacerbations during the 48 week treatment period treated with oral, inhaled or intravenous antibiotics. Results The mean pulmonary exacerbation rate (events/person-year) was 2.3 (95% CI, 2.0, 2.5) in the hypertonic saline group and 2.3 (95% CI, 2.1, 2.6) in the isotonic saline group; the rate ratio was 0.98 (95% CI, 0.84, 1.14)). Among participants with pulmonary exacerbations, the mean number of total antibiotic treatment days for a pulmonary exacerbation was 60 (95% CI 49, 70) in the hypertonic saline group and 52 (95% CI 43, 61) in the isotonic saline group. There was no significant difference in secondary endpoints including height, weight, respiratory rate, oxygen saturation, cough or respiratory symptom scores. Infant pulmonary function testing performed as an exploratory outcome in a subgroup (N=73, with acceptable measurements at 2 visits in 45) did not demonstrate significant differences between groups except for the mean change in forced expiratory volume in 0.5 seconds which was 38 ml greater (95% CI 1, 76) in the hypertonic saline group. Adherence by returned study drug ampoules was at least 75% in each group. Adverse event profiles were also similar, with the most common adverse event of moderate or severe severity in each group being cough (39% of hypertonic saline group, 38% of isotonic saline group). Conclusions Among infants and children with cystic fibrosis less than 6 years old, the use of inhaled hypertonic saline compared with isotonic saline did not reduce the rate of pulmonary exacerbations over 48 weeks of treatment. Trial Registration www.clinicaltrials.gov NCT00709280 PMID:22610452

Weaving Common Threads: Reaching out to American Indians and Native Peoples of Alaska and the Pacific Islands

NASA Technical Reports Server (NTRS)

2002-01-01

This report outlines the activities of the GLOBE (Global Learning and Observations to Benefit the Environment) Train-the-Trainer Workshop. Educators were introduced to the GLOBE protocols for Atmosphere, Hydrology, Soil and Land Cover. These protocols included measurement of pH, temperature, precipitation, salinity, and soil moisture content. Each topic included implementation plans and learning activities.

Gamma radiation in the reduction of S almonella spp. inoculated on minimally processed watercress ( Nasturtium officinalis)

NASA Astrophysics Data System (ADS)

Martins, C. G.; Behrens, J. H.; Destro, M. T.; Franco, B. D. G. M.; Vizeu, D. M.; Hutzler, B.; Landgraf, M.

2004-09-01

Consumer attitudes towards foods have changed in the last two decades increasing requirements for freshlike products. Consequently, less extreme treatments or additives are being required. Minimally processed foods have freshlike characteristics and satisfy this new consumer demand. Besides freshness, the minimally processing also provide convenience required by the market. Salad vegetables can be source of pathogen such as Salmonella, Escherichia coli O157:H7, Shigella spp. The minimal processing does not reduce the levels of pathogenic microorganisms to safe levels. Therefore, this study was carried out in order to improve the microbiological safety and the shelf-life of minimally processed vegetables using gamma radiation. Minimally processed watercress inoculated with a cocktail of Salmonella spp was exposed to 0.0, 0.2, 0.5, 0.7, 1.0, 1.2 and 1.5 kGy. Irradiated samples were diluted 1:10 in saline peptone water and plated onto tryptic soy agar that were incubated at 37°C/24 h. D 10 values for Salmonella spp. inoculated in watercress varied from 0.29 to 0.43 kGy. Therefore, a dose of 1.7 kGy will reduce Salmonella population in watercress by 4 log 10. The shelf-life was increased by 1 {1}/{2} day when the product was exposed to 1 kGy.

Competitive outcome of Daphnia-Simocephalus experimental microcosms: salinity versus priority effects.

PubMed

Loureiro, Cláudia; Pereira, Joana L; Pedrosa, M Arminda; Gonçalves, Fernando; Castro, Bruno B

2013-01-01

Competition is a major driving force in freshwaters, especially given the cyclic nature and dynamics of pelagic food webs. Competition is especially important in the initial species assortment during colonization and re-colonization events, which depends strongly on the environmental context. Subtle changes, such as saline intrusion, may disrupt competitive relationships and, thus, influence community composition. Bearing this in mind, our objective was to assess whether low salinity levels (using NaCl as a proxy) alter the competitive outcome (measured as the rate of population biomass increase) of Daphnia-Simocephalus experimental microcosms, taking into account interactions with priority effects (sequential species arrival order). With this approach, we aimed to experimentally demonstrate a putative mechanism of differential species sorting in brackish environments or in freshwaters facing secondary salinization. Experiments considered three salinity levels, regarding NaCl added (0.00, 0.75 and 1.50 g L(-1)), crossed with three competition scenarios (no priority, priority of Daphnia over Simocephalus, and vice-versa). At lower NaCl concentrations (0.00 and 0.75 g L(-1)), Daphnia was a significantly superior competitor, irrespective of the species inoculation order, suggesting negligible priority effects. However, the strong decrease in Daphnia population growth at 1.50 g L(-1) alleviated the competitive pressure on Simocephalus, causing an inversion of the competitive outcome in favour of Simocephalus. The intensity of this inversion depended on the competition scenario. This salinity-mediated disruption of the competitive outcome demonstrates that subtle environmental changes produce indirect effects in key ecological mechanisms, thus altering community composition, which may lead to serious implications in terms of ecosystem functioning (e.g. lake regime shifts due to reduced grazing) and biodiversity.

Advice for Travellers to Tropical and Subtropical Countries

PubMed Central

Lawee, David; Scapattura, Philip

1985-01-01

Travellers to areas where political instability, poverty or inadequate public health measures render them at risk of exposure to infections and diseases they are unlikely to meet at home should seek prophylaxis and protection before departure. Inoculation is the best form of protection for certain diseases; since protocols change frequently, a reliable local source of current information should be found. Prophylaxis against malaria involves both mechanical protection against mosquito bites, and appropriate chemoprophylaxis. Water safety must be considered where chlorinated water is not available; a protocol for decontamination is presented, and guidelines for avoiding gastrointestinal disorders are listed. PMID:21274125

Comparative effectiveness of Pseudomonas and Serratia sp. containing ACC-deaminase for improving growth and yield of wheat (Triticum aestivum L.) under salt-stressed conditions.

PubMed

Zahir, Zahir Ahmad; Ghani, Usman; Naveed, Muhammad; Nadeem, Sajid Mahmood; Asghar, Hafiz Naeem

2009-05-01

Ethylene synthesis is accelerated in response to various environmental stresses like salinity. Ten rhizobacterial strains isolated from wheat rhizosphere taken from different salt affected areas were screened for growth promotion of wheat under axenic conditions at 1, 5, 10 and 15 dS m(-1). Three strains, i.e., Pseudomonas putida (N21), Pseudomonas aeruginosa (N39) and Serratia proteamaculans (M35) showing promising performance under axenic conditions were selected for a pot trial at 1.63 (original), 5, 10 and 15 dS m(-1). Results showed that inoculation was effective even in the presence of higher salinity levels. P. putida was the most efficient strain compared to the other strains and significantly increased the plant height, root length, grain yield, 100-grain weight and straw yield up to 52, 60, 76, 19 and 67%, respectively, over uninoculated control at 15 dS m(-1). Similarly, chlorophyll content and K(+)/Na(+) of leaves also increased by P. putida over control. It is highly likely that under salinity stress, 1-aminocyclopropane-1-carboxylic acid-deaminase activity of these microbial strains might have caused reduction in the synthesis of stress (salt)-induced inhibitory levels of ethylene. The results suggested that these strains could be employed for salinity tolerance in wheat; however, P. putida may have better prospects in stress alleviation/reduction.

Activity and Safety of Inhaled Itraconazole Nanosuspension in a Model Pulmonary Aspergillus fumigatus Infection in Inoculated Young Quails.

PubMed

Wlaź, Piotr; Knaga, Sebastian; Kasperek, Kornel; Wlaź, Aleksandra; Poleszak, Ewa; Jeżewska-Witkowska, Grażyna; Winiarczyk, Stanisław; Wyska, Elżbieta; Heinekamp, Thorsten; Rundfeldt, Chris

2015-08-01

Pulmonary aspergillosis is frequently reported in parrots, falcons, and other birds held in captivity. Inhalation is the main route of infection for Aspergillus fumigatus, resulting in both acute and chronic disease conditions. Itraconazole (ITRA) is an antifungal commonly used in birds, but its administration requires repeated oral dosing, and the safety margin is narrow. To investigate the efficacy of inhaled ITRA, six groups of ten young quails (Coturnix japonica) were inoculated intratracheally with 5 × 10(6) spores (3 groups) or 5 × 10(7) spores (3 groups). Animals were exposed to nebulized ITRA nanosuspension as 10 % suspension or 4 % suspension, once daily for 30 min, starting 2 h after inoculation for 6 days. Control groups were exposed to nebulized saline for the same period of time. Survival and clinical scores were evaluated, and animals were subjected to gross pathology. In control animals, aspergillosis resulted in systemic disease without pulmonary or air sac granulomas. Animals died from multiple organ failure. Inhalation of 10 % ITRA nanosuspension blocked lethality and prevented disease-related symptoms in the quails exposed to the low dose of spores, while the disease course in quails inoculated with the high-spore dose was retarded. Inhalation of 4 % ITRA nanosuspension was less effective. Both inhalations were well tolerated, and gross pathology did not reveal signs of local toxicity. The data indicate that inhaled administration of 10 % ITRA nanosuspension is capable of alleviating an acute A. fumigatus infection in quails. A lower ITRA concentration may be only active in chronic pulmonary aspergillosis.

BCG vaccination in leprosy: final results of the trial in Karimui, Papua New Guinea, 1963-79.

PubMed Central

Bagshawe, A.; Scott, G. C.; Russell, D. A.; Wigley, S. C.; Merianos, A.; Berry, G.

1989-01-01

The efficacy of BCG vaccine in preventing the clinical manifestations of leprosy in a tuberculosis-free area of Papua New Guinea is reported. Between 1963 and 1966 a total of 5356 subjects, randomized to receive BCG or saline inoculations, were examined for leprosy before the vaccination and surveillance was continued until 1979. BCG afforded 48% protection against clinical leprosy, being most effective against borderline tuberculoid leprosy and in children vaccinated when under 15 years old. Protection was evident within 12 months in those vaccinated between the ages of 10 and 15 years but was delayed in other age groups. There was evidence for accelerated manifestations of tuberculoid leprosy in children vaccinated when under 5 years of age. Tuberculin sensitivity was more likely to be sustained following multiple BCG inoculations; vaccinees with sustained tuberculin sensitivity had the lowest incidence of leprosy, but protection was also evident in tuberculin-negative vaccinees. These results may have implications for ongoing trials of leprosy vaccine incorporating BCG. PMID:2680140

Arbuscular Mycorrhizal Symbiosis Modulates Antioxidant Response and Ion Distribution in Salt-Stressed Elaeagnus angustifolia Seedlings.

PubMed

Chang, Wei; Sui, Xin; Fan, Xiao-Xu; Jia, Ting-Ting; Song, Fu-Qiang

2018-01-01

Elaeagnus angustifolia L. is a drought-resistant species. Arbuscular mycorrhizal symbiosis is considered to be a bio-ameliorator of saline soils that can improve salinity tolerance in plants. The present study investigated the effects of inoculation with the arbuscular mycorrhizal fungus Rhizophagus irregularis on the biomass, antioxidant enzyme activities, and root, stem, and leaf ion accumulation of E. angustifolia seedlings grown during salt stress conditions. Salt-stressed mycorrhizal seedlings produced greater root, stem, and leaf biomass than the uninoculated stressed seedlings. In addition, the seedlings colonized by R. irregularis showed notably higher activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) in the leaves of the mycorrhizal seedlings in response to salinity compared to those of the non-mycorrhizal seedlings. Mycorrhizal seedlings not only significantly increased their ability to acquire K + , Ca 2+ , and Mg 2+ , but also maintained higher K + :Na + ratios in the leaves and lower Ca 2+ :Mg 2+ ratios than non-mycorrhizal seedlings during salt stress. These results suggest that the salt tolerance of E. angustifolia seedlings could be enhanced by R. irregularis. The arbuscular mycorrhizal symbiosis could be a promising method to restore and utilize salt-alkaline land in northern China.

Biodegradation of 3,5-dimethyl-2,4-dichlorophenol in saline wastewater by newly isolated Penicillium sp. yz11-22N2.

PubMed

Yan, Zhou; He, Huijun; Yang, Chunping; Zeng, Guangming; Luo, Le; Jiao, Panpan; Li, Huiru; Lu, Li

2017-07-01

In this study, the performance of 3,5-dimethyl-2,4-dichlorophenol (DCMX) degradation by a screened strain was investigated. 18S rDNA and the neighbor-joining method were used for identification of the isolated strain. The results of phylogenetic analysis and scanning electron micrographs showed that the most probable identity of the screened strain should be Penicillium sp. Growth characteristics of Penicillium sp. and degradation processes of DCMX were examined. Fourier transform infrared spectroscopy of the inoculated DCMX solution was recorded, which supported the capacity of DCMX degradation by the screened Penicillium sp. Under different salinity conditions, the highest growth rate and removal efficiency for DCMX were obtained at pH6.0. The removal efficiency decreased from 100% to 66% when the DCMX concentration increased from 5 to 60mg/L, respectively. Using a Box-Behnken design, the maximum DCMX removal efficiency was determined to be 98.4%. With acclimation to salinity, higher removal efficiency could be achieved. The results demonstrate that the screened Penicillium sp. has the capability for degradation of DCMX. Copyright © 2017. Published by Elsevier B.V.

Arbuscular Mycorrhizal Symbiosis Modulates Antioxidant Response and Ion Distribution in Salt-Stressed Elaeagnus angustifolia Seedlings

PubMed Central

Chang, Wei; Sui, Xin; Fan, Xiao-Xu; Jia, Ting-Ting; Song, Fu-Qiang

2018-01-01

Elaeagnus angustifolia L. is a drought-resistant species. Arbuscular mycorrhizal symbiosis is considered to be a bio-ameliorator of saline soils that can improve salinity tolerance in plants. The present study investigated the effects of inoculation with the arbuscular mycorrhizal fungus Rhizophagus irregularis on the biomass, antioxidant enzyme activities, and root, stem, and leaf ion accumulation of E. angustifolia seedlings grown during salt stress conditions. Salt-stressed mycorrhizal seedlings produced greater root, stem, and leaf biomass than the uninoculated stressed seedlings. In addition, the seedlings colonized by R. irregularis showed notably higher activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate peroxidase (APX) in the leaves of the mycorrhizal seedlings in response to salinity compared to those of the non-mycorrhizal seedlings. Mycorrhizal seedlings not only significantly increased their ability to acquire K+, Ca2+, and Mg2+, but also maintained higher K+:Na+ ratios in the leaves and lower Ca2+:Mg2+ ratios than non-mycorrhizal seedlings during salt stress. These results suggest that the salt tolerance of E. angustifolia seedlings could be enhanced by R. irregularis. The arbuscular mycorrhizal symbiosis could be a promising method to restore and utilize salt-alkaline land in northern China. PMID:29675008

Chemically induced disseminated pythiosis in BALB/c mice: A new experimental model for Pythium insidiosum infection.

PubMed

Tondolo, Juliana S M; Loreto, Érico S; Ledur, Pauline C; Jesus, Francielli P K; Silva, Taiara M; Kommers, Glaucia D; Alves, Sydney H; Santurio, Janio M

2017-01-01

Pythiosis is a severe and life-threatening disease that affects humans and various animal species. We report a model of vascular/disseminated pythiosis occurring after subcutaneous inoculation of 2 x 104 Pythium insidiosum zoospores/mL in immunocompromised BALB/c mice. For this model, we carried out two rounds of experiments. First, we evaluated two protocols of immunosuppression before inoculation: cyclophosphamide at 150 mg/kg (CYP group) and cyclophosphamide 200 mg/kg plus hydrocortisone acetate at 250 mg/kg (CYP+HCA group). It was not possible to obtain mortality in the CYP group; however, the combination of CYP+HCA altered disease outcomes, with mortality rates reaching 60%. Second, we used the CYP+HCA immunosuppression protocol to analyze the histological and immunological statuses triggered by disease. When we inoculated immunocompetent mice with P. insidiosum zoospores, self-healing occurred via increased levels of IL-2, IFN-γ and IL-17A, which are characteristic of the Th1/Th17 cytokine response. For infected and immunosuppressed mice, the cytokine profiles showed high levels of IL-10, IL-6 and TNF-α. Increased IL-10 values are related to fungal infection susceptibility and led us to speculate that infection may be established through suppression of the host immune response. In addition, histopathological evaluation of the kidneys and liver demonstrated the presence of hyphae and the cellular findings suggested an acute vascular inflammation that mimics vascular/disseminated pythiosis in humans. This is the first murine model for pythiosis that is useful both for understanding the pathogenesis of this disease and for evaluating new treatment approaches.

Metabolic responses of willow (Salix purpurea L.) leaves to mycorrhization as revealed by mass spectrometry and 1H NMR spectroscopy metabolite profiling

PubMed Central

Aliferis, Konstantinos A.; Chamoun, Rony; Jabaji, Suha

2015-01-01

The root system of most terrestrial plants form symbiotic interfaces with arbuscular mycorrhizal fungi (AMF), which are important for nutrient cycling and ecosystem sustainability. The elucidation of the undergoing changes in plants' metabolism during symbiosis is essential for understanding nutrient acquisition and for alleviation of soil stresses caused by environmental cues. Within this context, we have undertaken the task of recording the fluctuation of willow (Salix purpurea L.) leaf metabolome in response to AMF inoculation. The development of an advanced metabolomics/bioinformatics protocol employing mass spectrometry (MS) and 1H NMR analyzers combined with the in-house-built metabolite library for willow (http://willowmetabolib.research.mcgill.ca/index.html) are key components of the research. Analyses revealed that AMF inoculation of willow causes up-regulation of various biosynthetic pathways, among others, those of flavonoid, isoflavonoid, phenylpropanoid, and the chlorophyll and porphyrin pathways, which have well-established roles in plant physiology and are related to resistance against environmental stresses. The recorded fluctuation in the willow leaf metabolism is very likely to provide AMF-inoculated willows with a significant advantage compared to non-inoculated ones when they are exposed to stresses such as, high levels of soil pollutants. The discovered biomarkers of willow response to AMF inoculation and corresponding pathways could be exploited in biomarker-assisted selection of willow cultivars with superior phytoremediation capacity or genetic engineering programs. PMID:26042135

Nitrification at different salinities: Biofilm community composition and physiological plasticity.

PubMed

Gonzalez-Silva, Blanca M; Jonassen, Kjell Rune; Bakke, Ingrid; Østgaard, Kjetill; Vadstein, Olav

2016-05-15

This paper describes an experimental study of microbial communities of three moving bed biofilm reactors (MBBR) inoculated with nitrifying cultures originated from environments with different salinity; freshwater, brackish (20‰) and seawater. All reactors were run until they operated at a conversion efficiency of >96%. The microbial communities were profiled using 454-pyrosequencing of 16S rRNA gene amplicons. Statistical analysis was used to investigate the differences in microbial community structure and distribution of the nitrifying populations with different salinity environments. Nonmetric multidimensional scaling analysis (NMDS) and the PERMANOVA test based on Bray-Curtis similarities revealed significantly different community structure in the three reactors. The brackish reactor showed lower diversity index than fresh and seawater reactors. Venn diagram showed that 60 and 78% of the total operational taxonomic units (OTUs) in the ammonia-oxidizing bacteria (AOB) and nitrite-oxidizing bacteria (NOB) guild, respectively, were unique OTUs for a given reactor. Similarity Percentages (SIMPER) analysis showed that two-thirds of the total difference in community structure between the reactors was explained by 10 OTUs, indicating that only a small number of OTUs play a numerically dominant role in the nitrification process. Acute toxicity of salt stress on ammonium and nitrite oxidizing activities showed distinctly different patterns, reaching 97% inhibition of the freshwater reactor for ammonium oxidation rate. In the brackish culture, inhibition was only observed at maximal level of salinity, 32‰. In the fully adapted seawater culture, higher activities were observed at 32‰ than at any of the lower salinities. Copyright © 2016 Elsevier Ltd. All rights reserved.

Boron neutron capture therapy (BNCT) for the treatment of liver metastases: biodistribution studies of boron compounds in an experimental model.

PubMed

Garabalino, Marcela A; Monti Hughes, Andrea; Molinari, Ana J; Heber, Elisa M; Pozzi, Emiliano C C; Cardoso, Jorge E; Colombo, Lucas L; Nievas, Susana; Nigg, David W; Aromando, Romina F; Itoiz, Maria E; Trivillin, Verónica A; Schwint, Amanda E

2011-03-01

We previously demonstrated the therapeutic efficacy of different boron neutron capture therapy (BNCT) protocols in an experimental model of oral cancer. BNCT is based on the selective accumulation of (10)B carriers in a tumor followed by neutron irradiation. Within the context of exploring the potential therapeutic efficacy of BNCT for the treatment of liver metastases, the aim of the present study was to perform boron biodistribution studies in an experimental model of liver metastases in rats. Different boron compounds and administration conditions were assayed to determine which administration protocols would potentially be therapeutically useful in in vivo BNCT studies at the RA-3 nuclear reactor. A total of 70 BDIX rats were inoculated in the liver with syngeneic colon cancer cells DHD/K12/TRb to induce the development of subcapsular tumor nodules. Fourteen days post-inoculation, the animals were used for biodistribution studies. We evaluated a total of 11 administration protocols for the boron compounds boronophenylalanine (BPA) and GB-10 (Na(2)(10)B(10)H(10)), alone or combined at different dose levels and employing different administration routes. Tumor, normal tissue, and blood samples were processed for boron measurement by atomic emission spectroscopy. Six protocols proved potentially useful for BNCT studies in terms of absolute boron concentration in tumor and preferential uptake of boron by tumor tissue. Boron concentration values in tumor and normal tissues in the liver metastases model show it would be feasible to reach therapeutic BNCT doses in tumor without exceeding radiotolerance in normal tissue at the thermal neutron facility at RA-3. © Springer-Verlag 2010

Boron Neutron Capture Therapy (BCNT) for the Treatment of Liver Metastases: Biodistribution Studies of Boron Compounds in an Experimental Model

DOE Office of Scientific and Technical Information (OSTI.GOV)

Marcela A. Garabalino; Andrea Monti Hughes; Ana J. Molinari

2011-03-01

Abstract We previously demonstrated the therapeutic efficacy of different boron neutron capture therapy (BNCT) protocols in an experimental model of oral cancer. BNCT is based on the selective accumulation of 10B carriers in a tumor followed by neutron irradiation. Within the context of exploring the potential therapeutic efficacy of BNCT for the treatment of liver metastases, the aim of the present study was to perform boron biodistribution studies in an experimental model of liver metastases in rats. Different boron compounds and administration conditions were assayed to determine which administration protocols would potentially be therapeutically useful in in vivo BNCT studiesmore » at the RA-3 nuclear reactor. A total of 70 BDIX rats were inoculated in the liver with syngeneic colon cancer cells DHD/K12/TRb to induce the development of subcapsular tumor nodules. Fourteen days post-inoculation, the animals were used for biodistribution studies. We evaluated a total of 11 administration protocols for the boron compounds boronophenylalanine (BPA) and GB-10 (Na210B10H10), alone or combined at different dose levels and employing different administration routes. Tumor, normal tissue, and blood samples were processed for boron measurement by atomic emission spectroscopy. Six protocols proved potentially useful for BNCT studies in terms of absolute boron concentration in tumor and preferential uptake of boron by tumor tissue. Boron concentration values in tumor and normal tissues in the liver metastases model show it would be feasible to reach therapeutic BNCT doses in tumor without exceeding radiotolerance in normal tissue at the thermal neutron facility at RA-3.« less

The Clinical Urine Culture: Enhanced Techniques Improve Detection of Clinically Relevant Microorganisms

PubMed Central

Price, Travis K.; Dune, Tanaka; Hilt, Evann E.; Thomas-White, Krystal J.; Kliethermes, Stephanie; Brincat, Cynthia; Brubaker, Linda; Wolfe, Alan J.

2016-01-01

Enhanced quantitative urine culture (EQUC) detects live microorganisms in the vast majority of urine specimens reported as “no growth” by the standard urine culture protocol. Here, we evaluated an expanded set of EQUC conditions (expanded-spectrum EQUC) to identify an optimal version that provides a more complete description of uropathogens in women experiencing urinary tract infection (UTI)-like symptoms. One hundred fifty adult urogynecology patient-participants were characterized using a self-completed validated UTI symptom assessment (UTISA) questionnaire and asked “Do you feel you have a UTI?” Women responding negatively were recruited into the no-UTI cohort, while women responding affirmatively were recruited into the UTI cohort; the latter cohort was reassessed with the UTISA questionnaire 3 to 7 days later. Baseline catheterized urine samples were plated using both standard urine culture and expanded-spectrum EQUC protocols: standard urine culture inoculated at 1 μl onto 2 agars incubated aerobically; expanded-spectrum EQUC inoculated at three different volumes of urine onto 7 combinations of agars and environments. Compared to expanded-spectrum EQUC, standard urine culture missed 67% of uropathogens overall and 50% in participants with severe urinary symptoms. Thirty-six percent of participants with missed uropathogens reported no symptom resolution after treatment by standard urine culture results. Optimal detection of uropathogens could be achieved using the following: 100 μl of urine plated onto blood (blood agar plate [BAP]), colistin-nalidixic acid (CNA), and MacConkey agars in 5% CO2 for 48 h. This streamlined EQUC protocol achieved 84% uropathogen detection relative to 33% detection by standard urine culture. The streamlined EQUC protocol improves detection of uropathogens that are likely relevant for symptomatic women, giving clinicians the opportunity to receive additional information not currently reported using standard urine culture techniques. PMID:26962083

Efficacy of detergent and water versus bleach for disinfection of direct contact ophthalmic lenses.

PubMed

Abbey, Ashkan M; Gregori, Ninel Z; Surapaneni, Krishna; Miller, Darlene

2014-06-01

Although manufacturers recommend cleaning ophthalmic lenses with detergent and water and then with a specific disinfectant, disinfectants are rarely used in ophthalmic practices. The aim of this pilot study was to evaluate the efficacy of detergent and water versus that of bleach, a recommended disinfectant, to eliminate common ocular bacteria and viruses from ophthalmic lenses. Three bacterial strains (Staphylococcus epidermidis, Corynebacterium straitum, and methicillin-resistant Staphylococcus aureus and 2 viral strains (adenovirus and herpes simplex virus [HSV] type-1) were individually inoculated onto 20 gonioscopy and laser lenses. The lenses were washed with detergent and water and then disinfected with 10% bleach. All the lenses were cultured after inoculation, after washing with detergent and water, and after disinfecting with the bleach. Bacterial cultures in thioglycollate broth were observed for 3 weeks, and viral cultures were observed for 2 weeks. The presence of viruses was also detected by multiplex polymerase chain reaction (PCR). All 20 lenses inoculated with S. epidermidis, C. straitum, adenovirus, and HSV-1 showed growth after inoculation but no growth after washing with detergent/water and after disinfecting with the bleach. All lenses showed positive HSV and adenovirus PCR results after inoculation and negative PCR results after washing with detergent/water and after disinfecting with bleach. All methicillin-resistant S. aureus-contaminated lenses showed growth after inoculation and no growth after washing with detergent and water. However, 1 lens showed positive growth after disinfecting with bleach. Cleaning with detergent and water seemed to effectively eliminate bacteria and viruses from the surface of contaminated ophthalmic lenses. Further studies are warranted to design practical disinfection protocols that minimize lens damage.

Repercussions of salinity changes and osmotic stress in marine phytoplankton species

NASA Astrophysics Data System (ADS)

D'ors, A.; Bartolomé, M. C.; Sánchez-Fortún, S.

2016-06-01

The short-term effect of low salinity was studied using laboratory protocols on some coastal phytoplankton species such as chlorophycea Tetraselmis suecica, among diatom the strain Nitzschia N1c1 and dinoflagellates Alexandrium minutum and Prorocentrum lima. All of cultures were exposed to low salinities, and cell growth rate, photosynthetic quantum yield (ΦPSII), and gross photosynthesis (Pg) were analyzed. Growth rate inhibition was similar in all species, and all of them also tolerate short-term exposures to salinities in the range 5-35. There were no significant differences between ΦPSII and Pg endpoints from Tetraselmis suecica and Nitzschia sp., while Alexandrium minutum and Prorocentrum lima displayed a higher affectation rate on Pg than on ΦPSII activity. The influence of low salinity was higher on respiration in T. suecica, while both dinoflagellates had higher net photosynthesis. Nitzschia sp. exhibited similar involvement of the two photosynthetic parameters. Therefore, although the four phytoplankton monocultures studied are able to survive in internal areas of estuaries under low salinity conditions, the photosynthetic activity is more affected than the growth rate in all phytoplankton communities studied except in chlorophycea T. suecica, which has increased tolerance for this salinity decrease.

Acute toxic effects of single dose dacarbazine: hematological and histological changes in an animal model.

PubMed

Milijašević, B; Stefanović, D; Lalić-Popović, M; Tomić, Z; Kolarović, J; Lalošević, D; Mikov, M

2014-11-01

Treatment of advanced soft tissue sarcoma usually includes dacarbazine (DTIC), an alkylating agent that methylates DNA and is active during all phases of the cell cycle. Common side effects of DTIC include nausea, vomiting, impaired liver and kidney function, myelosuppression, and pneumonia. There are no accounts, however, of histological and hematological changes caused by DTIC. We investigated acute hematological and morphological changes in different organs and in tumors that were caused by a single dose of DTIC. Adult Syrian golden hamsters were inoculated with a suspension of tumorigenic baby hamster kidney (BHK) cells by subcutaneous injection. On day 14 after inoculation, doses of 1.4, 1.6, 1.8 or 2.0 g/m(2) DTIC were injected intraperitoneally into the hamsters. Hamsters in the control group were injected with physiological saline in the same way. Seven days after drug or saline injection the animals were sacrificed and samples of blood, heart, kidney, liver, lungs, spleen, small intestine and tumor were excised, processed and analyzed. Mitoses were counted using an ocular extension with engraved frame. Anemia, thrombocytopenia and leukocytosis were found in the control group of hamsters with fibrosarcoma, whereas animals with fibrosarcoma treated with DTIC developed anemia, thrombocytopenia and leukopenia. Severe pneumonia and moderate hepatitis were detected in all DTIC treated groups. Effects of DTIC on tumor cells included rounding and enlargement of nuclei and rarefaction of chromatin. The number of mitoses was reduced with increasing doses of DTIC. Hepatitis, myelosuppression, pneumonia, and dose-related inhibition of tumor cell proliferation were observed after a single dose of DTIC.

In vivo biodistribution of CNTs using a BALB/c mouse experimental model.

PubMed

Fufă, Mariana Oana Mihaela; Mihaiescu, Dan Eduard; Mogoantă, Laurenţiu; Bălşeanu, Tudor Adrian; Mogoşanu, George Dan; Grumezescu, Alexandru Mihai; Bolocan, Alexandra

2015-01-01

Due to their unique behaviors, carbon nanotubes (CNTs)-based systems meet essential requirements for modern applications, such as electronics, optics, photovoltaics, fuel cells, aerospace engineering, military and biomedical applications. CNTs biocompatibility and toxic effects were assessed both in vitro and in vivo, in terms of hemocompatibility, cytocompatibility, immunoreactions and genetic behavior. The aim of this paper is to evaluate the in vivo biodistribution and biocompatibility of carbon nanopowder synthesized by plasma processing, using a BALB/c mouse experimental model. Three months old BALB/c mice were aseptically injected with 100 μL of 1 mg/mL dispersions. The obtained carbon-based nano-systems were dispersed in saline solution and subsequently sterilized by using a 30 minutes treatment with UV irradiation. The reference mice were injected with 100 μL of saline. The mice were kept under standard conditions of light, temperature, humidity, food and water (ad libitum) before the vital organ harvest. The animal welfare was daily monitored. At two and 10 days after the inoculation, the animals were euthanized under general anesthesia, for the sampling of internal organs (brain, myocardium, pancreas, liver, lung, kidney and spleen). No animal died during the experiment. Brain, myocardium and pancreas were histologically normal, with no tissue damage, inflammatory infiltrate or inorganic deposits. CNTs were evidenced only in hepatic, renal, pulmonary and spleen tissue samples. Increased amounts of inorganic granular structures were reported after 10 days of treatment, when compared to the short-term (two days) inoculation. Our BALB/c mouse experimental model was found to be useful for the in vivo assessment of biodistribution and biocompatibility of CNTs.

An intra-laboratory cultural and real-time PCR method comparison and evaluation for the detection of subclinical paratuberculosis in dairy herds.

PubMed

Heuvelink, Annet; Hassan, Abdulwahed Ahmed; van Weering, Hilmar; van Engelen, Erik; Bülte, Michael; Akineden, Ömer

2017-05-01

Mycobacterium avium subsp. paratuberculosis (MAP) is a vigorous microorganism which causes incurable chronic enteritis, Johne's disease (JD) in cattle. A target of control programmes for JD is to accurately detect MAP-infected cattle early to reduce disease transmission. The present study evaluated the efficacy of two different cultural procedures and a TaqMan real-time PCR assay for detection of subclinical paratuberculosis in dairy herds. Therefore, sixty-one faecal samples were collected from two Dutch dairy herds (n = 40 and n = 21, respectively) which were known to be MAP-ELISA positive. All individual samples were assessed using two different cultural protocols in two different laboratories. The first cultural protocol (first laboratory) included a decontamination step with 0.75% hexadecylpyridinium chloride (HPC) followed by inoculation on Herrold's egg yolk media (HEYM). The second protocol (second laboratory) comprised of a decontamination step using 4% NaOH and malachite green-oxalic acid followed by inoculation on two media, HEYM and in parallel on modified Löwenstein-Jensen media (mLJ). For the TaqMan real-time PCR assay, all faecal samples were tested in two different laboratories using TaqMan® MAP (Johne's) reagents (Life Technologies). The cultural procedures revealed positive reactions in 1.64% of the samples for cultivation protocol 1 and 6.56 and 8.20% of the samples for cultivation protocol 2, respectively. The results of the TaqMan real-time PCR performed in two different laboratories yielded 13.11 and 19.76% positive reaction. The kappa test showed proportional agreement 0.54 between the mLJ media (second laboratory) and TaqMan® real-time PCR method (second laboratory). In conclusion, the TaqMan real-time PCR could be a strongly useful and efficient assay for the detection of subclinical paratuberculosis in dairy cattle leading to an improvement in the efficiency of MAP control strategies.

A Standardized Inoculation Protocol to Test Wheat Cultivars for Reaction to Head Blast caused by Magnaporthe oryzae (Triticum pathotype)

USDA-ARS?s Scientific Manuscript database

Wheat blast, caused by the Triticum pathotype of M. oryzae (MoT), poses a significant threat to wheat production worldwide. Because this pathotype does not occur in the U.S., it is important to prepare for its possible introduction. As part of this preparation, over 500 U.S. wheat cultivars were tes...

Choriodecidual Group B Streptococcal Inoculation Induces Fetal Lung Injury without Intra-Amniotic Infection and Preterm Labor in Macaca nemestrina

PubMed Central

Adams Waldorf, Kristina M.; Gravett, Michael G.; McAdams, Ryan M.; Paolella, Louis J.; Gough, G. Michael; Carl, David J.; Bansal, Aasthaa; Liggitt, H. Denny; Kapur, Raj P.; Reitz, Frederick B.; Rubens, Craig E.

2011-01-01

Background Early events leading to intrauterine infection and fetal lung injury remain poorly defined, but may hold the key to preventing neonatal and adult chronic lung disease. Our objective was to establish a nonhuman primate model of an early stage of chorioamnionitis in order to determine the time course and mechanisms of fetal lung injury in utero. Methodology/Principal Findings Ten chronically catheterized pregnant monkeys (Macaca nemestrina) at 118–125 days gestation (term = 172 days) received one of two treatments: 1) choriodecidual and intra-amniotic saline (n = 5), or 2) choriodecidual inoculation of Group B Streptococcus (GBS) 1×106 colony forming units (n = 5). Cesarean section was performed regardless of labor 4 days after GBS or 7 days after saline infusion to collect fetal and placental tissues. Only two GBS animals developed early labor with no cervical change in the remaining animals. Despite uterine quiescence in most cases, blinded review found histopathological evidence of fetal lung injury in four GBS animals characterized by intra-alveolar neutrophils and interstitial thickening, which was absent in controls. Significant elevations of cytokines in amniotic fluid (TNF-α, IL-8, IL-1β, IL-6) and fetal plasma (IL-8) were detected in GBS animals and correlated with lung injury (p<0.05). Lung injury was not directly caused by GBS, because GBS was undetectable in amniotic fluid (∼10 samples tested/animal), maternal and fetal blood by culture and polymerase chain reaction. In only two cases was GBS cultured from the inoculation site in low numbers. Chorioamnionitis occurred in two GBS animals with lung injury, but two others with lung injury had normal placental histology. Conclusions/Significance A transient choriodecidual infection can induce cytokine production, which is associated with fetal lung injury without overt infection of amniotic fluid, chorioamnionitis or preterm labor. Fetal lung injury may, thus, occur silently without symptoms and before the onset of the fetal systemic inflammatory response syndrome. PMID:22216148

Transmission of Salmonella to broilers by contaminated larval and adult lesser mealworms, Alphitobius diaperinus (Coleoptera: Tenebrionidae).

PubMed

Roche, A J; Cox, N A; Richardson, L J; Buhr, R J; Cason, J A; Fairchild, B D; Hinkle, N C

2009-01-01

The ability of the lesser mealworm, Alphitobius diaperinus (Panzer), commonly known as the darkling beetle, to transmit marker Salmonella Typhimurium to day-of-hatch broiler chicks was evaluated, as well as the spread to nonchallenged pen mates. In trial 1, day-of-hatch chicks were orally gavaged with 4 larval or 4 adult beetles that had been exposed to marker Salmonella-inoculated feed for 72 h. In addition, chicks were gavaged with the marker Salmonella in saline solution. These chicks were then placed into pens to serve as challenged broilers. In trial 2, all pens received 2 challenged chicks that were gavaged with larvae or beetles that had been exposed to marker Salmonella-inoculated feed for 24 h and then removed from the inoculated feed for a period of 7 d. At 3 wk of age, cecal samples from the marker Salmonella-challenged broilers and from 5 pen mates in trial 1, or 10 pen mates in trial 2, were evaluated for the presence of the marker Salmonella in their ceca, and at 6 wk of age, all remaining pen mates were sampled. To monitor the presence of the marker Salmonella within pens, stepped-on drag swab litter samples were taken weekly. For the Salmonella-saline pens, 29 to 33% of the broilers that had been challenged and 10 to 55% of the pen mates were positive at 3 wk of age, and only 2 to 6% had positive ceca at 6 wk. For the pens challenged with adult beetles, 0 to 57% of the challenged broilers and 20 to 40% of the pen mates had positive ceca at 3 wk, and 4 to 7% were positive at 6 wk. The pens challenged with larvae had the greatest percentage of marker Salmonella-positive broilers; 25 to 33% of the challenged broilers and 45 to 58% of pen mates were positive at 3 wk, and 11 to 27% were positive at 6 wk. These results demonstrated that ingestion of larval or adult beetles contaminated with a marker Salmonella could be a significant vector for transmission to broilers.

EXPERIMENTAL CHALLENGE STUDY OF FV3-LIKE RANAVIRUS INFECTION IN PREVIOUSLY FV3-LIKE RANAVIRUS INFECTED EASTERN BOX TURTLES (TERRAPENE CAROLINA CAROLINA) TO ASSESS INFECTION AND SURVIVAL.

PubMed

Hausmann, Jennifer C; Wack, Allison N; Allender, Matthew C; Cranfield, Mike R; Murphy, Kevin J; Barrett, Kevin; Romero, Jennell L; Wellehan, James F X; Blum, Stella A; Zink, M Christine; Bronson, Ellen

2015-12-01

The Maryland Zoo in Baltimore experienced an outbreak of Frog virus-3 (FV3)-like ranavirus during the summer of 2011, during which 14 of 27 (52%) of its captive eastern box turtles (Terrapene carolina carolina) survived. To assess survival, immunity, and viral shedding, an experimental challenge study was performed in which the surviving, previously infected turtles were reinfected with the outbreak strain of FV3-like ranavirus. Seven turtles were inoculated with virus intramuscularly and four control turtles received saline intramuscularly. The turtles were monitored for 8 wk with blood and oral swabs collected for quantitative polymerase chain reaction (qPCR). During that time, one of seven (14%) inoculated turtles and none of the controls (0%) died; there was no significant difference in survival. Clinical signs of the inoculated turtles, except for the turtle that died, were mild compared to the original outbreak. Quantitative PCR for FV3-like ranavirus on blood and oral swabs was positive for all inoculated turtles and negative for all controls. The turtle that died had intracytoplasmic inclusion bodies in multiple organs. Three inoculated and two control turtles were euthanized at the end of the study. No inclusion bodies were present in any of the organs. Quantitative PCR detected FV3-like ranavirus in the spleen of a control turtle, which suggested persistence of the virus. The surviving five turtles were qPCR-negative for FV3-like ranavirus from blood and oral swabs after brumation. Quantitative PCR for Terrapene herpesvirus 1 found no association between ranavirus infection and herpesvirus loads. In conclusion, previously infected eastern box turtles can be reinfected with the same strain of FV3-like ranavirus and show mild to no clinical signs but can shed the virus from the oral cavity.

Inoculation of α-synuclein preformed fibrils into the mouse gastrointestinal tract induces Lewy body-like aggregates in the brainstem via the vagus nerve.

PubMed

Uemura, Norihito; Yagi, Hisashi; Uemura, Maiko T; Hatanaka, Yusuke; Yamakado, Hodaka; Takahashi, Ryosuke

2018-05-11

Intraneuronal α-synuclein (α-Syn) aggregates known as Lewy bodies (LBs) and the loss of dopaminergic neurons in the substantia nigra pars compacta (SNpc) are the pathological hallmarks of Parkinson's disease (PD). Braak's hypothesis based on autopsy studies suggests that Lewy pathology initially occurs in the enteric nervous system (ENS) and then travels retrogradely to the dorsal motor nucleus of the vagus nerve (dmX), proceeding from there in a caudo-rostral direction. Recent evidence that α-Syn aggregates propagate between interconnected neurons supports this hypothesis. However, there is no direct evidence demonstrating this transmission from the ENS to the dmX and then to the SNpc. We inoculated α-Syn preformed fibrils (PFFs) or phosphate-buffered saline (PBS) into the mouse gastric wall and analyzed the progression of the pathology. The mice inoculated with α-Syn PFFs, but not with PBS, developed phosphorylated α-Syn (p-α-Syn)-positive LB-like aggregates in the dmX at 45 days postinoculation. This aggregate formation was completely abolished when vagotomy was performed prior to inoculation of α-Syn PFFs, suggesting that the aggregates in the dmX were retrogradely induced via the vagus nerve. Unexpectedly, the number of neurons containing p-α-Syn-positive aggregates in the dmX decreased over time, and no further caudo-rostral propagation beyond the dmX was observed up to 12 months postinoculation. P-α-Syn-positive aggregates were also present in the myenteric plexus at 12 months postinoculation. However, unlike in patients with PD, there was no cell-type specificity in neurons containing those aggregates in this model. These results indicate that α-Syn PFF inoculation into the mouse gastrointestinal tract can induce α-Syn pathology resembling that of very early PD, but other factors are apparently required if further progression of PD pathology is to be replicated in this animal model.

Antimicrobial Solution or Saline Solution in Maintaining Catheter Patency and Preventing Catheter-Related Blood Infections in Patients With Malignancies

ClinicalTrials.gov

2013-02-13

Chronic Myeloproliferative Disorders; Infection; Leukemia; Lymphoma; Lymphoproliferative Disorder; Multiple Myeloma and Plasma Cell Neoplasm; Myelodysplastic Syndromes; Myelodysplastic/Myeloproliferative Neoplasms; Unspecified Adult Solid Tumor, Protocol Specific

A high-throughput virus-induced gene silencing protocol identifies genes involved in multi-stress tolerance

PubMed Central

2013-01-01

Background Understanding the function of a particular gene under various stresses is important for engineering plants for broad-spectrum stress tolerance. Although virus-induced gene silencing (VIGS) has been used to characterize genes involved in abiotic stress tolerance, currently available gene silencing and stress imposition methodology at the whole plant level is not suitable for high-throughput functional analyses of genes. This demands a robust and reliable methodology for characterizing genes involved in abiotic and multi-stress tolerance. Results Our methodology employs VIGS-based gene silencing in leaf disks combined with simple stress imposition and effect quantification methodologies for easy and faster characterization of genes involved in abiotic and multi-stress tolerance. By subjecting leaf disks from gene-silenced plants to various abiotic stresses and inoculating silenced plants with various pathogens, we show the involvement of several genes for multi-stress tolerance. In addition, we demonstrate that VIGS can be used to characterize genes involved in thermotolerance. Our results also showed the functional relevance of NtEDS1 in abiotic stress, NbRBX1 and NbCTR1 in oxidative stress; NtRAR1 and NtNPR1 in salinity stress; NbSOS1 and NbHSP101 in biotic stress; and NtEDS1, NbETR1, NbWRKY2 and NbMYC2 in thermotolerance. Conclusions In addition to widening the application of VIGS, we developed a robust, easy and high-throughput methodology for functional characterization of genes involved in multi-stress tolerance. PMID:24289810

Inoculation of Brassica oxyrrhina with plant growth promoting bacteria for the improvement of heavy metal phytoremediation under drought conditions.

PubMed

Ma, Ying; Rajkumar, Mani; Zhang, Chang; Freitas, Helena

2016-12-15

The aim of this study was to investigate the effects of drought resistant serpentine rhizobacteria on plant growth and metal uptake by Brassica oxyrrhina under drought stress (DS) condition. Two drought resistant serpentine rhizobacterial strains namely Pseudomonas libanensis TR1 and Pseudomonas reactans Ph3R3 were selected based on their ability to stimulate seedling growth in roll towel assay. Further assessment on plant growth promoting (PGP) parameters revealed their ability to produce indole-3-acetic acid, siderophore and 1-aminocyclopropane-1-carboxylate deaminase. Moreover, both strains exhibited high resistance to various heavy metals, antibiotics, salinity and extreme temperature. Inoculation of TR1 and Ph3R3 significantly increased plant growth, leaf relative water and pigment content of B. oxyrrhina, whereas decreased concentrations of proline and malondialdehyde in leaves under metal stress in the absence and presence of DS. Regardless of soil water conditions, TR1 and Ph3R3 greatly improved organ metal concentrations, translocation and bioconcentration factors of Cu and Zn. The successful colonization and metabolic activities of P. libanensis TR1 and P. reactans Ph3R3 represented positive effects on plant development and metal phytoremediation under DS. These results indicate that these strains could be used as bio-inoculants for the improvement of phytoremediation of metal polluted soils under semiarid conditions. Copyright © 2016 Elsevier B.V. All rights reserved.

Dark septate endophyte decreases stress on rice plants.

PubMed

Santos, Silvana Gomes Dos; Silva, Paula Renata Alves da; Garcia, Andres Calderin; Zilli, Jerri Édson; Berbara, Ricardo Luis Louro

Abiotic stress is one of the major limiting factors for plant development and productivity, which makes it important to identify microorganisms capable of increasing plant tolerance to stress. Dark septate endophytes can be symbionts of plants. In the present study, we evaluated the ability of dark septate endophytes isolates to reduce the effects of water stress in the rice varieties Nipponbare and Piauí. The experiments were performed under gnotobiotic conditions, and the water stress was induced with PEG. Four dark septate endophytes were isolated from the roots of wild rice (Oryza glumaepatula) collected from the Brazilian Amazon. Plant height as well as shoot and root fresh and dry matter were measured. Leaf protein concentrations and antioxidant enzyme activity were also estimated. The dark septate endophytes were grown in vitro in Petri dishes containing culture medium; they exhibited different levels of tolerance to salinity and water stress. The two rice varieties tested responded differently to inoculation with dark septate endophytes. Endophytes promoted rice plant growth both in the presence and in the absence of a water deficit. Decreased oxidative stress in plants in response to inoculation was observed in nearly all inoculated treatments, as indicated by the decrease in antioxidant enzyme activity. Dark septate endophytes fungi were shown to increase the tolerance of rice plants to stress caused by water deficiency. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Impact of Actinomyces naeslundii on bisphosphonate-related osteonecrosis of the jaws in ovariectomized rats with periodontitis.

PubMed

Li, Chun Lei; Seneviratne, Chaminda Jayampath; Huo, Lei; Lu, Weijia William; Zheng, Li Wu

2015-10-01

Bisphosphonates-related osteonecrosis of the jaws (BRONJ) is a severe complication of BPs therapy with unknown pathogenesis. This study aimed to evaluate the impact of Actinomyces naeslundii (A. naeslundii) on the progression of BRONJ in ovariectomized (OVX) rat model with periodontal diseases. Sixty rats were randomly assigned into four groups. All rats underwent bilateral ovariectomy. Six weeks after surgery, animals with periodontitis induced by ligature placement were administrated with normal saline (NS), NS &A. naeslundii inoculation, zolecdronic acid (ZA) and ZA &A. naeslundii inoculation for 12 weeks, respectively. Loads of total bacteria and A. naeslundii in the mouth were assessed by real time PCR. After sacrifice, the mandibles were harvested for micro-computed tomography (micro-CT) and histological examination. Real-time PCR demonstrated that A. naeslundii was not routinely found in the rats and ZA treatment did not promote its accumulation. Micro-CT examination disclosed that ligature placement induced significant alveolar bone loss, which was greatly attenuated by ZA treatment and aggravated by A. naeslundii. Histological assessment demonstrated that ZA treatment increased the risk of developing BRONJ-like disease but this condition was not worsen with the presence of A. naeslundii. Our study suggested that oral A. naeslundii inoculation aggravated periodontal disease but not BRONJ in our animal model. Copyright © 2015 European Association for Cranio-Maxillo-Facial Surgery. Published by Elsevier Ltd. All rights reserved.

Microbial cooperation in the rhizosphere improves liquorice growth under salt stress

PubMed Central

Egamberdieva, Dilfuza; Wirth, Stephan; Li, Li; Abd-Allah, Elsayed Fathi

2017-01-01

ABSTRACT Liquorice (Glycyrrhiza uralensis Fisch.) is one of the most widely used plants in food production, and it can also be used as an herbal medicine or for reclamation of salt-affected soils. Under salt stress, inhibition of plant growth, nutrient acquisition and symbiotic interactions between the medicinal legume liquorice and rhizobia have been observed. We recently evaluated the interactions between rhizobia and root-colonizing Pseudomonas in liquorice grown in potting soil and observed increased plant biomass, nodule numbers and nitrogen content after combined inoculation compared to plants inoculated with Mesorhizobium alone. Several beneficial effects of microbes on plants have been reported; studies examining the interactions between symbiotic bacteria and root-colonizing Pseudomonas strains under natural saline soil conditions are important, especially in areas where a hindrance of nutrients and niches in the rhizosphere are high. Here, we summarize our recent observations regarding the combined application of rhizobia and Pseudomonas on the growth and nutrient uptake of liquorice as well as the salt stress tolerance mechanisms of liquorice by a mutualistic interaction with microbes. Our observations indicate that microbes living in the rhizosphere of liquorice can form a mutualistic association and coordinate their involvement in plant adaptations to stress tolerance. These results support the development of combined inoculants for improving plant growth and the symbiotic performance of legumes under hostile conditions. PMID:27780398

Serologic evaluation, efficacy, and safety of a commerical modified-live canine distemper vaccine in domestic ferrets.

PubMed

Wimsatt, J; Jay, M T; Innes, K E; Jessen, M; Collins, J K

2001-05-01

To determine efficacy and safety of a commercial modified-live canine distemper virus (CDV) vaccine used for prophylaxis in domestic ferrets. Sixteen 16-week-old neutered male ferrets. Equal groups of ferrets were inoculated subcutaneously at 16 and 20 weeks of age with saline (0.9% NaCl) solution or a vaccine derived from the Onderstepoort CDV strain and attenuated in a primate cell line. Live virulent CDV was administered to all ferrets intranasally and orally 3 weeks after the second inoculation. Clinical signs and body weights were monitored regularly during the study. Blood samples for serologic examination were drawn prior to each inoculation, before challenge exposure, and 10, 15, and 21 days after exposure. Blood samples for reverse transcriptase polymerase chain reaction (RT-PCR) were obtained 5 days after the first vaccination, and 5, 10, 15, and 21 days after challenge exposure. After challenge exposure, control ferrets had significantly more clinical signs and weight loss, compared with vaccinates. All vaccinated ferrets survived, whereas all control ferrets died. The RT-PCR assay was successful in detecting CDV in blood and fresh or formalin-fixed tissues from infected ferrets. Findings suggest that the vaccine when given SC to domestic ferrets as directed is safe and protective against challenge exposure with virulent CDV. The RT-PCR assay may simplify detection of CDV in fresh and fixed tissues.

Influence of capsaicin infusion on secondary peristalsis in patients with gastroesophageal reflux disease

PubMed Central

Yi, Chih-Hsun; Lei, Wei-Yi; Hung, Jui-Sheng; Liu, Tso-Tsai; Chen, Chien-Lin; Pace, Fabio

2016-01-01

AIM To determine whether capsaicin infusion could influence heartburn perception and secondary peristalsis in patients with gastroesophageal reflux disease (GERD). METHODS Secondary peristalsis was performed with slow and rapid mid-esophageal injections of air in 10 patients with GERD. In a first protocol, saline and capsaicin-containing red pepper sauce infusions were randomly performed, whereas 2 consecutive sessions of capsaicin-containing red pepper sauce infusions were performed in a second protocol. Tested solutions including 5 mL of red pepper sauce diluted with 15 mL of saline and 20 mL of 0.9% saline were infused into the mid-esophagus via the manometric catheter at a rate of 10 mL/min with a randomized and double-blind fashion. During each study protocol, perception of heartburn, threshold volumes and peristaltic parameters for secondary peristalsis were analyzed and compared between different stimuli. RESULTS Infusion of capsaicin significantly increased heartburn perception in patients with GERD (P < 0.001), whereas repeated capsaicin infusion significantly reduced heartburn perception (P = 0.003). Acute capsaicin infusion decreased threshold volume of secondary peristalsis (P = 0.001) and increased its frequency (P = 0.01) during rapid air injection. The prevalence of GERD patients with successive secondary peristalsis during slow air injection significantly increased after capsaicin infusion (P = 0.001). Repeated capsaicin infusion increased threshold volume of secondary peristalsis (P = 0.002) and reduced the frequency of secondary peristalsis (P = 0.02) during rapid air injection. CONCLUSION Acute esophageal exposure to capsaicin enhances heartburn sensation and promotes secondary peristalsis in gastroesophageal reflux disease, but repetitive capsaicin infusion reverses these effects. PMID:28018112

The Effect of Diode Laser on Planktonic Enterococcus faecalis in Infected Root Canals in an Ex Vivo Model.

PubMed

Cretella, Gilda; Lajolo, Carlo; Castagnola, Raffaella; Somma, Francesco; Inchingolo, MariaTeresa; Marigo, Luca

2017-04-01

This study examined the bactericidal effect of diode laser irradiation against intracanal Enterococcus faecalis. m total of 128 extracted single-rooted and single-canal teeth were treated with ProTaper instruments (Dentsply Maillefer, Ballaigues, Switzerland). A total of 120 root canals were inoculated with E. faecalis for 21 days, and the samples were randomly divided into five groups: Group 1 (n = 24) samples were irrigated with only saline solution (positive controls); Group 2 (n = 24) was treated with only 5.25% sodium hypochlorite; Group 3 (n = 24) was irrigated with saline solutions activated by diode laser; Group 4 (n = 24) was treated with 5.25% sodium hypochlorite activated by diode laser; and Group 5 (n = 24) was irrigated with saline solution with methylene blue dye activated by the diode laser Fox (Sweden & Martina, Padova, Italy); additionally, eight teeth were not contaminated and their canals were irrigated with saline solution and used as a negative control. The Uro-Quick system was used to determine the microbial residual charge. The data were analyzed using Pearson's chi-square test (p < 0.001). A statistically significant reduction in bacterial count was observed in Group 2 and Group 4 (p < 0.001). There were no statistically significant differences among the other groups (p > 0.001). Evidence indicates that the diode laser was not more effective than sodium hypochlorite in reducing free bacteria.

Plant Growth Promoting Rhizobacteria and Silicon Synergistically Enhance Salinity Tolerance of Mung Bean

PubMed Central

Mahmood, Sajid; Daur, Ihsanullah; Al-Solaimani, Samir G.; Ahmad, Shakeel; Madkour, Mohamed H.; Yasir, Muhammad; Hirt, Heribert; Ali, Shawkat; Ali, Zahir

2016-01-01

The present study explored the eco-friendly approach of utilizing plant-growth-promoting rhizobacteria (PGPR) inoculation and foliar application of silicon (Si) to improve the physiology, growth, and yield of mung bean under saline conditions. We isolated 18 promising PGPR from natural saline soil in Saudi Arabia, and screened them for plant-growth-promoting activities. Two effective strains were selected from the screening trial, and were identified as Enterobacter cloacae and Bacillus drentensis using matrix-assisted laser desorption ionization-time-of-flight mass spectrometry and 16S rRNA gene sequencing techniques, respectively. Subsequently, in a 2-year mung bean field trial, using a randomized complete block design with a split-split plot arrangement, we evaluated the two PGPR strains and two Si levels (1 and 2 kg ha−1), in comparison with control treatments, under three different saline irrigation conditions (3.12, 5.46, and 7.81 dS m−1). The results indicated that salt stress substantially reduced stomatal conductance, transpiration rate, relative water content (RWC), total chlorophyll content, chlorophyll a, chlorophyll b, carotenoid content, plant height, leaf area, dry biomass, seed yield, and salt tolerance index. The PGPR strains and Si levels independently improved all the aforementioned parameters. Furthermore, the combined application of the B. drentensis strain with 2 kg Si ha−1 resulted in the greatest enhancement of mung bean physiology, growth, and yield. Overall, the results of this study provide important information for the benefit of the agricultural industry. PMID:27379151

Comparison of U.S. Environmental Protection Agency and U.S. Composting Council microbial detection methods in finished compost and regrowth potential of Salmonella spp. and Escherichia coli O157:H7 in finished compost.

PubMed

Reynnells, Russell; Ingram, David T; Roberts, Cheryl; Stonebraker, Richard; Handy, Eric T; Felton, Gary; Vinyard, Bryan T; Millner, Patricia D; Sharma, Manan

2014-07-01

Bacterial pathogens may survive and regrow in finished compost due to incomplete thermal inactivation during or recontamination after composting. Twenty-nine finished composts were obtained from 19 U.S. states and were separated into three broad feedstock categories: biosolids (n=10), manure (n=4), and yard waste (n=15). Three replicates of each compost were inoculated with ≈ 1-2 log CFU/g of nonpathogenic Escherichia coli, Salmonella spp., and E. coli O157:H7. The U.S. Environmental Protection Agency's (EPA) protocols and U.S. Composting Council's (USCC) Test Methods for the Examination of Composting and Compost (TMECC) were compared to determine which method recovered higher percentages of inoculated E. coli (representing fecal coliforms) and Salmonella spp. from 400-g samples of finished composts. Populations of Salmonella spp. and E. coli O157:H7 were determined over 3 days while stored at 25°C and compared to physicochemical parameters to predict their respective regrowth potentials. EPA Method 1680 recovered significantly (p=0.0003) more inoculated E. coli (68.7%) than TMECC 07.01 (48.1%) due to the EPA method using more compost in the initial homogenate, larger transfer dilutions, and a larger most probable number scheme compared to TMECC 07.01. The recoveries of inoculated Salmonella spp. by Environmental Protection Agency Method 1682 (89.1%) and TMECC 07.02 (72.4%) were not statistically significant (p=0.44). The statistically similar recovery percentages may be explained by the use of a nonselective pre-enrichment step used in both methods. No physicochemical parameter (C:N, moisture content, total organic carbon) was able to serve as a sole predictor of regrowth of Salmonella spp. or E. coli O157:H7 in finished compost. However, statistical analysis revealed that the C:N ratio, total organic carbon, and moisture content all contributed to pathogen regrowth potential in finished composts. It is recommended that the USCC modify TMECC protocols to test larger amounts of compost in the initial homogenate to facilitate greater recovery of target organisms.

Capillary Electrophoresis Analysis of Organic Amines and Amino Acids in Saline and Acidic Samples Using the Mars Organic Analyzer

NASA Astrophysics Data System (ADS)

Stockton, Amanda M.; Chiesl, Thomas N.; Lowenstein, Tim K.; Amashukeli, Xenia; Grunthaner, Frank; Mathies, Richard A.

2009-11-01

The Mars Organic Analyzer (MOA) has enabled the sensitive detection of amino acid and amine biomarkers in laboratory standards and in a variety of field sample tests. However, the MOA is challenged when samples are extremely acidic and saline or contain polyvalent cations. Here, we have optimized the MOA analysis, sample labeling, and sample dilution buffers to handle such challenging samples more robustly. Higher ionic strength buffer systems with pKa values near pH 9 were developed to provide better buffering capacity and salt tolerance. The addition of ethylaminediaminetetraacetic acid (EDTA) ameliorates the negative effects of multivalent cations. The optimized protocol utilizes a 75 mM borate buffer (pH 9.5) for Pacific Blue labeling of amines and amino acids. After labeling, 50 mM (final concentration) EDTA is added to samples containing divalent cations to ameliorate their effects. This optimized protocol was used to successfully analyze amino acids in a saturated brine sample from Saline Valley, California, and a subcritical water extract of a highly acidic sample from the Río Tinto, Spain. This work expands the analytical capabilities of the MOA and increases its sensitivity and robustness for samples from extraterrestrial environments that may exhibit pH and salt extremes as well as metal ions.

Capillary electrophoresis analysis of organic amines and amino acids in saline and acidic samples using the Mars organic analyzer.

PubMed

Stockton, Amanda M; Chiesl, Thomas N; Lowenstein, Tim K; Amashukeli, Xenia; Grunthaner, Frank; Mathies, Richard A

2009-11-01

The Mars Organic Analyzer (MOA) has enabled the sensitive detection of amino acid and amine biomarkers in laboratory standards and in a variety of field sample tests. However, the MOA is challenged when samples are extremely acidic and saline or contain polyvalent cations. Here, we have optimized the MOA analysis, sample labeling, and sample dilution buffers to handle such challenging samples more robustly. Higher ionic strength buffer systems with pK(a) values near pH 9 were developed to provide better buffering capacity and salt tolerance. The addition of ethylaminediaminetetraacetic acid (EDTA) ameliorates the negative effects of multivalent cations. The optimized protocol utilizes a 75 mM borate buffer (pH 9.5) for Pacific Blue labeling of amines and amino acids. After labeling, 50 mM (final concentration) EDTA is added to samples containing divalent cations to ameliorate their effects. This optimized protocol was used to successfully analyze amino acids in a saturated brine sample from Saline Valley, California, and a subcritical water extract of a highly acidic sample from the Río Tinto, Spain. This work expands the analytical capabilities of the MOA and increases its sensitivity and robustness for samples from extraterrestrial environments that may exhibit pH and salt extremes as well as metal ions.

Bothrops jararaca envenomation: Pathogenesis of hemostatic disturbances and intravascular hemolysis.

PubMed

Senise, Luana V; Yamashita, Karine M; Santoro, Marcelo L

2015-11-01

To attain fully functional biological activity, vitamin-K dependent coagulation factors (VKDCF) are γ-carboxylated prior to secretion from liver. Warfarin impairs the γ-carboxylation, and consequently their physiological function. Bothrops jararaca snake venom (BjV) contains several activators of blood coagulation, especially procoagulant enzymes (prothrombin and factor X activators) and thrombin-like enzymes. In order to clarify the relative contribution of prothrombin and factor X activators to the hemostatic disturbances occurring during experimental B. jararaca envenomation, warfarin was used to deplete VKDCF, prior to BjV administration. Male Wistar rats were pretreated with saline (Sal) or warfarin (War) and inoculated subsequently with BjV or saline, thus forming four groups: Sal + Sal (negative control), Sal + BjV (positive control), War + Sal (warfarinization control), and War + BjV. Three hours after inoculation, prothrombin and factor X levels fell 40% and 50%, respectively; levels of both factors decreased more than 97% in the War + Sal and War + BjV groups. Platelet counts dropped 93% and 76% in Sal + BjV and War + BjV, respectively, and plasma fibrinogen levels decreased 86% exclusively in Sal + BjV. After 6 and 24 h, platelet counts and fibrinogen levels increased progressively. A dramatic augmentation in plasma hemoglobin levels and the presence of schizocytes and microcytes in the Sal + BjV group indicated the development of intravascular hemolysis, which was prevented by warfarin pretreatment. Our findings show that intravascular thrombin generation has the foremost role in the pathogenesis of coagulopathy and intravascular hemolysis, but not in the development of thrombocytopenia, in B. jararaca envenomation in rats; in addition, fibrinogenases (metalloproteinases) may contribute to coagulopathy more than thrombin-like enzymes. © 2015 by the Society for Experimental Biology and Medicine.

Effect of Passive Ultrasonic Irrigation on Enterococcus faecalis from Root Canals: An Ex Vivo Study.

PubMed

Guerreiro-Tanomaru, Juliane Maria; Chávez-Andrade, Gisselle Moraima; de Faria-Júnior, Norberto Batista; Watanabe, Evandro; Tanomaru-Filho, Mário

2015-01-01

Endodontic irrigation aims to clean and disinfect the root canal system. Passive ultrasonic irrigation (PUI) is based on the use of an ultrasound-activated instrument into the root canal filled with irrigant. The aim of this study was to evaluate, ex vivo, the effectiveness of PUI in eliminating Enterococcus faecalis from root canals. Seventy-five extracted human single-root teeth were used. After root canal preparation, specimens were inoculated with E. faecalis and incubated at 37 °C for 21 days. Specimens were distributed into five groups (n=15), according to the irrigation method: PUI + saline solution (PUI/SS); PUI + 1% NaOCl (PUI/NaOCl); conventional needle irrigation (CNI) + saline solution (CNI/SS); CNI + 1% NaOCl (CNI/NaOCl); No irrigation (control). Microbiological samples were collected at three time points: initial (21 days after inoculation), post-irrigation (immediately after irrigation), and final (7 days after irrigation). Data were obtained in CFU mL-1 and subjected to analysis by ANOVA and Tukey's tests at 5% significance level. The post-irrigation samples did not demonstrate statistical difference between PUI/SS and CNI/SS nor between PUI/NaOCl and CNI/NaOCl (p>0.05), but PUI/NaOCl and CNI/NaOCl had lower CFU mL-1 number than the other groups (p>0.05). Statistically significant difference was observed between the initial and post-irrigation samples and between the post-irrigation and final samples (p<0.05) in all groups, except in the control. The final samples of all groups presented bacterial counts similar to the initial samples. PUI or CNI with 1% NaOCl contribute to disinfection, but are unable to eradicate E. faecalis from the root canal system.

Acute aerocystitis in Nile tilapia bred in net cages and supplemented with chromium carbochelate and Saccharomyces cerevisiae.

PubMed

Castro, Marcello P; Claudiano, Gustavo S; Petrillo, Thalita R; Shimada, Marina Tie; Belo, Marco A A; Marzocchi-Machado, Cleni M; Moraes, Julieta R E; Manrique, G Wilson; Moraes, Flávio R

2014-01-01

Oreochromis niloticus bred in net cages were supplemented with cell wall of Saccharomyces cerevisiae (Sc) (0.3%) or chromium carbochelate (Cr) (18 mg/kg of feed) or in association (Sc + Cr), for 90 days. After this period, acute inflammation was induced in the swim bladder by inoculation of 3 × 10(8) CFU of inactivated Streptococcus agalactiae, and another group received 0.65% saline solution (control). Twelve, 24, and 48 h after stimulation, the inflammation was evaluated through total and differential counting of accumulated cells, and through leukocyte respiratory burst in the blood, cortisolemia, glycemia and serum lysozyme concentration. The results showed that there were greater total numbers of cells in the exudate of fish inoculated with inactivated bacterium than in those injected with saline solution, with predominance of lymphocytes, thrombocytes, macrophages and granulocytes. Tilapia supplemented with Cr presented increased total numbers of cells with significant accumulation of lymphocytes and reductions in cortisolemia and glycemia, but the different treatments did not have any influence on leukocyte respiratory burst or serum lysozyme concentration. Tilapia supplemented with Sc and the Cr + Sc association did not present significant changes to the variables evaluated, despite higher accumulation of lymphocytes in the inflammatory exudate from fish treated with Sc. The results indicate that tilapia bred in net cages and supplemented with Cr presented higher total accumulation of cells at the inflammatory focus, thus indicating an increase in the inflammatory response induced by the bacterium, probably due to the reduction in cortisolemia and higher glucose consumption. Thus, supplementation with Cr had beneficial action, which facilitated development of acute inflammation induced by the bacterium, but did not affect neither leukocyte respiratory burst in the blood nor serum lysozyme concentration. Copyright © 2013 Elsevier Ltd. All rights reserved.

Influence of arbuscular mycorrhizae on the root system of maize plants under salt stress.

PubMed

Sheng, Min; Tang, Ming; Chen, Hui; Yang, Baowei; Zhang, Fengfeng; Huang, Yanhui

2009-07-01

Salt stress has become a severe global problem, and salinity is one of the most important abiotic factors limiting plant growth and yield. It is known that arbuscular mycorrhizal (AM) fungi decrease plant yield losses under salinity. With the aim of determining whether AM inoculation would give an advantage to root development under salt stress, a greenhouse experiment was carried out with AM or without AM fungi. Maize plants were grown in a sand and soil mixture with 5 NaCl levels (0, 0.5, 1.0, 1.5, and 2.0 g/kg dry substrate) for 55 days, following 15 days of nonsaline pretreatment. At all salt levels, mycorrhizal plants had higher dry shoot and root mass, higher root activity, and lower root to shoot ratios than non-mycorrhizal plants. In salt-free soil, root length, root surface area, root volume, and number of root tips and forks were significantly larger in mycorrhizal plants than in non-mycorrhizal plants, whereas, under salt stress, average root diameter and root volume of mycorrhizal plants were larger than those of non-mycorrhizal plants. Regardless of the NaCl level, mycorrhizal plants had lower specific root length, lower percentage of root length in the 0-0.2 mm diameter class, and higher percentage of root length in both the 0.2-0.4 mm and 0.4-0.6 mm diameter classes, which suggests that the root system shows a significant shift towards a thicker root system when maize plants were inoculated with Glomus mosseae (Nicolson & Gerdemann). The results presented here indicate that the improvements in root activity and the coarse root system of mycorrhizal maize may help in alleviating salt stress on the plant.

Mitigation of NaCl Stress by Arbuscular Mycorrhizal Fungi through the Modulation of Osmolytes, Antioxidants and Secondary Metabolites in Mustard (Brassica juncea L.) Plants

PubMed Central

Sarwat, Maryam; Hashem, Abeer; Ahanger, Mohammad A.; Abd_Allah, Elsayed F.; Alqarawi, A. A.; Alyemeni, Mohammed N.; Ahmad, Parvaiz; Gucel, Salih

2016-01-01

Present work was carried out to investigate the possible role of arbuscular mycorrhizal fungi (AMF) in mitigating salinity-induced alterations in Brassica juncea L. Exposure to NaCl stress altered the morphological, physio-biochemical attributes, antioxidant activity, secondary metabolites and phytohormones in the mustard seedlings. The growth and biomass yield, leaf water content, and total chlorophyll content were decreased with NaCl stress. However, AMF-inoculated plants exhibited enhanced shoot and root length, elevated relative water content, enhanced chlorophyll content, and ultimately biomass yield. Lipid peroxidation and proline content were increased by 54.53 and 63.47%, respectively with 200 mM NaCl concentration. Further increase in proline content and decrease in lipid peroxidation was observed in NaCl-treated plants inoculated with AMF. The antioxidants, superoxide dismutase, ascorbate peroxidase, glutathione reductase, and reduced glutathione were increased by 48.35, 54.86, 43.85, and 44.44%, respectively, with 200 mM NaCl concentration. Further increase in these antioxidants has been observed in AMF-colonized plants indicating the alleviating role of AMF to salinity stress through antioxidant modulation. The total phenol, flavonoids, and phytohormones increase with NaCl treatment. However, NaCl-treated plants colonized with AMF showed further increase in the above parameters except ABA, which was reduced with NaCl+AMF treatment over the plants treated with NaCl alone. Our results demonstrated that NaCl caused negative effect on B. juncea seedlings; however, colonization with AMF enhances the NaCl tolerance by reforming the physio-biochemical attributes, activities of antioxidant enzymes, and production of secondary metabolites and phytohormones. PMID:27458462

Effects of acute change in salinity and moulting on the infection of white leg shrimp (Penaeus vannamei) with white spot syndrome virus upon immersion challenge.

PubMed

Van Thuong, K; Van Tuan, V; Li, W; Sorgeloos, P; Bossier, P; Nauwynck, H

2016-12-01

In the field, moulting and salinity drop in the water due to excessive rainfall have been mentioned to be risk factors for WSSV outbreaks. Therefore, in this study, the effect of an acute change in environmental salinity and shedding of the old cuticle shell on the susceptibility of Penaeus vannamei to WSSV was evaluated by immersion challenge. For testing the effect of abrupt salinity stress, early premoult shrimp that were acclimated to 35 g L -1 were subjected to salinities of 50 g L -1 , 35 g L -1 , 20 g L -1 , 10 g L -1 and 7 g L -1 or 5 g L -1 and simultaneously exposed to 10 5.5  SID 50 mL -1 of WSSV for 5 h, after which the salinity was brought back to 35 g L -1 . Shrimp that were transferred from 35 g L -1 to 50 g L -1 , 35 g L -1 and 20 g L -1 did not become infected with WSSV. Shrimp became infected with WSSV after an acute salinity drop from 35 g L -1 to 10 g L -1 and lower. The mortality in shrimp, subjected to a salinity change to 10 g L -1 , 7 g L -1 and 5 g L -1 , was 6.7%, 46.7% and 53.3%, respectively (P < 0.05) . For testing the effect of moulting, shrimp in early premoult, moulting and post-moult were immersed in sea water containing 10 5.5  SID 50 mL -1 of WSSV. The resulting mortality due to WSSV infection in shrimp inoculated during early premoult (0%), ecdysis (53.3%) and post-moult (26.72%) demonstrated that a significant difference exists in susceptibility of shrimp during the short moulting process (P < 0.05). The findings of this study indicate that during a drop in environmental salinity lower than 10 g L -1 and ecdysis, shrimp are at risk for a WSSV infection. These findings have important implications for WSSV control measures. © 2016 John Wiley & Sons Ltd.

Rhipicephalus microplus infected by Metarhizium: unveiling hemocyte quantification, GFP-fungi virulence, and ovary infection.

PubMed

de Paulo, Jéssica Fiorotti; Camargo, Mariana Guedes; Coutinho-Rodrigues, Caio Junior Balduino; Marciano, Allan Felipe; de Freitas, Maria Clemente; da Silva, Emily Mesquita; Gôlo, Patrícia Silva; Morena, Diva Denelle Spadacci; da Costa Angelo, Isabele; Bittencourt, Vânia Rita Elias Pinheiro

2018-06-01

Hemocytes, cells present in the hemocoel, are involved in the immune response of arthropods challenged with entomopathogens. The present study established the best methodology for harvesting hemocytes from Rhipicephalus microplus and evaluated the number of hemocytes in addition to histological analysis from ovaries of fungus-infected females and tested the virulence of GFP-fungi transformants. Different centrifugation protocols were tested, and the one in which presented fewer disrupted cells and higher cell recovery was applied for evaluating the effect of Metarhizium spp. on hemocytes against R. microplus. After processing, protocol number 1 (i.e., hemolymph samples were centrifuged at 500×g for 3 min at 4 °C) was considered more efficient, with two isolates used (Metarhizium robertsii ARSEF 2575 and Metarhizium anisopliae ARSEF 549), both wild types and GFP, to assess their virulence. In the biological assays, the GFP-fungi were as virulent as wild types, showing no significant differences. Subsequently, hemocyte quantifications were performed after inoculation, which exhibited notable changes in the number of hemocytes, reducing by approximately 80% in females previously treated with Metarhizium isolates in comparison to non-treated females. Complementarily, 48 h after inoculation, in which hemolymph could not be obtained, histological analysis showed the high competence of these fungi to colonize ovary from ticks. Here, for the first time, the best protocol (i.e., very low cell disruption and high cell recovery) for R. microplus hemocyte obtaining was established aiming to guide directions to other studies that involves cellular responses from ticks to fungi infection.

Use of Frankia and Actinorhizal Plants for Degraded Lands Reclamation

PubMed Central

Diagne, Nathalie; Ngom, Mariama; Nambiar-Veetil, Mathish; Franche, Claudine; Narayanan, Krishna Kumar; Laplaze, Laurent

2013-01-01

Degraded lands are defined by soils that have lost primary productivity due to abiotic or biotic stresses. Among the abiotic stresses, drought, salinity, and heavy metals are the main threats in tropical areas. These stresses affect plant growth and reduce their productivity. Nitrogen-fixing plants such as actinorhizal species that are able to grow in poor and disturbed soils are widely planted for the reclamation of such degraded lands. It has been reported that association of soil microbes especially the nitrogen-fixing bacteria Frankia with these actinorhizal plants can mitigate the adverse effects of abiotic and biotic stresses. Inoculation of actinorhizal plants with Frankia significantly improves plant growth, biomass, shoot and root N content, and survival rate after transplanting in fields. However, the success of establishment of actinorhizal plantation in degraded sites depends upon the choice of effective strains of Frankia. Studies related to the beneficial role of Frankia on the establishment of actinorhizal plants in degraded soils are scarce. In this review, we describe some examples of the use of Frankia inoculation to improve actinorhizal plant performances in harsh conditions for reclamation of degraded lands. PMID:24350296

Live-cell imaging of Salmonella Typhimurium interaction with zebrafish larvae after injection and immersion delivery methods.

PubMed

Varas, Macarena; Fariña, Alonso; Díaz-Pascual, Francisco; Ortíz-Severín, Javiera; Marcoleta, Andrés E; Allende, Miguel L; Santiviago, Carlos A; Chávez, Francisco P

2017-04-01

The zebrafish model has been used to determine the role of vertebrate innate immunity during bacterial infections. Here, we compare the in vivo immune response induced by GFP-tagged Salmonella Typhimurium inoculated by immersion and microinjection in transgenic zebrafish larvae. Our novel infection protocols in zebrafish allow live-cell imaging of Salmonella colonization. Copyright © 2017 Elsevier B.V. All rights reserved.

A regional assessment of salt marsh restoration and monitoring in the Gulf of Maine

USGS Publications Warehouse

Konisky, R.A.; Burdick, D.M.; Dionne, M.; Neckles, H.A.

2006-01-01

We compiled salt marsh monitoring datasets from 36 complete or imminent restoration projects in the Gulf of Maine to assess regional monitoring and restoration practices. Data were organized by functional indicators and restoration project types (culvert replacement, excavation works, or ditch plugging) then pooled to generate mean values for indicators before restoration, after restoration, and at reference sites. Monitoring data were checked against the regional standards of a voluntary protocol for the Gulf of Maine. Data inventories showed that vegetation and salinity indicators were most frequently collected (89 and 78% of sites, respectively), whereas nekton, bird, and hydrologic measures were collected at only about half of the sites. Reference conditions were monitored at 72% of sites. Indicators were analyzed to see if project sites were degraded relative to reference areas and to detect ecological responses to restoration activities. Results showed that compared to reference areas, prerestoration sites had smaller tidal ranges, reduced salinity levels, greater cover of brackish plants species, and lower cover of halophyte plants. Following restoration, physical factors rebounded rapidly with increased flood and salinity levels after about one year, especially for culvert projects. Biological responses were less definitive and occurred over longer time frames. Plant communities trended toward recovered halophytes and reduced brackish species at 3+ years following restoration. Nekton and avian indicators were indistinguishable among reference, impacted, and restored areas. The protocol was successful in demonstrating restoration response for the region, but results were limited by regional inconsistencies in field practices and relatively few multiyear datasets. To improve future assessment capabilities, we encourage greater adherence to the standard protocol throughout the Gulf of Maine salt marsh restoration community.

Effect of Simultaneous Inoculation with Yeast and Bacteria on Fermentation Kinetics and Key Wine Parameters of Cool-Climate Chardonnay

PubMed Central

Jussier, Delphine; Dubé Morneau, Amélie; Mira de Orduña, Ramón

2006-01-01

Inoculating grape musts with wine yeast and lactic acid bacteria (LAB) concurrently in order to induce simultaneous alcoholic fermentation (AF) and malolactic fermentation (MLF) can be an efficient alternative to overcome potential inhibition of LAB in wines because of high ethanol concentrations and reduced nutrient content. In this study, the simultaneous inoculation of yeast and LAB into must was compared with a traditional vinification protocol, where MLF was induced after completion of AF. For this, two suitable commercial yeast-bacterium combinations were tested in cool-climate Chardonnay must. The time courses of glucose and fructose, acetaldehyde, several organic acids, and nitrogenous compounds were measured along with the final values of other key wine parameters. Sensory evaluation was done after 12 months of storage. The current study could not confirm a negative impact of simultaneous AF/MLF on fermentation success and kinetics or on final wine parameters. While acetic acid concentrations were slightly increased in wines after simultaneous AF/MLF, the differences were of neither practical nor legal significance. No statistically significant differences were found with regard to the final values of pH or total acidity and the concentrations of ethanol, acetaldehyde, glycerol, citric and lactic acids, and the nitrogen compounds arginine, ammonia, urea, citrulline, and ornithine. Sensory evaluation by a semiexpert panel confirmed the similarity of the wines. However, simultaneous inoculation led to considerable reductions in overall fermentation durations. Furthermore, differences of physiological and microbiological relevance were found. Specifically, we report the vinification of “super-dry” wines devoid of glucose and fructose after simultaneous inoculation of yeast and bacteria. PMID:16391046

Multi-cropping edible truffles and sweet chestnuts: production of high-quality Castanea sativa seedlings inoculated with Tuber aestivum, its ecotype T. uncinatum, T. brumale, and T. macrosporum.

PubMed

Álvarez-Lafuente, Amaya; Benito-Matías, Luis F; Peñuelas-Rubira, Juan L; Suz, Laura M

2018-01-01

The plantation and management of sweet chestnut (Castanea sativa Mill.) orchards is a common and traditional land use system in many areas of Europe that offers the advantage of simultaneous production of nuts and timber. During the last decades, sweet chestnut has declined dramatically in many regions because of the profound social changes in rural areas coupled with pathogen attacks. Truffles, the hypogeous ascocarps of the ectomycorrhizal genus Tuber, are currently cultivated using host trees inoculated with these fungi for improving production in truffle orchards. The production of good forestry quality chestnut seedlings inoculated with European truffles in nurseries is essential for multi-cropping plantation establishment, but so far, it has not been implemented in agroforestry practices. Moreover, it is necessary to assess the physiological condition of the seedlings due to the high calcium amendment needed for the growth of Tuber spp. mycelium that can become toxic for the host plants. In this study, seedlings of C. sativa were inoculated with Tuber aestivum and its ecotypes T. uncinatum, T. brumale, and T. macrosporum and were grown in a greenhouse using culture conditions favorable for the production of high-quality plants for forestry purposes. At the end of the assay, levels of root colonization and morphological and physiological parameters of the seedlings were measured. The colonization of C. sativa with T. aestivum, its ecotype T. uncinatum, and T. brumale was successful, and the seedlings showed normal growth. Inoculation protocols with T. macrosporum need to be improved. Tuber species formed well-developed ectomycorrhizae on C. sativa in nursery conditions.

Natural infection of Lutzomyia neivai and Lutzomyia sallesi (Diptera: Psychodidae) by Leishmania infantum chagasi in Brazil.

PubMed

Saraiva, Lara; Carvalho, Gustavo M L; Gontijo, Célia M F; Quaresma, Patrícia F; Lima, Ana C V M R; Falcão, Alda L; Andrade Filho, José D

2009-09-01

Natural infections with Leishmania were found in females of the phlebotomine sand flies Lutzomyia neivai (Pinto) (= Nyssomyia neivai) and Lutzomyia sallesi (Galvão & Coutinho) (= Evandromyia sallesi) (Diptera: Psychodidae) from Lassance, in the Brazilian state of Minas Gerais. Promastigotes were found in the pyloric region of the former species and in the abdominal midgut of the latter species. Insects found to be infected by microscopic examination were macerated in saline solution and inoculated into hamsters. Subsequent analysis by polymerase chain reaction-restriction fragment length polymorphism revealed both isolates to belong to the species Leishmania infantum chagasi Cunha & Chagas.

Early surgical suction and washout for treatment of cytotoxic drug extravasations.

PubMed

Vandeweyer, E; Deraemaecker, R

2000-02-01

This case report is presented to assess safety and efficiency of early suction and saline washout of extravasated cytotoxic drugs. Through multiple small skin incisions, the area of extravasation is first suctioned and subsequently extensively washed out with saline. Incisions are left open and the arm is elevated for 24 hours. A complete healing was obtained in five days without any skin or soft tissue loss. No additional treatment was needed. Early referral and surgical treatment by suction and washout is a safe and reliable treatment protocol for major cytotoxic drug extravasation injuries.

Impact of Salinity Gradients on Ammonia Bioattenuation Processes in a Photosynthetic Wetland Biomat

NASA Astrophysics Data System (ADS)

Vega, M.; Jones, Z.; Sharp, J.

2017-12-01

Shallow, open water treatment wetlands may be able to offset challenges associated with the reclamation of impaired waters (e.g., membrane fouling, aeration costs, etc.) due to natural biogeochemical fluctuations produced by a benthic, photoactive biomat. This diatomaceous, redox-stratified biomat has demonstrated significant nitrate and trace organic removal from municipal wastewater streams and the microbial community has been thoroughly characterized. However, research is required to predict shifts in community structure and function in response to the excess salinity, ammonia, and metal gradients of impaired waters. Batch microcosm studies inoculating biomat from an active open water treatment wetland with incremental dilutions of hydraulic fracturing produced water were conducted in a light chamber with oscillating twelve-hour light and dark cycles to assess the effect of an impaired water matrix on biomat functionality. Diurnal photosynthetic signatures and ammonia removal kinetics were quantified in various experiments probing the effects of oscillating light conditions, biomat depth, water column isolation, nitrogen source, and salinity gradients in conjunction with phylogenetic profiles and morphological characterization. Diurnal pH and dissolved oxygen fluctuations were present at all produced water permutations, perhaps indicating stabilization of photosynthetic communities. Ammonia attenuation results suggest that the biomat is effective at removing ammonia, although first order rate constants decrease with increasing produced water abundance. Microbial community diversity appears to decrease with increasing salinity, and it is likely that these shifts correspond to variation in ecosystem function and thus treatment effectiveness. The application of shallow, open water treatment wetlands to remediate impaired waters has the potential to address societally relevant problems while discerning fundamental biogeochemical phenomena.

Effects of Bacillus subtilis on some physiological and biochemical parameters of Triticum aestivum L. (wheat) under salinity.

PubMed

Lastochkina, Oksana; Pusenkova, Ludmila; Yuldashev, Ruslan; Babaev, Marat; Garipova, Svetlana; Blagova, Dar'ya; Khairullin, Ramil; Aliniaeifard, Sasan

2017-12-01

Endophytic strain Bacillus subtilis (B. subtilis) 10-4, producing indole-3-acetic acid (IAA) and siderofores but not active in phosphate solubilization, exerted a protective effect on Triticum aestivum L. (wheat) plant grown under salinity (2% NaCl) stress. Exposure to salt stress resulted in an essential increase of proline (Pro) and malondialdehyde (MDA) level in the seedlings. At the same time the seedlings inoculated with B. subtilis 10-4 were characterized by decreased level of stress-induced Pro and MDA accumulation. It was revealed that both B. subtilis 10-4 and salinity caused increase in the content of endogenous salicylic acid (SA) in wheat seedlings as compared to SA content in the control, while B. subtilis 10-4 suppressed stress-induced SA accumulation. Water storage capacity (WSC) in leaf tissues was increased and stress-induced hydrolysis of statolite starch in root cap cells of the germinal roots was reduced by B. subtilis 10-4. The obtained data indicated that the activation of the defense reactions induced by B. subtilis 10-4 induced defense reactions may be connected with their ability to decrease the level of stress-induced oxidative and osmotic stress in seedlings and with the increase of endogenous SA level that can make a significant contribution to the implementation of the protective effect of B. subtilis 10-4 and is manifested in the improvement of plant growth, WSC of leaves and slowing down of the process of statolite starch hydrolysis under salinity. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Effects of preincubation of eggs and activation medium on the percentage of eyed embryos in ide (Leuciscus idus), an externally fertilizing fish.

PubMed

Siddique, Mohammad Abdul Momin; Linhart, Otomar; Krejszeff, Sławomir; Żarski, Daniel; Król, Jarosław; Butts, Ian Anthony Ernest

2016-03-15

Standardization of fertilization protocols is crucial for improving reproductive techniques for externally fertilizing fish in captive breeding. Therefore, the objectives of this study were to determine the effects of preincubation of eggs and activation medium on the percentage of eyed embryos for ide (Leuciscus idus). Pooled eggs from five females were preincubated in three different activating media for 0, 30, 60, 90, and 120 seconds and then fertilized by pooled sperm from five males. At the eyed-egg stage, the percentage of viable embryos was later calculated. Results showed that preincubation time was significant for the freshwater activation medium (P < 0.001), such that the percentage of eyed embryos declined across the preincubation time gradient. Additionally, there was an effect on the percentage of eyed embryos when eggs were incubated with Woynarovich solution (P < 0.001), such that a decline was detected at 90 seconds, whereas no effect was detected for the saline water medium. Activating medium had a significant effect on the percentage of eyed embryos for each preincubation time (P < 0.05). More precisely, freshwater produced the lowest percentage of eyed embryos at all preincubation times (ranged from 1.9% at 120 seconds to 43.6% at 0 seconds), whereas saline water and Woynarovich solution produced the highest percentage of eyed embryos at 0 seconds and 30 seconds before incubation. Woynarovich solution produced the highest percentage of eyed embryos at 60 seconds (65.26%), whereas saline water produced the highest percentage at 90 seconds (68.37%). No difference was detected between saline water and Woynarovich solution at 120 seconds. Examination of sperm traits showed no impact of activating medium on computer assisted sperm analysis parameters. Together, these results suggest that saline water or Woynarovich solution improve fertilization rate in ide during IVF; thus, these media are useful for standardizing fertilization protocols and controlled reproduction for this species. Copyright © 2016 Elsevier Inc. All rights reserved.

Lytic bacteriophages reduce Escherichia coli O157

PubMed Central

Ferguson, Sean; Roberts, Cheryl; Handy, Eric; Sharma, Manan

2013-01-01

The role of lytic bacteriophages in preventing cross contamination of produce has not been evaluated. A cocktail of three lytic phages specific for E. coli O157:H7 (EcoShield™) or a control (phosphate buffered saline, PBS) was applied to lettuce by either; (1) immersion of lettuce in 500 ml of EcoShield™ 8.3 log PFU/ml or 9.8 log PFU/ml for up to 2 min before inoculation with E. coli O157:H7; (2) spray-application of EcoShield™ (9.3 log PFU/ml) to lettuce after inoculation with E. coli O157:H7 (4.10 CFU/cm2) following exposure to 50 μg/ml chlorine for 30 sec. After immersion studies, lettuce was spot-inoculated with E. coli O157:H7 (2.38 CFU/cm2). Phage-treated, inoculated lettuce pieces were stored at 4°C for and analyzed for E. coli O157:H7 populations for up to 7 d. Immersion of lettuce in 9.8 log PFU/ml EcoShield™ for 2 min significantly (p < 0.05) reduced E. coli O157:H7 populations after 24 h when stored at 4°C compared with controls. Immersion of lettuce in suspensions containing high concentrations of EcoShield™ (9.8 log PFU/ml) resulted in the deposition of high concentrations (7.8 log log PFU/cm2) of bacteriophages on the surface of fresh cut lettuce, potentially contributing to the efficacy of the lytic phages on lettuce. Spraying phages on to inoculated fresh cut lettuce after being washed in hypochlorite solution was significantly more effective in reducing E. coli O157:H7 populations (2.22 log CFU/cm2) on day 0 compared with control treatments (4.10 log CFU/cm2). Both immersion and spray treatments provided protection from E. coli O157:H7 contamination on lettuce, but spray application of lytic bacteriophages to lettuce was more effective in immediately reducing E. coli O157:H7 populations fresh cut lettuce. PMID:23819106

Group A Rotavirus Infection and Age-Dependent Diarrheal Disease in Rats: a New Animal Model To Study the Pathophysiology of Rotavirus Infection

PubMed Central

Ciarlet, Max; Conner, Margaret E.; Finegold, Milton J.; Estes, Mary K.

2002-01-01

Group A rotaviruses are major pathogens causing acute gastroenteritis in children and animals. To determine if group A rotavirus replicates and induces disease in rats, antibody-negative Lewis neonatal or adult rats were inoculated orally with tissue culture-adapted human (Wa, WI61, and HAL1166), simian (rhesus rotavirus [RRV] and SA11), bovine (WC3), lapine (ALA), or porcine (OSU) rotavirus strains, wild-type murine (ECwt) rotavirus strain, or phosphate-buffered saline (PBS). Rotavirus infection in rats was evaluated by (i) clinical findings, (ii) virus antigen shedding or infectious virus titers in the feces or intestinal contents measured by enzyme-linked immunosorbent assay or fluorescent-focus assay, (iii) histopathological changes in the small intestine, (iv) distribution of rotavirus antigen in small-intestine sections by immunofluorescence, and (v) growth rate. Rotavirus infection of 5-day-old but not ≥21-day-old rats resulted in diarrhea that lasted from 1 to 10 days postinoculation. The severity of disease and spread of infection to naÏve littermates differed depending on the virus strain used for inoculation. The duration of virus antigen shedding following infection was considerably prolonged (up to 10 days) in neonatal rats compared to that in 21-day-old rats (1 or 2 days). Based on lack of virus antigen shedding and disease induction, the murine ECwt rotavirus was the only strain tested that did not infect rats. Histopathological changes in the small-intestine mucosa of 5-day-old RRV-inoculated rats but not of PBS-inoculated rats was limited to extensive enterocyte vacuolation in the ileum. In RRV-inoculated neonatal rats, rotavirus antigen was detected in the epithelial cells on the upper half of the intestinal villi of the jejunum and ileum. In addition, infection of neonatal rats with RRV but not with PBS resulted in reduced weight gain. Rats infected with group A rotaviruses provide a new animal model with unique features amenable to investigate rotavirus pathogenesis and the molecular mechanisms of intestinal development, including physiological factors that may regulate age-dependent rotavirus-induced diarrhea. PMID:11739670

Phenobarbital for acute alcohol withdrawal: a prospective randomized double-blind placebo-controlled study.

PubMed

Rosenson, Jonathan; Clements, Carter; Simon, Barry; Vieaux, Jules; Graffman, Sarah; Vahidnia, Farnaz; Cisse, Bitou; Lam, Joseph; Alter, Harrison

2013-03-01

Acute alcohol withdrawal syndrome (AAWS) is encountered in patients presenting acutely to the Emergency Department (ED) and often requires pharmacologic management. We investigated whether a single dose of intravenous (i.v.) phenobarbital combined with a standardized lorazepam-based alcohol withdrawal protocol decreases intensive care unit (ICU) admission in ED patients with acute alcohol withdrawal. This was a prospective, randomized, double-blind, placebo-controlled study. Patients were randomized to receive either a single dose of i.v. phenobarbital (10 mg/kg in 100 mL normal saline) or placebo (100 mL normal saline). All patients were placed on the institutional symptom-guided lorazepam-based alcohol withdrawal protocol. The primary outcome was initial level of hospital admission (ICU vs. telemetry vs. floor ward). There were 198 patients enrolled in the study, and 102 met inclusion criteria for analysis. Fifty-one patients received phenobarbital and 51 received placebo. Baseline characteristics and severity were similar in both groups. Patients that received phenobarbital had fewer ICU admissions (8% vs. 25%, 95% confidence interval 4-32). There were no differences in adverse events. A single dose of i.v. phenobarbital combined with a symptom-guided lorazepam-based alcohol withdrawal protocol resulted in decreased ICU admission and did not cause increased adverse outcomes. Copyright © 2013 Elsevier Inc. All rights reserved.

Dramatic reduction of culture time of Mycobacterium tuberculosis

NASA Astrophysics Data System (ADS)

Ghodbane, Ramzi; Raoult, Didier; Drancourt, Michel

2014-02-01

Mycobacterium tuberculosis culture, a critical technique for routine diagnosis of tuberculosis, takes more than two weeks. Here, step-by-step improvements in the protocol including a new medium, microaerophlic atmosphere or ascorbic-acid supplement and autofluorescence detection dramatically shortened this delay. In the best case, primary culture and rifampicin susceptibility testing were achieved in 72 hours when specimens were inoculated directly on the medium supplemented by antibiotic at the beginning of the culture.

[Effects of pretreatment of lipid, midazolam and propofol on ropivacaine-induced convulsion and LD50 in rats].

PubMed

Lü, Xiao-lan; Wan, Fu-hong; Zuo, Yun-xia

2012-09-04

To assess the effects of lipid on ropivacaine-induced convulsion and LD50 in rats and compare with those of the traditional anticonvulsants midazolam and propofol. Protocol 1: A total of 120 SD rats (60 males, 60 females), weighing 200-300 g, were randomly assigned into 4 groups with equal males and females: lipid (L), midazolam (M) and propofol (P) and control (C). Rats were pretreated with 10 ml/kg lipid intravenously in group L, saline and 0.23 mg/kg midazolam (10 ml/kg in volume) sequentially in group M, saline and 4 mg/kg propofol (10 ml/kg in volume) in group P and saline 10 ml/kg in group C. Then ropivacaine 44 mg/kg (0.75%) was injected intraperitoneally into each rat. The convulsion rate in each group and the time of convulsion after ropivacaine injection were observed. Meanwhile, the plasma concentration of ropivacaine at the time of convulsion was measured. Protocol 2: Additional 100 male SD rats were used for the measurements of ropivacaine LD50 with different pretreatments including lipid, midazolam, propofol and saline through the up-and-down method. Rats were randomly assigned into 4 groups similarly as protocol 1. The doses of ropivacaine in each group were determined according to our pilot study and 6 dosage levels with the same interval ratio 8.5 was applied in each group. The doses of these pretreatment drugs and administration methods were similarly as protocol 1. The convulsion rate after 44 mg/kg ropivacaine ip injection was 43.3% in group C, 0% in group M, 13.3% in group P and 70% in group L. Lipid increased the convulsion rate significantly. The plasma concentration of ropivacaine at the time of convulsion was 1.65 ± 0.30 µg/kg in group C, 1.73 ± 0.14 µg/kg in group P and 3.45 ± 0.26 µg/kg in group L. The LD50 of ropivacaine in group C was 64.39 mg/kg, 88.40 mg/kg in group M and 90.20 mg/kg in group P and 55.45 mg/kg in group L. Midazolam and propofol not only decrease the convulsion rate of ropivacaine, but also increase its LD50. Lipid not only increases the convulsion rate of ropivacaine, but also decreases its LD50. The application of lipid for the prevention of local anesthetic toxicity has potential risks.

Telomere maintenance in laser capture microdissection-purified Barrett's adenocarcinoma cells and effect of telomerase inhibition in vivo.

PubMed

Shammas, Masood A; Qazi, Aamer; Batchu, Ramesh B; Bertheau, Robert C; Wong, Jason Y Y; Rao, Manjula Y; Prasad, Madhu; Chanda, Diptiman; Ponnazhagan, Selvarangan; Anderson, Kenneth C; Steffes, Christopher P; Munshi, Nikhil C; De Vivo, Immaculata; Beer, David G; Gryaznov, Sergei; Weaver, Donald W; Goyal, Raj K

2008-08-01

The aims of this study were to investigate telomere function in normal and Barrett's esophageal adenocarcinoma (BEAC) cells purified by laser capture microdissection and to evaluate the effect of telomerase inhibition in cancer cells in vitro and in vivo. Epithelial cells were purified from surgically resected esophagi. Telomerase activity was measured by modified telomeric repeat amplification protocol and telomere length was determined by real-time PCR assay. To evaluate the effect of telomerase inhibition, adenocarcinoma cell lines were continuously treated with a specific telomerase inhibitor (GRN163L) and live cell number was determined weekly. Apoptosis was evaluated by Annexin labeling and senescence by beta-galactosidase staining. For in vivo studies, severe combined immunodeficient mice were s.c. inoculated with adenocarcinoma cells and following appearance of palpable tumors, injected i.p. with saline or GRN163L. Telomerase activity was significantly elevated whereas telomeres were shorter in BEAC cells relative to normal esophageal epithelial cells. The treatment of adenocarcinoma cells with telomerase inhibitor, GRN163L, led to loss of telomerase activity, reduction in telomere length, and growth arrest through induction of both the senescence and apoptosis. GRN163L-induced cell death could also be expedited by addition of the chemotherapeutic agents doxorubicin and ritonavir. Finally, the treatment with GRN163L led to a significant reduction in tumor volume in a subcutaneous tumor model. We show that telomerase activity is significantly elevated whereas telomeres are shorter in BEAC and suppression of telomerase inhibits proliferation of adenocarcinoma cells both in vitro and in vivo.

Real-time PCR assays for detection and quantification of aflatoxin-producing molds in foods.

PubMed

Rodríguez, Alicia; Rodríguez, Mar; Luque, M Isabel; Martín, Alberto; Córdoba, Juan J

2012-08-01

Aflatoxins are among the most toxic mycotoxins. Early detection and quantification of aflatoxin-producing species is crucial to improve food safety. In the present work, two protocols of real-time PCR (qPCR) based on SYBR Green and TaqMan were developed, and their sensitivity and specificity were evaluated. Primers and probes were designed from the o-methyltransferase gene (omt-1) involved in aflatoxin biosynthesis. Fifty-three mold strains representing aflatoxin producers and non-producers of different species, usually reported in food products, were used as references. All strains were tested for aflatoxins production by high-performance liquid chromatography-mass spectrometry (HPLC-MS). The functionality of the proposed qPCR method was demonstrated by the strong linear relationship of the standard curves constructed with the omt-1 gene copy number and Ct values for the different aflatoxin producers tested. The ability of the qPCR protocols to quantify aflatoxin-producing molds was evaluated in different artificially inoculated foods. A good linear correlation was obtained over the range 4 to 1 log cfu/g per reaction for all qPCR assays in the different food matrices (peanuts, spices and dry-fermented sausages). The detection limit in all inoculated foods ranged from 1 to 2 log cfu/g for SYBR Green and TaqMan assays. No significant effect was observed due to the different equipment, operator, and qPCR methodology used in the tests of repeatability and reproducibility for different foods. The proposed methods quantified with high efficiency the fungal load in foods. These qPCR protocols are proposed for use to quantify aflatoxin-producing molds in food products. Copyright © 2012 Elsevier Ltd. All rights reserved.

Arbuscular mycorrhizas enhance nutrient uptake in different wheat genotypes at high salinity levels under field and greenhouse conditions.

PubMed

Mardukhi, Baran; Rejali, Farhad; Daei, Gudarz; Ardakani, Mohammad Reza; Malakouti, Mohammad Javad; Miransari, Mohammad

2011-07-01

Since most experiments regarding the symbiosis between arbuscular mycorrhizal (AM) fungi and their host plants under salinity stress have been performed only under greenhouse conditions, this research work was also conducted under field conditions. The effects of three AM species including Glomus mosseae, G. etunicatum and G. intraradices on the nutrient uptake of different wheat cultivars (including Roshan, Kavir and Tabasi) under field and greenhouse (including Chamran and Line 9) conditions were determined. At field harvest, the concentrations of N, Ca, Mg, Fe, Cu, and Mn, and at greenhouse harvest, plant growth, root colonization and concentrations of different nutrients including N, K, P, Ca, Mg, Mn, Cu, Fe, Zn, Na and Cl were determined. The effects of wheat cultivars on the concentrations of N, Ca, and Mn, and of all nutrients were significant at field and greenhouse conditions, respectively. In both experiments, AM fungi significantly enhanced the concentrations of all nutrients including N, K, P, Ca, Mg, Mn, Cu, Fe, Zn, Na and Cl. The synergistic and enhancing effects of co-inoculation of AM species on plant growth and the inhibiting effect of AM species on Na(+) rather than on Cl(-) uptake under salinity are also among the important findings of this research work. Copyright © 2011 Académie des sciences. Published by Elsevier SAS. All rights reserved.

Ability of a haloalkaliphilic bacterium isolated from Soap Lake, Washington to generate electricity at pH 11.0 and 7% salinity.

PubMed

Paul, Varun G; Minteer, Shelley D; Treu, Becky L; Mormile, Melanie R

2014-01-01

A variety of anaerobic bacteria have been shown to transfer electrons obtained from organic compound oxidation to the surface of electrodes in microbial fuel cells (MFCs) to produce current. Initial enrichments for iron (III) reducing bacteria were set up with sediments from the haloalkaline environment of Soap Lake, Washington, in batch cultures and subsequent transfers resulted in a culture that grew optimally at 7.0% salinity and pH 11.0. The culture was used to inoculate the anode chamber of a MFC with formate as the electron source. Current densities up to 12.5 mA/m2 were achieved by this bacterium. Cyclic voltammetry experiments demonstrated that an electron mediator, methylene blue, was required to transfer electrons to the anode. Scanning electron microscopic imaging of the electrode surface did not reveal heavy colonization of bacteria, providing evidence that the bacterium may be using an indirect mode of electron transfer to generate current. Molecular characterization of the 16S rRNA gene and restriction fragment length profiles (RFLP) analysis showed that the MFC enriched for a single bacterial species with a 99% similarity to the 16S rRNA gene of Halanaerobium hydrogeniformans. Though modest, electricity production was achieved by a haloalkaliphilic bacterium at pH 11.0 and 7.0% salinity.

Adenosine receptor antagonist and augmented vasodilation during hypoxic exercise

PubMed Central

Madery, Brandon D.; Pike, Tasha L.; Eisenach, John H.; Dietz, Niki M.; Joyner, Michael J.; Wilkins, Brad W.

2009-01-01

We tested the hypothesis that adenosine contributes to augmented skeletal muscle vasodilation during hypoxic exercise. In separate protocols, subjects performed incremental rhythmic forearm exercise (10% and 20% of maximum) during normoxia and normocapnic hypoxia (80% arterial O2 saturation). In protocol 1 (n = 8), subjects received an intra-arterial administration of saline (control) and aminophylline (adenosine receptor antagonist). In protocol 2 (n = 10), subjects received intra-arterial phentolamine (α-adrenoceptor antagonist) and combined phentolamine and aminophylline administration. Forearm vascular conductance (FVC; in ml·min−1·100 mmHg−1) was calculated from forearm blood flow (in ml/min) and blood pressure (in mmHg). In protocol 1, the change in FVC (ΔFVC; change from normoxic baseline) during hypoxic exercise with saline was 172 ± 29 and 314 ± 34 ml·min−1·100 mmHg−1 (10% and 20%, respectively). Aminophylline administration did not affect ΔFVC during hypoxic exercise at 10% (190 ± 29 ml·min−1·100 mmHg−1, P = 0.4) or 20% (287 ± 48 ml·min−1·100 mmHg−1, P = 0.3). In protocol 2, ΔFVC due to hypoxic exercise with phentolamine infusion was 313 ± 30 and 453 ± 41 ml·min−1·100 mmHg−1 (10% and 20% respectively). ΔFVC was similar at 10% (352 ± 39 ml·min−1·100 mmHg−1, P = 0.8) and 20% (528 ± 45 ml·min−1·100 mmHg−1, P = 0.2) hypoxic exercise with combined phentolamine and aminophylline. In contrast, ΔFVC to exogenous adenosine was reduced by aminophylline administration in both protocols (P < 0.05 for both). These observations suggest that adenosine receptor activation is not obligatory for the augmented hyperemia during hypoxic exercise in humans. PMID:19661449

Simultaneous removal of organic matter and salt ions from coal gasification wastewater RO concentrate and microorganisms succession in a MBR.

PubMed

Jia, Shengyong; Han, Yuxing; Zhuang, Haifeng; Han, Hongjun; Li, Kun

2017-10-01

A lab-scale membrane bioreactor (MBR) with intermittent aeration was operated to treat the reverse osmosis concentrate derived from coal gasification wastewater. Results showed intermittent aeration represented slight effect on organic matter reduction but significant effect on nitrite and nitrate reduction, with 6h aeration and 6h non-aeration, removal efficiencies of organic matter, chloride, sulfate, nitrite and nitrate reached 48.35%, 40.91%, 34.28%, -36.05% and 64.34%, respectively. High-throughput sequencing showed a microorganisms succession from inoculated activated sludge (S1) to activated sludge in MBR (S2) with high salinity. Richness and diversity of microorganisms in S2 was lower than S1 and the community structure of S1 exhibited more even than S2. The most relative abundance of genus in S1 and S2 were unclassified_Desulfarculaceae (9.39%) and Roseibaca (62.1%), respectively. High salinity and intermittent aeration represented different influence on the denitrifying genus, and non-aeration phase provided feasible dissolved oxygen condition for denitrifying genera realizing denitrification. Copyright © 2017 Elsevier Ltd. All rights reserved.

Halophilic Microorganisms Are Responsible for the Rosy Discolouration of Saline Environments in Three Historical Buildings with Mural Paintings

PubMed Central

Ettenauer, Jörg D.; Jurado, Valme; Piñar, Guadalupe; Miller, Ana Z.; Santner, Markus; Saiz-Jimenez, Cesareo; Sterflinger, Katja

2014-01-01

A number of mural paintings and building materials from monuments located in central and south Europe are characterized by the presence of an intriguing rosy discolouration phenomenon. Although some similarities were observed among the bacterial and archaeal microbiota detected in these monuments, their origin and nature is still unknown. In order to get a complete overview of this biodeterioration process, we investigated the microbial communities in saline environments causing the rosy discolouration of mural paintings in three Austrian historical buildings using a combination of culture-dependent and -independent techniques as well as microscopic techniques. The bacterial communities were dominated by halophilic members of Actinobacteria, mainly of the genus Rubrobacter. Representatives of the Archaea were also detected with the predominating genera Halobacterium, Halococcus and Halalkalicoccus. Furthermore, halophilic bacterial strains, mainly of the phylum Firmicutes, could be retrieved from two monuments using special culture media. Inoculation of building materials (limestone and gypsum plaster) with selected isolates reproduced the unaesthetic rosy effect and biodeterioration in the laboratory. PMID:25084531

Development of molecular techniques for detection of lymphocystis disease virus in different marine fish species.

PubMed

Cano, I; Ferro, P; Alonso, M C; Bergmann, S M; Römer-Oberdörfer, A; Garcia-Rosado, E; Castro, D; Borrego, J J

2007-01-01

The development and evaluation of a protocol based on polymerase chain reaction (PCR) and nucleic acid hybridization techniques for the specific detection of lymphocystis disease virus (LCDV) in several marine fish species. The pair of primers for PCR, OBL3 and OBL4, was designed based on published nucleotide sequence (LCDV-1) and amplifies a fragment within the major capsid protein. The sensitivity was evaluated using DNA from purified viral particles, as well as from cells inoculated with several viral concentrations. The PCR combined with slot blot was the most sensitive methodology, detecting 2.5 ng of viral DNA. Using this methodology LCDV was detected at 5 days postinoculation from SAF-1 cells initially inoculated with 10(-5) TCID(50) ml(-1). The combination of PCR with membrane hybridization has also been proved to be adequate to detect LCDV from apparently healthy carriers by means of caudal fin sample analysis. This asymptomatic infection was also demonstrated by classical virological methods (cell culture and immunoblot). The protocol described in this study allows the specific detection of LCDV, both in cell cultures and in fin homogenates from asymptomatic fish. The detection of asymptomatic carriers by a rapid molecular method using caudal fin sampling, which does not imply animal killing, could be an important tool to control epizootics caused by LCDV, as fish could be analysed before their introduction and/or mobilization in farm facilities.

Role of adrenal vein sampling in primary aldosteronism: the Monash Health experience.

PubMed

Teng, J; Hutchinson, M E; Doery, J C G; Choy, K W; Chong, W; Fuller, P J; Yang, J

2015-11-01

Adrenal vein sampling (AVS) is useful for distinguishing unilateral versus bilateral hypersecretion in primary aldosteronism (PA), but is technically challenging. Furthermore, the use of adrenocorticotropic hormone (ACTH)-stimulation in AVS is controversial. We implemented a Monash Health-specific AVS protocol in 2010. The audit aimed to: (i) examine the impact of a dedicated protocol on success rates of AVS at a tertiary referral centre; (ii) evaluate the impact of AVS on sub-typing of PA; and (iii) assess the utility of ACTH stimulation in AVS. AVS was performed on patients with PA confirmed by positive saline suppression testing (aldosterone level >140 pmol/L post-saline infusion), with sequential sampling of adrenal and peripheral veins, pre- and post-ACTH infusion. Patients with unilateral aldosterone-producing adenoma diagnosed on successful AVS were referred for adrenalectomy. Between 2010 and 2014 inclusive, a total of 28 AVS procedures was performed, with complete pre- and post-ACTH data for 19 procedures. Bilateral successful cannulation rates improved post-implementation of our protocol (61% vs 41%). Of the patients, 32% had discordant imaging and AVS results: four patients with unilateral adenomas did not lateralise on AVS and were managed medically; four patients with bilateral or no adenomas on imaging, lateralised on AVS and had surgery. Overall, use of ACTH did not increase successful cannulation and tended to mask lateralisation. AVS is crucial in subtype classification of PA and should be performed by a dedicated radiologist with a standardised protocol. AVS outcomes were not improved with the use of ACTH stimulation. © 2015 Royal Australasian College of Physicians.

Recovery of Oesophagostomum dentatum from pigs by isolation of parasites migrating from large intestinal contents embedded in agar-gel.

PubMed

Slotved, H C; Barnes, E H; Bjørn, H; Christensen, C M; Eriksen, L; Roepstorff, A; Nansen, P

1996-06-01

Four groups with three pigs in each group were inoculated with Oesophagostomum dentatum larvae (L3 larvae). Groups 1 and 3 were inoculated with 20,000 larvae, and Groups 2 and 4 with 200,000 larvae. On Days 11 and 34, respectively, Groups 1 and 2 and Groups 3 and 4 were slaughtered, and the contents from the large intestines collected. Subsamples of intestinal contents were mixed with agar to a final concentration of 1% agar and allowed to set. The worms were allowed to migrate from the agar-gel into 38 degrees C 0.9% saline overnight. Then the worms were collected on a sieve (38 microns mesh) and counted. The worms retained in the agar-gel were counted after pouring the melted agar through a sieve (38 microns mesh). The results showed that more than 95% of the worms migrated out of the agar-gel, and subsequently were available for counting in an almost clean suspension. Additionally the method yielded a high worm recovery; all stages were recovered. The recovery percentage was not significantly affected by either the dose of parasites or the time interval from slaughtering to start of incubation (37-128 min).

The effects of Pantoea sp. strain Y4-4 on alfalfa in the remediation of heavy-metal-contaminated soil, and auxiliary impacts of plant residues on the remediation of saline-alkali soils.

PubMed

Li, Shuhuan; Wang, Jie; Gao, Nanxiong; Liu, Lizhu; Chen, Yahua

2017-04-01

The plant-growth-promoting rhizobacterium (PGPR) Y4-4 was isolated from plant rhizosphere soil and identified as Pantoea sp. by 16S rRNA sequence analysis. The effects of strain Y4-4 on alfalfa grown in heavy-metals-contaminated soil was investigated using a pot experiment. In a Cu-rich environment, the shoot dry mass and total dry mass of plants inoculated with strain Y4-4 increased by 22.6% and 21%, and Cu accumulation increased by 15%. In a Pb-Zn-rich environment, the shoot dry mass and total dry mass of plants inoculated with strain Y4-4 increased by 23.4% and 22%, and Zn accumulation increased by 30.3%. In addition, the salt tolerance and biomass of wheat seedlings could be improved by applying strain Y4-4 mixed with plant residue as a result of the Cu-rich plant residues providing copper nutrition to wheat. This study offers an efficient PGPR with strong salt tolerance and a safe strategy for the post-treatment of plant residue.

Methods for conducting bioassays using embryos and larvae of Pacific herring, Clupea pallasi.

PubMed

Dinnel, Paul A; Middaugh, Douglas P; Schwarck, Nathan T; Farren, Heather M; Haley, Richard K; Hoover, Richard A; Elphick, James; Tobiason, Karen; Marshall, Randall R

2011-02-01

The rapid decrease of several stocks of Pacific herring, Clupea pallasi, in Puget Sound, Washington, has led to concerns about the effects of industrial and nonpoint source contamination on the embryo and larval stages of this and related forage fish species. To address these concerns, the state of Washington and several industries have funded efforts to develop embryo and larval bioassay protocols that can be used by commercial laboratories for routine effluent testing. This article presents the results of research to develop herring embryo and larval bioassay protocols. Factors evaluated during protocol development included temperature, salinity, dissolved oxygen (DO), light intensity, photoperiod, larval feeding regimes, use of brine and artificial sea salts, gonad sources, collection methods, and egg quality.

Wetland plant species improve performance when inoculated with arbuscular mycorrhizal fungi: a meta-analysis of experimental pot studies.

PubMed

Ramírez-Viga, Thai Khan; Aguilar, Ramiro; Castillo-Argüero, Silvia; Chiappa-Carrara, Xavier; Guadarrama, Patricia; Ramos-Zapata, José

2018-06-04

The presence of arbuscular mycorrhizal fungi (AMF) in wetlands is widespread. Wetlands are transition ecosystems between aquatic and terrestrial systems, where shallow water stands or moves over the land surface. The presence of AMF in wetlands suggests that they are ecologically significant; however, their function is not yet clearly understood. With the aim of determining the overall magnitude and direction of AMF effect on wetland plants associated with them in pot assays, we conducted a meta-analysis of data extracted from 48 published studies. The AMF effect on their wetland hosts was estimated through different plant attributes reported in the studies including nutrient acquisition, photosynthetic activity, biomass production, and saline stress reduction. As the common metric, we calculated the standardized unbiased mean difference (Hedges' d) of wetland plant performance attributes in AMF-inoculated plants versus non-AMF-inoculated plants. Also, we examined a series of moderator variables regarding symbiont identity and experimental procedures that could influence the magnitude and direction of an AMF effect. Response patterns indicate that wetland plants significantly benefit from their association with AMF, even under flooded conditions. The beneficial AMF effect differed in magnitude depending on the plant attribute selected to estimate it in the published studies. The nature of these benefits depends on the identity of the host plant, phosphorus addition, and water availability in the soil where both symbionts develop. Our meta-analysis synthetizes the relationship of AMF with wetland plants in pot assays and suggests that AMF may be of comparable importance to wetland plants as to terrestrial plants.

[Effects of intrathecal administration of AM22-52 on mechanical allodynia and CCL2 expression in DRG in bone cancer rats].

PubMed

Chen, Ya-Juan; Huo, Yuan-Hui; Hong, Yanguo

2017-02-25

The pain peptide adrenomedullin (AM) plays a pivotal role in pathological pain. The present study was designed to investigate the effect of blockade of AM receptor on bone cancer pain (BCP) and its mechanism. BCP was developed by inoculation of Walker 256 mammary gland carcinoma cells in the tibia medullary cavity of Sprague Dawley rats. The selective AM receptor antagonist AM 22-52 was administered intrathecally on 15 d after the inoculation. Quantitative real-time PCR was used to detect mRNA level of CC chemokine ligand 2 (CCL2) in dorsal root ganglion (DRG). Double immunofluorescence staining was used to analyze the localizations of CCL2 and AM in DRG of normal rats. The results showed that, from 6 to15 d after the inoculation, the animals showed significant reduction in the mechanical pain threshold in the ipsilateral hindpaw, companied by the decline in bone density of tibia bone. The expression of CCL2 mRNA in DRG of BCP rats was increased by 3 folds (P < 0.001 vs saline group). Intrathecal administration of AM 22-52 abolished bone cancer-induced mechanical allodynia and increase of CCL2 mRNA level (P < 0.001). In normal rats, CCL2 was co-localized with AM in DRG neurons. These results suggest that AM may play a role in the pathogenesis of BCP. The increased AM bioactivity up-regulates CCL2 expression in DRG, which may contribute to the induction of pain hypersensitivity in bone cancer.

Evaluation of cell-mediated immune responses against porcine circovirus type 2 (PCV2) Cap and Rep proteins after vaccination with a commercial PCV2 sub-unit vaccine.

PubMed

Fort, Maria; Sibila, Marina; Nofrarías, Miquel; Pérez-Martín, Eva; Olvera, Alex; Mateu, Enric; Segalés, Joaquim

2012-11-15

This study investigated the development of cellular immunity to Porcine circovirus type 2 (PCV2) Cap and Rep proteins in pigs vaccinated with a commercial PCV2 genotype a (PCV2a) based sub-unit vaccine, before and after a heterologous challenge with a PCV2b isolate. At three weeks of age, 20 pigs were inoculated intramuscularly with either the vaccine product (V group, n=9) or phosphate buffered saline solution (PBS) (NV group, n=11). Three weeks after vaccination, pigs were challenged intranasally with PCV2b (V-C and NV-C groups) or PBS (V-NC and NV-NC groups). None of the pigs developed clinical signs during the whole experiment, but all NV-C and 3/5 V-C pigs developed viraemia. Vaccination induced the development IFN-γ-secreting cells in response to the Cap protein of PCV2, which appeared three weeks post-vaccination and increased after challenge. By that time, no significant differences were detected on PCV2 antibody titres between vaccinated and non-vaccinated pigs, although there were significant differences on day 7 post-challenge. PCV2-inoculation induced a cellular response against the Rep protein. Such response was significantly reduced or even absent in PCV2-inoculated pigs that were previously vaccinated (V-C group), presumably as a result of a lower PCV2 replication in vaccinated animals compared to non-vaccinated ones. Copyright © 2012 Elsevier B.V. All rights reserved.

Lack of soluble tumor necrosis factor alpha receptor 1 and 2 and interleukin-1beta compartmentalization in lungs of mice after a single intratracheal inoculation with live Porphyromonas gingivalis.

PubMed

Nemec, Ana; Pavlica, Zlatko; Svete, Alenka Nemec; Erzen, Damijan; Crossley, David A; Petelin, Milan

2009-09-01

Porphyromonas gingivalis aspiration pneumonia induces local and systemic cytokine responses, but the dynamic of the immune response following lung exposure to live P. gingivalis is poorly understood. Groups of 50 12-week-old male BALB/c mice were inoculated intratracheally with live P. gingivalis ATCC 33277 using low dose (2 x 10(5) colony-forming units [CFU]), high dose (2.9 x 10(9) CFU), or phosphate-buffered saline (PBS; sham-inoculated), and the 3 groups were sacrificed at 2, 6, 24, 72, 168 hours. Lung and serum samples were collected for tumor necrosis factor alpha (TNF-alpha), soluble TNF-alpha receptors (sTNFRs), interleukin (IL)-1beta, and IL-6 analysis and lung histology. Pneumonia, only observed in the high-dose group, was associated with an early increase in lung TNF-alpha, IL-1beta, and IL-6, whereas no significant changes were observed in lung sTNFRs. Serum sTNFRs were significantly increased in high-dose animals at all times. IL-1beta elevation occurred earlier in serum than in lungs. IL-1beta was also significantly elevated in serum from low-dose animals at 6 hours. Serum IL-6 and sTNFRs remained raised at 7 days, whereas all other measured cytokines returned to basal levels with resolution of pneumonia. Development of pneumonia is dependent on the P. gingivalis dose; however, part of the cytokine response is unique to the systemic compartment, even in animals that do not develop pneumonia.

Antenatal phosphodiesterase 4 inhibition restores postnatal growth and pulmonary development in a model of chorioamnionitis in rabbits.

PubMed

Homer, L; Launay, E; Joram, N; Jacqueline, C; Jarreau, P-H; Caillon, J; Moyon, T; Branger, B; Potel, G; Roze, J C; Méhats, C; Gras-Leguen, C

2012-03-01

Chorioamnionitis is implicated in the pathophysiology of bronchopulmonary disease, and the associated inflammatory response is responsible for adverse effects on alveolar development. The aim of this work was to analyze the effects of a phosphodiesterase 4 (PDE4)-selective inhibitor, rolipram (a modulator of the inflammatory response), in an experimental model of chorioamnionitis on pulmonary development and on the processes of infection and inflammation. Rabbit mothers were assigned to four groups: 1) saline serum inoculation (controls); 2) Escherichia coli intrauterine inoculation (C+); 3) rolipram infusion (R+); and 4) E. coli inoculation + rolipram infusion (C+R+). High rates of morbility and mortality were noticed in mothers and pups (5 of 13 pregnant rabbits in groups with rolipram). Alveolar development, inflammation, and infection were analyzed in pups at day 0 and day 5. At day 0, in the context of chorioamnionitis, rolipram significantly decreased birth weight (p < 0.01) relative to that of controls (p < 0.05). At day 5, weight normalized in group C+R+ but not in group C+ relative to controls (p < 0.001); moreover, alveolar airspace volume was preserved in group C+R+ but not in group C+ (p < 0.05). Interstitial volume decreased in group C+ versus controls (p < 0.05) but was preserved in group C+R+. Specific alveolar area was not significantly modified by rolipram. No significant difference was found concerning bronchoalveolar lavage cellularity, and all blood cultures remained sterile. In this model of impaired alveologenesis, rolipram significantly preserved specific alveolar density. However, PDE4 inhibition induced antenatal fetal demise and growth retardation.

Effects of Salinity on Leaf Spectral Reflectance and Biochemical Parameters of Nitrogen Fixing Soybean Plants (Glycine max L.)

NASA Astrophysics Data System (ADS)

Krezhova, Dora D.; Kirova, Elisaveta B.; Yanev, Tony K.; Iliev, Ilko Ts.

2010-01-01

Measurements of physiology and hyperspectral leaf reflectance were used to detect salinity stress in nitrogen fixing soybean plants. Seedlings were inoculated with suspension of Bradyrhizobium japonicum strain 273. Salinity was performed at the stage of 2nd-4th trifoliate expanded leaves by adding of NaCl in the nutrient solution of Helrigel in concentrations 40 mM and 80 mM. A comparative analysis was performed between the changes in the biochemical parameters - stress markers (phenols, proline, malondialdehyde, thiol groups), chlorophyll a and b, hydrogen peroxide, and leaf spectral reflectance in the spectral range 450-850 nm. The spectral measurements were carried out by an USB2000 spectrometer. The reflectance data of the control and treated plants in the red, green, red-edge and the near infrared ranges of the spectrum were subjected to statistical analysis. Statistically significant differences were found through the Student's t-criterion at the two NaCl concentrations in all of the ranges examined with the exception of the near infrared range at 40 mM NaCl concentration. Similar results were obtained through linear discriminant analysis. The tents of the phenols, malondialdehyde and chlorophyll a and b were found to decrease at both salinity treatments. In the spectral data this effect is manifested by decrease of the reflectance values in the green and red ranges. The contents of proline, hydrogen peroxide and thiol groups rose with the NaCl concentration increase. At 80 mM NaCl concentration the values of these markers showed a considerable increase giving evidence that the soybean plants were stressed in comparison with the control. This finding is in agreement with the results from the spectral reflectance analysis.

Trichoderma longibrachiatum Evx1 is a fungal biocatalyst suitable for the remediation of soils contaminated with diesel fuel and polycyclic aromatic hydrocarbons.

PubMed

Andreolli, Marco; Lampis, Silvia; Brignoli, Pierlorenzo; Vallini, Giovanni

2016-05-01

Trichoderma sp. strain Evx1 was isolated from a semi-deciduous forest soil in Southern Italy. It decolorizes polynuclear organic dyes and tolerates high concentrations of phenanthrene, anthracene, fluoranthene, and pyrene. The ability of this ascomycete fungus to degrade polycyclic aromatic hydrocarbons was verified in vitro and confirmed by its strong phenoloxidase activity in the presence of gallic acid. Phylogenetic characterization of Trichoderma sp. Evx1 positioned this strain within the species Trichoderma longibrachiatum. The potential use of this species for the bioremediation of contaminated environmental matrices was tested by inoculating diesel-spiked soil with a dense mycelial suspension. The biodegradation percentage of the C12-40 hydrocarbon fraction in the inoculated soil rose to 54.2 ± 1.6 %, much higher than that in non-inoculated soil or soil managed solely by a combination of watering and aeration. The survival and persistence of T. longibrachiatum Evx1 throughout the bioremediation trial was monitored by PCR-DGGE analysis. The fungal strain was still present in the soil 30 days after bioaugmentation. These findings indicate that T. longibrachiatum Evx1 may be a suitable inoculum in bioremediation protocols for the reclamation of soils contaminated by complex mixtures of hydrocarbons.

Signalling through NK1.1 triggers NK cells to die but induces NK T cells to produce interleukin-4.

PubMed

Asea, A; Stein-Streilein, J

1998-02-01

In vivo inoculation of specific antibody is an accepted protocol for elimination of specific cell populations. Except for anti-CD3 and anti-CD4, it is not known if the depleted cells are eliminated by signalling through the target molecule or through a more non-specific mechanism. C57BL/6 mice were inoculated with anti-natural killer (NK1.1) monoclonal antibody (mAb). Thereafter spleen cells were harvested, stained for both surface and intracellular markers, and analysed by flow cytometry. As early as 2 hr post inoculation, NK cells were signalled to become apoptotic while signalling through the NK1.1 molecule activated NK1.1+ T-cell receptor (TCR)+ (NK T) cells to increase in number, and produce interleukin-4 (IL-4). Anti NK1.1 mAb was less efficient at signalling apoptosis in NK cells when NK T-cell deficient [beta 2-microglobulin beta 2m-deficient] mice were used compared with wild type mice. Efficient apoptotic signalling was restored when beta 2m-deficient mice were reconstituted with NK T cells. NK-specific antibody best signals the apoptotic process in susceptible NK cells when resistant NK T cells are present, activated, and secrete IL-4.

Significance of Microbial Communities and Interactions in Safeguarding Reactive Mine Tailings by Ecological Engineering▿†

PubMed Central

N̆ancucheo, Ivan; Johnson, D. Barrie

2011-01-01

Pyritic mine tailings (mineral waste generated by metal mining) pose significant risk to the environment as point sources of acidic, metal-rich effluents (acid mine drainage [AMD]). While the accelerated oxidative dissolution of pyrite and other sulfide minerals in tailings by acidophilic chemolithotrophic prokaryotes has been widely reported, other acidophiles (heterotrophic bacteria that catalyze the dissimilatory reduction of iron and sulfur) can reverse the reactions involved in AMD genesis, and these have been implicated in the “natural attenuation” of mine waters. We have investigated whether by manipulating microbial communities in tailings (inoculating with iron- and sulfur-reducing acidophilic bacteria and phototrophic acidophilic microalgae) it is possible to mitigate the impact of the acid-generating and metal-mobilizing chemolithotrophic prokaryotes that are indigenous to tailing deposits. Sixty tailings mesocosms were set up, using five different microbial inoculation variants, and analyzed at regular intervals for changes in physicochemical and microbiological parameters for up to 1 year. Differences between treatment protocols were most apparent between tailings that had been inoculated with acidophilic algae in addition to aerobic and anaerobic heterotrophic bacteria and those that had been inoculated with only pyrite-oxidizing chemolithotrophs; these differences included higher pH values, lower redox potentials, and smaller concentrations of soluble copper and zinc. The results suggest that empirical ecological engineering of tailing lagoons to promote the growth and activities of iron- and sulfate-reducing bacteria could minimize their risk of AMD production and that the heterotrophic populations could be sustained by facilitating the growth of microalgae to provide continuous inputs of organic carbon. PMID:21965397

Changes in Leaf Anatomical Traits Enhanced Photosynthetic Activity of Soybean Grown in Hydroponics with Plant Growth-Promoting Microorganisms.

PubMed

Paradiso, Roberta; Arena, Carmen; De Micco, Veronica; Giordano, Maria; Aronne, Giovanna; De Pascale, Stefania

2017-01-01

The use of hydroponic systems for cultivation in controlled climatic conditions and the selection of suitable genotypes for the specific environment help improving crop growth and yield. We hypothesized that plant performance in hydroponics could be further maximized by exploiting the action of plant growth-promoting organisms (PGPMs). However, the effects of PGPMs on plant physiology have been scarcely investigated in hydroponics. Within a series of experiments aimed to identify the best protocol for hydroponic cultivation of soybean [ Glycine max (L.) Merr.], we evaluated the effects of a PGPMs mix, containing bacteria, yeasts, mycorrhiza and trichoderma beneficial species on leaf anatomy, photosynthetic activity and plant growth of soybean cv. 'Pr91m10' in closed nutrient film technique (NFT). Plants were grown in a growth chamber under semi-aseptic conditions and inoculated at seed, seedling and plant stages, and compared to non-inoculated (control) plants. Light and epi-fluorescence microscopy analyses showed that leaves of inoculated plants had higher density of smaller stomata (297 vs. 247 n/mm 2 ), thicker palisade parenchyma (95.0 vs. 85.8 μm), and larger intercellular spaces in the mesophyll (57.5% vs. 52.2%), compared to non-inoculated plants. The modifications in leaf functional anatomical traits affected gas exchanges; in fact starting from the reproductive phase, the rate of leaf net photosynthesis (NP) was higher in inoculated compared to control plants (8.69 vs. 6.13 μmol CO 2 m -2 s -1 at the beginning of flowering). These data are consistent with the better maximal PSII photochemical efficiency observed in inoculated plants (0.807 vs. 0.784 in control); conversely no difference in leaf chlorophyll content was found. The PGPM-induced changes in leaf structure and photosynthesis lead to an improvement of plant growth (+29.9% in plant leaf area) and seed yield (+36.9%) compared to control. Our results confirm that PGPMs may confer benefits in photosynthetic traits of soybean plants even in hydroponics (i.e., NFT), with positive effects on growth and seed production, prefiguring potential application of beneficial microorganisms in plant cultivation in hydroponics.

Changes in Leaf Anatomical Traits Enhanced Photosynthetic Activity of Soybean Grown in Hydroponics with Plant Growth-Promoting Microorganisms

PubMed Central

Paradiso, Roberta; Arena, Carmen; De Micco, Veronica; Giordano, Maria; Aronne, Giovanna; De Pascale, Stefania

2017-01-01

The use of hydroponic systems for cultivation in controlled climatic conditions and the selection of suitable genotypes for the specific environment help improving crop growth and yield. We hypothesized that plant performance in hydroponics could be further maximized by exploiting the action of plant growth-promoting organisms (PGPMs). However, the effects of PGPMs on plant physiology have been scarcely investigated in hydroponics. Within a series of experiments aimed to identify the best protocol for hydroponic cultivation of soybean [Glycine max (L.) Merr.], we evaluated the effects of a PGPMs mix, containing bacteria, yeasts, mycorrhiza and trichoderma beneficial species on leaf anatomy, photosynthetic activity and plant growth of soybean cv. ‘Pr91m10’ in closed nutrient film technique (NFT). Plants were grown in a growth chamber under semi-aseptic conditions and inoculated at seed, seedling and plant stages, and compared to non-inoculated (control) plants. Light and epi-fluorescence microscopy analyses showed that leaves of inoculated plants had higher density of smaller stomata (297 vs. 247 n/mm2), thicker palisade parenchyma (95.0 vs. 85.8 μm), and larger intercellular spaces in the mesophyll (57.5% vs. 52.2%), compared to non-inoculated plants. The modifications in leaf functional anatomical traits affected gas exchanges; in fact starting from the reproductive phase, the rate of leaf net photosynthesis (NP) was higher in inoculated compared to control plants (8.69 vs. 6.13 μmol CO2 m-2 s-1 at the beginning of flowering). These data are consistent with the better maximal PSII photochemical efficiency observed in inoculated plants (0.807 vs. 0.784 in control); conversely no difference in leaf chlorophyll content was found. The PGPM-induced changes in leaf structure and photosynthesis lead to an improvement of plant growth (+29.9% in plant leaf area) and seed yield (+36.9%) compared to control. Our results confirm that PGPMs may confer benefits in photosynthetic traits of soybean plants even in hydroponics (i.e., NFT), with positive effects on growth and seed production, prefiguring potential application of beneficial microorganisms in plant cultivation in hydroponics. PMID:28529515

Chemotherapy Extravasation Management: 21-Year Experience.

PubMed

Onesti, Maria Giuseppina; Carella, Sara; Fioramonti, Paolo; Scuderi, Nicolò

2017-11-01

Chemotherapy extravasation may result in serious damage to patients, with irreversible local injures and disability. Evidence-based standardization on extravasation management is lacking and many institutions do not practice adequate procedures to prevent the severer damages. Our aim was to explore the prevention and treatment of extravasation injuries, proposing a standard therapeutic protocol together with a review of the literature. From January 1994 to December 2015, 545 cases were reviewed (age range, 5-87 years; 282 men and 263 women). Our therapeutic protocol consisted of local infiltration of saline solution and topical occlusive applications of corticosteroids. The infiltrations were administrated 3 to 6 times a week depending on damage severity. Our protocol allowed us to prevent ulceration in 373 cases. Only 27 patients required surgery (escarectomy, skin graft, regional, and free flap). Numerous treatments have been proposed in literature. The antidotes have been discussed controversially and are not considered standard methods for treatment, especially when polychemotherapy is administrated and the identification of the responsible drug is not possible. We proposed the use of saline solution injection to dilute rapidly the drug, thus reducing its local toxic effects. This method is easy to use and always reproducible even when the drug is not known or when it is administrated in combination with other drugs. It is possible to perform it in ambulatory regimen, and, overall, it represents a standard method.

[Hydrogen-rich saline attenuates hyperalgesia and reduces cytokines in rats with post-herpetic neuralgia via activating autophagy].

PubMed

Ma, Hongtao; Chen, Hongguang; Dong, Aili; Wang, Yanyan; Bian, Yingxue; Xie, Keliang

2017-02-01

To investigate the role of autophagy in hydrogen-rich saline attenuating post-herpetic neuralgia( PHN) in rats. A total of 100 male SD rats were randomly divided into the five groups( n = 20) : control group,PHN group,PHN group treated with hydrogen-rich saline( PHN-H2group),PHN group treated with hydrogen-rich saline and3-MA( PHN-H2-3-MA group),PHN group treated with hydrogen-rich saline and rapamycin( PHN-H2-Rap group). PHN models were established by varicella-zoster virus( VZV) inoculation. After modeling,15 mg / kg 3-MA or 10 mg / kg rapamycin were intraperitoneally injected in corresponding rats with PHN once two days for 3 times. Hydrogen-rich saline( 10 m L / kg)was injected intraperitoneally twice a day for 7 consecutive days in PHN-H2 group,PHN-H2-3-MA group and PHN-H2-Rap group after VZV injection. The paw withdrawal thresholds( PWT) of 50 rats were detected at 3,7,14 and 21 days after modeling. Spinal cord enlargements of the other 50 rats were collected to examine tumor necrosis factor α( TNF-α),interleukine 1β( IL-1β) and IL-6 by ELISA and autophagy protein microtubule-associated protein 1 light chain 3( LC3),beclin 1and P62 by Western blotting. Compared with the control group,the rats in the PHN group presented with decreased PWT,increased levels of TNF-α,IL-1β,IL-6,LC3Ⅱ and beclin 1,and down-regulated P62 expression. Compared with PHN group,the rats in the PHN-H2 group and PHN-H2-Rap group showed increased PWT,decreased levels of TNF-α,IL-1β and IL-6,further up-regulated expressions of LC3 and beclin 1 as wel as P62 expression. Compared with PHN-H2 group,the rats in the PHN-H2-3-MA group had reduced PWT,elevated expressions of TNF-α,IL-1β and IL-6,suppressed expressions of LC3 and beclin 1,and enhanced p62 expression. Hydrogen-rich saline attenuated PWT and inhibited the release of cytokines TNF-α,IL-1β,IL-6 in rats with PHN via activating autophagy.

[Hydrogen-rich saline attenuates hyperalgesia and reduces cytokines in rats with post-herpetic neuralgia via activating autophagy].

PubMed

Ma, Hongtao; Chen, Hongguang; Dong, Aili; Wang, Yanyan; Bian, Yingxue; Xie, Keliang

2017-02-01

Objective To investigate the role of autophagy in hydrogen-rich saline attenuating post-herpetic neuralgia (PHN) in rats. Methods A total of 100 male SD rats were randomly divided into the five groups (n=20): control group, PHN group, PHN group treated with hydrogen-rich saline (PHN-H 2 group), PHN group treated with hydrogen-rich saline and 3-MA (PHN-H 2 -3-MA group), PHN group treated with hydrogen-rich saline and rapamycin (PHN-H 2 -Rap group). PHN models were established by varicella-zoster virus (VZV) inoculation. After modeling, 15 mg/kg 3-MA or 10 mg/kg rapamycin were intraperitoneally injected in corresponding rats with PHN once two days for 3 times. Hydrogen-rich saline (10 mL/kg) was injected intraperitoneally twice a day for 7 consecutive days in PHN-H 2 group, PHN-H 2 -3-MA group and PHN-H 2 -Rap group after VZV injection. The paw withdrawal thresholds (PWT) of 50 rats were detected at 3, 7, 14 and 21 days after modeling. Spinal cord enlargements of the other 50 rats were collected to examine tumor necrosis factor α (TNF-α), interleukine 1β (IL-1β) and IL-6 by ELISA and autophagy protein microtubule-associated protein 1 light chain 3 (LC3), beclin 1 and P62 by Western blotting. Results Compared with the control group, the rats in the PHN group presented with decreased PWT, increased levels of TNF-α, IL-1β, IL-6, LC3II and beclin 1, and down-regulated P62 expression. Compared with PHN group, the rats in the PHN-H 2 group and PHN-H 2 -Rap group showed increased PWT, decreased levels of TNF-α, IL-1β and IL-6, further up-regulated expressions of LC3 and beclin 1 as well as P62 expression. Compared with PHN-H 2 group, the rats in the PHN-H 2 -3-MA group had reduced PWT, elevated expressions of TNF-α, IL-1β and IL-6, suppressed expressions of LC3 and beclin 1, and enhanced p62 expression. Conclusion Hydrogen-rich saline attenuated PWT and inhibited the release of cytokines TNF-α, IL-1β, IL-6 in rats with PHN via activating autophagy.

First service pregnancy rates following post-AI use of HCG in Ovsynch and Heatsynch programmes in lactating dairy cows.

PubMed

Shabankareh, H Karami; Zandi, M; Ganjali, M

2010-08-01

Lactating dairy cows (n = 667) at random stages of the oestrous cycle were assigned to either ovsynch (O, n = 228), heatsynch (H, n = 252) or control (C, n = 187) groups. Cows in O and H groups received 100 microg of GnRH agonist, i.m. (day 0) starting at 44 +/- 3 days in milk (DIM), and 500 microg of cloprostenol, i.m. (day 7). In O group, cows received 100 microg of GnRH (day 9) and were artificially inseminated without oestrus detection 16-20 h later. In H group, cows received 1 mg oestradiol benzoate (EB) i.m., 24 h after the cloprostenol injection and were artificially inseminated without oestrus detection 48-52 h after the EB injection. Cows in C group were inseminated at natural oestrus. On the day of artificial insemination (AI), cows in all groups were assigned to subgroups as follows: human Chorionic Gonadotrophin (O-hCG) (n = 112), O-saline (n = 116), H-hCG (n = 123), H-saline (n = 129), C-hCG (n = 94) and C-saline (n = 93) subgroups. Cows in hCG and saline subgroups received 3000 IU hCG i.m. and or 10 ml saline at day 5 post-AI (day 15), respectively. Pregnancy status was assessed by palpation per rectum at days 40 to 45 after AI. The logistic regression model using just main effects of season (summer and winter), parity (primiparous and pluriparous), method(1) (O, H and C) and method(2) (hCG and saline) showed that all factors, except method(1), were significant. Significant effects of season (p < 0.01), hCG and parity (p < 0.01), and a trend of parity and season (p < 0.1) were detected. A clear negative effect of warm period on first service pregnancy rate was noted (p < 0.01). The pregnancy rate was the lowest in the H protocol during warm period (p < 0.05). Treatment with hCG 5 days after AI significantly improved pregnancy rates in those cows that were treated with the H protocol compared with saline treatments (41.5% vs 24.8%; p < 0.01). O and H were more effective in primiparous than in pluriparous cows (46.1% vs 29.9%; p < 0.1 and 43.6% vs 24.6%; p < 0.01). First service pregnancy rates were higher in primiparous hCG-treated than in pluriparous hCG-treated cows (57.9% vs 32.3%; p < 0.01). The pregnancy rate was higher for the hCG-treated cows compared with saline-treated cows during warm period (37.9% vs 23.6%; p < 0.001).

Right-to-left shunt detection sensitivity with air-saline and air-succinil gelatin transcranial Doppler.

PubMed

Puledda, Francesca; Toscano, Massimiliano; Pieroni, Alessio; Veneroso, Gabriele; Di Piero, Vittorio; Vicenzini, Edoardo

2016-02-01

Air-saline transcranial Doppler is nowadays the first-choice examination to identify right-to-left shunt. To increase right-to-left shunt detection in echocardiography, cardiologists also use air-gelatin mixtures, which are more stable, more echogenic, and easier to be prepared. We assessed the sensitivity of air-gelatin compared with air-saline for transcranial Doppler right-to-left shunt detection. Air-saline transcranial Doppler, during unilateral middle cerebral artery monitoring at rest and after Valsalva maneuver, was performed in patients referred to our neurosonology laboratory for right-to-left shunt detection. The same transcranial Doppler protocol was repeated with air-gelatin. To consider transcranial Doppler positive for cardiac right-to-left shunt, at least one embolic signal had to be detected within 20″ from contrast injection. Later signals were interpreted of pulmonary origin. Trans-thoracic echocardiography was repeated with both air-saline and air-gelatin. A total of 97 patients were enrolled; 46 had negative transcranial Doppler for cardiac right-to-left shunt with both air-saline and air-gelatin; out of these, four patients with air-saline plus two more patients with air-gelatin presented late, isolated microemboli, slightly more numerous with air-gelatin: these were interpreted as pulmonary shunts and confirmed with trans-thoracic echocardiography. In 28 patients with already early positive air-saline transcranial Doppler at rest, air-gelatin induced a marked right-to-left shunt increase, facilitating its visualization at trans-thoracic echocardiography. In 23 patients in whom air-saline transcranial Doppler was negative at rest and positive for cardiac right-to-left shunt only after Valsalva maneuver, air-gelatin was able to reveal shunt also at rest. Air-gelatin increases right-to-left shunt detection sensitivity with transcranial Doppler in particular at rest, even in patients in whom air-saline mixture fails to identify the shunt. The choice of air-gelatin mixture should be considered for multicentric, clinical, and research trials. © 2016 World Stroke Organization.

Perioperative antimicrobial prophylaxis in neurosurgery: clinical trial of systemic flomoxef administration and saline containing gentamicin for irrigation.

PubMed

Yamamoto, M; Jimbo, M; Ide, M; Tanaka, N; Umebara, Y; Hagiwara, S

1996-06-01

The efficacy of a new protocol consisting of a prophylactic antibiotic regimen of peri- and postoperative intravenous administration of flomoxef and irrigation of the operative field with saline containing gentamicin was assessed by comparing infection rates in two consecutive series of patients who underwent neurosurgical procedures. Group A received postoperative flomoxef administration, with saline containing no antibiotics for irrigation, from July 1988 to December 1989. Group B received the new protocol from January 1990 to December 1991. For further evaluation, this protocol was continued in most patients who underwent surgery from January 1992 through December 1993 (Group C). Only adult or adolescent patients who underwent clean neurosurgical procedures were included. The number of patients and procedures in each group were: 76 patients (97 procedures) in Group A, 103 (133) in Group B, and 107 (137) in Group C. There were no significant differences between Groups A and B in age, sex, clinical category, coexistent disease, clinical outcome, surgical procedures, general anesthesia, emergency operation, steroid administration, and the timing (season), duration, and frequency of surgery. Meningitis developed in three patients and subcutaneous infection in one in Group A. None of the patients in Group B experienced postoperative infection. This difference in infection rates (4.1% vs. 0%) was statistically significant (p = 0.0305). Furthermore, no postoperative infections developed in the Group C patients. The most appropriate interval for multiple dose administration was determined by analyzing intraoperative time-related changes in the serum flomoxef concentration during surgery in 21 recent patients. Serum flomoxef concentrations fell below therapeutic levels (3.0 micrograms/ml) by the 6th post-administration hour in 70% of patients. We conclude that this antibiotic regimen significantly reduces the postoperative infection rate following neurosurgical procedures. Multiple dose administration of flomoxef is recommended when the duration of surgery is 6 hours or more.

Arbuscular mycorrhizal symbiosis mitigates the negative effects of salinity on durum wheat.

PubMed

Fileccia, Veronica; Ruisi, Paolo; Ingraffia, Rosolino; Giambalvo, Dario; Frenda, Alfonso Salvatore; Martinelli, Federico

2017-01-01

Arbuscular mycorrhizal (AM) symbiosis is generally considered to be effective in ameliorating the plant tolerance to salt stress. Unfortunately, the comprehension of the mechanisms implicated in salinity stress alleviation by AM symbiosis is far from being complete. Thus, an experiment was performed by growing durum wheat (Triticum durum Desf.) plants under salt-stress conditions to evaluate the influence of AM symbiosis on both the plant growth and the regulation of a number of genes related to salt stress and nutrient uptake. Durum wheat plants were grown outdoors in pots in absence or in presence of salt stress and with or without AM fungi inoculation. The inoculum consisted of a mixture of spores of Rhizophagus irregularis (formerly Glomus intraradices) and Funneliformis mosseae (formerly G. mosseae). Results indicate that AM symbiosis can alleviate the detrimental effects of salt stress on the growth of durum wheat plants. In fact, under salt stress conditions mycorrhizal plants produced more aboveground and root biomass, had higher N uptake and aboveground N concentration, and showed greater stability of plasma membranes compared to non-mycorrhizal plants. Inoculation with AM fungi had no effect on the expression of the N transporter genes AMT1.1, AMT1.2, and NAR2.2, either under no-stress or salt stress conditions, probably due to the fact that plants were grown under optimal N conditions; on the contrary, NRT1.1 was always upregulated by AM symbiosis. Moreover, the level of expression of the drought stress-related genes AQP1, AQP4, PIP1, DREB5, and DHN15.3 observed in the mycorrhizal stressed plants was markedly lower than that observed in the non-mycorrhizal stressed plants and very close to that observed in the non-stressed plants. Our hypothesis is that, in the present study, AM symbiosis did not increase the plant tolerance to salt stress but instead generated a condition in which plants were subjected to a level of salt stress lower than that of non-mycorrhizal plants.

[Influence of the root canal instrumentation size on the disinfection of intracanal microbe by Er:YAG laser].

PubMed

Xu, Xue-song; Zhu, Ya-qin

2015-12-01

To evaluate the influence of different root canal instrumentation size on disinfection of intracanal microbe of dental root canal. 368 extracted human anterior teeth with single straight root were randomly divided into 8 groups of 46 roots in each. They were instrumented with K3 Ni-Ti files as follows: group A1 and group B1(#25/0.06), group A2 and group B2(#30/0.06), group A3 and group B3(#35/0.06), group A4 and group B4(#40/0.06). After being prepared and sterilized by autoclaving, group A was inoculated with Enterococcus faecalis, and group B was inoculated with Candida albicans. All groups were irrigated with Er:YAG laser combination of 3% NaOCl, 17% EDTA and 0.9% saline, and then the numbers of microbe on the surface of root canal walls were counted after the treatment, the absolute reduction of counting colony forming units(CFUs) and the relative residual rate of CFUs in the individual group was determined. The Date was analyzed with GraphPad Prism 5.02 software package by one-way analysis of variance. Levels of disinfection on E.faecalis and C.albicans increased when root canals were enlarged; #40/0.06 showed the best disinfection, #35/0.06 showed a significantly better disinfection than #30/0.06 and #25/0.06. Substantial reduction of microbe was obtained in #35/0.06 and #40/0.06 compared with #25/0.06 and #30/0.06(P<0.05). Within the root canal size of #25/0.06-#40/0.06, under the conditions of Er:YAG laser combination of 3% NaOCl, 17% EDTA and 0.9% saline, it was concluded that the reduction of E.faecalis and C.albicans of the anterior straight root canals could be predicted by increasing the root canal instrumentation size large than #30/0.06.

Cardiopulmonary and arterial baroreceptor unloading during passive hyperthermia does not contribute to hyperthermia-induced hyperventilation

PubMed Central

Lucas, Rebekah A. I.; Pearson, James; Schlader, Zachary J.; Crandall, Craig G.

2016-01-01

This study tested the hypothesis that baroreceptor unloading during passive hyperthermia contributes to increases in ventilation and decreases in end-tidal partial pressure of carbon dioxide (PET,CO2) during that exposure. Two protocols were performed, in which healthy subjects underwent passive hyperthermia (increasing intestinal temperature by ~1.8°C) to cause a sustained increase in ventilation and reduction in PET,CO2. Upon attaining hyperthermic hyperventilation, in protocol 1 (n = 10; three females) a bolus (19 ± 2 ml kg−1) of warm (~38°C) isotonic saline was rapidly (5–10 min) infused intravenously to restore reductions in central venous pressure, whereas in protocol 2 (n = 11; five females) phenylephrine was infused intravenously (60–120 μg min−1) to return mean arterial pressure to normothermic levels. In protocol 1, hyperthermia increased ventilation (by 2.2 ± 1.7 l min−1, P < 0.01), while reducing PET,CO2 (by 4 ± 3 mmHg, P = 0.04) and central venous pressure (by 5 ± 1 mmHg, P <0.01). Saline infusion increased central venous pressure by 5 ± 1 mmHg (P < 0.01), restoring it to normothermic values, but did not change ventilation or PET,CO2 (P > 0.05). In protocol 2, hyperthermia increased ventilation (by 5.0 ± 2.7l min−1, P <0.01) and reduced PET ,CO2 (by 5 ± 2 mmHg, P < 0.01) and mean arterial pressure (by 9 ± 7 mmHg, P <0.01). Phenylephrine infusion increased mean arterial pressure by 12 ± 3 mmHg (P < 0.01), restoring it to normothermic values, but did not change ventilation or PET,CO2 (P > 0.05). The absence of a reduction in ventilation upon reloading the cardiopulmonary and arterial baroreceptors to pre-hyperthermic levels indicates that baroreceptor unloading with hyperthermia is unlikely to contribute to hyperthermic hyperventilation in humans. PMID:26299270

Engineering the gut microbiota to treat hyperammonemia.

PubMed

Shen, Ting-Chin David; Albenberg, Lindsey; Bittinger, Kyle; Chehoud, Christel; Chen, Ying-Yu; Judge, Colleen A; Chau, Lillian; Ni, Josephine; Sheng, Michael; Lin, Andrew; Wilkins, Benjamin J; Buza, Elizabeth L; Lewis, James D; Daikhin, Yevgeny; Nissim, Ilana; Yudkoff, Marc; Bushman, Frederic D; Wu, Gary D

2015-07-01

Increasing evidence indicates that the gut microbiota can be altered to ameliorate or prevent disease states, and engineering the gut microbiota to therapeutically modulate host metabolism is an emerging goal of microbiome research. In the intestine, bacterial urease converts host-derived urea to ammonia and carbon dioxide, contributing to hyperammonemia-associated neurotoxicity and encephalopathy in patients with liver disease. Here, we engineered murine gut microbiota to reduce urease activity. Animals were depleted of their preexisting gut microbiota and then inoculated with altered Schaedler flora (ASF), a defined consortium of 8 bacteria with minimal urease gene content. This protocol resulted in establishment of a persistent new community that promoted a long-term reduction in fecal urease activity and ammonia production. Moreover, in a murine model of hepatic injury, ASF transplantation was associated with decreased morbidity and mortality. These results provide proof of concept that inoculation of a prepared host with a defined gut microbiota can lead to durable metabolic changes with therapeutic utility.

Deliberate Establishment of Asymptomatic Bacteriuria-A Novel Strategy to Prevent Recurrent UTI.

PubMed

Wullt, Björn; Svanborg, Catharina

2016-07-29

We have established a novel strategy to reduce the risk for recurrent urinary tract infection (UTI), where rapidly increasing antibiotic resistance poses a major threat. Epidemiologic studies have demonstrated that asymptomatic bacteriuria (ABU) protects the host against symptomatic infections with more virulent strains. To mimic this protective effect, we deliberately establish ABU in UTI-prone patients, who are refractory to conventional therapy. The patients are inoculated with Escherichia coli (E. coli) 83972, now widely used as a prototype ABU strain. Therapeutic efficacy has been demonstrated in a placebo-controlled trial, supporting the feasibility of using E. coli 83972 as a tool to prevent recurrent UTI and, potentially, to outcompete antibiotic-resistant strains from the human urinary tract. In addition, the human inoculation protocol offers unique opportunities to study host-parasite interaction in vivo in the human urinary tract. Here, we review the clinical evidence for protection using this approach as well as some molecular insights into the pathogenesis of UTI that have been gained during these studies.

Genetic diversity and symbiotic compatibility among rhizobial strains and Desmodium incanum and Lotus spp. plants.

PubMed

Granada, Camille E; Strochein, Marcos; Vargas, Luciano K; Bruxel, Manuela; de Sá, Enilson Luiz Saccol; Passaglia, Luciane M P

2014-06-01

This work aimed to evaluate the symbiotic compatibility and nodulation efficiency of rhizobia isolated from Desmodium incanum, Lotus corniculatus, L. subbiflorus, L. uliginosus and L. glaber plants by cross-inoculation. Twelve reference strains and 21 native isolates of rhizobia were genetically analyzed by the BOX-PCR technique, which showed a high genetic diversity among the rhizobia studied. The isolates were also characterized based on their production of indolic compounds and siderophores, as well as on their tolerance to salinity. Fifteen of the 33 rhizobia analyzed were able to produce indolic compounds, whereas 13 produced siderophores. All the tested rhizobia were sensitive to high salinity, although some were able to grow in solutions of up to 2% NaCl. Most of the native rhizobia isolated from L. uliginosus were able to induce nodulation in all plant species studied. In a greenhouse experiment using both D. incanum and L. corniculatus plants, the rhizobia isolate UFRGS Lu2 promoted the greatest plant growth. The results demonstrate that there are native rhizobia in the soils of southern Brazil that have low host specificity and are able to induce nodulation and form active nodules in several plant species.

Genetic diversity and symbiotic compatibility among rhizobial strains and Desmodium incanum and Lotus spp. plants

PubMed Central

Granada, Camille E.; Strochein, Marcos; Vargas, Luciano K.; Bruxel, Manuela; de Sá, Enilson Luiz Saccol; Passaglia, Luciane M.P.

2014-01-01

This work aimed to evaluate the symbiotic compatibility and nodulation efficiency of rhizobia isolated from Desmodium incanum, Lotus corniculatus, L. subbiflorus, L. uliginosus and L. glaber plants by cross-inoculation. Twelve reference strains and 21 native isolates of rhizobia were genetically analyzed by the BOX-PCR technique, which showed a high genetic diversity among the rhizobia studied. The isolates were also characterized based on their production of indolic compounds and siderophores, as well as on their tolerance to salinity. Fifteen of the 33 rhizobia analyzed were able to produce indolic compounds, whereas 13 produced siderophores. All the tested rhizobia were sensitive to high salinity, although some were able to grow in solutions of up to 2% NaCl. Most of the native rhizobia isolated from L. uliginosus were able to induce nodulation in all plant species studied. In a greenhouse experiment using both D. incanum and L. corniculatus plants, the rhizobia isolate UFRGS Lu2 promoted the greatest plant growth. The results demonstrate that there are native rhizobia in the soils of southern Brazil that have low host specificity and are able to induce nodulation and form active nodules in several plant species. PMID:25071405

Antibacterial properties and healing effects of Melipona scutellaris honey in MRSA-infected wounds of rats.

PubMed

Medeiros, Vanessa de Fátima Lima Paiva; Azevedo, Ítalo Medeiros; Rêgo, Amália Cínthia Meneses; Egito, Eryvaldo Sócrates Tabosa do; Araújo-Filho, Irami; Medeiros, Aldo Cunha

2016-05-01

To investigate the antimicrobial, immunological and healing effects of Melipona scutellaris honey on infected wounds of rat skin. Twenty four Wistar rats were distributed in four groups (6-each). The uninfected skin wounds of group I rats were treated daily with saline for 7 days. Uninfected wounds (group II) rats were treated with honey. In group III (treated with saline) and group IV (treated with honey) wounds were inoculated with MRSA ATTC43300. The first bacterial culture was performed 24 hours later. In the 7th day new culture was done, and wound biopsies were used for cytokines dosage and histopathology. In group I and III rats the CFU/g count of S. aureus in wounds was zero. In group II rats the CFU/g counts in the wound tissue were significantly higher than in wounds of group IV rats. The density histopathological parameters and the expression of TNF-α, IL1-β, Il-6 were significantly higher on wounds of group IV then in the other groups. Honey of Melipona scutellaris was effective in the management of infected wounds, by significant bacterial growth inhibition, enhancement of cytokine expression, and positively influenced the wound repair.

Detection of influenza A virus nucleoprotein antibodies in oral fluid specimens from pigs infected under experimental conditions using a blocking ELISA.

PubMed

Panyasing, Y; Goodell, C K; Wang, C; Kittawornrat, A; Prickett, J R; Schwartz, K J; Ballagi, A; Lizano, S; Zimmerman, J J

2014-04-01

In commercial swine populations, influenza is an important component of the porcine respiratory disease complex (PRDC) and a pathogen with major economic impact. Previously, a commercial blocking ELISA (FlockChek(™) Avian Influenza Virus MultiS-Screen(®) Antibody Test Kit, IDEXX Laboratories, Inc., Westbrook, ME, USA) designed to detect influenza A nucleoprotein (NP) antibodies in avian serum was shown to accurately detect NP antibodies in swine serum. The purpose of this study was to determine whether this assay could detect NP antibodies in swine oral fluid samples. Initially, the procedure for performing the NP-blocking ELISA on oral fluid was modified from the serum testing protocol by changing sample dilution, sample volume, incubation time and incubation temperature. The detection of NP antibody was then evaluated using pen-based oral fluid samples (n = 182) from pigs inoculated with either influenza A virus subtype H1N1 or H3N2 under experimental conditions and followed for 42 days post inoculation (DPI). NP antibodies in oral fluid were detected from DPI 7 to 42 in all inoculated groups, that is, the mean sample-to-negative (S/N) ratio of influenza-inoculated pigs was significantly different (P < 0.0001) from uninoculated controls (unvaccinated or vaccinated-uninoculated groups) through this period. Oral fluid versus serum S/N ratios from the same pen showed a correlation of 0.796 (Pearson's correlation coefficient, P < 0.0001). The results showed that oral fluid samples from influenza virus-infected pigs contained detectable levels of NP antibodies for ≥42 DPI. Future research will be required to determine whether this approach could be used to monitor the circulation of influenza virus in commercial pig populations. © 2012 Blackwell Verlag GmbH.

Pilot-scale continuous recycling of growth medium for the mass culture of a halotolerant Tetraselmis sp. in raceway ponds under increasing salinity: a novel protocol for commercial microalgal biomass production.

PubMed

Fon Sing, S; Isdepsky, A; Borowitzka, M A; Lewis, D M

2014-06-01

The opportunity to recycle microalgal culture medium for further cultivation is often hampered by salinity increases from evaporation and fouling by dissolved and particulate matter. In this study, the impact of culture re-use after electro-flocculation of seawater-based medium on growth and biomass productivity of the halotolerant green algal strain Tetraselmis sp., MUR 233, was investigated in pilot-scale open raceway ponds over 5months. Despite a salinity increase from 5.5% to 12% (w/v) NaCl, Tetraselmis MUR 233 grown on naturally DOC-enriched recycled medium produced 48-160% more ash free dry weight (AFDW) biomass daily per unit pond area than when grown on non-recycled medium. A peak productivity of 37.5±3.1gAFDWm(-2)d(-1) was reached in the recycled medium upon transition from ∼14% to ∼7% NaCl. The combination of high biomass-yielding mixotrophic growth under high salinity has been proven to be a successful sustainable cultivation strategy. Copyright © 2014 Elsevier Ltd. All rights reserved.

Suppression of humoral response during the course of Candida albicans infection in mice.

PubMed

Valdez, J C; Meson, O E; de Valdez, G A; Sirena, A

1984-10-30

This paper aims at demonstrating the non-specific immunosuppression as regards thyme-dependent antigens sheep erythrocytes (SRBC) during the course of Candida albicans systemic infection. Three lots of syngeneic/BALB/c mice, 8-12 weeks of age, were used. The first normal lot was inoculated via the intraperitoneal route with a (SRBC) suspension (4 X 10(8) cells ml) in a Hank's balanced saline solution. The primary response of antibodies formed by splenic cells was measured from 4 to 8 days after inoculation using the direct plaque forming cells technique. The second lot was infected by the same route with a suspension of Candida albicans (1 X 10(7) cells). Positive retrocultures from the blood and kidneys of these infected mice were obtained. These yeasts cultivated in a Sabouraud medium were harvested after 20 h at 37 degrees C. Following the same methodology the immune response to SRBC was determined. The serum obtained from infected mice was transferred to a third lot of mice at different intervals during the course of the infection. The immune response to SRBC was done by the direct plaque-forming cells technique. Controls were carried out using normal donors and recipients. A suppression of the immune response was obtained as from the 2nd day of inoculation up to the 28th day. It was not possible to transfer such suppression passively by means of the serum. These results suggest that the systemic infection by Candida albicans induce a non-specific immunosuppression in the organism, already demonstrated in viral infections, bacteria, protozoaria and metazoaria in mammals. In some way, this will contribute to explain the mechanisms of immune response to Candida albicans.

The ability of lipopolysaccharide (LPS) of Pasteurella multocida B:2 to induce clinical and pathological lesions in the nervous system of buffalo calves following experimental inoculation.

PubMed

Marza, Ali Dhiaa; Jesse Abdullah, Faez Firdaus; Ahmed, Ihsan Muneer; Teik Chung, Eric Lim; Ibrahim, Hayder Hamzah; Zamri-Saad, Mohd; Omar, Abdul Rahman; Abu Bakar, Md Zuki; Saharee, Abdul Aziz; Haron, Abdul Wahid; Alwan, Mohammed Jwaid; Mohd Lila, Mohd Azmi

2017-03-01

Lipopolysaccharide (LPS) of P. multocida B:2, a causative agent of haemorrhagic septicaemia (HS) in cattle and buffaloes, is considered as the main virulence factor and contribute in the pathogenesis of the disease. Recent studies provided evidences about the involvement of the nervous system in pathogenesis of HS. However, the role of P. multocida B:2 immunogens, especially the LPS is still uncovered. Therefore, this study was designed to investigate the role of P. multocida B:2 LPS to induce pathological changes in the nervous system. Nine eight-month-old, clinically healthy buffalo calves were used and distributed into three groups. Calves of Group 1 and 2 were inoculated orally and intravenously with 10 ml of LPS broth extract represent 1 × 10 12  cfu/ml of P. multocida B:2, respectively, while calves of Group 3 were inoculated orally with 10 ml of phosphate buffer saline as a control. Significant differences were found in the mean scores for clinical signs, post mortem and histopathological changes especially in Group 2, which mainly affect different anatomic regions of the nervous system, mainly the brain. On the other hand, lower scores have been recorded for clinical signs, gross and histopathological changes in Group 1. These results provide for the first time strong evidence about the ability of P. multocida B:2 LPS to cross the blood brain barrier and induce pathological changes in the nervous system of the affected buffalo calves. Copyright © 2017 Elsevier Ltd. All rights reserved.

Duration of growth depression and pathogen shedding in experimentally reproduced poult enteritis syndrome.

PubMed

Jindal, Naresh; Patnayak, Devi P; Ziegler, Andre F; Lago, Alfonso; Goyal, Sagar M

2009-12-01

An experimental study was conducted to determine the duration of growth depression and virus shedding in turkey poults after oral inoculation with intestinal contents from birds affected with poult enteritis syndrome (PES). Poults at day 14 of age were divided into four groups (groups A, B, C, and D) of 40 poults each and inoculated orally with unfiltered supernatant, filtered supernatant, sediment suspended in phosphate-buffered saline (PBS), or PBS alone (control), respectively. The poults were observed daily for clinical signs, and their growth response, pathology, and pathogen shedding were examined at 10, 20, 30, 40, and 50 days postinoculation (DPI). Body weights of eight poults in each group were recorded at each of these intervals followed by euthanasia. Dullness, depression, and diarrhea were observed in birds inoculated with supernatant or sediment suspension. All three treatments significantly reduced body weight gain of poults compared with the control group; average weight loss was 14%. Gross pathologic changes consisted of pale distended intestines with watery contents and distended ceca with frothy and watery contents. Astrovirus and rotavirus were detected in the inoculum by reverse transcription (RT)-PCR, whereas Salmonella was identified on bacterial isolation. Both viruses were detected in treated poults by RT-PCR for up to 10 and 40 DPI, respectively. Of the three treatments, sediment suspension caused maximal decrease in weight gain as well as greatest pathologic lesions followed by unfiltered supernatant and filtered supernatant. These findings suggest a role for bacteria in increasing the severity of PES. Lower weight gain in treated poults (compared with controls) at 9 wk of age also indicates that PES-affected poults may not reach normal weight at marketing, leading to economic losses for the producer.

Detection of Infectious Bovine Rhinotracheitis and Bovine Viral Diarrhea Viruses in the Nasal Epithelial Cells by the Direct Immunofluorescence Technique

PubMed Central

Silim, A.; Elazhary, M.A.S.Y.

1983-01-01

Nasal epithelial cells were collected by cotton swabs for the diagnosis in experimental and field cases of infectious bovine rhinotracheitis and field cases of bovine viral diarrhea in calves. A portion of the cells was washed twice in phosphate buffered saline and a 25 µL drop was placed on microscope slides. The cells were dried, fixed and stained according to the direct fluorescent antibody technique. Another portion of the same specimen was inoculated onto primary bovine skin cell cultures for virus isolation. In the experimental studies for infectious bovine rhinotracheitis, 29/35 specimens were positive by fluorescent antibody technique and 32/35 by cell culture and in the field cases, 22/119 were positive by fluorescent antibody technique and 19/119 by cell culture. In the field cases of bovine viral diarrhea, 28/69 samples were positive by fluorescent antibody technique and 14/69 by cell culture. When fluorescent antibody technique was performed on inoculated cell cultures a total of 24/69 specimens were positive for bovine viral diarrhea. The sensitivity of fluorescent antibody technique was thus comparable to that of cell culture method for infectious bovine rhinotracheitis and bovine viral diarrhea. ImagesFig. 1.Fig. 2.Fig. 3. PMID:6299484

THE INFLUENCE OF THE FACTORS OF SEX, AGE, AND METHOD OF INOCULATION UPON THE COURSE OF EXPERIMENTAL SYPHILIS IN THE RABBIT.

PubMed

Chesney, A M

1923-10-31

For convenience the results are summarized in tabular form. Table I shows a comparison of the primary reaction in the groups of female and male rabbits inoculated intradermally. In Table II the incidence of metastatic lesions is shown in the various groups of animals. In Table III are shown the character and date of appearance of the generalized lesions (exclusive of orchitis) in the various groups of animals. The results presented in Tables I, II, and III, together with a study of the individual protocols, permit the following general statements to be made relative to the effect of the factors studied upon the course of experimental syphilis in the rabbit. Sex.-In the group of females inoculated intradermally the lesions at the site of inoculation were in general less marked than in the group of males similarly inoculated. Moreover, they attained their maximum size earlier and began to recede earlier than was the case in the males. In none of the females was there clinical evidence of the production of generalized lesions, while in the corresponding group of males metastatic lesions of skin or bones detectible by clinical examination occurred in one instance, or 14.3 per cent. In both groups there was wide variation in the character of the initial reaction in individual rabbits and in the time required for it to reach its maximum size. See PDF for Structure Age.-In the five young males inoculated intratesticularly the disease was not greatly different from that observed in the older males similarly inoculated. The initial reactions were slightly slower in making their appearance, but the magnitude of the reaction, as judged by enlargement of the testicles, was somewhat greater. The percentage incidence of metastatic orchitis was almost the same in the two groups. The younger animals showed a slightly greater incidence of generalized lesions involving structures other than the testicles, although the difference was insignificant, but the lesions were smaller, fewer in number, and confined entirely to the skin, and there was no instance of severe generalized lesions. Moreover, the occurrence of metastatic lesions (skin) in the younger animals was definitely delayed. Method of Injection.-In the animals inoculated by the intratesticular route the development of the primary reaction and the incidence of generalized lesions involving skin and bone were much greater than in See PDF for Structure the animals inoculated intradermally. Thus, of the latter group only one, or 14.3 per cent, showed a secondary metastatic lesion (skin), while in the group inoculated by the testicular route the incidence of metastatic skin and bone lesions was as high as 71.4 per cent. The difference in the character of the disease exhibited by these two groups of animals inoculated in a different manner is quite striking. Intratesticular inoculation produced a much more violent local reaction and a much greater incidence of generalized lesions than did intradermal inoculation. Effect of Castration.-In the group of animals with unilateral orchitis in which the infection was allowed to run its course without any attempt at suppression by removal of the initial focus, metastatic lesions appeared more promptly and in slightly greater incidence than See PDF for Structure in the group of animals in which the initial local reaction was suppressed by removal of the focus itself through castration. The percentage differences are not very great, but they are constantly in favor of the first group, as is the time interval of appearance. This experiment is not in strict accord with similar experiments observed by Brown and Pearce (3) in the past, and while the reasons for this difference are not clear the fact may be noted that the incidence of generalized lesions in uncastrated animals is somewhat higher than that reported by these authors, also that there was a high percentage of recurrence of primary lesions.

Validation of a Nylon-Flocked-Swab Protocol for Efficient Recovery of Bacterial Spores from Smooth and Rough Surfaces▿

PubMed Central

Probst, Alexander; Facius, Rainer; Wirth, Reinhard; Moissl-Eichinger, Christine

2010-01-01

In order to meet planetary-protection requirements, culturable bacterial spore loads are measured representatively for the total microbial contamination of spacecraft. However, the National Aeronautics and Space Administration's (NASA's) cotton swab protocols for spore load determination have not changed for decades. To determine whether a more efficient alternative was available, a novel swab was evaluated for recovery of different Bacillus atrophaeus spore concentrations on stainless steel and other surfaces. Two protocols for the nylon-flocked swab (NFS) were validated and compared to the present NASA standard protocol. The results indicate that the novel swab protocols recover 3- to 4-fold more (45.4% and 49.0% recovery efficiency) B. atrophaeus spores than the NASA standard method (13.2%). Moreover, the nylon-flocked-swab protocols were superior in recovery efficiency for spores of seven different Bacillus species, including Bacillus anthracis Sterne (recovery efficiency, 20%). The recovery efficiencies for B. atrophaeus spores from different surfaces showed a variation from 5.9 to 62.0%, depending on the roughness of the surface analyzed. Direct inoculation of the swab resulted in a recovery rate of about 80%, consistent with the results of scanning electron micrographs that allowed detailed comparisons of the two swab types. The results of this investigation will significantly contribute to the cleanliness control of future life detection missions and will provide significant improvement in detection of B. anthracis contamination for law enforcement and security efforts. PMID:20543054

Unravelling the contribution of lactic acid bacteria and acetic acid bacteria to cocoa fermentation using inoculated organisms.

PubMed

Ho, Van Thi Thuy; Fleet, Graham H; Zhao, Jian

2018-08-20

Cocoa beans (Theobroma cacao L.) are the raw material for chocolate production. Fermentation of the bean pulp by microorganisms is essential for developing the precursors of chocolate flavour. Currently, the cocoa fermentation is still conducted by an uncontrolled traditional process via a consortium of indigenous species of yeasts, lactic acid bacteria and acetic acid bacteria. Although the essential contribution of yeasts to the production of good quality beans and, typical chocolate character is generally agreed, the roles of lactic acid bacteria and acetic acid bacteria are less certain. The objective of this study was to investigate the contribution of LAB and AAB in cocoa bean fermentation by conducting small scale laboratory fermentations under aseptic conditions, inoculated with different groups of microorganisms previously isolated from spontaneous cocoa fermentations. The inoculation protocols were: (1) four yeasts Hanseniaspora guilliermondii, Pichia kudriavzevii, Kluyveromyces marxianus and Saccharomyces cerevisiae; (2) four yeasts plus the lactic acid bacteria Lactobacillus plantarum and Lactobacillus fermentum; (3) four yeasts plus the acetic acid bacteria Acetobacter pasteurianus and Gluconobacter frateuri and (4) four yeasts plus two lactic acid bacteria and two acetic acid bacteria. Only the inoculated species were detected in the microbiota of their respective fermentations. Beans from the inoculated fermentations showed no significant differences in colour, shell weights and concentrations of residual sugars, alcohols and esters (p>0.05), but they were slightly different in contents of lactic acid and acetic acid (p<0.05). All beans were fully brown and free of mould. Residual sugar levels were less than 2.6 mg/g while the shell contents and ethanol were in the range of 11-13.4% and 4.8-7 mg/g, respectively. Beans fermented in the presence of LAB contained higher levels of lactic acid (0.6-1.2 mg/g) whereas higher concentrations of acetic acid (1.8-2.2 mg/g) were detected in beans inoculated with AAB. Triangle and hedonic sensory evaluations of chocolates prepared from beans taken from the three fermentations showed no significant differences (p > 0.05). It was concluded that the growth of lactic acid bacteria and acetic acid bacteria may not be essential for the fermentation of cocoa beans. Copyright © 2018 Elsevier B.V. All rights reserved.

The combination of arbuscular mycorrhizal fungi inoculation (Glomus versiforme) and 28-homobrassinolide spraying intervals improves growth by enhancing photosynthesis, nutrient absorption, and antioxidant system in cucumber (Cucumis sativus L.) under salinity.

PubMed

Ahmad, Husain; Hayat, Sikandar; Ali, Muhammad; Liu, Tao; Cheng, Zhihui

2018-06-01

Salinity is one of the major obstacles in the agriculture industry causing huge losses in productivity. Several strategies such as plant growth regulators with arbuscular mycorrhizal fungi (AMF) have been used to decrease the negative effects of salt stress. In our experiment, 28-homobrassinolide (HBL) with spraying intervals was combined with AMF ( Glomus versiforme ) in cucumber cultivars Jinyou 1 # (salt sensitive) and (Changchun mici, in short, CCMC, salt tolerant) under NaCl (100 mmol/L). Studies have documented that the foliar application of HBL and AMF colonization can enhance tolerance to plants under stress conditions. However, the mechanism of the HBL spraying intervals after 15 and 30 days in combination with AMF in cucumber under salt stress is still unknown. Our results revealed that the HBL spraying interval after 15 days in combination with AMF resulted in improved growth, photosynthesis, and decreased sodium toxicity under NaCl. Moreover, the antioxidant enzymes SOD (superoxide dismutase; EC 1.15.1.1) and POD activity (peroxidase; EC 1.11.1.7) showed a gradual increase after every 10 days, while the CAT (catalase; EC 1.11.1.6) increased after 30 days of salt treatments in both cultivars. This research suggests that the enhanced tolerance to salinity was mainly related to elevated levels of antioxidant enzymes and lower uptake of Na + , which lowers the risk of ion toxicity and decreases cell membrane damage.

Plant growth-promoting bacteria Bacillus amyloliquefaciens NBRISN13 modulates gene expression profile of leaf and rhizosphere community in rice during salt stress.

PubMed

Nautiyal, Chandra Shekhar; Srivastava, Suchi; Chauhan, Puneet Singh; Seem, Karishma; Mishra, Aradhana; Sopory, Sudhir Kumar

2013-05-01

Growth and productivity of rice and soil inhabiting microbial population is negatively affected by soil salinity. However, some salt resistant, rhizosphere competent bacteria improve plant health in saline stress. Present study evaluated the effect of salt tolerant Bacillus amyloliquefaciens NBRISN13 (SN13) inoculation on rice plants in hydroponic and soil conditions exposed to salinity. SN13 increased plant growth and salt tolerance (NaCl 200 mM) and expression of at least 14 genes under hydroponic and soil conditions in rice. Among these 14 genes 4 (NADP-Me2, EREBP, SOSI, BADH and SERK1) were up-regulated and 2 (GIG and SAPK4) repressed under salt stress in hydroponic condition. In greenhouse experiment, salt stress resulted in accumulation of MAPK5 and down-regulation of the remaining 13 transcripts was observed. SN13 treatment, with or without salt gave similar expression for all tested genes as compared to control. Salt stress caused changes in the microbial diversity of the rice rhizosphere and stimulated population of betaine-, sucrose-, trehalose-, and glutamine-utilizing bacteria in salt-treated rice rhizosphere (SN13 + salt). The observations imply that SN13 confers salt tolerance in rice by modulating differential transcription in a set of at least 14 genes. Stimulation of osmoprotectant utilizing microbial population as a mechanism of inducing salt tolerance in rice is reported for the first time in this study to the best of our knowledge. Copyright © 2013 Elsevier Masson SAS. All rights reserved.

Laboratory grown subaerial biofilms on granite: application to the study of bioreceptivity.

PubMed

Vázquez-Nion, Daniel; Silva, Benita; Troiano, Federica; Prieto, Beatriz

2017-01-01

Simulated environmental colonisation of granite was induced under laboratory conditions in order to develop an experimental protocol for studying bioreceptivity. The experimental set-up proved suitable for producing subaerial biofilms by inoculating granite blocks with planktonic multi-species phototrophic cultures derived from natural biofilms. The ability of four different cultures to form biofilms was monitored over a three-month growth period via colour measurements, quantification of photosynthetic pigments and EPS, and CLSM observations. One of the cultures under study, which comprised several taxa including Bryophyta, Charophyta, Chlorophyta and Cyanobacteria, was particularly suitable as an inoculum, mainly because of its microbial richness, its rapid adaptability to the substratum and its high colonisation capacity. The use of this culture as an inoculum in the proposed experimental set-up to produce subaerial biofilms under laboratory conditions will contribute to standardising the protocols involved, thus enabling more objective assessment of the bioreceptivity of granite in further experiments.

Isolation and in vitro culture of trypanosomes from Leptodactylus ocellatus from the Atlantic Forest in a new experimental culture medium.

PubMed

Lemos, M; Souza, C S F; da Costa, S C Gonçalves; Souto-Padrón, T; D'Agosto, M

2013-02-01

The purpose of this study was to verify the in vitro development of Trypanosoma sp. isolated from Leptodactylus ocellatus frogs under a new protocol using a biphasic medium composed of Novy, McNeal, and Nicolle (NNN) blood agar medium as a solid phase and liver infusion, brain heart infusion, and tryptose (LIBHIT) medium as a liquid phase. Blood forms, collected by cardiac puncture or after the maceration of different organs, were inoculated in culture tubes containing the biphasic medium composed by NNN and LIBHIT. Trypanosomes were observed 4 days postinoculation; most bloodstream trypomastigotes had differentiated into epimastigotes and amastigotes by this time. Trypomastigotes were again observed in older cultures (7 days). Parasites were successfully subcultured for 8 mo in this medium and successfully cryopreserved. The present study provides a new protocol medium for the isolation and culture of anuran trypanosomes.

Reduction in Nontuberculous Mycobacteria at a Tuberculosis Hospital Following a Quality Assurance Intervention

PubMed Central

Kizilbash, Quratulain; Jost, Kenneth; Armitige, Lisa; Griffith, David E; Dunbar, Denise; Seaworth, Barbara

2017-01-01

Abstract Background Non tuberculous mycobacteria (NTM) are widely distributed in soil and water. NTM/Mycobacterium tuberculosis complex (MTBC) mixes may yield positive AFB smears falsely attributed to tuberculosis (TB) and false-resistance profiles for TB due to contaminated diagnostic samples. This as well as isolation of NTM may pose diagnostic and management problems. Texas Center for Infectious Disease (TCID) is a hospital for patients with confirmed TB. After a cluster of isolates of Mycobacterium gordonae was identified, a quality assurance review found inadequate protocols which included eating and drinking prior to collection. Changes made to the sputum collection protocol included reeducation of respiratory therapists and a sterile saline rinse intervention prior to sputum collection. Methods All sputa collected for AFB culture from diagnosed TB patients at TCID from January 1st, 2014 to December 31st, 2014 prior to the intervention and from August 1st, 2016 to January 31st, 2017, the 6 months following the quality assurance intervention were included. Sputum samples were processed at the Texas Department of State and Health Services (DSHS) Laboratory. Results A total of 1,853 sputum samples were processed; 1,288 from 2014 and 565 following the intervention. NTM decreased from 56 (4.3%) to 7 (1.2%) after the quality assurance intervention was instituted for a NTM decrease of 75.0%. M. gordonae decreased by 78.6%. No patients had evidence of NTM disease. Conclusion A breach in sputum collection protocols at TCID accounted for the increase in NTM isolation in 2014, half of which were M. gordonae. The reeducation of respiratory therapy staff and initiation of sterile saline rinse prior to sputum collection resulted in a significant reduction in the overall NTM rate. M. gordonae was isolated only three times following the intervention. At TCID, a location where tap water and bottled water contains NTM, drinking these prior to sputum collection possibly contributed to the cluster for NTM, especially M. gordonae. We recommend rinsing the mouth with sterile saline or water prior to sputum collection to decrease isolation of rarely pathogenic NTM. Disclosures All authors: No reported disclosures.

Does Prebiopsy, Nonsterile Ultrasonography Gel Affect Biopsy-Site Asepsis?

DOE Office of Scientific and Technical Information (OSTI.GOV)

Gurel, Kamil; Karabay, Oguz; Gurel, Safiye

2008-01-15

Purpose. The purpose of this study was to determine the extent to which the use of nonsterile gel, prior to antiseptic procedures in ultrasonography (US)-guided percutaneous biopsies, results in contamination of the biopsy site. Materials and Methods. Patients referred for US-guided percutaneous biopsies were included in this study. Transmission material used for US evaluation before biopsy-site antiseptic procedures were performed was either nonsterile gel or sterile saline. Patients were randomly assigned to two groups: nonsterile gel (n = 30) and sterile saline (n = 30). Before the transmission material was used and after antiseptic procedures were performed, microbial swabs ofmore » a 10-cm{sup 2}-diameter area were obtained at the biopsy site. Swabs were also obtained from the gel, saline, and povidine-iodine. Inoculated specimen plates were incubated at 37{sup o}C under aerobic conditions, and the numbers of colony-forming units recorded. Nominal logistic regression analysis was used to calculate the odds of postantisepsis bacterial growth (after antiseptic procedures were performed) based on group, gender, coincidental disease (diabetes, chronic renal failure, and malignancy), biopsy-site location (head and neck or breast and abdomen), and local factors (skin fold, skin tag, and hair). Results. The following odds ratios (adjusted for the other variables) and their 95% confidence intervals were calculated: (1) group (2.9 [0.8-11.1]; p = 0.10); (2) gender (1.2 [0.3-5.2]; p = 0.78); (3) coincidental disease (7.6 [0.9-166.7]; p = 0.09); (4) biopsy site location (6.2 [1.4-31.3]; p = 0.02); and (5) local factors (7.0 [1.6-36.0]; p = 0.01). No bacterial growth occurred with swabs obtained from gel, povidine-iodine, or saline. Conclusion. We conclude that nonsterile gel used prior to percutaneous biopsy does not affect biopsy-site asepsis.« less

Augmented Quadruple-Phase Contrast Media Administration and Triphasic Scan Protocol Increases Image Quality at Reduced Radiation Dose During Computed Tomography Urography.

PubMed

Saade, Charbel; Mohamad, May; Kerek, Racha; Hamieh, Nadine; Alsheikh Deeb, Ibrahim; El-Achkar, Bassam; Tamim, Hani; Abdul Razzak, Farah; Haddad, Maurice; Abi-Ghanem, Alain S; El-Merhi, Fadi

The aim of this article was to investigate the opacification of the renal vasculature and the urogenital system during computed tomography urography by using a quadruple-phase contrast media in a triphasic scan protocol. A total of 200 patients with possible urinary tract abnormalities were equally divided between 2 protocols. Protocol A used the conventional single bolus and quadruple-phase scan protocol (pre, arterial, venous, and delayed), retrospectively. Protocol B included a quadruple-phase contrast media injection with a triphasic scan protocol (pre, arterial and combined venous, and delayed), prospectively. Each protocol used 100 mL contrast and saline at a flow rate of 4.5 mL. Attenuation profiles and contrast-to-noise ratio of the renal arteries, veins, and urogenital tract were measured. Effective radiation dose calculation, data analysis by independent sample t test, receiver operating characteristic, and visual grading characteristic analyses were performed. In arterial circulation, only the inferior interlobular arteries in both protocols showed a statistical significance (P < 0.05). Venously, the inferior vena cava, proximal and distal renal veins demonstrated a significant opacification reduction in protocol B than in protocol A (P < 0.001). Protocol B showed a significantly higher mean contrast-to-noise ratio than protocol A (protocol B: 22.68 ± 13.72; protocol A: 14.75 ± 5.76; P < 0.001). Radiation dose was significantly reduced in protocol B (7.38 ± 2.22 mSv) than in protocol A (12.28 ± 2.72 mSv) (P < 0.001). Visual grading characteristic (P < 0.027) and receiver operating characteristic (P < 0.0001) analyses demonstrated a significant preference for protocol B. In computed tomography urography, augmented quadruple-phase contrast media and triphasic scan protocol usage increases the image quality at a reduced radiation dose.

Effect of water extract of Turkish propolis on tuberculosis infection in guinea-pigs.

PubMed

Yildirim, Zeki; Hacievliyagil, Süleyman; Kutlu, Nurettin Onur; Aydin, Nasuhi Engin; Kurkcuoglu, Mine; Iraz, Mustafa; Durmaz, Riza

2004-03-01

Mycobacterium tuberculosis (H(37)R(v))-infected guinea-pig model was used to investigate the effect of water extract of propolis (WEP). After subcutaneous inoculation of tubercle bacilli, each animal received oral WEP (n=9), isoniazid (n=5) or saline (n=6) as placebo and were sacrificed 30 days later. Formation of necrosis was less prominent in the group treated with WEP, but was not statistically significant (P>0.05). The granuloma formation in the same group was more prominent than the placebo and isoniazid groups; however, this finding failed to reach statistical significance by the Kruskal-Wallis test (P>0.05). These findings suggest that Turkish WEP may have a limited effect on the development of tuberculosis infection in this guinea-pig model.

Resuscitation from severe hemorrhagic shock after traumatic brain injury using saline, shed blood, or a blood substitute.

PubMed

Gibson, Jeffrey B; Maxwell, Robert A; Schweitzer, John B; Fabian, Timothy C; Proctor, Kenneth G

2002-03-01

The original purpose of this study was to compare initial resuscitation of hemorrhagic hypotension after traumatic brain injury (TBI) with saline and shed blood. Based on those results, the protocol was modified and saline was compared to a blood substitute, diaspirin cross-linked hemoglobin (DCLHb). Two series of experiments were performed in anesthetized and mechanically ventilated (FiO2 = 0.4) pigs (35-45 kg). In Series 1, fluid percussion TBI (6-8 ATM) was followed by a 30% hemorrhage. At 120 min post-TBI, initial resuscitation consisted of either shed blood (n = 7) or a bolus of 3x shed blood volume as saline (n = 13). Saline supplements were then administered to all pigs to maintain a systolic arterial blood pressure (SAP) of >100 mmHg and a heart rate (HR) of <110 beats/min. In Series 2, TBI (4-5 ATM) was followed by a 35% hemorrhage. At 60 min post-TBI, initial resuscitation consisted of either 500 mL of DCLHb (n = 6) or 500 mL of saline (n = 5). This was followed by saline supplements to all pigs to maintain a SAP of >100 mmHg and a HR of <110 beats/min. In Series 1, most systemic markers of resuscitation (e.g., SAP, HR, cardiac output, filling pressures, lactate, etc.) were normalized, but there were 0/7 vs. 5/13 deaths within 5 h (P = 0.058) with blood vs. saline. At constant arterial O2 saturation (SaO2), mixed venous O2 saturation (SvO2), cerebral perfusion pressure (CPP), and cerebral venous O2 saturation (ScvO2) were all higher, intracranial pressure (ICP) was lower, and CO2 reactivity was preserved with blood vs. saline (all P < 0.05). In Series 2, SAP, ICP, CPP, and lactate were higher with DCLHb vs. saline (all P< 0.05). Cardiac output was lower even though filling pressure was markedly elevated with DCLHb vs. saline (both P< 0.05). Neither SvO2 nor cerebrovascular CO2 reactivity were improved, and ScvO2 was lower with DCLHb vs. saline (P < 0.05). All survived at least 72 h with neuropathologic changes that included sub-arachnoid hemorrhage, midline cerebellar necrosis, and diffuse axonal injury. These changes were similar with DCLHb vs. saline. Thus, whole blood was more effective than saline for resuscitation of TBI, whereas DCLHb was no more, and according to many variables, less effective than saline resuscitation. These experimental results are comparable to those in a recent multicenter trial using DCLHb for the treatment of severe traumatic shock. Further investigations in similar experimental models might provide some plausible explanations why DCLHb unexpectedly increased mortality in patients.

Nitric oxide plays a role in the regulation of adrenal blood flow and adrenocorticomedullary functions in the llama fetus

PubMed Central

Riquelme, Raquel A; Sánchez, Gina; Liberona, Leonel; Sanhueza, Emilia M; Giussani, Dino A; Blanco, Carlos E; Hanson, Mark A; Llanos, Aníbal J

2002-01-01

The hypothesis that nitric oxide plays a key role in the regulation of adrenal blood flow and plasma concentrations of cortisol and catecholamines under basal and hypoxaemic conditions in the llama fetus was tested. At 0.6-0.8 of gestation, 11 llama fetuses were surgically prepared for long-term recording under anaesthesia with vascular and amniotic catheters. Following recovery all fetuses underwent an experimental protocol based on 1 h of normoxaemia, 1 h of hypoxaemia and 1 h of recovery. In nine fetuses, the protocol occurred during fetal i.v. infusion with saline and in five fetuses during fetal i.v. treatment with the nitric oxide synthase inhibitor l-NAME. Adrenal blood flow was determined by the radiolabelled microsphere method during each of the experimental periods during saline infusion and treatment with l-NAME. Treatment with l-NAME during normoxaemia led to a marked fall in adrenal blood flow and a pronounced increase in plasma catecholamine concentrations, but it did not affect plasma ACTH or cortisol levels. In saline-infused fetuses, acute hypoxaemia elicited an increase in adrenal blood flow and in plasma ACTH, cortisol, adrenaline and noradrenaline concentrations. Treatment with l-NAME did not affect the increase in fetal plasma ACTH, but prevented the increments in adrenal blood flow and in plasma cortisol and adrenaline concentrations during hypoxaemia in the llama fetus. In contrast, l-NAME further enhanced the increase in fetal plasma noradrenaline. These data support the hypothesis that nitric oxide has important roles in the regulation of adrenal blood flow and adrenal corticomedullary functions during normoxaemia and hypoxaemia functions in the late gestation llama fetus. PMID:12356897

Effectiveness of Rack Sanitation Procedures for Elimination of Bacteria from Automatic Watering Manifolds.

PubMed

Costello, Terry; Watkins, Linda; Straign, Mike; Bean, William; Toth, Linda; Rehg, Jerold

1998-03-01

An important responsibility of animal care programs is to protect research animals from exposure to potentially pathogenic microorganisms. To validate the need for steam sterilization of rodent automatic watering racks, we evaluate the post-sanitation microbial contamination of experimentally inoculated racks and of racks that had been used to house conventional mice. We tested three sanitation protocols: rack-washer sanitation without manifold flush, sanitation that included manifold flush, and sanitation that included manifold flush followed by autoclaving. Rack sanitation, with or without manifold flush, did not reliably eliminate microbial flora from the lixits or manifold drainage water. A total of 43% of all non-autoclaved racks were positive for bacterial contamination after sanitation, and racks that had been used for conventional animal housing were more frequently positive than were experimentally inoculated racks (79% vs 18%). These data indicate that steam sterilization is necessary for eliminating bacteria from automatic watering systems. These observations are particularly important in light of increasing numbers of immune-impaired rodents that may be inadvertently and unnecessarily exposed to microbial pathogens via the automatic watering system.

Parameters for determining inoculated pack/challenge study protocols.

PubMed

2010-01-01

The National Advisory Committee on Microbiological Criteria for Foods developed guidelines for conducting challenge studies on pathogen inhibition and inactivation studies in a variety of foods. The document is intended for use by the food industry, including food processors, food service operators, and food retailers; federal, state, and local food safety regulators; public health officials; food testing laboratories; and process authorities. The document is focused on and limited to bacterial inactivation and growth inhibition and does not make specific recommendations with respect to public health. The Committee concluded that challenge studies should be designed considering the most current advances in methodologies, current thinking on pathogens of concern, and an understanding of the product preparation, variability, and storage conditions. Studies should be completed and evaluated under the guidance of an expert microbiologist in a qualified laboratory and should include appropriate statistical design and data analyses. This document provides guidelines for choice of microorganisms for studies, inoculum preparation, inoculum level, methods of inoculation, incubation temperatures and times, sampling considerations, and interpreting test results. Examples of appropriately designed growth inhibition and inactivation studies are provided.

Survival of bacteria of laboratory animal origin on cage bedding and inactivation by hydrogen peroxide vapour.

PubMed

Benga, Laurentiu; Benten, W Peter M; Engelhardt, Eva; Gougoula, Christina; Schulze-Röbbecke, Roland; Sager, Martin

2017-08-01

This study aims to determine the ability of laboratory animal bacteria to resist desiccation and inactivation by hydrogen peroxide vapour (HPV) on paper bedding pieces. Bedding pieces were saturated with bacterial suspensions in water or 2% (w/v) bovine serum albumin (BSA) in water, and held in a mouse facility. Viable counts showed variable survival rates over time for the bacterial species used ([ Pasteurella] pneumotropica, Muribacter muris, Pseudomonas aeruginosa, Acinetobacter redioresistens, Escherichia coli, Klebsiella oxytoca, Bordetella bronchiseptica, Bordetella hinzii, Enterococcus faecalis, β-haemolytic Streptococcus spp., Staphylococcus aureus and Staphylococcus xylosus). Overall, BSA increased bacterial survival in the bedding pieces. The survival rates of Bacillus safensis were not influenced by BSA but depended on sporulation. When bedding pieces and Petri dishes inoculated with E. coli, P. aeruginosa and S. aureus were subjected to HPV disinfection, all bacterial species on the bedding pieces inoculated with bacterial suspensions in water were readily inactivated. By contrast, S. aureus and P. aeruginosa, but not E. coli cells survived HPV treatment in high numbers when inoculated on bedding pieces as a BSA suspension. Notably, all three bacterial species were readily inactivated by HPV even in the presence of BSA when smeared on smooth surfaces. In conclusion, the suspension medium and the carrier can influence the environmental survival and susceptibility of bacterial species to HPV. Our results may help to develop standard protocols that can be used to ensure the microbiological quality of experimental rodent housing.

Revisiting the Life Cycle of Dung Fungi, Including Sordaria fimicola.

PubMed

Newcombe, George; Campbell, Jason; Griffith, David; Baynes, Melissa; Launchbaugh, Karen; Pendleton, Rosemary

2016-01-01

Dung fungi, such as Sordaria fimicola, generally reproduce sexually with ascospores discharged from mammalian dung after passage through herbivores. Their life cycle is thought to be obligate to dung, and thus their ascospores in Quaternary sediments have been interpreted as evidence of past mammalian herbivore activity. Reports of dung fungi as endophytes would seem to challenge the view that they are obligate to dung. However, endophyte status is controversial because surface-sterilization protocols could fail to kill dung fungus ascospores stuck to the plant surface. Thus, we first tested the ability of representative isolates of three common genera of dung fungi to affect plant growth and fecundity given that significant effects on plant fitness could not result from ascospores merely stuck to the plant surface. Isolates of S. fimicola, Preussia sp., and Sporormiella sp. reduced growth and fecundity of two of three populations of Bromus tectorum, the host from which they had been isolated. In further work with S. fimicola we showed that inoculations of roots of B. tectorum led to some colonization of aboveground tissues. The same isolate of S. fimicola reproduced sexually on inoculated host plant tissues as well as in dung after passage through sheep, thus demonstrating a facultative rather than an obligate life cycle. Finally, plants inoculated with S. fimicola were not preferred by sheep; preference had been expected if the fungus were obligate to dung. Overall, these findings make us question the assumption that these fungi are obligate to dung.

Revisiting the Life Cycle of Dung Fungi, Including Sordaria fimicola

PubMed Central

Newcombe, George; Campbell, Jason; Griffith, David; Baynes, Melissa; Launchbaugh, Karen; Pendleton, Rosemary

2016-01-01

Dung fungi, such as Sordaria fimicola, generally reproduce sexually with ascospores discharged from mammalian dung after passage through herbivores. Their life cycle is thought to be obligate to dung, and thus their ascospores in Quaternary sediments have been interpreted as evidence of past mammalian herbivore activity. Reports of dung fungi as endophytes would seem to challenge the view that they are obligate to dung. However, endophyte status is controversial because surface-sterilization protocols could fail to kill dung fungus ascospores stuck to the plant surface. Thus, we first tested the ability of representative isolates of three common genera of dung fungi to affect plant growth and fecundity given that significant effects on plant fitness could not result from ascospores merely stuck to the plant surface. Isolates of S. fimicola, Preussia sp., and Sporormiella sp. reduced growth and fecundity of two of three populations of Bromus tectorum, the host from which they had been isolated. In further work with S. fimicola we showed that inoculations of roots of B. tectorum led to some colonization of aboveground tissues. The same isolate of S. fimicola reproduced sexually on inoculated host plant tissues as well as in dung after passage through sheep, thus demonstrating a facultative rather than an obligate life cycle. Finally, plants inoculated with S. fimicola were not preferred by sheep; preference had been expected if the fungus were obligate to dung. Overall, these findings make us question the assumption that these fungi are obligate to dung. PMID:26839959

Use of one-ply composite tissues in an automated optical assay for recovery of Listeria from food contact surfaces and poultry-processing environments.

PubMed

Yan, Zhinong; Vorst, Keith L; Zhang, Lei; Ryser, Elliot T

2007-05-01

A novel one-ply composite tissue (CT) method using the Soleris (formerly BioSys) optical analysis system was compared with the conventional U.S. Department of Agriculture (USDA) environmental sponge enrichment method for recovery of Listeria from food contact surfaces and poultry-processing environments. Stainless steel and high-density polyethylene plates were inoculated to contain a six-strain L. monocytogenes cocktail at 10(4), 10(2), and 10 CFU per plate, whereas samples from naturally contaminated surfaces and floor drains from a poultry-processing facility were collected with CTs and environmental sponges. CT samples were transferred into Soleris system vials, and presumptive-positive samples were further confirmed. Sponge samples were processed for Listeria using the USDA culture method. L. monocytogenes recovery rates from inoculated stainless steel and polyethylene surfaces were then compared for the two methods in terms of sensitivity, specificity, and positive and negative predictive values. No significant differences (P > 0.05) were found between the two methods for recovery of L. monocytogenes from any of the inoculated stainless steel and polyethylene surfaces or environmental samples. Sensitivity, specificity, and overall accuracy of the CT-Soleris for recovery of Listeria from environmental samples were 83, 97, and 95%, respectively. Listeria was detected 2 to 3 days sooner with the CT-Soleris method than with the USDA culture method, thus supporting the increased efficacy of this new protocol for environmental sampling.

Fast filtration sampling protocol for mammalian suspension cells tailored for phosphometabolome profiling by capillary ion chromatography - tandem mass spectrometry.

PubMed

Kvitvang, Hans F N; Bruheim, Per

2015-08-15

Capillary ion chromatography (capIC) is the premium separation technology for low molecular phosphometabolites and nucleotides in biological extracts. Removal of excessive amounts of salt during sample preparation stages is a prerequisite to enable high quality capIC separation in combination with reproducible and sensitive MS detection. Existing sampling protocols for mammalian cells used for GC-MS and LC-MS metabolic profiling can therefore not be directly applied to capIC separations. Here, the development of a fast filtration sampling protocol for mammalian suspension cells tailored for quantitative profiling of the phosphometabolome on capIC-MS/MS is presented. The whole procedure from sampling the culture to transfer of filter to quenching and extraction solution takes less than 10s. To prevent leakage it is critical that a low vacuum pressure is applied, and satisfactorily reproducibility was only obtained by usage of a vacuum pressure controlling device. A vacuum of 60mbar was optimal for filtration of multiple myeloma Jjn-3 cell cultures through 5μm polyvinylidene (PVDF) filters. A quick deionized water (DI-water) rinse step prior to extraction was tested, and significantly higher metabolite yields were obtained during capIC-MS/MS analyses in this extract compared to extracts prepared by saline and reduced saline (25%) washing steps only. In addition, chromatographic performance was dramatically improved. Thus, it was verified that a quick DI-water rinse is tolerated by the cells and can be included as the final stage during filtration. Over 30 metabolites were quantitated in JJN-3 cell extracts by using the optimized sampling protocol with subsequent capIC-MS/MS analysis, and up to 2 million cells can be used in a single filtration step for the chosen filter and vacuum pressure. The technical set-up is also highly advantageous for microbial metabolome filtration protocols after optimization of vacuum pressure and washing solutions, and the reduced salt content of the extract will also improve the quality of LC-MS analysis due to lower salt adduct ion formation. Copyright © 2015 Elsevier B.V. All rights reserved.

Isolation and identification of halotolerant soil bacteria from coastal Patenga area.

PubMed

Rahman, Shafkat Shamim; Siddique, Romana; Tabassum, Nafisa

2017-10-30

Halotolerant bacteria have multiple uses viz. fermentation with lesser sterility control and industrial production of bioplastics. Moreover, it may increase the crop productivity of coastal saline lands in Bangladesh by transferring the salt tolerant genes into the plants. The study focused on the isolation and identification of the halotolerant bacteria from three soil samples, collected from coastal Patenga area. The samples were inoculated in nutrient media containing a wide range of salt concentrations. All the samples showed 2, 4 and 6% (w/v) salt tolerance. The isolates from Patenga soil (4, 6%) and beach soil (2%) showed catalase activity and all the isolates showed negative results for oxidase activity, indole production, lactose and motility. All the samples provided positive results for dextrose fermentation. Other tests provided mixed results. Based on the morphological characteristics, biochemical tests and ABIS software analysis the isolates fall within the Enterobacteriaceae, Clostridium and Corynebacterium, with a predominance of Vibrios. Overall the isolates can be considered as mild halotolerant, with the best growth observed at lower salinities and no halophilism detected. Among many possibilities, the genes responsible for the salt tolerant trait in these species can be identified, extracted and inserted into the crop plants to form a transgenic plant to result in higher yield for the rest of the year.

Forced diuresis with matched hydration in reducing acute kidney injury during transcatheter aortic valve implantation (Reduce-AKI): study protocol for a randomized sham-controlled trial.

PubMed

Arbel, Yaron; Ben-Assa, Eyal; Halkin, Amir; Keren, Gad; Schwartz, Arie Lorin; Havakuk, Ofer; Leshem-Rubinow, Eran; Konigstein, Maayan; Steinvil, Arie; Abramowitz, Yigal; Finkelstein, Ariel; Banai, Shmuel

2014-07-02

Acute kidney injury (AKI) is observed in up to 41% of patients undergoing transcatheter aortic valve implantation (TAVI) and is associated with increased risk for mortality. The aim of the present study is to evaluate whether furosemide-induced diuresis with matched isotonic intravenous hydration using the RenalGuard system reduces AKI in patients undergoing TAVI. Reduce-AKI is a randomized sham-controlled study designed to examine the effect of an automated matched hydration system in the prevention of AKI in patients undergoing TAVI. Patients will be randomized in a 1:1 fashion to the RenalGuard system (active group) versus non-matched saline infusion (sham-controlled group). Both arms receive standard overnight saline infusion and N-acetyl cysteine before the procedure. The Reduce-AKI trial will investigate whether the use of automated forced diuresis with matched saline infusion is an effective therapeutic tool to reduce the occurrence of AKI in patients undergoing TAVI. Clinicaltrials.gov: NCT01866800, 30 April 30 2013.

Forced diuresis with matched hydration in reducing acute kidney injury during transcatheter aortic valve implantation (Reduce-AKI): study protocol for a randomized sham-controlled trial

PubMed Central

2014-01-01

Background Acute kidney injury (AKI) is observed in up to 41% of patients undergoing transcatheter aortic valve implantation (TAVI) and is associated with increased risk for mortality. The aim of the present study is to evaluate whether furosemide-induced diuresis with matched isotonic intravenous hydration using the RenalGuard system reduces AKI in patients undergoing TAVI. Methods/Design Reduce-AKI is a randomized sham-controlled study designed to examine the effect of an automated matched hydration system in the prevention of AKI in patients undergoing TAVI. Patients will be randomized in a 1:1 fashion to the RenalGuard system (active group) versus non-matched saline infusion (sham-controlled group). Both arms receive standard overnight saline infusion and N-acetyl cysteine before the procedure. Discussion The Reduce-AKI trial will investigate whether the use of automated forced diuresis with matched saline infusion is an effective therapeutic tool to reduce the occurrence of AKI in patients undergoing TAVI. Trial registration Clinicaltrials.gov: NCT01866800, 30 April 30 2013. PMID:24986373

Limitation of Infarct Size and No-Reflow by Intracoronary Adenosine Depends Critically on Dose and Duration.

PubMed

Yetgin, Tuncay; Uitterdijk, André; Te Lintel Hekkert, Maaike; Merkus, Daphne; Krabbendam-Peters, Ilona; van Beusekom, Heleen M M; Falotico, Robert; Serruys, Patrick W; Manintveld, Olivier C; van Geuns, Robert-Jan M; Zijlstra, Felix; Duncker, Dirk J

2015-12-28

In the absence of effective clinical pharmacotherapy for prevention of reperfusion-mediated injury, this study re-evaluated the effects of intracoronary adenosine on infarct size and no-reflow in a porcine model of acute myocardial infarction using clinical bolus and experimental high-dose infusion regimens. Despite the clear cardioprotective effects of adenosine, when administered prior to ischemia, studies on cardioprotection by adenosine when administered at reperfusion have yielded contradictory results in both pre-clinical and clinical settings. Swine (54 ± 1 kg) were subjected to a 45-min mid-left anterior descending artery occlusion followed by 2 h of reperfusion. In protocol A, an intracoronary bolus of 3 mg adenosine injected over 1 min (n = 5) or saline (n = 10) was administered at reperfusion. In protocol B, an intracoronary infusion of 50 μg/kg/min adenosine (n = 15) or saline (n = 21) was administered starting 5 min prior to reperfusion and continued throughout the 2-h reperfusion period. In protocol A, area-at-risk, infarct size, and no-reflow were similar between groups. In protocol B, risk zones were similar, but administration of adenosine resulted in significant reductions in infarct size from 59 ± 3% of the area-at-risk in control swine to 46 ± 4% (p = 0.02), and no-reflow from 49 ± 6% of the infarct area to 26 ± 6% (p = 0.03). During reperfusion, intracoronary adenosine can limit infarct size and no-reflow in a porcine model of acute myocardial infarction. However, protection was only observed when adenosine was administered via prolonged high-dose infusion, and not via short-acting bolus injection. These findings warrant reconsideration of adenosine as an adjuvant therapy during early reperfusion. Copyright © 2015 American College of Cardiology Foundation. Published by Elsevier Inc. All rights reserved.

Alfalfa (Medicago sativa L.).

PubMed

Fu, Chunxiang; Hernandez, Timothy; Zhou, Chuanen; Wang, Zeng-Yu

2015-01-01

Alfalfa (Medicago sativa L.) is a high-quality forage crop widely grown throughout the world. This chapter describes an efficient protocol that allows for the generation of large number of transgenic alfalfa plants by sonication-assisted Agrobacterium-mediated transformation. Binary vectors carrying different selectable marker genes that confer resistance to phosphinothricin (bar), kanamycin (npt II), or hygromycin (hph) were used to generate transgenic alfalfa plants. Intact trifoliates collected from clonally propagated plants in the greenhouse were sterilized with bleach and then inoculated with Agrobacterium strain EHA105. More than 80 % of infected leaf pieces could produce rooted transgenic plants in 4-5 months after Agrobacterium-mediated transformation.

Genetic Transformation of Switchgrass

NASA Astrophysics Data System (ADS)

Xi, Yajun; Ge, Yaxin; Wang, Zeng-Yu

Switchgrass (Panicum virgatum L.) is a highly productive warm-season C4 species that is being developed into a dedicated biofuel crop. This chapter describes a protocol that allows the generation of transgenic switchgrass plants by Agrobacterium tumefaciens-mediated transformation. Embryogenic calluses induced from caryopses or inflorescences were used as explants for inoculation with A. tumefaciens strain EHA105. Hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin was used as the selection agent. Calluses resistant to hygromycin were obtained after 5-6 weeks of selection. Soil-grown switchgrass plants were regenerated about 6 months after callus induction and Agrobacterium-mediated transformation.

Genetic transformation of switchgrass.

PubMed

Xi, Yajun; Ge, Yaxin; Wang, Zeng-Yu

2009-01-01

Switchgrass (Panicum virgatum L.) is a highly productive warm-season C4 species that is being developed into a dedicated biofuel crop. This chapter describes a protocol that allows the generation of transgenic switchgrass plants by Agrobacterium tumefaciens-mediated transformation. Embryogenic calluses induced from caryopses or inflorescences were used as explants for inoculation with A. tumefaciens strain EHA105. Hygromycin phosphotransferase gene (hph) was used as the selectable marker and hygromycin was used as the selection agent. Calluses resistant to hygromycin were obtained after 5-6 weeks of selection. Soil-grown switchgrass plants were regenerated about 6 months after callus induction and Agrobacterium-mediated transformation.

Application of Plant-Growth-Promoting Fungi Trichoderma longibrachiatum T6 Enhances Tolerance of Wheat to Salt Stress through Improvement of Antioxidative Defense System and Gene Expression

PubMed Central

Zhang, Shuwu; Gan, Yantai; Xu, Bingliang

2016-01-01

Soil salinity is a serious problem worldwide that reduces agricultural productivity. Trichoderma longibrachiatum T6 (T6) has been shown to promote wheat growth and induce plant resistance to parasitic nematodes, but whether the plant-growth-promoting fungi T6 can enhance plant tolerance to salt stress is unknown. Here, we determined the effect of plant-growth-promoting fungi T6 on wheat seedlings’ growth and development under salt stress, and investigated the role of T6 in inducing the resistance to NaCl stress at physiological, biochemical, and molecular levels. Wheat seedlings were inoculated with the strain of T6 and then compared with non-inoculated controls. Shoot height, root length, and shoot and root weights were measured on 15 days old wheat seedlings grown either under 150 mM NaCl or in a controlled setting without any NaCl. A number of colonies were re-isolated from the roots of wheat seedlings under salt stress. The relative water content in the leaves and roots, chlorophyll content, and root activity were significantly increased, and the accumulation of proline content in leaves was markedly accelerated with the plant growth parameters, but the content of leaf malondialdehyde under saline condition was significantly decreased. The antioxidant enzymes-superoxide dismutase (SOD), peroxidase (POD), and catalase (CAT) in wheat seedlings were increased by 29, 39, and 19%, respectively, with the application of the strain of T6 under salt stress; the relative expression of SOD, POD, and CAT genes in these wheat seedlings were significantly up-regulated. Our results indicated that the strain of T6 ameliorated the adverse effects significantly, protecting the seedlings from salt stress during their growth period. The possible mechanisms by which T6 suppresses the negative effect of NaCl stress on wheat seedling growth may be due to the improvement of the antioxidative defense system and gene expression in the stressed wheat plants. PMID:27695475

Mouse model of plasma cell mastitis.

PubMed

Yu, Jian-jun; Bao, Shan-lin; Yu, Sheng-lin; Zhang, Da-Qing; Loo, Wings T Y; Chow, Louis W C; Su, Li; Cui, Zhen; Chen, Kai; Ma, Li-Qiong; Zhang, Ning; Yu, Hui; Yang, Yun-Zhen; Dong, Yu; Yip, Adrian Y S; Ng, Elizabeth L Y

2012-09-19

Plasma cell mastitis is distinct from the common form of mastitis and clinically resembles breast carcinoma. The lesion occurs in non-lactating young women, and the incidence rate is rising. Surgical resection is the main treatment, but cannot prevent recurrence of the disease. Disfigurement or removal of breast after the operations can cause marked physical and psychological distress. The etiology of plasma cell mastitis is unclear up till now. It is therefore necessary to investigate further the underlying immunological changes of the disease. The lesions of plasma cell mastitis removed from patients through aseptic operation were mixed with normal saline into homogenate tube machine (homogenate tubes were disinfected and sterilized prior to treatment). The mixture was homogenized at medium speed and grinded in ultrasonic cell disruptor. The homogenate obtained was made into oil emulsion with Freund's adjuvant. Thirty female BALB/c mice (6 weeks after sexual maturity) were divided into five groups A-E: group A was blank control; group B was normal saline control; group C was inoculated with 0.02 ml water-in-oil emulsion; group D was inoculated with 0.04 ml water-in-oil emulsion; group E was complete Freund's adjuvant control. Pathology results showed that mouse mammary gland acinar cells remained integral without any abnormal changes observed in control groups A and B. Experimental groups C and D showed dilation of mouse mammary ductal tissue with a large number of epithelial cells and debris in the lumen, and fibrosis around ducts accompanied by large duct cells, neutrophils, lymphocytes, and especially plasma cell infiltration. Pathological changes were observed in 3 (50%) mice and 5 (83.3%) mice in group C and D respectively. In group E, neutrophil infiltration in mammary gland was observed in 5 mice, but neither infiltration of plasma cells nor other abnormal pathological changes were observed. The lesions of patient with plasma cell mastitis could make the female BALB/c mice experience the similar clinical and pathological manifestation. High-dose group can successfully establish a mouse model of plasma cell mastitis.

Arbuscular mycorrhizal symbiosis mitigates the negative effects of salinity on durum wheat

PubMed Central

Ingraffia, Rosolino; Giambalvo, Dario; Frenda, Alfonso Salvatore

2017-01-01

Arbuscular mycorrhizal (AM) symbiosis is generally considered to be effective in ameliorating the plant tolerance to salt stress. Unfortunately, the comprehension of the mechanisms implicated in salinity stress alleviation by AM symbiosis is far from being complete. Thus, an experiment was performed by growing durum wheat (Triticum durum Desf.) plants under salt-stress conditions to evaluate the influence of AM symbiosis on both the plant growth and the regulation of a number of genes related to salt stress and nutrient uptake. Durum wheat plants were grown outdoors in pots in absence or in presence of salt stress and with or without AM fungi inoculation. The inoculum consisted of a mixture of spores of Rhizophagus irregularis (formerly Glomus intraradices) and Funneliformis mosseae (formerly G. mosseae). Results indicate that AM symbiosis can alleviate the detrimental effects of salt stress on the growth of durum wheat plants. In fact, under salt stress conditions mycorrhizal plants produced more aboveground and root biomass, had higher N uptake and aboveground N concentration, and showed greater stability of plasma membranes compared to non-mycorrhizal plants. Inoculation with AM fungi had no effect on the expression of the N transporter genes AMT1.1, AMT1.2, and NAR2.2, either under no-stress or salt stress conditions, probably due to the fact that plants were grown under optimal N conditions; on the contrary, NRT1.1 was always upregulated by AM symbiosis. Moreover, the level of expression of the drought stress-related genes AQP1, AQP4, PIP1, DREB5, and DHN15.3 observed in the mycorrhizal stressed plants was markedly lower than that observed in the non-mycorrhizal stressed plants and very close to that observed in the non-stressed plants. Our hypothesis is that, in the present study, AM symbiosis did not increase the plant tolerance to salt stress but instead generated a condition in which plants were subjected to a level of salt stress lower than that of non-mycorrhizal plants. PMID:28877207

Demonstration and Commercialization of the Sediment Ecosystem Assessment Protocol: Project ER-201130 Environmental Restoration Project

DTIC Science & Technology

2017-01-01

65 5-15. Dissolved oxygen and temperature data from T = 34 month (2015) post-remedy...Continuous measurements of dissolved oxygen and temperature in SEA Ring chambers placed at 3-meter depth at Chollas Creek mouth (CC1-B) and adjacent to...which the organisms would be exposed, such as salinity and temperature . This action provided valuable data to determine if any effects observed were

Contrast-enhanced magnetic resonance angiography: first-pass arterial enhancement as a function of gadolinium-chelate concentration, and the saline chaser volume and injection rate.

PubMed

Husarik, Daniela B; Bashir, Mustafa R; Weber, Paul W; Nichols, Eli B; Howle, Laurens E; Merkle, Elmar M; Nelson, Rendon C

2012-02-01

To evaluate the effect of the contrast medium (CM) concentration and the saline chaser volume and injection rate on first-pass aortic enhancement characteristics in contrast-enhanced magnetic resonance angiography using a physiologic flow phantom. Imaging was performed on a 3.0-T magnetic resonance system (MAGNETOM Trio, Siemens Healthcare Solutions, Inc, Erlangen, Germany) using a 2-dimensional fast low angle shot T1-weighted sequence (repetition time, 500 milliseconds; echo time, 1.23 milliseconds; flip angle, 8 degrees; 1 frame/s × 60 seconds). The following CM concentrations injected at 2 mL/s were used with 3 different contrast agents (gadolinium [Gd]-BOPTA, Gd-HP-DO3A, Gd-DTPA): 20 mL of undiluted CM (100%) and 80%, 40%, 20%, 10%, 5%, and 2.5% of the full amount, all diluted in saline to a volume of 20 mL to ensure equal bolus volume. The CM was followed by saline chasers of 20 to 60 mL injected at 2 mL/s and 6 mL/s. Aortic signal intensity (SI) was measured, and normalized SI versus time (SI/Tn) curves were generated. The maximal SI (SI(max)), bolus length, and areas under the SI/Tn curve were calculated. Decreasing the CM concentration from 100% to 40% resulted in a decrease of SI(max) to 86.1% (mean). Further decreasing the CM concentration to 2.5% decreased SI(max) to 5.1% (mean). Altering the saline chaser volume had no significant effect on SI(max). Increasing the saline chaser injection rate had little effect (mean increase, 2.2%) on SI(max) when using ≥40% of CM. There was a larger effect (mean increase, 19.6%) when ≤20% of CM were used. Bolus time length was significantly shorter (P < 0.001), and area under the SI/T(n) curve was significantly smaller (P < 0.01) for the CM protocols followed by a saline chaser injected at 6 mL/s compared with a saline chaser injected at 2 mL/s. With 40% of CM and a fast saline chaser, SImax close to that with undiluted CM can be achieved. An increased saline chaser injection rate has a more pronounced effect on aortic enhancement characteristics at lower CM concentrations than at higher CM concentrations.

Soil and fertilizer amendments and edge effects on the floral succession of pulverized fuel ash

DOE Office of Scientific and Technical Information (OSTI.GOV)

Shaw, P.

2009-01-15

Plots of fresh pulverized fuel ash (PFA, an industrial waste) were inoculated with soils from existing PFA sites and fertilizers in a factorial design, then left unmanaged for 12 years during which time the floral development and soil chemistry were monitored annually. For the first 3 years, the site supported a sparse mix of chenopods (including the scarce Chenopodium glaucum) and halophytes. As salinity declined, ruderals, legumes, and grasses plus the fire-site moss Funaria hygrometrica colonized, followed by Festuca arundinacea grassland (NVC community MG12) and Hippophae rhamnoides scrub. Dactylorhiza incarnata (orchidacea) appeared after 7 years, but only in plots thatmore » had received soil from existing orchid colonies. Four years later, a larger second generation of Dactylorhiza appeared, but only in the central zone of the site where vegetation was thinnest. By year 12, the site was dominated by coarse grasses and scrub, with early successional species persisting only in the sparsely vegetated center, where nitrate levels were lowest. This edge effect is interpreted as centripetal encroachment, a process of potentially wider concern for the conservation of low-fertility habitat patches. Overall, seed bank inoculation seems to have introduced few but desirable species (D. incarnata, Pyrola rotundifolia, some halophytes, and annuals), whereas initial application of organic fertilizer had long-lasting ({ge} 10 years) effects on cover and soil composition.« less

Role of the Dermonecrotic Toxin of Bordetella bronchiseptica in the Pathogenesis of Respiratory Disease in Swine

PubMed Central

Brockmeier, Susan L.; Register, Karen B.; Magyar, Tibor; Lax, Alistair J.; Pullinger, Gillian D.; Kunkle, Robert A.

2002-01-01

Bordetella bronchiseptica is one of the etiologic agents causing atrophic rhinitis and pneumonia in swine. It produces several purported virulence factors, including the dermonecrotic toxin (DNT), which has been implicated in the turbinate atrophy seen in cases of atrophic rhinitis. The purpose of these experiments was to clarify the role of this toxin in respiratory disease by comparing the pathogenicity in swine of two isogenic dnt mutants to their virulent DNT+ parent strains. Two separate experiments were performed, one with each of the mutant-parent pairs. One-week-old cesarean-derived, colostrum-deprived pigs were inoculated intranasally with the parent strain, the dnt mutant strain, or phosphate-buffered saline. Weekly nasal washes were performed to monitor colonization of the nasal cavity, and the pigs were euthanized 4 weeks after inoculation to determine colonization of tissues and to examine the respiratory tract for pathology. There was evidence that colonization of the upper respiratory tract, but not the lower respiratory tract, was slightly greater for the parent strains than for the dnt mutants. Moderate turbinate atrophy and bronchopneumonia were found in most pigs given the parent strains, while there was no turbinate atrophy or pneumonia in pigs challenged with the dnt mutant strains. Therefore, production of DNT by B. bronchiseptica is necessary to produce the lesions of turbinate atrophy and bronchopneumonia in pigs infected with this organism. PMID:11796573

Expression of animal anti-apoptotic gene Ced-9 enhances tolerance during Glycine max L.-Bradyrhizobium japonicum interaction under saline stress but reduces nodule formation.

PubMed

Robert, Germán; Muñoz, Nacira; Melchiorre, Mariana; Sánchez, Federico; Lascano, Ramiro

2014-01-01

The mechanisms by which the expression of animal cell death suppressors in economically important plants conferred enhanced stress tolerance are not fully understood. In the present work, the effect of expression of animal antiapoptotic gene Ced-9 in soybean hairy roots was evaluated under root hairs and hairy roots death-inducing stress conditions given by i) Bradyrhizobium japonicum inoculation in presence of 50 mM NaCl, and ii) severe salt stress (150 mM NaCl), for 30 min and 3 h, respectively. We have determined that root hairs death induced by inoculation in presence of 50 mM NaCl showed characteristics of ordered process, with increased ROS generation, MDA and ATP levels, whereas the cell death induced by 150 mM NaCl treatment showed non-ordered or necrotic-like characteristics. The expression of Ced-9 inhibited or at least delayed root hairs death under these treatments. Hairy roots expressing Ced-9 had better homeostasis maintenance, preventing potassium release; increasing the ATP levels and controlling the oxidative damage avoiding the increase of reactive oxygen species production. Even when our results demonstrate a positive effect of animal cell death suppressors in plant cell ionic and redox homeostasis under cell death-inducing conditions, its expression, contrary to expectations, drastically inhibited nodule formation even under control conditions.

The in vitro and in vivo evaluation of ddC as a topical antiviral for ocular adenovirus infections.

PubMed

Romanowski, Eric G; Yates, Kathleen A; Gordon, Y Jerold

2009-11-01

To evaluate the antiviral activity of 2', 3'-dideoxycytidine (ddC) in vitro against a panel of ocular adenovirus serotypes and in vivo in the ocular Ad5/NZW rabbit replication model. In vitro, the 50% inhibitory concentrations (IC(50)) of ddC and cidofovir were determined using standard plaque-reduction assays. In vivo, 40 rabbits were topically inoculated in both eyes with Ad5 after corneal scarification. On day 1, the rabbits were equally divided into four topical treatment groups: 3% ddC; 2% ddC; 0.5% cidofovir; and saline. ddC and saline eyes were treated four times daily for 7 days, and cidofovir-treated eyes were treated twice daily for 7 days. Eyes were cultured for virus a multiple times over 2 weeks. The in vitro IC(50) for ddC ranged from 0.18 to 1.85 microg/mL, whereas those for cidofovir ranged from 0.018 to 5.47 microg/mL. ddC was more potent than cidofovir for seven of nine serotypes. In vivo, 3% ddC, 2% ddC, and 0.5% cidofovir significantly reduced the number of Ad5-positive cultures per total (days 1-14), mean Ad5 ocular titer (days 1-5), and duration of shedding (among other outcome measures) compared with the saline control. The 3% and 2% ddC treatments were significantly more efficacious than the 0.5% cidofovir treatment in the parameters listed above. ddC demonstrated potent antiadenovirus activity in vitro and in vivo. Systemic safety studies after topical ocular administration are needed to evaluate ddC as a topical antiviral treatment for adenoviral ocular infections in the target population.

Relaxivity of Ferumoxytol at 1.5 T and 3.0 T.

PubMed

Knobloch, Gesine; Colgan, Timothy; Wiens, Curtis N; Wang, Xiaoke; Schubert, Tilman; Hernando, Diego; Sharma, Samir D; Reeder, Scott B

2018-05-01

The aim of this study was to determine the relaxation properties of ferumoxytol, an off-label alternative to gadolinium-based contrast agents, under physiological conditions at 1.5 T and 3.0 T. Ferumoxytol was diluted in gradually increasing concentrations (0.26-4.2 mM) in saline, human plasma, and human whole blood. Magnetic resonance relaxometry was performed at 37°C at 1.5 T and 3.0 T. Longitudinal and transverse relaxation rate constants (R1, R2, R2*) were measured as a function of ferumoxytol concentration, and relaxivities (r1, r2, r2*) were calculated. A linear dependence of R1, R2, and R2* on ferumoxytol concentration was found in saline and plasma with lower R1 values at 3.0 T and similar R2 and R2* values at 1.5 T and 3.0 T (1.5 T: r1saline = 19.9 ± 2.3 smM; r1plasma = 19.0 ± 1.7 smM; r2saline = 60.8 ± 3.8 smM; r2plasma = 64.9 ± 1.8 smM; r2*saline = 60.4 ± 4.7 smM; r2*plasma = 64.4 ± 2.5 smM; 3.0 T: r1saline = 10.0 ± 0.3 smM; r1plasma = 9.5 ± 0.2 smM; r2saline = 62.3 ± 3.7 smM; r2plasma = 65.2 ± 1.8 smM; r2*saline = 57.0 ± 4.7 smM; r2*plasma = 55.7 ± 4.4 smM). The dependence of relaxation rates on concentration in blood was nonlinear. Formulas from second-order polynomial fittings of the relaxation rates were calculated to characterize the relationship between R1blood and R2 blood with ferumoxytol. Ferumoxytol demonstrates strong longitudinal and transverse relaxivities. Awareness of the nonlinear relaxation behavior of ferumoxytol in blood is important for ferumoxytol-enhanced magnetic resonance imaging applications and for protocol optimization.

A randomized comparison of 1-h sodium bicarbonate hydration versus standard peri-procedural saline hydration in patients with chronic kidney disease undergoing intravenous contrast-enhanced computerized tomography.

PubMed

Kooiman, Judith; Sijpkens, Yvo W J; de Vries, Jean-Paul P M; Brulez, Harald F H; Hamming, Jaap F; van der Molen, Aart J; Aarts, Nico J M; Cannegieter, Suzanne C; Putter, Hein; Swarts, Renate; van den Hout, Wilbert B; Rabelink, Ton J; Huisman, Menno V

2014-05-01

Guidelines recommend saline hydration for prophylaxis of contrast-induced acute kidney injury (CI-AKI) in patients with chronic kidney disease (CKD) undergoing intravenous contrast media-enhanced CT (CE-CT). The safety and efficacy of a brief hydration protocol using sodium bicarbonate in this population is unknown. We analysed whether 1-h sodium bicarbonate hydration prior to CE-CT is non-inferior to saline hydration prior to and after CE-CT in CKD patients. We performed an open-label multicentre randomized trial. Patients were randomized to 250 mL of 1.4% sodium bicarbonate hydration prior to CE-CT or 1000 mL of 0.9% saline hydration prior to and, once again, after CE-CT. Primary outcome was the relative increase in serum creatinine 48-96 h post-CE-CT. Secondary outcomes were incidence of CI-AKI [serum creatinine increase >25%/>44 µmol/L (0.5 mg/dL)], recovery of renal function, the need for dialysis and 2-month hospital costs. Five hundred and seventy adult CKD patients undergoing CE-CT were randomized between 2010 and 2012, of whom 548 were included in the intention-to-treat population. Mean relative serum creatinine increase was 1.2% for sodium bicarbonate and 1.5% for saline (mean difference -0.3%; 95% confidence interval -2.7 to 2.1, P-value for non-inferiority <0.0001). CI-AKI occurred in 22 patients (4.1%); 8 (3.0%) randomized to sodium bicarbonate versus 14 (5.1%) to saline (P = 0.23). Renal function recovered in 75 and 69% of CI-AKI patients, respectively (P = 0.81). No patients developed a need for dialysis. Mean hydration costs per patient were €224 for the sodium bicarbonate and €683 for the saline regime (P < 0.001). Other healthcare costs were similar. Short hydration with sodium bicarbonate prior to CE-CT was non-inferior to peri-procedural saline hydration with respect to renal safety and may result in healthcare savings. [Netherlands Trial Register (http://www.trialregister.nl/trialreg/index.asp), Nr 2149, date of registration 23 December 2009.].

Salinity control in a clay soil beneath an orchard irrigated with treated waste water in the presence of a high water table: A numerical study

NASA Astrophysics Data System (ADS)

Russo, David; Laufer, Asher; Bardhan, Gopali; Levy, Guy J.

2015-12-01

A citrus orchard planted on a structured, clay soil associated with a high water table, irrigated by drip irrigation system using treated waste water (TWW) and local well water (LWW) was considered here. The scope of the present study was to analyze transport of mixed-ion, interacting salts in a combined vadose zone-groundwater flow system focusing on the following issues: (i) long-term effects of irrigation with TWW on the response of the flow system, identifying the main factors (e.g., soil salinity, soil sodicity) that control these effects, and (ii) salinity control aiming at improving both crop productivity and groundwater quality. To pursue this two-fold goal, 3-D numerical simulations of field-scale flow and transport were performed for an extended period of time, considering realistic features of the soil, water table, crop, weather and irrigation, and the coupling between the flow and the transport through the dependence of the soil hydraulic functions, K(ψ) and θ(ψ), on soil solution concentration C, and sodium adsorption ratio, SAR. Results of the analyses suggest that in the case studied, the long-term effect of irrigation with TWW on the response of the flow system is attributed to the enhanced salinity of the TWW, and not to the increase in soil sodicity. The latter findings are attributed to: (i) the negative effect of soil salinity on water uptake, and the tradeoff between water uptake and drainage flux, and, concurrently, solute discharge below the root zone; and, (ii) the tradeoff between the effects of C and SAR on K(ψ) and θ(ψ). Furthermore, it was demonstrated that a data-driven protocol for soil salinity control, based on alternating irrigation water quality between TWW and desalinized water, guided by the soil solution salinity at the centroid of the soil volume active in water uptake, may lead to a substantial increase in crop yield, and to a substantial decrease in the salinity load in the groundwater.

Experimental evidence for carbonate precipitation and CO 2 degassing during sea ice formation

NASA Astrophysics Data System (ADS)

Papadimitriou, S.; Kennedy, H.; Kattner, G.; Dieckmann, G. S.; Thomas, D. N.

2004-04-01

Chemical and stable carbon isotopic modifications during the freezing of artificial seawater were measured in four 4 m 3 tank incubations. Three of the four incubations were inoculated with a nonaxenic Antarctic diatom culture. The 18 days of freezing resulted in 25 to 27 cm thick ice sheets overlying the residual seawater. The ice phase was characterized by a decrease in temperature from -1.9 to -2.2°C in the under-ice seawater down to -6.7°C in the upper 4 cm of the ice sheet, with a concurrent increase in the salinity of the under-ice seawater and brine inclusions of the ice sheet as a result of physical concentration of major dissolved salts by expulsion from the solid ice matrix. Measurements of pH, total dissolved inorganic carbon (C T) and its stable isotopic composition (δ 13C T) all exhibited changes, which suggest minimal effect by biological activity during the experiment. A systematic drop in pH and salinity-normalized C T by up to 0.37 pH SWS units and 376 μmol C kg -1 respectively at the lowest temperature and highest salinity part of the ice sheet were coupled with an equally systematic 13C enrichment of the C T. Calculations based on the direct pH and C T measurements indicated a steady increase in the in situ concentration of dissolved carbon dioxide (CO 2(aq)) with time and increasing salinity within the ice sheet, partly due to changes in the dissociation constants of carbonic acid in the low temperature-high salinity range within sea ice. The combined effects of temperature and salinity on the solubility of CO 2 over the range of conditions encountered during this study was a slight net decrease in the equilibrium CO 2(aq) concentration as a result of the salting-out overriding the increase in solubility with decreasing temperature. Hence, the increase in the in situ CO 2(aq) concentration lead to saturation or supersaturation of the brine inclusions in the ice sheet with respect to atmospheric pCO 2 (≈3.5 × 10 -4 atm). When all physico-chemical processes are considered, we expect CO 2 degassing and carbonate mineral precipitation from the brine inclusions of the ice sheet, which were saturated or highly supersaturated with respect to both the anhydrous (calcite, aragonite, vaterite) and hydrated (ikaite) carbonate minerals.

Cardiopulmonary and arterial baroreceptor unloading during passive hyperthermia does not contribute to hyperthermia-induced hyperventilation.

PubMed

Lucas, Rebekah A I; Pearson, James; Schlader, Zachary J; Crandall, Craig G

2015-11-01

What is the central question of this study? Does baroreceptor unloading during passive hyperthermia contribute to increases in ventilation and decreases in end-tidal carbon dioxide during that exposure? What is the main finding and its importance? Hyperthermic hyperventilation is not mitigated by expanding central blood volume and reloading the cardiopulmonary baroreceptors via rapid saline infusion or by reloading the arterial baroreceptors via phenylephrine administration. The absence of a reduction in ventilation upon reloading the baroreceptors to pre-hyperthermic levels indicates that cardiopulmonary and arterial baroreceptor unloading with hyperthermia is unlikely to contribute to hyperthermic hyperventilation in humans. This study tested the hypothesis that baroreceptor unloading during passive hyperthermia contributes to increases in ventilation and decreases in end-tidal partial pressure of carbon dioxide (P ET ,CO2) during that exposure. Two protocols were performed, in which healthy subjects underwent passive hyperthermia (increasing intestinal temperature by ∼1.8°C) to cause a sustained increase in ventilation and reduction in P ET ,CO2. Upon attaining hyperthermic hyperventilation, in protocol 1 (n = 10; three females) a bolus (19 ± 2 ml kg(-1) ) of warm (∼38°C) isotonic saline was rapidly (5-10 min) infused intravenously to restore reductions in central venous pressure, whereas in protocol 2 (n = 11; five females) phenylephrine was infused intravenously (60-120 μg min(-1) ) to return mean arterial pressure to normothermic levels. In protocol 1, hyperthermia increased ventilation (by 2.2 ± 1.7 l min(-1) , P < 0.01), while reducing P ET ,CO2 (by 4 ± 3 mmHg, P = 0.04) and central venous pressure (by 5 ± 1 mmHg, P <0.01). Saline infusion increased central venous pressure by 5 ± 1 mmHg (P < 0.01), restoring it to normothermic values, but did not change ventilation or P ET ,CO2 (P > 0.05). In protocol 2, hyperthermia increased ventilation (by 5.0 ± 2.7 l min(-1) , P <0.01) and reduced P ET ,CO2 (by 5 ± 2 mmHg, P < 0.01) and mean arterial pressure (by 9 ± 7 mmHg, P <0.01). Phenylephrine infusion increased mean arterial pressure by 12 ± 3 mmHg (P < 0.01), restoring it to normothermic values, but did not change ventilation or P ET ,CO2 (P > 0.05). The absence of a reduction in ventilation upon reloading the cardiopulmonary and arterial baroreceptors to pre-hyperthermic levels indicates that baroreceptor unloading with hyperthermia is unlikely to contribute to hyperthermic hyperventilation in humans. © 2015 The Authors. Experimental Physiology © 2015 The Physiological Society.

Quantitative real-time PCR method with internal amplification control to quantify cyclopiazonic acid producing molds in foods.

PubMed

Rodríguez, Alicia; Werning, María L; Rodríguez, Mar; Bermúdez, Elena; Córdoba, Juan J

2012-12-01

A quantitative TaqMan real-time PCR (qPCR) method that includes an internal amplification control (IAC) to quantify cyclopiazonic acid (CPA)-producing molds in foods has been developed. A specific primer pair (dmaTF/dmaTR) and a TaqMan probe (dmaTp) were designed on the basis of dmaT gene which encodes the enzyme dimethylallyl tryptophan synthase involved in the biosynthesis of CPA. The IAC consisted of a 105 bp chimeric DNA fragment containing a region of the hly gene of Listeria monocytogenes. Thirty-two mold reference strains representing CPA producers and non-producers of different mold species were used in this study. All strains were tested for CPA production by high-performance liquid chromatography-mass spectrometry (HPLC-MS). The functionality of the designed qPCR method was demonstrated by the high linear relationship of the standard curves relating to the dmaT gene copy numbers and the Ct values obtained from the different CPA producers tested. The ability of the qPCR protocol to quantify CPA-producing molds was evaluated in different artificially inoculated foods. A good linear correlation was obtained over the range 1-4 log cfu/g in the different food matrices. The detection limit in all inoculated foods ranged from 1 to 2 log cfu/g. This qPCR protocol including an IAC showed good efficiency to quantify CPA-producing molds in naturally contaminated foods avoiding false negative results. This method could be used to monitor the CPA producers in the HACCP programs to prevent the risk of CPA formation throughout the food chain. Copyright © 2012 Elsevier Ltd. All rights reserved.

Evaluation of various cleaning methods to remove bacillus spores from spacecraft hardware materials

NASA Technical Reports Server (NTRS)

Venkateswaran, Kasthuri; Chung, Shirley; Allton, Judith; Kern, Roger

2004-01-01

A detailed study was made of the biological cleaning effectiveness, defined in terms of the ability to remove bacterial spores, of a number of methods used to clean hardware surfaces. Aluminum (Al 6061) and titanium (Ti 6Al-4V) were chosen for the study as they were deemed the two materials most likely to be used in spacecraft extraterrestrial sampler construction. Metal coupons (1 cm x 2.5 cm) were precleaned and inoculated with 5.8 x 10(3) cultivable Bacillus subtilis spores, which are commonly found on spacecraft surfaces and in the assembly environments. The inoculated coupons were subsequently cleaned using: (1) 70% isopropyl alcohol wipe; (2) water wipe; (3) multiple-solvent flight-hardware cleaning procedures used at the Jet Propulsion Laboratory (JPL); (4) Johnson Space Center-developed ultrapure water rinse; and (5) a commercial, semi-aqueous, multiple-solvent (SAMS) cleaning process. The biological cleaning effectiveness was measured by agar plate assay, sterility test (growing in liquid media), and epifluorescent microscopy. None of the cleaning protocols tested completely removed viable spores from the surface of the aluminum. In contrast, titanium was capable of being cleaned to sterility by two methods, the JPL standard and the commercial SAMS cleaning process. Further investigation showed that the passivation step employed in the JPL standard method is an effective surface sterilant on both metals but not compatible with aluminum. It is recommended that titanium (Ti 6Al-4V) be considered superior to aluminum (Al 6061) for use in spacecraft sampling hardware, both for its potential to be cleaned to sterilization and for its ability to withstand the most effective cleaning protocols.

Evaluation of Various Cleaning Methods to Remove Bacillus Spores from Spacecraft Hardware Materials

NASA Astrophysics Data System (ADS)

Venkateswaran, Kasthuri; Chung, Shirley; Allton, Judith; Kern, Roger

2004-09-01

A detailed study was made of the biological cleaning effectiveness, defined in terms of the ability to remove bacterial spores, of a number of methods used to clean hardware surfaces. Aluminum (Al 6061) and titanium (Ti 6Al-4V) were chosen for the study as they were deemed the two materials most likely to be used in spacecraft extraterrestrial sampler construction. Metal coupons (1 cm × 2.5 cm) were precleaned and inoculated with 5.8 × 103 cultivable Bacillus subtilis spores, which are commonly found on spacecraft surfaces and in the assembly environments. The inoculated coupons were subsequently cleaned using: (1) 70% isopropyl alcohol wipe; (2) water wipe; (3) multiple-solvent flight-hardware cleaning procedures used at the Jet Propulsion Laboratory (JPL); (4) Johnson Space Center-developed ultrapure water rinse; and (5) a commercial, semi-aqueous, multiple-solvent (SAMS) cleaning process. The biological cleaning effectiveness was measured by agar plate assay, sterility test (growing in liquid media), and epifluorescent microscopy. None of the cleaning protocols tested completely removed viable spores from the surface of the aluminum. In contrast, titanium was capable of being cleaned to sterility by two methods, the JPL standard and the commercial SAMS cleaning process. Further investigation showed that the passivation step employed in the JPL standard method is an effective surface sterilant on both metals but not compatible with aluminum. It is recommended that titanium (Ti 6Al-4V) be considered superior to aluminum (Al 6061) for use in spacecraft sampling hardware, both for its potential to be cleaned to sterilization and for its ability to withstand the most effective cleaning protocols.

Evaluation of various cleaning methods to remove bacillus spores from spacecraft hardware materials.

PubMed

Venkateswaran, Kasthuri; Chung, Shirley; Allton, Judith; Kern, Roger

2004-01-01

A detailed study was made of the biological cleaning effectiveness, defined in terms of the ability to remove bacterial spores, of a number of methods used to clean hardware surfaces. Aluminum (Al 6061) and titanium (Ti 6Al-4V) were chosen for the study as they were deemed the two materials most likely to be used in spacecraft extraterrestrial sampler construction. Metal coupons (1 cm x 2.5 cm) were precleaned and inoculated with 5.8 x 10(3) cultivable Bacillus subtilis spores, which are commonly found on spacecraft surfaces and in the assembly environments. The inoculated coupons were subsequently cleaned using: (1) 70% isopropyl alcohol wipe; (2) water wipe; (3) multiple-solvent flight-hardware cleaning procedures used at the Jet Propulsion Laboratory (JPL); (4) Johnson Space Center-developed ultrapure water rinse; and (5) a commercial, semi-aqueous, multiple-solvent (SAMS) cleaning process. The biological cleaning effectiveness was measured by agar plate assay, sterility test (growing in liquid media), and epifluorescent microscopy. None of the cleaning protocols tested completely removed viable spores from the surface of the aluminum. In contrast, titanium was capable of being cleaned to sterility by two methods, the JPL standard and the commercial SAMS cleaning process. Further investigation showed that the passivation step employed in the JPL standard method is an effective surface sterilant on both metals but not compatible with aluminum. It is recommended that titanium (Ti 6Al-4V) be considered superior to aluminum (Al 6061) for use in spacecraft sampling hardware, both for its potential to be cleaned to sterilization and for its ability to withstand the most effective cleaning protocols.

Treatment of experimentally induced pneumonic pasteurellosis of young calves with tilmicosin.

PubMed Central

Morck, D W; Merrill, J K; Gard, M S; Olson, M E; Nation, P N

1997-01-01

Twenty four (24) healthy male Holstein calves (< 70 kg) were each experimentally infected by intrabronchial inoculation of 4.0 x 10(9) viable cells of Pasteurella haemolytica-AI (B122) at Time = 0 h. At 1 h following inoculation animals received either: 1) Sham treatment with sterile 0.85% saline SC (n = 12); or 2) a single injection of 10 mg tilmicosin per kg body weight (n = 12). Calves that were non-infected and tilmicosin-treated were also included for determining tilmicosin concentrations in serum and lung tissue at 1, 2, 4, 6, 8, 24, 48, and 72 h (n = 3-per time). In the infected calves, response to therapy was monitored clinically. Serum samples were collected for determination of tilmicosin concentrations using HPLC. Any animal becoming seriously ill was humanely killed. Complete necropsy examinations were performed on all animals and included gross pathologic changes, bacteriologic analysis, histopathology, and determination of pulmonary concentrations of tilmicosin. Tilmicosin treated animals responded significantly better to therapy than saline-treated control calves. Clinical assessment of calves during the study indicated that tilmicosin-treated calves had significantly improved by T = 8 h compared to satine-treated animals (P < 0.05). At necropsy tilmicosin-treated calves had significantly less severe gross and histological lesions (P < 0.05) of the pulmonary tissue. Of the 12 saline-treated calves, 92% (11/12) had Pasteurella haemolytica-A1 in lung tissue, while of the tilmicosin-treated calves 0% (0/12) cultured positive for P. haemolytica. Mean (+/- standard error) serum tilmicosin concentrations in infected calves peaked at 1 h post-injection (1.10 +/- 0.06 micrograms/mL) and rapidly decreased to 0.20 +/- 0.03 microgram/mL, well below the MIC of 0.50 microgram/mL for P. haemolytica-A1 (B122), by 12 h. These serum concentrations were very similar to serum concentrations of tilmicosin in non-infected tilmicosin-treated calves. Lung tissue concentrations of the antibiotic were comparatively high, even at 72 h post-infection (6.50 +/- 0.75 ppm). Lung tissue concentrations at 72 h were significantly higher in experimentally infected calves than in non-infected tilmicosin-treated animals (P < 0.05). These data demonstrate that tilmicosin was effective in treating experimentally-induced pneumonic pasteurellosis as determined by alleviation of clinical signs, pathological findings at post mortem, and presence of viable bacteria from the lung. Concentrations substantially above MIC for P. haemolytica were present in lung tissue even at 72 h following a single subcutaneous injection of 10 mg tilmicosin per kg body weight. Images Figure 2A. Figure 2B. PMID:9242998

Nasal Irrigation for Chronic Rhinosinusitis and Fatigue in Patients with Gulf War Syndrome

DTIC Science & Technology

2014-07-01

xylitol has continued to provide daily use packets at no cost as indicated in the protocol and letter of support in the initial grant application...Pharmaceutical Research Center (PRC) which will be housing and dispensing all study medications ( Xylitol and Saline). We have final approval on all...recruitment strategies  Procurement of xylitol  Recruitment and enrollment of subjects 6 REPORTABLE OUTCOMES:  UW HS IRB continuing review

Hypertonic Saline Dextran (HSD) in a Complex Military Injury - A Preclinical Study

DTIC Science & Technology

2010-04-01

applicable to civilian settings e.g. after terrorist bombings where there is often disruption to infrastructure and security issues relating to secondary... systemic inflammatory response that develops after trauma and which is thought to be part of the aetiology underlying later complications [19- 22...consisted of end-tidal CO2, pulse oximetry via a tail probe and skin surface electrocardiogram electrodes (Propac 106EL, Protocol Systems Inc., Oregon

Port in oncology practice: 3-monthly locking with normal saline for catheter maintenance, a preliminary report.

PubMed

Solinas, Gianfranca; Platini, Francesca; Trivellato, Maurizio; Rigo, Carla; Alabiso, Oscar; Galetto, Alessandra S

2017-07-14

Patients with cancer need stable venous access using central vascular devices like central venous ports and peripherally inserted central catheters that can be used for a wide range of indications. Numerous flushing protocols exist including different frequencies for catheter locking to maintain catheter patency. The aim of this retrospective study was to evaluate the incidence of lumen occlusion of central venous ports in a group of adult cancer patients, adopting a policy of locking with normal saline every three months. This is a single-center retrospective observational study. During follow-up, we analyzed adult cancer patients who had undergone port insertion from January 1st, 2007 to August 31st, 2014. Flushing and locking were performed every three months with a syringe containing normal saline. We collected data from 381 patients with ports inserted in subclavian vein (379 patients) and in the right jugular vein (2 patients). Locking was performed during 3-monthly follow-up visits. Median follow-up was 810 days (90-2700 days). Among 381 ports, 59 were removed; the reasons for removal were: end of use (45 cases), catheter rupture (9 cases), dislocation (3 cases) and catheter-related bloodstream infection (2 cases). We had no reports of lumen occlusion. Our data suggest that locking ports with normal saline every three months is not associated with an increased risk of lumen occlusion.

Reversal of rocuronium-induced neuromuscular blockade by sugammadex allows for optimization of neural monitoring of the recurrent laryngeal nerve.

PubMed

Lu, I-Cheng; Wu, Che-Wei; Chang, Pi-Ying; Chen, Hsiu-Ya; Tseng, Kuang-Yi; Randolph, Gregory W; Cheng, Kuang-I; Chiang, Feng-Yu

2016-04-01

The use of neuromuscular blocking agent may effect intraoperative neuromonitoring (IONM) during thyroid surgery. An enhanced neuromuscular-blockade (NMB) recovery protocol was investigated in a porcine model and subsequently clinically applied during human thyroid neural monitoring surgery. Prospective animal and retrospective clinical study. In the animal experiment, 12 piglets were injected with rocuronium 0.6 mg/kg and randomly allocated to receive normal saline, sugammadex 2 mg/kg, or sugammadex 4 mg/kg to compare the recovery of laryngeal electromyography (EMG). In a subsequent clinical application study, 50 patients who underwent thyroidectomy with IONM followed an enhanced NMB recovery protocol-rocuronium 0.6 mg/kg at anesthesia induction and sugammadex 2 mg/kg at the operation start. The train-of-four (TOF) ratio was used for continuous quantitative monitoring of neuromuscular transmission. In our porcine model, it took 49 ± 15, 13.2 ± 5.6, and 4.2 ± 1.5 minutes for the 80% recovery of laryngeal EMG after injection of saline, sugammadex 2 mg/kg, and sugammadex 4 mg/kg, respectively. In subsequent clinical human application, the TOF ratio recovered from 0 to >0.9 within 5 minutes after administration of sugammadex 2 mg/kg at the operation start. All patients had positive and high EMG amplitude at the early stage of the operation, and intubation was without difficulty in 96% of patients. Both porcine modeling and clinical human application demonstrated that sugammadex 2 mg/kg allows effective and rapid restoration of neuromuscular function suppressed by rocuronium. Implementation of this enhanced NMB recovery protocol assures optimal conditions for tracheal intubation as well as IONM in thyroid surgery. NA. © 2016 The American Laryngological, Rhinological and Otological Society, Inc.

Results of the Intravascular Cooling in the Treatment of Acute Stroke 2 Trial (ICTuS-2)

PubMed Central

Lyden, Patrick; Hemmen, Thomas; Grotta, James; Rapp, Karen; Ernstrom, Karin; Rzesiewicz, Teresa; Parker, Stephanie; Concha, Mauricio; Syed, Hussain; Agarwal, Sachin; Meyer, Brett; Jurf, Julie; Altafullah, Irfan; Raman, Rema

2016-01-01

Background and Purpose Therapeutic hypothermia (TH) is a potent neuroprotectant approved for cerebral protection after neonatal hypoxia-ischemia and cardiac arrest. TH for acute ischemic stroke is safe and feasible in pilot trials. We designed a study protocol to provide safer, faster TH in stroke patients. Methods Safety procedures and 4°C saline infusions for faster cooling were added to the Intravascular Cooling Treatment in Acute Stroke (ICTuS) trial protocol. A femoral venous intravascular cooling catheter following intravenous rt-PA in eligible patients provided 24 hours cooling followed by a 12 hour re-warm. Serial safety assessments and imaging were performed. The primary endpoint was 3-month modified Rankin score 0,1. Results Of the intended 1600 subjects, 120 were enrolled before the study was stopped. Randomly, 63 were to receive hypothermia (HY) plus anti-shivering treatment and 57 normothermia (NT). Compared to prior studies, cooling rates were improved with a cold saline bolus, without fluid overload. The intention-to-treat primary outcome of 90-day mRS 0,1 occurred in 33% HY and 38% NT subjects, OR (95% CL) of 0.81 (0.36, 1.85). Serious adverse events occurred equally. Mortality was 15.9% HY and 8.8% NT subjects, OR (95% CL) of 1.95 (0.56, 7.79). Pneumonia occurred in 19% HY vs. 10.5% in NT subjects, OR (95% CL) of 1.99 (0.63, 6.98). Conclusion Intravascular TH was confirmed to be safe and feasible in rt-PA treated acute ischemic stroke patients. Protocol changes designed to reduce pneumonia risk appeared to fail, although the sample is small. Clinical trial registration clinicaltrials.gov NCT 01123161. PMID:27834742

Carvedilol has stronger anti-inflammation and anti-virus effects than metoprolol in murine model with coxsackievirus B3-induced viral myocarditis.

PubMed

Wang, Dan; Chen, Yiming; Jiang, Jianbin; Zhou, Aihua; Pan, Lulu; Chen, Qi; Qian, Yan; Chu, Maoping; Chen, Chao

2014-09-01

This study aims to compare the effects of carvedilol and metoprolol in alleviating viral myocarditis (VMC) induced by coxsackievirus B3 (CVB3) in mice. A total of 116 Balb/c mice were included in this study. Ninety-six mice were inoculated intraperitoneally with CVB3 to induce VMC. The CVB3 inoculated mice were evenly divided into myocarditis group (n=32), carvedilol group (n=32) and metoprolol group (n=32). Twenty mice (control group) were inoculated intraperitoneally with normal saline. Hematoxylin and eosin staining and histopathologic scoring were used to investigate the effects of carvedilol and metoprolol on myocardial histopathologic changes on days 3 and 5. In addition, serum cTn-I levels, cytokine levels and virus titers were determined using chemiluminescence immunoassay, enzyme-linked immunosorbent assay and plaque assay, respectively, on days 3 and 5. Finally, the levels of phosphorylated p38MAPK were studied using immunohistochemical staining and Western blotting on day 5. Carvedilol had a stronger effect than metoprolol in reducing the pathological scores of VMC induced by CVB3. Both carvedilol and metoprolol reduced the levels of cTn-I, but the effect of carvedilol was stronger. Carvedilol and metoprolol decreased the levels of myocardial pro-inflammatory cytokines and increased the expression of anti-inflammatory cytokine, with the effects of carvedilol being stronger than those of metoprolol. Carvedilol had a stronger effect in reducing myocardial virus concentration compared with metoprolol. Carvedilol was stronger than metoprolol in decreasing the levels of myocardial phosphorylated p38MAPK. In conclusion, carvedilol was more potent than metoprolol in ameliorating myocardial lesions in VMC, probably due to its stronger modulation of the balance between pro- and anti-inflammatory cytokines by inhibiting the activation of p38MAPK pathway through β1- and β2-adrenoreceptors. Copyright © 2014 Elsevier B.V. All rights reserved.

The growth-inhibitory and apoptosis-inducing effect of deferoxamine combined with arsenic trioxide on HL-60 xenografts in nude mice.

PubMed

Yu, Runhong; Wang, Dao; Ren, Xiuhua; Zeng, Li; Liu, Yufeng

2014-09-01

The aim of this study is to investigate the growth-inhibitory and apoptosis-inducing effect of deferoxamine (DFO) combined with arsenic trioxide (ATO) on the human HL-60 xenografts in nude mice and its mechanism. The highly tumorigenic leukemia cell line HL-60 cells were inoculated subcutaneously into nude mice to establish a human leukemia xenograft model. The HL-60 xenograft nude mice models were randomly divided into four groups: control (Normal saline, NS), 50mg/kg DFO, 3mg/kg ATO, the combined treatment (50mg/kg DFO+1.5mg/kg ATO) once HL-60 cells were inoculated. Tumor sizes, growth curves, inhibitory rates, cell apoptosis, and the expression of apoptosis related markers were measured to evaluate the tumor growth. Xenografted tumors were observed in all nude mice since the 5th day of inoculation. The inhibitory rates of tumor weight were 2.67%, 10.69%, and 25.57% in DFO, ATO and combination therapy groups, respectively. The combination of DFO with ATO induces significantly more tumor cell apoptosis than either agent alone (p<0.05). The expression of NF-κBp65 and survivin proteins decreased significantly while the expression of Caspase-3 and Bax increased in the combination therapy group (p<0.05). Double immunofluorescence for Caspase-3 and NFκBp65 demonstrated an inverse relationship between Caspase-3-positive areas and NFκBp65-positive areas, as well as the co-localization of Bax and survivin in xenografted tumor cells. Combination of DFO and ATO has synergistic effects on tumor growth inhibition and apoptosis-inducing in vivo with no significant side effects. The DFO and ATO can up-regulate the expression of Caspase-3 and Bax, and down-regulate the expression of NF-κBp65 and survivin, especially for their combination. Copyright © 2014 Elsevier Ltd. All rights reserved.

Rapid Diagnosis of Tuberculosis by Real-Time High-Resolution Imaging of Mycobacterium tuberculosis Colonies.

PubMed

Ghodbane, Ramzi; Asmar, Shady; Betzner, Marlena; Linet, Marie; Pierquin, Joseph; Raoult, Didier; Drancourt, Michel

2015-08-01

Culture remains the cornerstone of diagnosis for pulmonary tuberculosis, but the fastidiousness of Mycobacterium tuberculosis may delay culture-based diagnosis for weeks. We evaluated the performance of real-time high-resolution imaging for the rapid detection of M. tuberculosis colonies growing on a solid medium. A total of 50 clinical specimens, including 42 sputum specimens, 4 stool specimens, 2 bronchoalveolar lavage fluid specimens, and 2 bronchial aspirate fluid specimens were prospectively inoculated into (i) a commercially available Middlebrook broth and evaluated for mycobacterial growth indirectly detected by measuring oxygen consumption (standard protocol) and (ii) a home-made solid medium incubated in an incubator featuring real-time high-resolution imaging of colonies (real-time protocol). Isolates were identified by Ziehl-Neelsen staining and matrix-assisted laser desorption ionization-time of flight mass spectrometry. Use of the standard protocol yielded 14/50 (28%) M. tuberculosis isolates, which is not significantly different from the 13/50 (26%) M. tuberculosis isolates found using the real-time protocol (P = 1.00 by Fisher's exact test), and the contamination rate of 1/50 (2%) was not significantly different from the contamination rate of 2/50 (4%) using the real-time protocol (P = 1.00). The real-time imaging protocol showed a 4.4-fold reduction in time to detection, 82 ± 54 h versus 360 ± 142 h (P < 0.05). These preliminary data give the proof of concept that real-time high-resolution imaging of M. tuberculosis colonies is a new technology that shortens the time to growth detection and the laboratory diagnosis of pulmonary tuberculosis. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Effect of intermittent oral administration of ponazuril on experimental Sarcocystis neurona infection of horses.

PubMed

Mackay, Robert J; Tanhauser, Susan T; Gillis, Karen D; Mayhew, Ian G; Kennedy, Tom J

2008-03-01

To evaluate the effect of intermittent oral administration of ponazuril on immunoconversion against Sarcocystis neurona in horses inoculated intragastrically with S neurona sporocysts. 20 healthy horses that were seronegative for S neurona-specific IgG. 5 control horses were neither inoculated with sporocysts nor treated. Other horses (5 horses/group) each received 612,500 S neurona sporocysts via nasogastric tube (day 0) and were not treated or were administered ponazuril (20 mg/kg, PO) every 7 days (beginning on day 5) or every 14 days (beginning on day 12) for 12 weeks. Blood and CSF samples were collected on day - 1 and then every 14 days after challenge for western blot assessment of immunoconversion. Clinical signs of equine protozoal myeloencephalitis (EPM) were monitored, and tissues were examined histologically after euthanasia. Sera from all challenged horses yielded positive western blot results within 56 days. Immunoconversion in CSF was detected in only 2 of 5 horses that were treated weekly; all other challenged horses immunoconverted within 84 days. Weekly administration of ponazuril significantly reduced the antibody response against the S neurona 17-kd antigen in CSF. Neurologic signs consistent with EPM did not develop in any group; likewise, histologic examination of CNS tissue did not reveal protozoa or consistent degenerative or inflammatory changes. Administration of ponazuril every 7 days, but not every 14 days, significantly decreased intrathecal anti-S neurona antibody responses in horses inoculated with S neurona sporocysts. Protocols involving intermittent administration of ponazuril may have application in prevention of EPM.

Ancient microbes from halite fluid inclusions: optimized surface sterilization and DNA extraction.

PubMed

Sankaranarayanan, Krithivasan; Timofeeff, Michael N; Spathis, Rita; Lowenstein, Tim K; Lum, J Koji

2011-01-01

Fluid inclusions in evaporite minerals (halite, gypsum, etc.) potentially preserve genetic records of microbial diversity and changing environmental conditions of Earth's hydrosphere for nearly one billion years. Here we describe a robust protocol for surface sterilization and retrieval of DNA from fluid inclusions in halite that, unlike previously published methods, guarantees removal of potentially contaminating surface-bound DNA. The protocol involves microscopic visualization of cell structures, deliberate surface contamination followed by surface sterilization with acid and bleach washes, and DNA extraction using Amicon centrifugal filters. Methods were verified on halite crystals of four different ages from Saline Valley, California (modern, 36 ka, 64 ka, and 150 ka), with retrieval of algal and archaeal DNA, and characterization of the algal community using ITS1 sequences. The protocol we developed opens up new avenues for study of ancient microbial ecosystems in fluid inclusions, understanding microbial evolution across geological time, and investigating the antiquity of life on earth and other parts of the solar system.

Assessing viability of extracorporeal preserved muscle transplants using external field stimulation: a novel tool to improve methods prolonging bridge-to-transplantation time

PubMed Central

Taeger, Christian D.; Friedrich, Oliver; Dragu, Adrian; Weigand, Annika; Hobe, Frieder; Drechsler, Caroline; Geppert, Carol I.; Arkudas, Andreas; Münch, Frank; Buchholz, Rainer; Pollmann, Charlotte; Schramm, Axel; Birkholz, Torsten; Horch, Raymund E.; Präbst, Konstantin

2015-01-01

Preventing ischemia-related cell damage is a priority when preserving tissue for transplantation. Perfusion protocols have been established for a variety of applications and proven to be superior to procedures used in clinical routine. Extracorporeal perfusion of muscle tissue though cumbersome is highly desirable since it is highly susceptible to ischemia-related damage. To show the efficacy of different perfusion protocols external field stimulation can be used to immediately visualize improvement or deterioration of the tissue during active and running perfusion protocols. This method has been used to show the superiority of extracorporeal perfusion using porcine rectus abdominis muscles perfused with heparinized saline solution. Perfused muscles showed statistically significant higher ability to exert force compared to nonperfused ones. These findings can be confirmed using Annexin V as marker for cell damage, perfusion of muscle tissue limits damage significantly compared to nonperfused tissue. The combination of extracorporeal perfusion and external field stimulation may improve organ conservation research. PMID:26145230

Comparison of Two Methods of RNA Extraction from Formalin-Fixed Paraffin-Embedded Tissue Specimens

PubMed Central

Gouveia, Gisele Rodrigues; Ferreira, Suzete Cleusa; Ferreira, Jerenice Esdras; Siqueira, Sheila Aparecida Coelho; Pereira, Juliana

2014-01-01

The present study aimed to compare two different methods of extracting RNA from formalin-fixed paraffin-embedded (FFPE) specimens of diffuse large B-cell lymphoma (DLBCL). We further aimed to identify possible influences of variables—such as tissue size, duration of paraffin block storage, fixative type, primers used for cDNA synthesis, and endogenous genes tested—on the success of amplification from the samples. Both tested protocols used the same commercial kit for RNA extraction (the RecoverAll Total Nucleic Acid Isolation Optimized for FFPE Samples from Ambion). However, the second protocol included an additional step of washing with saline buffer just after sample rehydration. Following each protocol, we compared the RNA amount and purity and the amplification success as evaluated by standard PCR and real-time PCR. The results revealed that the extra washing step added to the RNA extraction process resulted in significantly improved RNA quantity and quality and improved success of amplification from paraffin-embedded specimens. PMID:25105117

Reverse transcription multiplex PCR for differentiation between polio- and enteroviruses from clinical and environmental samples.

PubMed

Egger, D; Pasamontes, L; Ostermayer, M; Bienz, K

1995-06-01

For the rapid detection of polioviruses and their differentiation from nonpoliovirus enteroviruses, we developed a protocol in which clinical or environmental specimens are first inoculated onto cell cultures in tubes. After overnight incubation, the cultures are subjected to reverse transcription multiplex PCR with a primer pair which detects all enteroviruses (T. Hyypiä, P. Auvinen, and M. Maaronen, J. Gen. Virol. 70:3261-3268 1989) and two newly designed primer pairs specific for all 36 poliovirus strains tested. The PCR products can unequivocally be identified by their lengths in agarose gels, whereas the genetic heterogeneity of the poliovirus strains precludes identification by back-hybridization with internal probes. The proposed protocol is highly insensitive to the inhibitory effects of substances in the sample (stool, sewage). It allows for the detection of polioviruses and for polioviruses to be distinguished from nonpoliovirus enteroviruses within 24 h, and it allows for the concomitant isolation of a viable strain suitable for further typing.

An evaluation of different steam disinfection protocols for cystic fibrosis nebulizers.

PubMed

Hohenwarter, K; Prammer, W; Aichinger, W; Reychler, G

2016-01-01

Contamination is a key element in cystic fibrosis. For this reason, nebulizer hygiene is an important, but complex and time-consuming task for cystic fibrosis patients. The aim of this study was to compare different steam disinfection and drying protocols. One hundred nebulizer parts were inoculated with cystic fibrosis-related bacteria in high concentrations (Burkholderia multivorans 3.9 × 10(10)/ml, Staphylococcus aureus 8.9 × 10(8/)ml and Pseudomonas aeruginosa 2.1 × 10(9)/ml). Tubes with Mycobacterium abscessus complex were additionally tested. Six steam disinfectors were compared. Different methods of drying were examined. All tested bacteria were efficiently killed by the different steam disinfectors tested. The risk of contamination depended on the method of drying. Steam disinfection is a safe disinfection method. It is better to leave the nebulizers wet after steam disinfection than to manipulate them by active drying, which seems to be a source of recontamination. Copyright © 2015 European Cystic Fibrosis Society. Published by Elsevier B.V. All rights reserved.

A Method of Managing Severe Traumatic Brain Injury in the Absence of Intracranial Pressure Monitoring: The Imaging and Clinical Examination Protocol.

PubMed

Chesnut, Randall M; Temkin, Nancy; Dikmen, Sureyya; Rondina, Carlos; Videtta, Walter; Petroni, Gustavo; Lujan, Silvia; Alanis, Victor; Falcao, Antonio; de la Fuenta, Gustavo; Gonzalez, Luis; Jibaja, Manuel; Lavarden, Arturo; Sandi, Freddy; Mérida, Roberto; Romero, Ricardo; Pridgeon, Jim; Barber, Jason; Machamer, Joan; Chaddock, Kelley

2018-01-01

The imaging and clinical examination (ICE) algorithm used in the Benchmark Evidence from South American Trials: Treatment of Intracranial Pressure (BEST TRIP) randomized controlled trial is the only prospectively investigated clinical protocol for traumatic brain injury management without intracranial pressure (ICP) monitoring. As the default literature standard, it warrants careful evaluation. We present the ICE protocol in detail and analyze the demographics, outcome, treatment intensity, frequency of intervention usage, and related adverse events in the ICE-protocol cohort. The 167 ICE protocol patients were young (median 29 years) with a median Glasgow Coma Scale motor score of 4 but with anisocoria or abnormal pupillary reactivity in 40%. This protocol produced outcomes not significantly different from those randomized to the monitor-based protocol (favorable 6-month extended Glasgow Outcome Score in 39%; 41% mortality rate). Agents commonly employed to treat suspected intracranial hypertension included low-/moderate-dose hypertonic saline (72%) and mannitol (57%), mild hyperventilation (adjusted partial pressure of carbon dioxide 30-35 mm Hg in 73%), and pressors to maintain cerebral perfusion (62%). High-dose hyperosmotics or barbiturates were uncommonly used. Adverse event incidence was low and comparable to the BEST TRIP monitored group. Although this protocol should produce similar/acceptable results under circumstances comparable to those in the trial, influences such as longer pre-hospital times and non-specialist transport personnel, plus an intensive care unit model of aggressive physician-intensive care by small groups of neurotrauma-focused intensivists, which differs from most high-resource models, support caution in expecting the same results in dissimilar settings. Finally, this protocol's ICP-titration approach to suspected intracranial hypertension (vs. crisis management for monitored ICP) warrants further study.

Acute kidney injury after primary angioplasty: effect of different hydration treatments.

PubMed

Manari, Antonio; Magnavacchi, Paolo; Puggioni, Enrico; Vignali, Luigi; Fiaccadori, Enrico; Menozzi, Mila; Tondi, Stefano; Robotti, Stefano; Ferrari, Duilio; Valgimigli, Marco

2014-01-01

We evaluated the effect of different dose hydration protocols, with normal saline or bicarbonate, on the incidence of contrast-induced acute kidney injury (CI-AKI) in patients with ST-elevation myocardial infarction (STEMI) treated with primary percutaneous coronary intervention (PPCI). We considered 592 STEMI patients treated with PPCI in 5 Italian centers. Patients were randomized to receive standard or high-dose infusions of normal saline or sodium bicarbonate started immediately before contrast medium administration and continued for the following 12 h. The cumulative incidence of CI-AKI was 18.1% without any difference among treatment groups. Shock, age, ejection fraction 35% or less, and basal serum creatinine were significantly associated with an increased risk of CI-AKI. Follow-up at 12 months was complete in 573 patients. Overall, 25 out of 573 patients died (4.3%). We observed higher short-term mortality rates in patients receiving high-volume hydration. Otherwise, only age, shock and CI-AKI were significantly associated with 1-year mortality. In patients with STEMI undergoing PPCI, high-volume hydration with normal saline or sodium bicarbonate administrated at the time of contrast media administration was not associated with any significant advantage in terms of CI-AKI prevention.

Does Osmotic Stress Affect Natural Product Expression in Fungi?

PubMed

Overy, David; Correa, Hebelin; Roullier, Catherine; Chi, Wei-Chiung; Pang, Ka-Lai; Rateb, Mostafa; Ebel, Rainer; Shang, Zhuo; Capon, Rob; Bills, Gerald; Kerr, Russell

2017-08-13

The discovery of new natural products from fungi isolated from the marine environment has increased dramatically over the last few decades, leading to the identification of over 1000 new metabolites. However, most of the reported marine-derived species appear to be terrestrial in origin yet at the same time, facultatively halo- or osmotolerant. An unanswered question regarding the apparent chemical productivity of marine-derived fungi is whether the common practice of fermenting strains in seawater contributes to enhanced secondary metabolism? To answer this question, a terrestrial isolate of Aspergillus aculeatus was fermented in osmotic and saline stress conditions in parallel across multiple sites. The ex-type strain of A. aculeatus was obtained from three different culture collections. Site-to-site variations in metabolite expression were observed, suggesting that subculturing of the same strain and subtle variations in experimental protocols can have pronounced effects upon metabolite expression. Replicated experiments at individual sites indicated that secondary metabolite production was divergent between osmotic and saline treatments. Titers of some metabolites increased or decreased in response to increasing osmolite (salt or glycerol) concentrations. Furthermore, in some cases, the expression of some secondary metabolites in relation to osmotic and saline stress was attributed to specific sources of the ex-type strains.

Use of hygiene protocols to control the spread of viruses in a hotel.

PubMed

Sifuentes, Laura Y; Koenig, David W; Phillips, Ronnie L; Reynolds, Kelly A; Gerba, Charles P

2014-09-01

The goals of this study were to observe the spread of viruses in a hotel setting and to assess the effectiveness of a hygiene intervention in reducing their spread. Selected fomites in one hotel room were inoculated with bacteriophage ϕx-174, and fomites in a conference center within the same hotel were inoculated using bacteriophage MS2. Cleaning of the contaminated room resulted in the spread of viruses to other rooms by the housekeeping staff. Furthermore, viruses were transferred by hotel guests to the conference center and a communal kitchen area. Additionally, conference attendees transferred viruses from the conference center to their hotel rooms and a communal kitchen area. This study demonstrated how viruses can be spread throughout a hotel setting by both housekeepers and guests. A hygiene intervention, which included providing hand hygiene products and facial tissues to the guests and disinfecting solutions with disposable wipes to the housekeeping staff, was successful in reducing the spread of viruses between the hotel guest rooms and conference center. The hygiene intervention resulted in significantly reduced transfer of the ϕx-174 between the contaminated hotel room and other hotel rooms, communal areas, and the conference center (p = 0.02).

High Concentration of Benzyladenine Solution Stimulates Anthers for Inducing Callus in Ricinus Communis L.

NASA Astrophysics Data System (ADS)

Liu, Ying; Yan, Shuying; Yang, Fuguang; Li, Dongliang; Tang, Jianian; Liu, Guoxuan; Lin, Shiwan; Niu, Sufang; Yang, Yali

2017-12-01

An high-frequency protocol for induction of callus from anther explants of Ricinus communis was described. When anther explants of R. communis was cultured directly onto medium containing 6-benzylaminopurine (BA) induced formation of only poor quality callus that had a low induction frequency of anther callus (10.67%). However, treating the anther explants with high concentrations (7.5-120 mg/L) of BA solution for short time periods (5-80 min) helped to improve the induction frequency and enhance the quality of the callus formation significantly. The best callus induction (41.25%) was observed when anther explants were treated with 15 mg/L BA solution for 10 min before being inoculated onto hormone-free Murashige and Skoog (MS) medium for 30 days. In order to further optimize the culture system, after treated with 15 mg/L BA for 10 min, anther explants were inoculated on the hormone-free MS medium contained concentrations of sodium nitroprusside (SNP). The results showed that SNP significantly promoted the response of callus induction, especially when 8 mg/L SNP was applied, the the highest percentage of callus induction (60.37%) were gained.

Evaluation of the VIDAS Listeria (LIS) immunoassay for the detection of Listeria in foods using demi-Fraser and Fraser enrichment broths, as modification of AOAC Official Method 999.06 (AOAC Official Method 2004.06).

PubMed

Silbernagel, Karen M; Jechorek, Robert P; Kaufer, Amanda L; Johnson, Ronald L; Aleo, V; Brown, B; Buen, M; Buresh, J; Carson, M; Franklin, J; Ham, P; Humes, L; Husby, G; Hutchins, J; Jechorek, R; Jenkins, J; Kaufer, A; Kexel, N; Kora, L; Lam, L; Lau, D; Leighton, S; Loftis, M; Luc, S; Martin, J; Nacar, I; Nogle, J; Park, J; Schultz, A; Seymore, D; Smith, C; Smith, J; Thou, P; Ulmer, M; Voss, R; Weaver, V

2005-01-01

A multilaboratory study was conducted to compare the VIDAS LIS immunoassay with the standard cultural methods for the detection of Listeria in foods using an enrichment modification of AOAC Official Method 999.06. The modified enrichment protocol was implemented to harmonize the VIDAS LIS assay with the VIDAS LMO2 assay. Five food types--brie cheese, vanilla ice cream, frozen green beans, frozen raw tilapia fish, and cooked roast beef--at 3 inoculation levels, were analyzed by each method. A total of 15 laboratories representing government and industry participated. In this study, 1206 test portions were tested, of which 1170 were used in the statistical analysis. There were 433 positive by the VIDAS LIS assay and 396 positive by the standard culture methods. A Chi-square analysis of each of the 5 food types, at the 3 inoculation levels tested, was performed. The resulting average Chi square analysis, 0.42, indicated that, overall, there are no statistical differences between the VIDAS LIS assay and the standard methods at the 5% level of significance.

Experimental Clostridium perfringens type D enterotoxemia in goats.

PubMed

Uzal, F A; Kelly, W R

1998-03-01

The effects of intraduodenal administration of Clostridium perfringens cultures and culture products in goats were evaluated to develop a reliable experimental model of enterotoxemia in this species. Five conventionally reared, 11-16-week-old Angora goat kids were dosed intraduodenally with whole cultures of C. perfringens type D; five similar animals were dosed with C. perfringens type D filtered culture supernatant; and a third group of five kids was dosed with C. perfringens type D washed cells. Two kids were used as controls and received sterile, nontoxic culture medium intraduodenally. All animals received starch solution into the abomasum. All five kids inoculated with whole culture and three of five dosed with culture supernatant and with washed cells developed central nervous system signs. Diarrhea was observed in two of five kids inoculated with whole culture, in all five of those dosed with culture supernatant, and in three of five of those that received washed cells. The most striking postmortem findings consisted of lung edema, necrotizing pseudomembranous colitis, and cerebral vasogenic edema. The protocol thus provided a reasonable model of naturally occurring enterotoxemia in goats, producing a range of clinical signs and postmortem changes similar to those observed in the natural disease.

Use of GDNF Releasing Nanofiber Nerve Guide Conduits for the Repair of Conus Medullaris/Cauda Equina Injury in the Nonhuman Primate

DTIC Science & Technology

2015-02-01

repair of conus medullaris/cauda equina injury in rhesus macaques using a biodegradable bridging graft that releasing the trophic factor, GDNF. All...and biodegradable nerve guidance channels as bridging grafts. The follow section describes the methods and protocols for laminectomy, ventral root...in saline until being grafted, 8. For use of a bridging biodegradable NGC segment, the NGCs were fabricated and comprised of electrospun

Nasal Irrigation for Chronic Rhinosinusitis and Fatigue in Patients with Gulf War Syndrome

DTIC Science & Technology

2013-07-01

materials (neti pots, xylitol , saline), and subject recruitment and enrollment. The human subjects research approval process required protocol submission...application submitted on 11/25/11 was also approved on 7/10/12. The company that provided xylitol (Danisco) is now owned by a private company (DuPont) a...regarding continuity of study as well as procurement and packaging of xylitol packets (sachets) for individual use by participants in the xylitol arm of

Combined calcitriol and menadione reduces experimental murine triple negative breast tumor.

PubMed

Bohl, Luciana; Guizzardi, Solange; Rodríguez, Valeria; Hinrichsen, Lucila; Rozados, Viviana; Cremonezzi, David; Tolosa de Talamoni, Nori; Picotto, Gabriela

2017-10-01

Calcitriol (D) or 1,25(OH) 2 D 3 inhibits the growth of several tumor cells including breast cancer cells, by activating cell death pathways. Menadione (MEN), a glutathione-depleting compound, may be used to potentiate the antiproliferative actions of D on cancer cells. We have previously shown in vitro that MEN improved D-induced growth arrest on breast cancer cell lines, inducing oxidative stress and DNA damage via ROS generation. Treatment with MEN+D resulted more effective than D or MEN alone. To study the in vivo effect of calcitriol, MEN or their combination on the development of murine transplantable triple negative breast tumor M-406 in its syngeneic host. Tumor M-406 was inoculated s.c., and when tumors reached the desired size, animals were randomly assigned to one of four groups receiving daily i.p. injections of either sterile saline solution (controls, C), MEN, D, or both (MEN+D). Body weight and tumor volume were recorded three times a week. Serum calcium was determined before and at the end of the treatment, at which time tumor samples were obtained for histological examination. None of the drugs, alone or in combination, affected mice body weight in the period studied. The combined treatment reduced tumor growth rate (C vs. MEN+D, P<0.05) and the corresponding histological sections exhibited small remaining areas of viable tumor only in the periphery. A concomitant DNA fragmentation was observed in all treated groups and MEN potentiated the calcitriol effect on tumor growth. As previously observed in vitro, treatment with MEN and D delayed tumor growth in vivo more efficiently than the individual drugs, with evident signals of apoptosis induction. Our results propose an alternative protocol to treat triple negative breast cancer, using GSH depleting drugs together with calcitriol, which would allow lower doses of the steroid to maintain the antitumor effect while diminishing its adverse pharmacological effects. Copyright © 2017. Published by Elsevier Masson SAS.

Resin-modified glass ionomer cements: fluoride release and influence on Streptococcus mutans growth.

PubMed

Friedl, K H; Schmalz, G; Hiller, K A; Shams, M

1997-02-01

The aims of the present study were to measure the fluoride release of 1 glass ionomer cement, 1 cermet, 3 resin-modified glass ionomer cements and 1 compomer, and to determine the influence of each material on bacterial growth. Test specimens were eluted in saline for 180 days. Every 2 days, the specimens were transferred into fresh saline and the fluoride content of the solution was measured. Furthermore, 48-h, 14-d, 90-d, and 180-d eluates were inoculated with Streptococcus mutans and bacterial growth was recorded nephelometrically. Fluoride release dropped significantly over time for each material with values between 6.2 (Ketac-Silver) and 29.3 (Photac-Fil) ppm after 48 h to values between 0.6 (Ketac-Silver) and 1.7 (Ketac-Fil, Vitremer) ppm after 180 days. Each material reduced bacterial growth at each time of examination, but the effect decreased significantly over time with a maximum growth of 71.7% (Ketac-Fil) to 85.6% (Ketac-Silver) after 48 h and 94.7 (Vitremer) to 99.0% (Ketac-Silver) after 180 days (growth control = 100%). Both Ketac-Silver and Dyract showed a significantly lower inhibiting effect on bacterial growth than the other materials. The tested materials showed a good correlation between fluoride release and influence on bacterial growth. However, both effects dropped dramatically over the 180-days period.

Describing the physiological responses of different rice genotypes to salt stress using sigmoid and piecewise linear functions.

PubMed

Radanielson, Ando M; Angeles, Olivyn; Li, Tao; Ismail, Abdelbagi M; Gaydon, Donald S

2018-05-01

Rice is the staple food for almost half of the world population. In South and South East Asia, about 40% of rice production is from deltaic regions that are vulnerable to salt stress. A quantitative approach was developed for characterizing genotypic variability in biomass production, leaf transpiration rate and leaf net photosynthesis responses to salinity during the vegetative stage, with the aim of developing efficient screening protocols to accelerate breeding varieties adapted to salt-affected areas. Three varieties were evaluated in pots under greenhouse conditions and in the field, with average soil salinity ranging from 2 to 12 dS m -1 . Plant biomass, net photosynthesis rate, leaf transpiration rate and leaf conductance were measured at regular intervals. Crop responses were fitted using a logistic function with three parameters: 1) maximum rate under control conditions (Y max ), 2) salinity level for 50% of reduction (b), and 3) rate of reduction ( a) . Variation in the three parameters correlated significantly with variation in plant biomass production under increasing salinity. Salt stress levels that caused 50% reduction in net leaf photosynthesis and transpiration rates were higher in the tolerant genotype BRRI Dhan47 (16.5 dS m -1 and 14.3 dS m -1 , respectively) than the sensitive genotype IR29 (11.1 dS m -1 and 6.8 dS m -1 ). In BRRI Dhan47, the threshold beyond which growth was significantly reduced was above 5 dS m -1 and the rate of growth reduction beyond this threshold was as low as 4% per unit increase in salinity. This quantitative approach to screening for salinity tolerance in rice offers a means to better understand rice growth under salt stress and, using simulation modelling, can provide an improved tool for varietal characterization.

[Local reactions after diphtheria-tetanus-acellular pertussis vaccines in mice; changes in histopathology at the injection site].

PubMed

Nagaoka, Chiharu; Katsuta, Tomohiro; Honjo, Ayako; Tateyama, Satoshi; Tokutake, Tadaomi; Arimoto, Yutaka; Nakajima, Natsuki; Goshima, Toshiro; Kato, Tatsuo

2006-03-01

Diphtheria-tetanus-acellular pertussis vaccine (DTaP) developed in Japan is now widely used worldwide. DTaP is safer than the diphtheria-tetanus-whole-cell pertussis vaccine (DTwP) and has fewer severe side effects, but local reactions such as redness, swelling, and induration are still reported. The pathophysiological mechanism of these reactions is controversial. To clarify the cause of local reactions, we conducted studies using the mouse model. After administering either one or two abdominal subcutaneous DTaP inoculations, we observed changes in histopathology at the injection site at 24h, 48h, and 7 days. The control group, inoculated with physiologic saline, showed no significant changes either pathologically or with the naked eye. All mice after DTaP vaccination showed indurations at the injection site. Pathologically, we watched leukocyte invasion into or around the site, especially neutrophils and eosinophils. After the first vaccination, the extent of the invasion was strong 24h and 7 days later. At 24h following the second vaccination, a dramatic leukocyte invasion seen persisted at 7days. At 7 days after the first vaccination, peripheral fibrosis had begun, and when a second vaccination was administered, it began even earlier at the second site. These histopathological changes show that local reactions are caused by both inflammatory and allergic responses. Because this mouse study resulted in the same pattern of reactions observed in humans, this method will be useful for studies focusing on local reactions.

FT-IR microspectroscopy in rapid identification of bacteria in pure and mixed culture

NASA Astrophysics Data System (ADS)

Fontoura, Inglid; Belo, Ricardo; Sakane, Kumiko; Cardoso, Maria Angélica Gargione; Khouri, Sônia; Uehara, Mituo; Raniero, Leandro; Martin, Airton A.

2010-02-01

In recent years FT-IR microspectroscopy has been developed for microbiology analysis and applied successfully in pure cultures of microorganisms to rapidly identify strains of bacteria, yeasts and fungi. The investigation and characterization of microorganism mixed cultures is also of growing importance, especially in hospitals where it is common to poly-microbial infections. In this work, the rapid identification of bacteria in pure and mixed cultures was studied. The bacteria were obtained from the Institute Oswaldo Cruz culture collection at Brazil. Escherichia coli ATCC 10799 and Staphylococcus aureus ATCC 14456 were analyzed, 3 inoculations were examined in triplicate: Escherichia coli, Staphylococcus aureus and a mixed culture of them. The inoculations were prepared according to McFarland 0.5, incubated at 37 ° C for 6 hours, diluted in saline, placed in the CaF2 window and store for one hour at 50°C to obtain thin film. The measurement was performed by Spectrum Spotlight 400 (Perkin-Elmer) equipment in the range of 4000-900 cm-1, with 32 scans using a transmittance technique with point and image modes. The data were processed (baseline, normalization, calculation of first derivate followed by smoothing with 9 point using a Savitzky-Golay algorithm) and a cluster analysis were done by Ward's algorithm and an excellent discrimination between pure and mixed culture was obtained. Our preliminary results indicate that the FT-IR microspectroscopy associated with cluster analysis can be used to discriminate between pure and mixed culture.

Sanitation of wallboard colonized with Stachybotrys chartarum.

PubMed

Price, D L; Ahearn, D G

1999-07-01

Sections (8 cm2) of unused, nonsterile gypsum wallboard (dry wall) were inoculated with varying densities (10(4) to approximately 10(8)/ml) of conidia from 14- to 21-day cultures of Stachybotrys chartarum grown on cellulose agar. The sections were permitted to air dry and were placed into vessels with 86% or 92% RH and incubated at 22-25 degrees C for up to 12 weeks. The moisture content of the dryboard increased from near 10% to over 35%. Selected sections with confluent surface growth, mainly of S. chartarum, were obtained within 3 weeks. Sections were cleaned with a quaternary or quaternary and chlorine dioxide or a concentrated oxygen-saline solution and treated, in some cases, with a preservative system and returned to humidity vessels. Reemergence of S. chartarum from inoculated and treated surfaces occurred within 5 weeks only with sections treated with the quaternary alone. Other fungi, mostly species of Aspergillus, Chaetomium and Penicillium, slowly colonized (between 9-12 weeks) at least some areas of most treated surfaces and most uninoculated control surfaces. Stachybotrys chartarum was also found on several sections of uninoculated controls. Sections treated with a quaternary/acrylic and placed in a dynamic challenging chamber remained visually free of colonized fungi for over 90 days. These studies indicate that control samples of uninstalled wallboard, available from local distributors, can contain a baseline bioburden, including S. chartarum, that will colonize surfaces under high humidity conditions. Sanitation and preservation treatment of the wallboard can markedly delay regrowth of these fungi, particularly of S. chartarum.

Development of a Simple Method for Concentrating Enteroviruses from Oysters

PubMed Central

Sobsey, Mark D.; Wallis, Craig; Melnick, Joseph L.

1975-01-01

The development of a simple method for concentrating enteroviruses from oysters is described. In this method viruses in homogenized oyster tissues are efficiently adsorbed to oyster solids at pH 5.5 and low salt concentration. After low-speed centrifugation, the supernatant is discarded and viruses are eluted from the sedimented oyster solids by resuspending them in pH 3.5 glycine-buffered saline. The solids are then removed by low-speed centrifugation, and the virus-containing supernatant is filtered through a 0.2-μm porosity filter to remove bacteria and other small particulates without removing viruses. The virus-containing filtrate is then concentrated to a volume of a few milliliters by ultrafiltration, and the concentrate obtained is inoculated directly into cell cultures for virus assay. When tested with pools of oysters experimentally contaminated with small amounts of different enteroviruses, virus recovery efficiency averaged 63%. PMID:234154

Development of a simple method for concentrating enteroviruses from oysters.

PubMed

Sobsey, M D; Wallis, C; Melnick, J L

1975-01-01

The development of a simple method for concentrating enteroviruses from oysters is described. In this method viruses in homogenized oyster tissues are efficiently absorbed to oyster solids at pH 5.5 and low salt concentration. After low-speed centrifugation, the supernatant is discarded and viruses are eluted from the sedimented oyster solids by resuspending them in pH 3.5 glycine-buffered saline. The solids are then removed by low-speed centrifugation, and the virus-containing supernatant is filtered through a 0.2-micronm porosity filter to remove bacteria and other small particulates without removing viruses. The virus-containing filtrate is then concentrated to a volume of a few milliliters by ultrafiltration, and the concentrate obtained is inoculated directly into cell cultures for virus assay. When tested with pools of oysters experimentally contaminated with small amounts of different enteroviruses, virus recovery efficiency averaged 63%.

Is bacteriostatic saline superior to normal saline as an echocardiographic contrast agent?

PubMed

Cardozo, Shaun; Gunasekaran, Prasad; Patel, Hena; McGorisk, Timothy; Toosi, Mehrdad; Faraz, Haroon; Zalawadiya, Sandip; Alesh, Issa; Kottam, Anupama; Afonso, Luis

2014-12-01

Objective data on the performance characteristics and physical properties of commercially available saline formulations [normal saline (NS) vs. bacteriostatic normal saline (bNS)] are sparse. This study sought to compare the in vitro physical properties and in vivo characteristics of two commonly employed echocardiographic saline contrast agents in an attempt to assess superiority. Nineteen patients undergoing transesophageal echocardiograms were each administered agitated regular NS and bNS injections in random order and in a blinded manner according to a standardized protocol. Video time-intensity (TI) curves were constructed from a representative region of interest, placed paraseptally within the right atrium, in the bicaval view. TI curves were analyzed for maximal plateau acoustic intensity (Vmax, dB) and dwell time (DT, s), defined as time duration between onset of Vmax and decay of video intensity below clinically useful levels, reflecting the duration of homogenous opacification of the right atrium. To further characterize the physical properties of the bubbles in vitro, fixed aliquots of similarly agitated saline were injected into a glass well slide-cover slip assembly and examined using an optical microscope to determine bubble diameter in microns (µm) and concentration [bubble count/high power field (hpf)]. A higher acoustic intensity (a less negative dB level), higher bubble concentration and longer DT were considered properties of a superior contrast agent. For statistical analysis, a paired t test was conducted to evaluate the differences in means of Vmax and DT. Compared to NS, bNS administration was associated with superior opacification (video intensity -8.69 ± 4.7 vs. -10.46 ± 4.1 dB, P = 0.002), longer DT (17.3 ± 6.1 vs. 10.2 ± 3.7 s) in vivo and smaller mean bubble size (43.4 vs. 58.6 μm) and higher bubble concentration (1,002 vs. 298 bubble/hpf) in vitro. bNS provides higher intensity and more sustained opacification of the right atrium compared to NS. Higher bubble concentration and stability appear to be additional desirable rheological characteristics favoring bNS as a contrast agent.

Preovulatory uterine flushing with saline as a treatment for unexplained infertility: a randomised controlled trial protocol.

PubMed

Maheux-Lacroix, Sarah; Dodin, Sylvie; Moore, Lynne; Bujold, Emmanuel; Lefebvre, Jessica; Bergeron, Marie-Ève

2016-01-06

In vitro fertilisation (IVF) is the treatment of choice for unexplained infertility. Preovulatory uterine flushing could reduce intrauterine debris and inflammatory factors preventing pregnancy and constitute an alternative to IVF. Our objective is to assess the efficacy of preovulatory uterine flushing with physiological saline for the treatment of unexplained infertility. We will perform a randomised controlled trial based on consecutive women aged between 18 and 37 years consulting for unexplained infertility for at least 1 year. On the day of their luteinising hormone surge, 192 participants will be randomised in two equal groups to either receive 20 mL of physiological saline by an intrauterine catheter or 10 mL of saline intravaginally. We will assess relative risk of live birth (primary outcome), as well as pregnancy (secondary outcome) over one cycle of treatment. We will report the side effects, complications and acceptability of the intervention. This project was approved by the Ethics committee of the Centre Hospitatlier Universitaire de Quebec (no 2015-1146). Uterine flushing is usually well tolerated by women and would constitute a simple, affordable and minimally invasive treatment for unexplained infertility. We plan to communicate the results of the review by presenting research abstracts at conferences and by publishing the results in a peer-reviewed journal. NCT02539290; Pre-results. Published by the BMJ Publishing Group Limited. For permission to use (where not already granted under a licence) please go to http://www.bmj.com/company/products-services/rights-and-licensing/

Experimental dissociation of the effects of prostaglandins on renal sodium and water reabsorption by cyclo-oxygenase inhibitors in the rat.

PubMed

Bartoli, E; Branca, G F; Faedda, R; Olmeo, N A; Satta, A; Soggia, G

1982-07-01

1 The relative importance of the effect of prostaglandins on renal sodium and water reabsorption was assessed in rats. 2 Clearance experiments were performed on 24 anaesthetized rats divided into 3 groups. Each group was infused throughout either with Ringer solution at 9 ml/h (Protocol I), or at 3 ml/h (Protocol II) or with hypotonic fluid at 5 ml/h (Protocol III). Clearance periods were performed before and after intravenous injection of indomethacin (5 mg/kg) and then of aspirin (20 mg/kg). The natriuretic response to different degrees of volume expansion was not modified during the action of the inhibitors. 3 When baseline urine osmolality (Uosm) was high (Protocol II) no further increase occurred in the presence of prostaglandin inhibition. Conversely, Uosm rose from 771 +/- 134 to 1356 +/- 414 and from 575 +/- 245 to 841 +/- 407 mosm/kg (P less than 0.05) in Protocol I and Protocol III respectively, when antidiuretic hormone secretion was inhibited by the higher degree of volume expansion. 4 There was a significant correlation between the change in urine flow rate induced by cyclooxygenase inhibitors and the attendant variations in Na excretion, r = 0.42, n = 41, P less than 0.01. 5 Thus, prostaglandins affect Na loss during saline load as a side effect of their action on water permeability. They could play an important role in volume depletion by counterbalancing the large secretion rate of renal vasoconstrictors.

Experimental dissociation of the effects of prostaglandins on renal sodium and water reabsorption by cyclo-oxygenase inhibitors in the rat.

PubMed Central

Bartoli, E.; Branca, G. F.; Faedda, R.; Olmeo, N. A.; Satta, A.; Soggia, G.

1982-01-01

1 The relative importance of the effect of prostaglandins on renal sodium and water reabsorption was assessed in rats. 2 Clearance experiments were performed on 24 anaesthetized rats divided into 3 groups. Each group was infused throughout either with Ringer solution at 9 ml/h (Protocol I), or at 3 ml/h (Protocol II) or with hypotonic fluid at 5 ml/h (Protocol III). Clearance periods were performed before and after intravenous injection of indomethacin (5 mg/kg) and then of aspirin (20 mg/kg). The natriuretic response to different degrees of volume expansion was not modified during the action of the inhibitors. 3 When baseline urine osmolality (Uosm) was high (Protocol II) no further increase occurred in the presence of prostaglandin inhibition. Conversely, Uosm rose from 771 +/- 134 to 1356 +/- 414 and from 575 +/- 245 to 841 +/- 407 mosm/kg (P less than 0.05) in Protocol I and Protocol III respectively, when antidiuretic hormone secretion was inhibited by the higher degree of volume expansion. 4 There was a significant correlation between the change in urine flow rate induced by cyclooxygenase inhibitors and the attendant variations in Na excretion, r = 0.42, n = 41, P less than 0.01. 5 Thus, prostaglandins affect Na loss during saline load as a side effect of their action on water permeability. They could play an important role in volume depletion by counterbalancing the large secretion rate of renal vasoconstrictors. PMID:6809089

Saline versus balanced crystalloids for intravenous fluid therapy in the emergency department: study protocol for a cluster-randomized, multiple-crossover trial.

PubMed

Self, Wesley H; Semler, Matthew W; Wanderer, Jonathan P; Ehrenfeld, Jesse M; Byrne, Daniel W; Wang, Li; Atchison, Leanne; Felbinger, Matthew; Jones, Ian D; Russ, Stephan; Shaw, Andrew D; Bernard, Gordon R; Rice, Todd W

2017-04-13

Prior studies in critically ill patients suggest the supra-physiologic chloride concentration of 0.9% ("normal") saline may be associated with higher risk of renal failure and death compared to physiologically balanced crystalloids. However, the comparative effects of 0.9% saline and balanced fluids are largely unexamined among patients outside the intensive care unit, who represent the vast majority of patients treated with intravenous fluids. This study, entitled Saline Against Lactated Ringer's or Plasma-Lyte in the Emergency Department (SALT-ED), is a pragmatic, cluster, multiple-crossover trial at a single institution evaluating clinical outcomes of adults treated with 0.9% saline versus balanced crystalloids for intravenous fluid resuscitation in the emergency department. All adults treated in the study emergency department receiving at least 500 mL of isotonic crystalloid solution during usual clinical care and subsequently hospitalized outside the intensive care unit are included. Treatment allocation of 0.9% saline versus balanced crystalloids is assigned by calendar month, with study patients treated during the same month assigned to the same fluid type. The first month (January 2016) was randomly assigned to balanced crystalloids, with each subsequent month alternating between 0.9% saline and balanced crystalloids. For balanced crystalloid treatment, clinicians can choose either Lactated Ringer's or Plasma-Lyte A©. The study period is set at 16 months, which will result in an anticipated estimated sample size of 15,000 patients. The primary outcome is hospital-free days to day 28, defined as the number of days alive and out of the hospital from the index emergency department visit until 28 days later. Major secondary outcomes include proportion of patients who develop acute kidney injury by creatinine measurements; major adverse kidney events by hospital discharge or day 30 (MAKE30), which is a composite outcome of death, new renal replacement therapy, and persistent creatinine elevation >200% of baseline; and in-hospital mortality. This ongoing pragmatic trial will provide the most comprehensive evaluation to date of clinical outcomes associated with 0.9% saline compared to physiologically balanced fluids in patients outside the intensive care unit. ClinicalTrials.gov, NCT02614040 . Registered on 18 November 2015.

Inoculation effects on root-colonizing arbuscular mycorrhizal fungal communities spread beyond directly inoculated plants

PubMed Central

Krak, Karol; Vosátka, Miroslav; Püschel, David; Štorchová, Helena

2017-01-01

Inoculation with arbuscular mycorrhizal fungi (AMF) may improve plant performance at disturbed sites, but inoculation may also suppress root colonization by native AMF and decrease the diversity of the root-colonizing AMF community. This has been shown for the roots of directly inoculated plants, but little is known about the stability of inoculation effects, and to which degree the inoculant and the inoculation-induced changes in AMF community composition spread into newly emerging seedlings that were not in direct contact with the introduced propagules. We addressed this topic in a greenhouse experiment based on the soil and native AMF community of a post-mining site. Plants were cultivated in compartmented pots with substrate containing the native AMF community, where AMF extraradical mycelium radiating from directly inoculated plants was allowed to inoculate neighboring plants. The abundances of the inoculated isolate and of native AMF taxa were monitored in the roots of the directly inoculated plants and the neighboring plants by quantitative real-time PCR. As expected, inoculation suppressed root colonization of the directly inoculated plants by other AMF taxa of the native AMF community and also by native genotypes of the same species as used for inoculation. In the neighboring plants, high abundance of the inoculant and the suppression of native AMF were maintained. Thus, we demonstrate that inoculation effects on native AMF propagate into plants that were not in direct contact with the introduced inoculum, and are therefore likely to persist at the site of inoculation. PMID:28738069

Inoculation effects on root-colonizing arbuscular mycorrhizal fungal communities spread beyond directly inoculated plants.

PubMed

Janoušková, Martina; Krak, Karol; Vosátka, Miroslav; Püschel, David; Štorchová, Helena

2017-01-01

Inoculation with arbuscular mycorrhizal fungi (AMF) may improve plant performance at disturbed sites, but inoculation may also suppress root colonization by native AMF and decrease the diversity of the root-colonizing AMF community. This has been shown for the roots of directly inoculated plants, but little is known about the stability of inoculation effects, and to which degree the inoculant and the inoculation-induced changes in AMF community composition spread into newly emerging seedlings that were not in direct contact with the introduced propagules. We addressed this topic in a greenhouse experiment based on the soil and native AMF community of a post-mining site. Plants were cultivated in compartmented pots with substrate containing the native AMF community, where AMF extraradical mycelium radiating from directly inoculated plants was allowed to inoculate neighboring plants. The abundances of the inoculated isolate and of native AMF taxa were monitored in the roots of the directly inoculated plants and the neighboring plants by quantitative real-time PCR. As expected, inoculation suppressed root colonization of the directly inoculated plants by other AMF taxa of the native AMF community and also by native genotypes of the same species as used for inoculation. In the neighboring plants, high abundance of the inoculant and the suppression of native AMF were maintained. Thus, we demonstrate that inoculation effects on native AMF propagate into plants that were not in direct contact with the introduced inoculum, and are therefore likely to persist at the site of inoculation.

Complex conductivity of organic-rich shales

NASA Astrophysics Data System (ADS)

Woodruff, W. F.; Revil, A.; Torres-Verdin, C.

2013-12-01

We can accurately determine the intrinsic anisotropy and material properties in the laboratory, providing empirical evidence of transverse isotropy and the polarization of the organic and metallic fractions in saturated and unsaturated shales. We develop two distinct approaches to obtain the complex conductivity tensor from spectral induced polarization (SIP) measurements. Experimental results indicate clear anisotropy, and characterize the effects of thermal maturation, TOC, and pyrite, aiding in the calibration and interpretation of geophysical data. SIP is a non-intrusive measurement, sensitive to the surface conductance of mineral grains, frequency-dependent polarization of the electrical double layer, and bulk conductivity of the pore water. The in-phase and quadrature components depend upon parameters of principal importance in unconventional shale formation evaluation (e.g., the distribution of pore throat sizes, formation factor, permeability, salinity and cation exchange capacity (CEC), fluid saturation and wettability). In addition to the contribution of the electrical double layer of non-conducting minerals to surface conductivity, we have observed a clear relaxation associated with kerogen pyrolysis, pyrite distribution, and evidence that the CEC of the kerogen fraction may also contribute, depending on thermal maturation history. We utilize a recent model for anisotropic complex conductivity, and rigorous experimental protocols to quantify the role of kerogen and pyrolysis on surface and quadrature conductivity in mudrocks. The complex conductivity tensor σ* describes the directional dependence of electrical conduction in a porous medium, and accounts for both conduction and polarization. The complex-valued tensor components are given as σ*ij , where σ'ij represents in-phase and σ"ij denotes quadrature conductivities. The directional dependence of the complex conductivity tensor is relegated to the textural properties of the material. The components of the formation factor and connectivity (tortuosity) tensors Fij and Tij (affecting the bulk and surface conductivity, respectively) are correlated as Fij=TijΦ. Both conductivity and connectivity tensors share the same eigenvectors; the anisotropy ratio is equivalent in TI media. At high pore water salinity, surface and quadrature conductivity share the same bulk tortuosity; when surface conductivity dominates (low salinity), conductivity is controlled by the surface conductance, and the tortuosity of electrical current along mineral surfaces usually higher than that of the pore water. We developed two distinct SIP measurement protocols to obtain the tensor: (1) azimuthal sampling and inversion of phasor potentials through the full-field solution of the Laplace equation; (2) direct measurement of complex conductivity eigenvalues by polarized, single-component stimulus current. Experiments also include unsaturated and saturated measurements with three brines of known salinity and pH, at log-distributed frequencies ranging 1 mHz to 45 kHz. Both azimuthal spectra and eigenvalue spectra validate the theoretical model and illustrate the effectiveness of the protocols themselves. We obtain the textural tensors and invert key parameters including Archie exponents and CEC, and characterize the relaxation phenomena associated with kerogen content and maturity for multiphase fluid systems.

Agrobacterium tumefaciens-mediated transformation of blueberry (Vaccinium corymbosum L.).

PubMed

Song, Guo-Qing; Sink, K C

2004-12-01

Transient expression studies using blueberry leaf explants and monitored by beta-glucuronidase (GUS) assays indicated Agrobacterium tumefaciens strain EHA105 was more effective than LBA4404 or GV3101; and the use of acetosyringone (AS) at 100 microM for inoculation and 6 days co-cultivation was optimum compared to 2, 4, 8, 10 or 12 days. Subsequently, explants of the cultivars Aurora, Bluecrop, Brigitta, and Legacy were inoculated with strain EHA105 containing the binary vector pBISN1 with the neomycin phosphotransferase gene (nptII) and an intron-interrupted GUS gene directed by the chimeric super promoter (Aocs)3AmasPmas. Co-cultivation was for 6 days on modified woody plant medium (WPM) plus 100 microM AS. Explants were then placed on modified WPM supplemented with 1.0 mg l(-1) thidiazuron, 0.5 mg l(-1) alpha-naphthaleneacetic, 10 mg l(-1) kanamycin (Km), and 250 mg l(-1) cefotaxime. Selection for Km-resistant shoots was carried out in the dark for 2 weeks followed by culture in the light at 30 microE m(-2) s(-1) at 25 degrees C. After 12 weeks, selected shoots that were both Km resistant and GUS positive were obtained from 15.3% of the inoculated leaf explants of cultivar Aurora. Sixty-eight independent clones derived from such shoots all tested positive by the polymerase chain reaction using a nptII primer. Eight of eight among these 68 clones tested positive by Southern hybridization using a gusA gene derived probe. The transformation protocol also yielded Km-resistant, GUS-positive shoots that were also PCR positive at frequencies of 5.0% for Bluecrop, 10.0% for Brigitta and 5.6% for Legacy.

Elimination of Porcine Epidemic Diarrhea Virus in an Animal Feed Manufacturing Facility.

PubMed

Huss, Anne R; Schumacher, Loni L; Cochrane, Roger A; Poulsen, Elizabeth; Bai, Jianfa; Woodworth, Jason C; Dritz, Steve S; Stark, Charles R; Jones, Cassandra K

2017-01-01

Porcine Epidemic Diarrhea Virus (PEDV) was the first virus of wide scale concern to be linked to possible transmission by livestock feed or ingredients. Measures to exclude pathogens, prevent cross-contamination, and actively reduce the pathogenic load of feed and ingredients are being developed. However, research thus far has focused on the role of chemicals or thermal treatment to reduce the RNA in the actual feedstuffs, and has not addressed potential residual contamination within the manufacturing facility that may lead to continuous contamination of finished feeds. The purpose of this experiment was to evaluate the use of a standardized protocol to sanitize an animal feed manufacturing facility contaminated with PEDV. Environmental swabs were collected throughout the facility during the manufacturing of a swine diet inoculated with PEDV. To monitor facility contamination of the virus, swabs were collected at: 1) baseline prior to inoculation, 2) after production of the inoculated feed, 3) after application of a quaternary ammonium-glutaraldehyde blend cleaner, 4) after application of a sodium hypochlorite sanitizing solution, and 5) after facility heat-up to 60°C for 48 hours. Decontamination step, surface, type, zone and their interactions were all found to impact the quantity of detectable PEDV RNA (P < 0.05). As expected, all samples collected from equipment surfaces contained PEDV RNA after production of the contaminated feed. Additionally, the majority of samples collected from non-direct feed contact surfaces were also positive for PEDV RNA after the production of the contaminated feed, emphasizing the potential role dust plays in cross-contamination of pathogen throughout a manufacturing facility. Application of the cleaner, sanitizer, and heat were effective at reducing PEDV genomic material (P < 0.05), but did not completely eliminate it.

Comparing rapid methods for detecting Listeria in seafood and environmental samples using the most probably number (MPN) technique.

PubMed

Cruz, Cristina D; Win, Jessicah K; Chantarachoti, Jiraporn; Mutukumira, Anthony N; Fletcher, Graham C

2012-02-15

The standard Bacteriological Analytical Manual (BAM) protocol for detecting Listeria in food and on environmental surfaces takes about 96 h. Some studies indicate that rapid methods, which produce results within 48 h, may be as sensitive and accurate as the culture protocol. As they only give presence/absence results, it can be difficult to compare the accuracy of results generated. We used the Most Probable Number (MPN) technique to evaluate the performance and detection limits of six rapid kits for detecting Listeria in seafood and on an environmental surface compared with the standard protocol. Three seafood products and an environmental surface were inoculated with similar known cell concentrations of Listeria and analyzed according to the manufacturers' instructions. The MPN was estimated using the MPN-BAM spreadsheet. For the seafood products no differences were observed among the rapid kits and efficiency was similar to the BAM method. On the environmental surface the BAM protocol had a higher recovery rate (sensitivity) than any of the rapid kits tested. Clearview™, Reveal®, TECRA® and VIDAS® LDUO detected the cells but only at high concentrations (>10(2) CFU/10 cm(2)). Two kits (VIP™ and Petrifilm™) failed to detect 10(4) CFU/10 cm(2). The MPN method was a useful tool for comparing the results generated by these presence/absence test kits. There remains a need to develop a rapid and sensitive method for detecting Listeria in environmental samples that performs as well as the BAM protocol, since none of the rapid tests used in this study achieved a satisfactory result. Copyright © 2011 Elsevier B.V. All rights reserved.

Heritable, De Novo Resistance to Leaf Rust and Other Novel Traits in Selfed Descendants of Wheat Responding to Inoculation with Wheat Streak Mosaic Virus

PubMed Central

Seifers, Dallas L.; Haber, Steve; Martin, Terry J.; McCallum, Brent D.

2014-01-01

Stable resistance to infection with Wheat streak mosaic virus (WSMV) can be evolved de novo in selfing bread wheat lines subjected to cycles of WSMV inoculation and selection of best-performing plants or tillers. To learn whether this phenomenon might be applied to evolve resistance de novo to pathogens unrelated to WSMV, we examined the responses to leaf rust of succeeding generations of the rust- and WSMV-susceptible cultivar ‘Lakin’ following WSMV inoculation and derived rust-resistant sublines. After three cycles of the iterative protocol five plants, in contrast to all others, expressed resistance to leaf and stripe rust. A subset of descendant sublines of one of these, ‘R1’, heritably and uniformly expressed the new trait of resistance to leaf rust. Such sublines, into which no genes from a known source of resistance had been introgressed, conferred resistance to progeny of crosses with susceptible parents. The F1 populations produced from crosses between, respectively, susceptible and resistant ‘Lakin’ sublines 4-3-3 and 4-12-3 were not all uniform in their response to seedling inoculation with race TDBG. In seedling tests against TDBG and MKPS races the F2s from F1 populations that were uniformly resistant had 3∶1 ratios of resistant to susceptible individuals but the F2s from susceptible F1 progenitors were uniformly susceptible. True-breeding lines derived from resistant individuals in F2 populations were resistant to natural stripe and leaf rust inoculum in the field, while the ‘Lakin’ progenitor was susceptible. The next generation of six of the ‘Lakin’-derived lines exhibited moderate to strong de novo resistance to stem rust races TPMK, QFCS and RKQQ in seedling tests while the ‘Lakin’ progenitor was susceptible. These apparently epigenetic effects in response to virus infection may help researchers fashion a new tool that expands the range of genetic resources already available in adapted germplasm. PMID:24497941

SEE locomotor behavior test discriminates C57BL/6J and DBA/2J mouse inbred strains across laboratories and protocol conditions.

PubMed

Kafkafi, Neri; Lipkind, Dina; Benjamini, Yoav; Mayo, Cheryl L; Elmer, Gregory I; Golani, Ilan

2003-06-01

Conventional tests of behavioral phenotyping frequently have difficulties differentiating certain genotypes and replicating these differences across laboratories and protocol conditions. This study explores the hypothesis that automated tests can be designed to quantify ethologically relevant behavior patterns that more readily characterize heritable and replicable phenotypes. It used SEE (Strategy for the Exploration of Exploration) to phenotype the locomotor behavior of the C57BL/6 and DBA/2 mouse inbred strains across 3 laboratories. The 2 genotypes differed in 15 different measures of behavior, none of which had a significant genotype-laboratory interaction. Within the same laboratory, most of these differences were replicated in additional experiments despite the test photoperiod phase being changed and saline being injected. Results suggest that well-designed tests may considerably enhance replicability across laboratories.

Cast Iron Inoculation Enhanced by Supplementary Oxy-sulfides Forming Elements

NASA Astrophysics Data System (ADS)

Riposan, Iulian; Stan, Stelian; Uta, Valentin; Stefan, Ion

2017-09-01

Inoculation is one of the most important metallurgical treatments applied to the molten cast iron immediately prior to casting, to promote solidification without excessive eutectic undercooling, which favors carbides formation usually with undesirable graphite morphologies. The paper focused on the separate addition of an inoculant enhancer alloy [S, O, oxy-sulfides forming elements] with a conventional Ca-FeSi alloy, in the production of gray and ductile cast irons. Carbides formation tendency decreased with improved graphite characteristics as an effect of the [Ca-FeSi + Enhancer] inoculation combination, when compared to other Ca/Ca, Ba/Ca, RE-FeSi alloy treatments. Adding an inoculant enhancer greatly enhances inoculation, lowers inoculant consumption up to 50% or more and avoids the need to use more costly inoculants, such as a rare earth bearing alloy. The Inoculation Specific Factor [ISF] was developed as a means to more realistically measure inoculant treatment efficiency. It compares the ratio between the improved characteristic level and total inoculant consumption for this effect. Addition of any of the commercial inoculants plus the inoculant enhancer offered outstanding inoculation power [increased ISF] even at higher solidification cooling rates, even though the total enhancer addition was at a small fraction of the amount of commercial inoculant used.

Petunia (Petunia hybrida).

PubMed

Lutke, W Kevin

2006-01-01

Petunia hybrida genetic transformation continues to be a valuable tool for genetic research into biochemical pathways and gene expression, as well as generating commercial products with varying floral colors. In this chapter, we describe a simple and reproducible genetic transformation protocol for generating transgenic petunia plants harboring a gene of interest and selectable marker. The system utilizes Agrobacterium tumefaciens for transgene integration with plant recovery via shoot organogenesis from leaf explant material. Selection for transgenic plants is achieved using the bar gene conferring resistance to glufosinate or nptII gene for resistance to kanamycin. Transformation efficiencies of around 10% are achievable with shoots being recovered about 8 wk after transgene insertion and rooted plants transferred to the greenhouse about twelve weeks after inoculation.

Autoimmune Myocarditis, Valvulitis, and Cardiomyopathy

PubMed Central

Myers, Jennifer M.; Cunningham, Madeleine W.; Fairweather, DeLisa; Huber, Sally A.

2013-01-01

Cardiac myosin-induced autoimmune myocarditis (EAM) is a model of inflammatory heart disease initiated by CD4+ T cells (Smith and Allen 1991; Li, Heuser et al. 2004). It is a paradigm of the immune-mediated cardiac damage believed to play a role in the pathogenesis of a subset of postinfectious human cardiomyopathies (Rose, Herskowitz et al. 1993). Myocarditis is induced in susceptible mice by immunization with purified cardiac myosin (Neu, Rose et al. 1987) or specific peptides derived from cardiac myosin (Donermeyer, Beisel et al. 1995; Pummerer, Luze et al. 1996) (see Basic Protocol 1), or by adoptive transfer of myosin-reactive T cells (Smith and Allen 1991) (see Alternate Protocol). Myocarditis has been induced in Lewis rats by immunization with purified rat or porcine cardiac myosin (Kodama, Matsumoto et al. 1990; Li, Heuser et al. 2004) (see Basic Protocol 2) or S2-16 peptide (Li, Heuser et al. 2004), or by adoptive transfer of T cells stimulated by specific peptides derived from cardiac myosin (Wegmann, Zhao et al. 1994). Myocarditis begins 12 to 14 days after the first immunization, and is maximal after 21 days. Other animal models commonly used to study myocarditis development include the pathogen-induced models in which disease is initiated by viral infection. The first murine model of acute viral myocarditis causes sudden death via viral damage to cardiomyocytes (Huber, Gauntt et al. 1998; Horwitz, La Cava et al. 2000; Fong 2003; Fuse, Chan et al. 2005; Fairweather and Rose 2007; Cihakova and Rose 2008) whereas the second model is based on inoculation with heart-passaged coxsackievirus B3 (CVB3) that includes damaged heart proteins (Fairweather, Frisancho-Kiss et al. 2004; Fairweather D 2004; Fairweather and Rose 2007; Cihakova and Rose 2008) In addition to the protocols used to induce EAM in mice and rats, support protocols are included for preparing purified cardiac myosin using mouse or rat heart tissue (see Support Protocol 1), preparing purified cardiac myosin for injection (see Support Protocol 2), and collecting and assessing hearts by histopathological means (see Support Protocol 3). PMID:23564686

Accessing inoculation methods of maize and wheat with Azospirillum brasilense.

PubMed

Fukami, Josiane; Nogueira, Marco Antonio; Araujo, Ricardo Silva; Hungria, Mariangela

2016-03-01

The utilization of inoculants containing Azospirillum is becoming more popular due to increasing reports of expressive gains in grain yields. However, incompatibility with pesticides used in seed treatments represents a main limitation for a successful inoculation. Therefore, in this study we searched for alternatives methods for seed inoculation of maize and wheat, aiming to avoid the direct contact of bacteria with pesticides. Different doses of inoculants containing Azospirillum brasilense were employed to perform inoculation in-furrow, via soil spray at sowing and via leaf spray after seedlings had emerged, in comparison to seed inoculation. Experiments were conducted first under greenhouse controlled conditions and then confirmed in the field at different locations in Brazil. In the greenhouse, most parameters measured responded positively to the largest inoculant dose used in foliar sprays, but benefits could also be observed from both in-furrow and soil spray inoculation. However, our results present evidence that field inoculation with plant-growth promoting bacteria must consider inoculant doses, and point to the need of fine adjustments to avoid crossing the threshold of growth stimulation and inhibition. All inoculation techniques increased the abundance of diazotrophic bacteria in plant tissues, and foliar spray improved colonization of leaves, while soil inoculations favored root and rhizosphere colonization. In field experiments, inoculation with A. brasilense allowed for a 25 % reduction in the need for N fertilizers. Our results have identified alternative methods of inoculation that were as effective as the standard seed inoculation that may represent an important strategy to avoid the incompatibility between inoculant bacteria and pesticides employed for seed treatment.

Tranexamic acid attenuates oleic-acid-induced pulmonary extravasation.

PubMed

Moriuchi, H; Arai, I; Yuizono, T

1995-12-01

Activation of fibrinolysis is implicated in the development of vascular injury in certain lung injuries. It has yet to be reported that activation of plasmin is involved in extravasation caused by oleic acid (OA). We examined whether or not plasmin is involved in pulmonary extravasation by OA. Prospective trial. University laboratory. A total of 78 guinea pigs (498.9 +/- 10.6 g). Evans blue (EB) was administered to anesthetized guinea pigs. Subsequently four protocols were followed: (1) After 1 min, 60 micro l/kg of OA was injected. Perfusion was performed 30, 60 or 90 min after OA injection to wash out intravascular EB. (2) After 1 min, 15, 30 or 60 micro l/kg of OA was injected. (3) Tranexamic acid (TA) (2 g/kg) or saline was administered 30 min before OA (15 micro l/kg) injection. (4) Diphenhydramine hydrochloride (2.9 mg/kg) or saline was administered 7 min before OA (15 micro l/kg) injection. Except in protocol 1, the chest cavity was opened 90 min after OA injection. Perfusion was then performed. Airway was separated into four parts from trachea to distal bronchus. EB was extracted from the tissues and measured. OA caused an extravasation throughout airways in a time- and dose-dependent manner. Extravasation was more conspicuous in peripheral tissues. TA significantly attenuated extravasation, while diphenhydramine hydrochloride did not. It is suggested that plasmin, but not histamine, is involved in extravasation by OA. Inhibition of plasmin can be an effective strategy for treatment of this kind of lung injury.

The addition of albumin improves Schwann cells viability in nerve cryopreservation.

PubMed

González Porto, Sara Alicia; Domenech, Nieves; González Rodríguez, Alba; Avellaneda Oviedo, Edgar Mauricio; Blanco, Francisco J; Arufe Gonda, María C; Álvarez Jorge, Ángel; Sánchez Ibañez, Jacinto; Rendal Vázquez, Esther

2018-04-26

The purpose of the current study was to establish a valid protocol for nerve cryopreservation, and to evaluate if the addition of albumin supposed any advantage in the procedure. We compared a traditional cryopreservation method that uses dimethyl sulfoxide (DMSO) as cryoprotectant, to an alternative method that uses DMSO and albumin. Six Wistar Lewis rats were used to obtain twelve 20 mm fragments of sciatic nerve. In the first group, six fragments were cryopreserved in 199 media with 10% DMSO, with a temperature decreasing rate of 1 °C per minute. In the second group, six fragments were cryopreserved adding 4% human albumin. The unfreezing process consisted of sequential washings with saline in the first group, and saline and 20% albumin in the second group at 37 °C until the crioprotectant was removed. Structural evaluation was performed through histological analysis and electronic microscopy. The viability was assessed with the calcein-AM (CAM) and 4',6-diamino-2-fenilindol (DAPI) staining. Histological results showed a correct preservation of peripheral nerve architecture and no significant differences were found between the two groups. However, Schwann cells viability showed in the CAM-DAPI staining was significantly superior in the albumin group. The viability of Schwann cells was significantly increased when albumin was added to the nerve cryopreservation protocol. However, no significant structural differences were found between groups. Further studies need to be performed to assess the cryopreserved nerve functionality using this new method.

Does Osmotic Stress Affect Natural Product Expression in Fungi?

PubMed Central

Overy, David; Chi, Wei-Chiung; Pang, Ka-Lai; Rateb, Mostafa; Shang, Zhuo; Capon, Rob; Bills, Gerald; Kerr, Russell

2017-01-01

The discovery of new natural products from fungi isolated from the marine environment has increased dramatically over the last few decades, leading to the identification of over 1000 new metabolites. However, most of the reported marine-derived species appear to be terrestrial in origin yet at the same time, facultatively halo- or osmotolerant. An unanswered question regarding the apparent chemical productivity of marine-derived fungi is whether the common practice of fermenting strains in seawater contributes to enhanced secondary metabolism? To answer this question, a terrestrial isolate of Aspergillus aculeatus was fermented in osmotic and saline stress conditions in parallel across multiple sites. The ex-type strain of A. aculeatus was obtained from three different culture collections. Site-to-site variations in metabolite expression were observed, suggesting that subculturing of the same strain and subtle variations in experimental protocols can have pronounced effects upon metabolite expression. Replicated experiments at individual sites indicated that secondary metabolite production was divergent between osmotic and saline treatments. Titers of some metabolites increased or decreased in response to increasing osmolite (salt or glycerol) concentrations. Furthermore, in some cases, the expression of some secondary metabolites in relation to osmotic and saline stress was attributed to specific sources of the ex-type strains. PMID:28805714

An Electrochemistry Study of Cryoelectrolysis in Frozen Physiological Saline.

PubMed

Manuel, Thomas J; Munnangi, Pujita; Rubinsky, Boris

2017-07-01

Cryoelectrolysis is a new minimally invasive tissue ablation surgical technique that combines the processes of electrolysis and solid/liquid phase transformation (freezing). This study investigated this new technique by measuring the pH front propagation and the changes in resistance in a tissue simulant made of physiological saline gel with a pH dye as a function of the sample temperature in the high subzero range above the eutectic. Results demonstrated that effective electrolysis can occur in a high subzero freezing milieu and that the propagation of the pH front is only weakly dependent on temperature. These observations are consistent with a mechanism involving ionic movement through the concentrated saline solution channels between ice crystals at subfreezing temperatures above the eutectic. Moreover, results suggest that Joule heating in these microchannels may cause local microscopic melting, the observed weak dependence of pH front propagation on temperature, and the large changes in resistance with time. A final insight provided by the results is that the pH front propagation from the anode is more rapid than from the cathode, a feature indicative of the electro-osmotic flow from the cathode to the anode. The findings in this paper may be critical for designing future cryoelectrolytic ablation surgery protocols.

Myocardial protection induced by fentanyl in pigs exposed to high-dose adrenaline.

PubMed

da Luz, Vinicius Fernando; Otsuki, Denise Aya; Gonzalez, Maria Margarita Castro; Negri, Elnara Marcia; Caldini, Elia Garcia; Damaceno-Rodrigues, Nilsa Regina; Malbouisson, Luiz Marcelo Sá; Viana, Bruno Gonçalves; Vane, Matheus Fachini; Carmona, Maria Jose Carvalho

2015-10-01

The use of high doses of adrenaline is common in critical patients, especially during cardiac arrest. During these situations, myocardial dysfunction can be a result of multiple factors, including adrenaline use. In addition, opioids have been shown to have anti-arrhythmic and anti-ischemic mechanisms that may confer cardiac protection. This study aimed to evaluate the effects of fentanyl on myocardial function in pigs exposed to high-dose adrenaline. After institutional ethics committee approval, 26 pigs were randomly allocated to receive either 20 μg/kg fentanyl (n = 10; fentanyl group) administered 5 min before five doses of adrenaline (20 μg/kg), equivalent-volume saline (n = 10; saline group) using the same adrenaline dosing protocol, or neither fentanyl nor adrenaline (n = 6; sham group). The fentanyl group showed lower levels of troponin at the end of the sixth hour compared with the saline group (1.91 ± 1.47 vs 5.44 ± 5.35 ng/mL, P = 0.019). Transmission electron microscopy and immunohistochemistry also showed less myocardial injury in the fentanyl group. The conclusion was reached that fentanyl attenuates myocardial injury caused by high-dose adrenaline without blunting the hemodynamic effect of adrenaline. © 2015 Wiley Publishing Asia Pty Ltd.

Prediction of Greenhouse Gas (GHG) Fluxes from Coastal Salt Marshes using Artificial Neural Network

NASA Astrophysics Data System (ADS)

Ishtiaq, K. S.; Abdul-Aziz, O. I.

2017-12-01

Coastal salt marshes are among the most productive ecosystems on earth. Given the complex interactions between ambient environment and ecosystem biological exchanges, it is difficult to predict the salt marsh greenhouse gas (GHG) fluxes (CO2 and CH4) from their environmental drivers. In this study, we developed an artificial neural network (ANN) model to robustly predict the salt marsh GHG fluxes using a limited number of input variables (photosynthetically active radiation, soil temperature and porewater salinity). The ANN parameterization involved an optimized 3-layer feed forward Levenberg-Marquardt training algorithm. Four tidal salt marshes of Waquoit Bay, MA — incorporating a gradient in land-use, salinity and hydrology — were considered as the case study sites. The wetlands were dominated by native Spartina Alterniflora, and characterized by high salinity and frequent flooding. The developed ANN model showed a good performance (training R2 = 0.87 - 0.96; testing R2 = 0.84 - 0.88) in predicting the fluxes across the case study sites. The model can be used to estimate wetland GHG fluxes and potential carbon balance under different IPCC climate change and sea level rise scenarios. The model can also aid the development of GHG offset protocols to set monitoring guidelines for restoration of coastal salt marshes.

[Effect of ectomycorrhizae on the growth of Picea koraiensis seedlings].

PubMed

Song, Rui-Qing; Wu, Ke

2005-12-01

Basidioscarps of Agaricales in different Picea koraiensis forest plantations were collected during August-October, 2000. 36 isolaters of species of Agaricales were obtained by isolating and culturing to the basidioscarps. Through indoor inoculation test on seedlings of Picea koraiensis, 6 ectomycorrhizae fungi cultures were obtained from 36 isolaters. The inoculation results show that the period for ectomycorrhizae inoculation to 1-year seedlings of Picea koraiensis should be about 30 days after seedlings emerging, the suitable temperature for ectomycorrhizae forming is about 20 degrees C. 6 ectomycorrhizae strains all have growth-promoting effect to the seedlings of Picea koraiensis. The contents of chlorophyll a of the seedlings inoculated strains of Agaricus silvaticus, 031 and L15 were significantly higher than other strains and control. The contents of chlorophyll b in the seedlings inoculated strains 009, 004, Agaricus silvaticus and L15 were significantly higher than other strains and control. The weights of seedlings which inoculated strains 009, 025, 031, Agaricus silvaticus and L15 were significantly different to control, the weight of seedlings inoculated strains of Agaricus silvaticus and L15 are 19.23% and 23.08% more than control; The heights of the seedlings inoculated 6 strains all have significant difference to control, the weight of seedlings inoculated strains of Agaricus silvaticus and L15 are 17.83% and 16.37% more than control. The results of outdoor inoculation show that the seedlings inoculated Agaricus silvaticus grow best on height, 9.25% more than control after inoculated 70 days; the seedlings inoculated strain L15 grow best on collar diameter, 9.92% more than control after inoculated 70 days; the lateral root numbers of seedlings inoculated strain 009 is largest, 51.91% more than control after inoculated 70 days; the main roots of seedlings inoculated strain 009 are longest, 3.36% more than control after inoculated 70 days; the ectomycorrhizae fungi infestation percentage of seedlings inoculated strain 031 is highest, 13.90% more than control after inoculated 70 days; the photosynthesis of seedlings inoculated strain 031 is more powerful, 20.02% more than control after inoculated 70 days. To sum up, strain 009, strain 031 and strain L15 should be fine strains to seedlings of Picea koraiensis.

Inexpensive metagenomic DNA extraction protocol with high quality from marine sediments contaminated by petroleum hydrocarbons.

PubMed

García-Bautista, I; Toledano-Thompson, T; Dantán-González, E; González-Montilla, J; Valdez-Ojeda, R

2017-09-21

Marine environments are a reservoir of relevant information on dangerous contaminants such as hydrocarbons, as well as microbial communities with probable degradation skills. However, to access microbial diversity, it is necessary to obtain high-quality DNA. An inexpensive, reliable, and effective metagenomic DNA (mgDNA) extraction protocol from marine sediments contaminated with petroleum hydrocarbons was established in this study from modifications to Zhou's protocol. The optimization included pretreatment of sediment with saline solutions for the removal of contaminants, a second precipitation and enzymatic degradation of RNA, followed by purification of mgDNA extracted by electroelution. The results obtained indicated that the modifications applied to 12 sediments with total petroleum hydrocarbon (TPH) concentrations from 22.6-174.3 (µg/g dry sediment) yielded 20.3-321.3 ng/µL mgDNA with A 260 /A 280 and A 260 /A 230 ratios of 1.75 ± 0.08 and 1.19 ± 0.22, respectively. The 16S rRNA amplification confirmed the purity of the mgDNA. The suitability of this mgDNA extraction protocol lies in the fact that all chemical solutions utilized are common in all molecular biology laboratories, and the use of dialysis membrane does not require any sophisticated or expensive equipment, only an electrophoretic chamber.

Analytical platform for metabolome analysis of microbial cells using methyl chloroformate derivatization followed by gas chromatography-mass spectrometry.

PubMed

Smart, Kathleen F; Aggio, Raphael B M; Van Houtte, Jeremy R; Villas-Bôas, Silas G

2010-09-01

This protocol describes an analytical platform for the analysis of intra- and extracellular metabolites of microbial cells (yeast, filamentous fungi and bacteria) using gas chromatography-mass spectrometry (GC-MS). The protocol is subdivided into sampling, sample preparation, chemical derivatization of metabolites, GC-MS analysis and data processing and analysis. This protocol uses two robust quenching methods for microbial cultures, the first of which, cold glycerol-saline quenching, causes reduced leakage of intracellular metabolites, thus allowing a more reliable separation of intra- and extracellular metabolites with simultaneous stopping of cell metabolism. The second, fast filtration, is specifically designed for quenching filamentous micro-organisms. These sampling techniques are combined with an easy sample-preparation procedure and a fast chemical derivatization reaction using methyl chloroformate. This reaction takes place at room temperature, in aqueous medium, and is less prone to matrix effect compared with other derivatizations. This protocol takes an average of 10 d to complete and enables the simultaneous analysis of hundreds of metabolites from the central carbon metabolism (amino and nonamino organic acids, phosphorylated organic acids and fatty acid intermediates) using an in-house MS library and a data analysis pipeline consisting of two free software programs (Automated Mass Deconvolution and Identification System (AMDIS) and R).

Design and performance testing of a real-time PCR assay for sensitive and reliable direct quantification of Brettanomyces in wine.

PubMed

Tessonnière, H; Vidal, S; Barnavon, L; Alexandre, H; Remize, F

2009-02-28

Because the yeast Brettanomyces produces volatile phenols and acetic acid, it is responsible for wine spoilage. The uncontrolled accumulation of these molecules in wine leads to sensorial defects that compromise wine quality. The need for a rapid, specific, sensitive and reliable method to detect this spoilage yeast has increased over the last decade. All these requirements are met by real-time PCR. We here propose improvements of existing methods to enhance the robustness of the assay. Six different protocols to isolate DNA from a wine and three PCR mix compositions were tested, and the best method was selected. Insoluble PVPP addition during DNA extraction by a classical phenol:chloroform protocol succeeded in the relief of PCR inhibitors from wine. We developed an internal control which was efficient to avoid false negative results due to decreases in the efficiency of DNA isolation and/or amplification. The method was evaluated by an intra-laboratory study for its specificity, linearity, repeatability and reproducibility. A standard curve was established from 14 different wines artificially inoculated. The quantification limit was 31 cfu/mL.

Ghrelin Pre-treatment Attenuates Local Oxidative Stress and End Organ Damage During Cardiopulmonary Bypass in Anesthetized Rats

PubMed Central

Sukumaran, Vijayakumar; Tsuchimochi, Hirotsugu; Fujii, Yutaka; Hosoda, Hiroshi; Kangawa, Kenji; Akiyama, Tsuyoshi; Shirai, Mikiyasu; Tatsumi, Eisuke; Pearson, James T.

2018-01-01

Cardiopulmonary bypass (CPB) induced systemic inflammation significantly contributes to the development of postoperative complications, including respiratory failure, myocardial, renal and neurological dysfunction and ultimately can lead to failure of multiple organs. Ghrelin is a small endogenous peptide with wide ranging physiological effects on metabolism and cardiovascular regulation. Herein, we investigated the protective effects of ghrelin against CPB-induced inflammatory reactions, oxidative stress and acute organ damage. Adult male Sprague Dawley rats randomly received vehicle (n = 5) or a bolus of ghrelin (150 μg/kg, sc, n = 5) and were subjected to CPB for 4 h (protocol 1). In separate rats, ghrelin pre-treatment (protocol 2) was compared to two doses of ghrelin (protocol 3) before and after CPB for 2 h followed by recovery for 2 h. Blood samples were taken prior to CPB, and following CPB at 2 h and 4 h. Organ nitrosative stress (3-nitrotyrosine) was measured by Western blotting. CPB induced leukocytosis with increased plasma levels of tumor necrosis factor-α and interleukin-6 indicating a potent inflammatory response. Ghrelin treatment significantly reduced plasma organ damage markers (lactate dehydrogenase, aspartate aminotransferase, alanine aminotransferase) and protein levels of 3-nitrotyrosine, particularly in the brain, lung and liver, but only partly suppressed inflammatory cell invasion and did not reduce proinflammatory cytokine production. Ghrelin partially attenuated the CPB-induced elevation of epinephrine and to a lesser extent norepinephrine when compared to the CPB saline group, while dopamine levels were completely suppressed. Ghrelin treatment sustained plasma levels of reduced glutathione and decreased glutathione disulphide when compared to CPB saline rats. These results suggest that even though ghrelin only partially inhibited the large CPB induced increase in catecholamines and organ macrophage infiltration, it reduced oxidative stress and subsequent cell damage. Pre-treatment with ghrelin might provide an effective adjunct therapy for preventing widespread CPB induced organ injury. PMID:29593559

Standardization and validation of real time PCR assays for the diagnosis of histoplasmosis using three molecular targets in an animal model.

PubMed

López, Luisa F; Muñoz, César O; Cáceres, Diego H; Tobón, Ángela M; Loparev, Vladimir; Clay, Oliver; Chiller, Tom; Litvintseva, Anastasia; Gade, Lalitha; González, Ángel; Gómez, Beatriz L

2017-01-01

Histoplasmosis is considered one of the most important endemic and systemic mycoses worldwide. Until now few molecular techniques have been developed for its diagnosis. The aim of this study was to develop and evaluate three real time PCR (qPCR) protocols for different protein-coding genes (100-kDa, H and M antigens) using an animal model. Fresh and formalin-fixed and paraffin-embedded (FFPE) lung tissues from BALB/c mice inoculated i.n. with 2.5x106 Histoplasma capsulatum yeast or PBS were obtained at 1, 2, 3, 4, 8, 12 and 16 weeks post-infection. A collection of DNA from cultures representing different clades of H. capsulatum (30 strains) and other medically relevant pathogens (36 strains of related fungi and Mycobacterium tuberculosis) were used to analyze sensitivity and specificity. Analytical sensitivity and specificity were 100% when DNAs from the different strains were tested. The highest fungal burden occurred at first week post-infection and complete fungal clearance was observed after the third week; similar results were obtained when the presence of H. capsulatum yeast cells was demonstrated in histopathological analysis. In the first week post-infection, all fresh and FFPE lung tissues from H. capsulatum-infected animals were positive for the qPCR protocols tested except for the M antigen protocol, which gave variable results when fresh lung tissue samples were analyzed. In the second week, all qPCR protocols showed variable results for both fresh and FFPE tissues. Samples from the infected mice at the remaining times post-infection and uninfected mice (controls) were negative for all protocols. Good agreement was observed between CFUs, histopathological analysis and qPCR results for the 100-kDa and H antigen protocols. We successfully standardized and validated three qPCR assays for detecting H. capsulatum DNA in fresh and FFPE tissues, and conclude that the 100-kDa and H antigen molecular assays are promising tests for diagnosing this mycosis.

Duplex real-time polymerase chain reaction reveals competition between Erwinia amylovora and E. pyrifoliae on pear blossoms.

PubMed

Lehman, Susan M; Kim, Won-Sik; Castle, Alan J; Svircev, Antonet M

2008-06-01

Erwinia amylovora and E. pyrifoliae are the causative agents of fire blight and Asian pear blight, respectively. The pathogens are closely related, with overlapping host ranges. Data are unavailable on the current distribution of E. pyrifoliae and on the interaction between the two species when they are present together on the same host. In this study, a duplex real-time polymerase chain reaction (PCR) protocol was developed to monitor the population dynamics of E. amylovora and E. pyrifoliae on the surface of Bartlett pear blossoms. Bacterial cells washed from blossoms were used directly as the PCR template without DNA extraction. Primers and a probe based on the E. amylovora levansucrase gene detected all E. amylovora strains. All E. pyrifoliae strains, including the Japanese Erwinia strains previously described as E. amylovora, were detected with a primer and probe combination based on the E. pyrifoliae hrpW gene. Disease development and severity were not significantly different in blossoms inoculated with individual Erwinia species or with a mixture of the two species. However, E. amylovora grew to greater population sizes than did E. pyrifoliae in both single species inoculations and in mixtures, suggesting that E. amylovora has a greater competitive fitness on Bartlett pear blossoms than E. pyrifoliae.

A DNA vaccine against dolphin morbillivirus is immunogenic in bottlenose dolphins.

PubMed

Vaughan, Kerrie; Del Crew, Jason; Hermanson, Gary; Wloch, Mary K; Riffenburgh, Robert H; Smith, Cynthia R; Van Bonn, William G

2007-12-15

The immunization of exotic species presents considerable challenges. Nevertheless, for facilities like zoos, animal parks, government facilities and non-profit conservation groups, the protection of valuable and endangered species from infectious disease is a growing concern. The rationale for immunization in these species parallels that for human and companion animals; to decrease the incidence of disease. The U.S. Navy Marine Mammal Program, in collaboration with industry and academic partners, has developed and evaluated a DNA vaccine targeting a marine viral pathogen - dolphin morbillivirus (DMV). The DMV vaccine consists of the fusion (F) and hemagglutinin (H) genes of DMV. Vaccine constructs (pVR-DMV-F and pVR-DMV-H) were evaluated for expression in vitro and then for immunogenicity in mice. Injection protocols were designed for application in Atlantic bottlenose dolphins (Tursiops truncatus) to balance vaccine effectiveness with clinical utility. Six dolphins were inoculated, four animals received both pDMV-F and pDMV-H and two animals received a mock vaccine (vector alone). All animals received an inoculation week 0, followed by two booster injections weeks 8 and 14. Vaccine-specific immune responses were documented in all four vaccinated animals. To our knowledge, this is the first report of pathogen-specific immunogenicity to a DNA vaccine in an aquatic mammal species.

Agrobacterium tumefaciens-mediated transformation of Narcissus tazzeta var. chinensis.

PubMed

Lu, Gang; Zou, Qingcheng; Guo, Deping; Zhuang, Xiaoying; Yu, Xiaolin; Xiang, Xun; Cao, Jiashu

2007-09-01

Phytoene synthase (PSY), as a key regulatory enzyme for carotene biosynthesis, plays an important role in regulating color formation in many species. In the present study, a protocol was developed for the transformation of Narcissus tazzeta var chinensis using Agrobacterium tumefaciens strain LBA4404 harboring a binary vector pCAMBIA1301 plasmid which contained an antisense phytoene synthase gene, a reporter beta-glucuronidase gene and a selectable marker hygromycin phosphotransferase gene. Effects of some factors on efficiency of transformation and regeneration were examined. Preculture of the explants for 6 days before inoculation enhanced the transient GUS expression. The addition of acetosyringone (AS) at 100 micromol l(-1) for inoculation and a period of 3 days co-cultivation yielded efficient transient GUS expression. Transformants were obtained through selection on MS medium containing 5 mg l(-1) 6-benzylaminopurine (BA), 0.1 mg l(-1)alpha-naphthalene acetic acid (NAA) and 40 mg l(-1) hygromycin. The transformation frequency was 1.24% based on PCR analysis of gus gene. One or two copies of transgene were demonstrated in different transformations by Southern blotting analyses. Northern blotting results confirmed that the transcription of the endogenous psy gene in transgenic plants was inhibited or silenced. The method reported here provides new opportunities for improvement of quality traits of Narcissus tazzeta via genetic transformation.

Cerebral Blood Flow and Cerebral Edema in Rats With Diabetic Ketoacidosis

PubMed Central

Yuen, Natalie; Anderson, Steven E.; Glaser, Nicole; Tancredi, Daniel J.; O'Donnell, Martha E.

2008-01-01

OBJECTIVE— Cerebral edema (CE) is a potentially life-threatening complication of diabetic ketoacidosis (DKA) in children. Osmotic fluctuations during DKA treatment have been considered responsible, but recent data instead suggest that cerebral hypoperfusion may be involved and that activation of cerebral ion transporters may occur. Diminished cerebral blood flow (CBF) during DKA, however, has not been previously demonstrated. We investigated CBF and edema formation in a rat model of DKA and determined the effects of bumetanide, an inhibitor of Na-K-Cl cotransport. RESEARCH DESIGN AND METHODS— Juvenile rats with streptozotocin-induced DKA were treated with intravenous saline and insulin, similar to human treatment protocols. CBF was determined by magnetic resonance (MR) perfusion–weighted imaging before and during treatment, and CE was assessed by determining apparent diffusion coefficients (ADCs) using MR diffusion–weighted imaging. RESULTS— CBF was significantly reduced in DKA and was responsive to alterations in pCO2. ADC values were reduced, consistent with cell swelling. The reduction in ADCs correlated with dehydration, as reflected in blood urea nitrogen concentrations. Bumetanide caused a rapid rise in ADCs of DKA rats without significantly changing CBF, while saline/insulin caused a rapid rise in CBF and a gradual rise in ADCs. DKA rats treated with bumetanide plus saline/insulin showed a trend toward more rapid rise in cortical ADCs and a larger rise in striatal CBF than those observed with saline/insulin alone. CONCLUSIONS— These data demonstrate that CE in DKA is accompanied by cerebral hypoperfusion before treatment and suggest that blocking Na-K-Cl cotransport may reduce cerebral cell swelling. PMID:18633109

Stable expression of mtlD gene imparts multiple stress tolerance in finger millet.

PubMed

Hema, Ramanna; Vemanna, Ramu S; Sreeramulu, Shivakumar; Reddy, Chandrasekhara P; Senthil-Kumar, Muthappa; Udayakumar, Makarla

2014-01-01

Finger millet is susceptible to abiotic stresses, especially drought and salinity stress, in the field during seed germination and early stages of seedling development. Therefore developing stress tolerant finger millet plants combating drought, salinity and associated oxidative stress in these two growth stages is important. Cellular protection through osmotic adjustment and efficient free radical scavenging ability during abiotic stress are important components of stress tolerance mechanisms in plants. Mannitol, an osmolyte, is known to scavenge hydroxyl radicals generated during various abiotic stresses and thereby minimize stress damage in several plant species. In this study transgenic finger millet plants expressing the mannitol biosynthetic pathway gene from bacteria, mannitol-1-phosphate dehydrogenase (mtlD), were developed through Agrobacterium tumefaciens-mediated genetic transformation. mtlD gene integration in the putative transgenic plants was confirmed by Southern blot. Further, performance of transgenic finger millet under drought, salinity and oxidative stress was studied at plant level in T1 generation and in T1 and T2 generation seedlings. Results from these experiments showed that transgenic finger millet had better growth under drought and salinity stress compared to wild-type. At plant level, transgenic plants showed better osmotic adjustment and chlorophyll retention under drought stress compared to the wild-type. However, the overall increase in stress tolerance of transgenics for the three stresses, especially for oxidative stress, was only marginal compared to other mtlD gene expressing plant species reported in the literature. Moreover, the Agrobacterium-mediated genetic transformation protocol developed for finger millet in this study can be used to introduce diverse traits of agronomic importance in finger millet.

Effect of actinobacteria agent inoculation methods on cellulose degradation during composting based on redundancy analysis.

PubMed

Zhao, Yue; Lu, Qian; Wei, Yuquan; Cui, Hongyang; Zhang, Xu; Wang, Xueqin; Shan, Si; Wei, Zimin

2016-11-01

In this study, actinobacteria agent including Streptomyces sp. and Micromonospora sp. were inoculated during chicken manure composting by different inoculation methods. The effect of different treatments on cellulose degradation and the relationship between inoculants and indigenous actinobacteria were investigated during composting. The results showed that inoculation in different stages of composting all improved the actinobacteria community diversity particularly in the cooling stage of composting (M3). Moreover, inoculation could distinctly accelerate the degradation of organic matters (OM) especially celluloses. Redundancy analysis indicated that the correlation between indigenous actinobacteria and degradation of OM and cellulose were regulated by inoculants and there were significant differences between different inoculation methods. Furthermore, synergy between indigenous actinobacteria and inoculants for degradation of OM and cellulose in M3 was better than other treatments. Conclusively, we suggested an inoculation method to regulate the indigenous actinobacteria based on the relationship between inoculants and indigenous actinobacteria and degradation content. Copyright © 2016 Elsevier Ltd. All rights reserved.

Antigenicity, Immunogenicity and Protective Efficacy of Three Proteins Expressed in the Promastigote and Amastigote Stages of Leishmania infantum against Visceral Leishmaniasis

PubMed Central

Martins, Vivian Tamietti; Chávez-Fumagalli, Miguel Angel; Lage, Daniela Pagliara; Duarte, Mariana Costa; Garde, Esther; Costa, Lourena Emanuele; da Silva, Viviane Gomes; Oliveira, Jamil Silvano; de Magalhães-Soares, Danielle Ferreira; Teixeira, Santuza Maria Ribeiro; Fernandes, Ana Paula; Soto, Manuel; Tavares, Carlos Alberto Pereira; Coelho, Eduardo Antonio Ferraz

2015-01-01

In the present study, two Leishmania infantum hypothetical proteins present in the amastigote stage, LiHyp1 and LiHyp6, were combined with a promastigote protein, IgE-dependent histamine-releasing factor (HRF); to compose a polyproteins vaccine to be evaluated against L. infantum infection. Also, the antigenicity of the three proteins was analyzed, and their use for the serodiagnosis of canine visceral leishmaniasis (CVL) was evaluated. The LiHyp1, LiHyp6, and HRF DNA coding sequences were cloned in prokaryotic expression vectors and the recombinant proteins were purified. When employed in ELISA assays, all proteins were recognized by sera from visceral leishmaniasis (VL) dogs, and presented no cross-reactivity with either sera from dogs vaccinated with a Brazilian commercial vaccine, or sera of Trypanosoma cruzi-infected or Ehrlichia canis-infected animals. In addition, the antigens were not recognized by antibodies from non-infected animals living in endemic or non-endemic areas for leishmaniasis. The immunogenicity and protective efficacy of the three proteins administered in the presence of saponin, individually or in combination (composing a polyproteins vaccine), were evaluated in a VL murine model: BALB/c mice infected with L. infantum. Spleen cells from mice inoculated with the individual proteins or with the polyproteins vaccine plus saponin showed a protein-specific production of IFN-γ, IL-12, and GM-CSF after an in vitro stimulation, which was maintained after infection. These animals presented significant reductions in the parasite burden in different evaluated organs, when compared to mice inoculated with saline or saponin. The decrease in parasite burden was associated with an IL-12-dependent production of IFN-γ against parasite total extracts (produced mainly by CD4+ T cells), correlated to the induction of parasite proteins-driven NO production. Mice inoculated with the recombinant protein-based vaccines showed also high levels of parasite-specific IgG2a antibodies. The polyproteins vaccine administration induced a more pronounced Th1 response before and after challenge infection than individual vaccines, which was correlated to a higher control of parasite dissemination to internal organs. PMID:26367128

Effect of toad skin extracts on the pain behavior of cancer model mice and its peripheral mechanism of action.

PubMed

Chen, Tao; Yuan, Shen-Jun; Yu, Xue-Qin; Jiao, Liang-Bo; Hu, Wei; Chen, Wang-Long; Xie, Bo

2017-01-01

The changes in thermal and mechanical hyperalgesia in paw cancer pain model mice and the action mechanism of toad skin extracts (TSE) was investigated. Eighty female mice were subcutaneously injected with saline or inoculated with H22 hepatoma cells in the right hind paw and administration with saline, vehicle, morphine and TSE. The pain behavior was recorded before treatment and at 0.5, 1.0, 1.5, 3 and 6h after initial administration, and thereafter on the 2nd, 4th, 6th, and 8th day after administration. On the last day, samples were collected after the euthanasia for the detection of β-END, CRF, IL-1β, POMC, μ-OR, CD3+, CD8+ and CD4+ in sera and the tumor tissues. The results showed that TSE significantly increased the thresholds of thermal pain and mechanical pain, and upregulated the expressions of β-END, CRF, POMC, CD3+, CD8+ and μ-OR, and downregulated the expression of CD4+. These results indicate that TSE significantly relieved pain in cancer pain model mice and raised their pain threshold. In addition, TSE seems to play a prominent role in promoting the activity of tumor infiltrating lymphocytes (TILs, CD3+ and CD8+ T cells), and this immune-cell-derived peripheral analgesic pathway might have widespread potential for clinical use. Copyright © 2016 Elsevier B.V. All rights reserved.

Safety of refrigerated storage of admixed parenteral fluids.

PubMed Central

Weil, D C; Arnow, P M

1988-01-01

Many hospital pharmacies are reluctant to store admixed parenteral fluids longer than 24 to 48 h because of concern about possible microbial contamination. We evaluated the safety of prolonged refrigerated storage of admixtures by culturing mixtures in 471 bags prepared routinely in a hospital pharmacy and stored at 4 degrees C for up to 15 days. Low-level contamination (1 CFU per bag) was found in 3 of 253 solutions of saline and/or glucose into which nonantibiotic additives had been injected. None of 171 saline and/or glucose solutions with antibiotic additives or 47 parenteral nutrition fluids was culture positive. The risk of contamination did not increase with duration of storage, and none of 107 bags stored greater than or equal to 5 days was culture positive. Laboratory studies to measure growth of bacteria and fungi in glucose infusate and parenteral nutrition solutions confirmed that storage at 4 degrees C suppresses growth. Eight of twelve bacterial isolates grew in glucose solutions at 25 degrees C, while none grew at 4 degrees C. Of 13 species of bacteria and fungi inoculated in parenteral nutrition fluids, 8 proliferated at 25 degrees C and none proliferated at 4 degrees C. We conclude that refrigerated storage of parenteral fluids for up to 1 week following admixture, as permitted by Centers for Disease Control guidelines, does not appear to increase the risk of microbial contamination when standard aseptic procedures for admixing and storage are followed. PMID:3183025

Preliminary in vivo evaluation of [131I]-2-iodo-D-phenylalanine as a potential radionuclide therapeutic agent in R1M-fluc rhabdomyosarcoma tumor-bearing NuNu mice using bioluminescent imaging.

PubMed

Bauwens, Matthias; Wimana, Lena; Keyaerts, Marleen; Peleman, Cindy; Lahoutte, Tony; Kersemans, Ken; Snykers, Sarah; Vinken, Mathieu; Mertens, John; Bossuyt, Axel

2010-04-01

Carrier-added [(123)I]-2-iodo-D-phenylalanine (CA [(123)I]-2-I-D-Phe) was previously found to have a preferential retention in tumors with a high tumor background contrast in animal models. A previous human dosimetry study demonstrated a favorable biodistribution and radiation burden in human subjects. The aim of this study was to investigate the potential of CA [(131)I]-2-I-D-Phe as an agent for radionuclide therapy. Sixty (60) nude athymic mice were inoculated subcutaneously with firefly luciferase-transduced R1M rhabdomyosarcoma cells. The mice in the therapy group were injected intravenously (i.v.) with 148 MBq [(131)I]-2-I-D-Phe (432 GBq/mmol) in kit solution. Controls were injected with kit solution without radioactivity, with physiological saline, or with 148 MBq [(131)I](-) in physiological saline. Tumor growth was quantified using bioluminescent imaging and caliper measurements. [(131)I]-2-I-D-Phe clearly reduced tumor growth in the treated mice compared with the control groups. A tumor growth-rate reduction of at least 33% was found for mice receiving a therapeutic dose. There were no serious adverse side-effects of the therapy. In conclusion, i.v. injection of CA 148 MBq [(131)I]-2-I-D-Phe specifically reduces tumor growth in athymic nude mice without relevant side-effects on the animals' health.

Systemic immunity influences hearing preservation in cochlear implantation.

PubMed

Souter, Melanie; Eastwood, Hayden; Marovic, Paul; Kel, Gordana; Wongprasartsuk, Sarin; Ryan, Allen F; O'Leary, Stephen John

2012-06-01

To determine whether a systemic immune response influences hearing thresholds and tissue response after cochlear implantation of hearing guinea pigs. Guinea pigs were inoculated with sterile antigen (Keyhole limpet hemocyanin) 3 weeks before cochlear implantation. Pure-tone auditory brainstem response thresholds were performed before implantation and 1 and 4 weeks later. Dexamethasone phosphate 20% was adsorbed onto a hyaluronic acid carboxymethylcellulose sponge and was applied to the round window for 30 minutes before electrode insertion. Normal saline was used for controls. Cochlear histology was performed at 4 weeks after implantation to assess the tissue response to implantation. To control for the effect of keyhole limpet hemocyanin priming, a group of unprimed animals underwent cochlear implantation with a saline-soaked pledget applied to the round window. Keyhole limpet hemocyanin priming had no significant detrimental effect on thresholds without implantation. Thresholds were elevated after implantation across all frequencies tested (2-32 kHz) in primed animals but only at higher frequencies (4-32 kHz) in unprimed controls. In primed animals, dexamethasone treatment significantly reduced threshold shifts at 2 and 8 kHz. Keyhole limpet hemocyanin led to the more frequent observation of lymphocytes in the tissue response to the implant. Systemic immune activation at the time of cochlear implantation broadened the range of frequencies experiencing elevated thresholds after implantation. Local dexamethasone provides partial protection against this hearing loss, but the degree and extent of protection are less compared to previous studies with unprimed animals.

Detection of the Entomopathogenic Fungus Beauveria bassiana in the Rhizosphere of Wound-Stressed Zea mays Plants

PubMed Central

McKinnon, Aimee C.; Glare, Travis R.; Ridgway, Hayley J.; Mendoza-Mendoza, Artemio; Holyoake, Andrew; Godsoe, William K.; Bufford, Jennifer L.

2018-01-01

Entomopathogenic fungi from the genus Beauveria (Vuillemin) play an important role in controlling insect populations and have been increasingly utilized for the biological control of insect pests. Various studies have reported that Beauveria bassiana (Bals.), Vuill. also has the ability to colonize a broad range of plant hosts as endophytes without causing disease but while still maintaining the capacity to infect insects. Beauveria is often applied as an inundative spore application, but little research has considered how plant colonization may alter the ability to persist in the environment. The aim of this study was to investigate potential interactions between B. bassiana and Zea mays L. (maize) in the rhizosphere following inoculation, in order to understand the factors that may affect environmental persistence of the fungi. The hypothesis was that different isolates of B. bassiana have the ability to colonize maize roots and/or rhizosphere soil, resulting in effects to the plant microbiome. To test this hypothesis, a two-step nested PCR protocol was developed to find and amplify Beauveria in planta or in soil; based on the translation elongation factor 1-alpha (ef1α) gene. The nested protocol was also designed to enable Beauveria species differentiation by sequence analysis. The impact of three selected B. bassiana isolates applied topically to roots on the rhizosphere soil community structure and function were consequently assessed using denaturing gradient gel electrophoresis (DGGE) and MicroRespTM techniques. The microbial community structure and function were not significantly affected by the presence of the isolates, however, retention of the inocula in the rhizosphere at 30 days after inoculation was enhanced when plants were subjected to intensive wounding of foliage to crudely simulate herbivory. The plant defense response likely changed under wound stress resulting in the apparent recruitment of Beauveria in the rhizosphere, which may be an indirect defensive strategy against herbivory and/or the result of induced systemic susceptibility in maize enabling plant colonization. PMID:29942287

White adipose tissue cells and the progression of cachexia: inflammatory pathways

PubMed Central

Neves, Rodrigo X.; Rosa‐Neto, José Cesar; Yamashita, Alex S.; Matos‐Neto, Emidio M.; Riccardi, Daniela M. R.; Lira, Fabio S.; Batista, Miguel L.

2015-01-01

Abstract Background Cachexia is a systemic syndrome leading to body wasting, systemic inflammation, and to metabolic chaos. It is a progressive condition, and little is known about its dynamics. Detection of the early signs of the disease may lead to the attenuation of the associated symptoms. The white adipose tissue is an organ with endocrine functions, capable of synthesising and secreting a plethora of proteins, including cytokines, chemokines, and adipokines. It is well established that different adipose tissue depots demonstrate heterogeneous responses to physiological and pathological stimuli. The present study aimed at providing insight into adipocyte involvement in inflammation along the progression of cachexia. Methods Eight‐weeks‐old male rats were subcutaneously inoculated with a Walker 256 carcinosarcoma cell suspension (2 × 107 cells in 1.0 mL; tumour‐bearing, T) or Phosphate‐buffered saline (control, C). The retroperitoneal, epididymal, and mesenteric adipose pads were excised on Days 0, 7, and 14 post‐tumour cell injection, and the adipocytes were isolated. Results Mesenteric and epididymal adipocytes showed up‐regulation of IL‐1β protein expression and activation of the inflammasome pathway, contributing for whole tissue inflammation. The stromal vascular fraction of the retroperitoneal adipose tissue, on the other hand, seems to be the major contributor for the inflammation in this specific pad. Conclusion Adipocytes seem to play a relevant role in the establishment of white adipose tissue inflammation, through the activation of the NF‐κB and inflammasome pathways. In epididymal adipocytes, induction of the inflammasome may be detected already on Day 7 post‐tumour cell inoculation. PMID:27493872

PRE-INOCULATION OF URINARY CATHETERS WITH ESCHERICHIA COLI 83972 INHIBITS CATHETER COLONIZATION BY ENTEROCOCCUS FAECALIS

PubMed Central

TRAUTNER, BARBARA W.; DAROUICHE, RABIH O.; HULL, RICHARD A.; HULL, SHEILA; THORNBY, JOHN I.

2010-01-01

Purpose The capacity of a preexisting coating of Escherichia coli 83972 to reduce catheter colonization by Enterococcus faecalis 210 was investigated. Enterococcus was chosen for these trials since it is a common urinary pathogen in patients with an indwelling urinary catheter. Materials and Methods Each experiment tested 3 growth conditions. Group 1 or E. coli plus Enterococcus catheters were exposed to E. coli 83972 for 24 hours and then to Enterococcus for 30 minutes. Group 2 or E. coli alone catheters were incubated in E. coli for 24 hours and then in sterile broth for 30 minutes. Group 3 or Enterococcus alone catheters did not undergo the initial incubation with E. coli before the 30-minute incubation with Enterococcus: All catheters were then incubated in sterile human urine for 24 hours. Catheters were washed with saline and cut into 5, 1 cm. segments. Each segment was sonicated and the sonication fluid was diluted and plated. The results of each of the 5 segments were averaged and the set of experiments was repeated 7 times. Results A preexisting coating of E. coli 83972 reduced catheter colonization by E. faecalis 210 more than 10-fold. Enterococcus alone catheters had a median of 9.7 × 105 enterococci per cm., whereas E. coli plus Enterococcus catheters had a median of 0.38 × 105 enterococci per cm. (p = 0.016). Conclusions Pre-inoculating urinary catheters with E. coli 83972 significantly impedes catheter colonization by Enterococcus: These promising in vitro results prompt the clinical investigation of this particular application of bacterial interference. PMID:11743359

Evaluation of the use of a needle-free injection syringe as a cause of non-specific reactions in the intradermal tuberculin test used for the diagnosis of bovine tuberculosis.

PubMed

Díez-Guerrier, A; Roy, A; de la Cruz, M L; Sáez, J L; Sanz, C; Boschiroli, M L; Romero, B; de Juan, L; Domínguez, L; Bezos, J

2018-05-24

The objective of the study was to elucidate whether the use of the needle-free Dermojet syringe, which is based on a high pressure inoculation and is used to inject tuberculin in cattle in several countries, may, in itself, cause skin reactions that can be interpreted as positive reactions to the intradermal tests that are not, in fact, related to the real infection status of the animals. Forty-four cattle from an officially tuberculosis-free (OTF) herd were selected, and four single intradermal tuberculin (SIT) tests were performed on each animal, two on each side of the neck. Three different Dermojet (D1, D2 and D3) and one McLintock (M4) syringes were used to carry out sterile phosphate buffer saline (PBS) with 10% of glycerol and bovine PPD injections. No positive reactions to the SIT test were observed when using the D1-D3 syringes in the case of either bovine PPD or PBS. With regard to M4 (PBS), all the tests were negative when using a standard interpretation but three were positive in the case of the severe interpretation. Significant differences (p < 0.05) in the skin fold thickness measured were found only between certain Dermojet and McLintock syringes at certain inoculation sites. The results showed that the needle-free Dermojet syringe used for PPD intradermal testing in cattle did not cause significant reactions that could be misunderstood as positives. Copyright © 2018 Elsevier Ltd. All rights reserved.

Synergy of subgroup J avian leukosis virus and Eimeria tenella to increase pathogenesis in specific-pathogen-free chickens.

PubMed

Cui, Ning; Wang, Qi; Shi, Wenyan; Han, Linzhen; Wang, Jiazhong; Ma, Xingjiang; Li, Hongmei; Wang, Fangkun; Su, Shuai; Zhao, Xiaomin

2016-09-01

To investigate the effects of co-infections of subgroup J avian leukosis virus (ALV-J) and Eimeria tenella on the pathogenesis in specific-pathogen-free (SPF) white leghorn chickens, groups of chickens were infected with ALV-J strain NX0101 at one day of age or with E. tenella at 14 days of age or both. The control group was left uninfected and was mock-inoculated with phosphate buffer saline (PBS). Mortality rates, body weights, cecal lesions, and viremia of infected chickens in each group were evaluated. Immune status was evaluated by measuring several parameters: immune organ weight/body weight index, specific humoral responses to inactivated NDV vaccine and to inoculated E. tenella, proportions of blood CD3+CD4+ and CD3+CD8α+ lymphocytes and transcriptional levels of cytokines in blood and cecal tonsils. The results show that co-infections of ALV-J and E. tenella induced a higher mortality rate and a lower body weight in SPF chickens compared to single-pathogen infection. In co-infected chickens, ALV-J accelerated the disease symptoms induced by E. tenella, and the E. tenella extended the ALV-J viremia. Thymus atrophy, decrease in the humoral response levels to pathogens and the NDV vaccine, modifications in the blood lymphocyte sub-populations and transcriptional cytokine disorders were found in co-infected chickens compared to chickens infected with one pathogen alone and to controls. We underline a synergy between ALV-J and E. tenella that results in increasing pathogenesis in SPF chickens. Copyright © 2016 Elsevier B.V. All rights reserved.

Transcatheter Arterial Embolization of Renal VX-2 Carcinoma: Ethiodol-Ethanol Capillary Embolization Combined with Carboplatin

PubMed Central

Choi, Byung Gil; Van Pelt, Carolyn S.; Wright, Kenneth C.

2007-01-01

Objective We wanted to determine whether transcatheter Ethiodol-based capillary embolization in combination with carboplatin could improve the efficiency of a 1:1 Ethiodol-ethanol mixture (EEM) to ablate kidneys that been inoculated with VX-2 carcinoma. Materials and Methods The right kidney in 34 New Zealand white rabbits were inoculated with fresh VX-2 tumor fragments. One week later, the kidneys were subjected to transarterial treatment (4-5 rabbits/group): Saline infusion (Group 1); carboplatin infusion (5 or 10 mg, Groups 2A and 2B); carboplatin-Ethiodol (CE) alone (Group 3) and followed by main renal artery occlusion with ethanol (RAO) (Group 4); carboplatin-EEM (C-EEM) followed by RAO (Group 5); carboplatin infusion followed by EEM plus RAO (Group 6); and EEM followed by RAO (Group 7). The animals were followed for up to 3-weeks. The treated kidneys were evaluated angiographically and macroscopically. The kidneys that showed successful embolization macroscopically were entirely cut into serial sections, and these were examined microscopically. Histologically, the kidneys were evaluated on the basis of the residual tumor found in the serial sections. Results The results obtained with carboplatin infusion alone (Groups 2A and 2B) and CE without RAO (Group 3) were similar to those of the control animals (Group 1). Kidneys from Groups 4-7 demonstrated macroscopically successful embolization with histologically proven complete renal parenchyma infarction; however, some residual tumor was evident in all but one animal. Conclusion None of the Ethiodol-based modalities combined with locoregional carboplatin were more efficacious for tumor ablation than EEM alone. PMID:17420631

Changes in the microbial community during bioremediation of gasoline-contaminated soil.

PubMed

Leal, Aline Jaime; Rodrigues, Edmo Montes; Leal, Patrícia Lopes; Júlio, Aline Daniela Lopes; Fernandes, Rita de Cássia Rocha; Borges, Arnaldo Chaer; Tótola, Marcos Rogério

We aimed to verify the changes in the microbial community during bioremediation of gasoline-contaminated soil. Microbial inoculants were produced from successive additions of gasoline to municipal solid waste compost (MSWC) previously fertilized with nitrogen-phosphorous. To obtain Inoculant A, fertilized MSWC was amended with gasoline every 3 days during 18 days. Inoculant B received the same application, but at every 6 days. Inoculant C included MSWC fertilized with N-P, but no gasoline. The inoculants were applied to gasoline-contaminated soil at 10, 30, or 50g/kg. Mineralization of gasoline hydrocarbons in soil was evaluated by respirometric analysis. The viability of the inoculants was evaluated after 103 days of storage under refrigeration or room temperature. The relative proportions of microbial groups in the inoculants and soil were evaluated by FAME. The dose of 50g/kg of inoculants A and B led to the largest CO 2 emission from soil. CO 2 emissions in treatments with inoculant C were inversely proportional to the dose of inoculant. Heterotrophic bacterial counts were greater in soil treated with inoculants A and B. The application of inoculants decreased the proportion of actinobacteria and increased of Gram-negative bacteria. Decline in the density of heterotrophic bacteria in inoculants occurred after storage. This reduction was bigger in inoculants stored at room temperature. The application of stored inoculants in gasoline-contaminated soil resulted in a CO 2 emission twice bigger than that observed in uninoculated soil. We concluded that MSWC is an effective material for the production of microbial inoculants for the bioremediation of gasoline-contaminated soil. Copyright © 2016 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

Automatic inoculating apparatus. [includes movable carraige, drive motor, and swabbing motor

NASA Technical Reports Server (NTRS)

Wilkins, J. R.; Mills, S. M. (Inventor)

1974-01-01

An automatic inoculating apparatus for agar trays is described and using a simple inoculating element, such as a cotton swab or inoculating loop. The apparatus includes a movable carriage for supporting the tray to be inoculated, a drive motor for moving the tray along a trackway, and a swabbing motor for automatically swabbing the tray during the movement. An actuator motor controls lowering of the inoculating element onto the tray and lifting of the inoculating element. An electrical control system, including limit microswitches, enables automatic control of the actuator motor and return of the carriage to the initial position after inoculating is completed.

DOE Office of Scientific and Technical Information (OSTI.GOV)

Poston, Ted M.; Thrall, Karla D.; Penner, Jocelyn D.

I was asked to submit a response to the Protocol Review forum for the journal LabAnimal by Dr. Jerald Silverman. This forum request views and opinions on issues that are faced by research animal welfare committees. The particular issue is: Dr. John Smith, a distinguished investigator at Great Eastern University, had spent his career searching for a treatment for hearing loss. After 20 years of research, he developed a drug that potentially could improve auditory function after damage to the inner ear. The drug was administered by injection into the middle ear with the use of a fine needle insertedmore » through the eardrum. Smith submitted a protocol to the IACUC to conduct a small study on cats, proposing to treat one group of animals with the drug and another with saline, to determine if it prevented the changes in the anatomy of the inner ear that normally occur with aging.« less

Cryoelectrolysis—electrolytic processes in a frozen physiological saline medium

PubMed Central

Lugnani, Franco; Macchioro, Matteo

2017-01-01

Background Cryoelectrolysis is a new minimally invasive tissue ablation surgical technique that combines the ablation techniques of electrolytic ablation with cryosurgery. The goal of this study is to examine the hypothesis that electrolysis can take place in a frozen aqueous saline solution. Method To examine the hypothesis we performed a cryoelectrolytic ablation protocol in which electrolysis and cryosurgery are delivered simultaneously in a tissue simulant made of physiological saline gel with a pH dye. We measured current flow, voltage and extents of freezing and pH dye staining. Results Using optical measurements and measurements of currents, we have shown that electrolysis can occur in frozen physiological saline, at high subzero freezing temperatures, above the eutectic temperature of the frozen salt solution. It was observed that electrolysis occurs when the tissue resides at high subzero temperatures during the freezing stage and essentially throughout the entire thawing stage. We also found that during thawing, the frozen lesion temperature raises rapidly to high subfreezing values and remains at those values throughout the thawing stage. Substantial electrolysis occurs during the thawing stage. Another interesting finding is that electro-osmotic flows affect the process of cryoelectrolysis at the anode and cathode, in different ways. Discussion The results showing that electrical current flow and electrolysis occur in frozen saline solutions imply a mechanism involving ionic movement in the fluid concentrated saline solution channels between ice crystals, at high subfreezing temperatures. Temperatures higher than the eutectic are required for the brine to be fluid. The particular pattern of temperature and electrical currents during the thawing stage of frozen tissue, can be explained by the large amounts of energy that must be removed at the outer edge of the frozen lesion because of the solid/liquid phase transformation on that interface. Conclusion Electrolysis can occur in a frozen domain at high subfreezing temperature, probably above the eutectic. It appears that the most effective period for delivering electrolytic currents in cryoelectrolysis is during the high subzero temperatures stage while freezing and immediately after cooling has stopped, throughout the thawing stage. PMID:28123904

Cryoelectrolysis-electrolytic processes in a frozen physiological saline medium.

PubMed

Lugnani, Franco; Macchioro, Matteo; Rubinsky, Boris

2017-01-01

Cryoelectrolysis is a new minimally invasive tissue ablation surgical technique that combines the ablation techniques of electrolytic ablation with cryosurgery. The goal of this study is to examine the hypothesis that electrolysis can take place in a frozen aqueous saline solution. To examine the hypothesis we performed a cryoelectrolytic ablation protocol in which electrolysis and cryosurgery are delivered simultaneously in a tissue simulant made of physiological saline gel with a pH dye. We measured current flow, voltage and extents of freezing and pH dye staining. Using optical measurements and measurements of currents, we have shown that electrolysis can occur in frozen physiological saline, at high subzero freezing temperatures, above the eutectic temperature of the frozen salt solution. It was observed that electrolysis occurs when the tissue resides at high subzero temperatures during the freezing stage and essentially throughout the entire thawing stage. We also found that during thawing, the frozen lesion temperature raises rapidly to high subfreezing values and remains at those values throughout the thawing stage. Substantial electrolysis occurs during the thawing stage. Another interesting finding is that electro-osmotic flows affect the process of cryoelectrolysis at the anode and cathode, in different ways. The results showing that electrical current flow and electrolysis occur in frozen saline solutions imply a mechanism involving ionic movement in the fluid concentrated saline solution channels between ice crystals, at high subfreezing temperatures. Temperatures higher than the eutectic are required for the brine to be fluid. The particular pattern of temperature and electrical currents during the thawing stage of frozen tissue, can be explained by the large amounts of energy that must be removed at the outer edge of the frozen lesion because of the solid/liquid phase transformation on that interface. Electrolysis can occur in a frozen domain at high subfreezing temperature, probably above the eutectic. It appears that the most effective period for delivering electrolytic currents in cryoelectrolysis is during the high subzero temperatures stage while freezing and immediately after cooling has stopped, throughout the thawing stage.

Balanced crystalloids versus saline in the intensive care unit: study protocol for a cluster-randomized, multiple-crossover trial.

PubMed

Semler, Matthew W; Self, Wesley H; Wang, Li; Byrne, Daniel W; Wanderer, Jonathan P; Ehrenfeld, Jesse M; Stollings, Joanna L; Kumar, Avinash B; Hernandez, Antonio; Guillamondegui, Oscar D; May, Addison K; Siew, Edward D; Shaw, Andrew D; Bernard, Gordon R; Rice, Todd W

2017-03-16

Saline, the intravenous fluid most commonly administered to critically ill adults, contains a high chloride content, which may be associated with acute kidney injury and death. Whether using balanced crystalloids rather than saline decreases the risk of acute kidney injury and death among critically ill adults remains unknown. The Isotonic Solutions and Major Adverse Renal Events Trial (SMART) is a pragmatic, cluster-level allocation, cluster-level crossover trial being conducted between 1 June 2015 and 30 April 2017 in five intensive care units at Vanderbilt University Medical Center in Nashville, TN, USA. SMART compares saline (0.9% sodium chloride) with balanced crystalloids (clinician's choice of lactated Ringer's solution or Plasma-Lyte A®). Each intensive care unit is assigned to provide either saline or balanced crystalloids each month, with the assigned crystalloid alternating monthly over the course of the trial. All adults admitted to participating intensive care units during the study period are enrolled and followed until hospital discharge or 30 days after enrollment. The anticipated enrollment is approximately 14,000 patients. The primary outcome is Major Adverse Kidney Events within 30 days-the composite of in-hospital death, receipt of new renal replacement therapy, or persistent renal dysfunction (discharge creatinine ≥200% of baseline creatinine). Secondary clinical outcomes include in-hospital mortality, intensive care unit-free days, ventilator-free days, vasopressor-free days, and renal replacement therapy-free days. Secondary renal outcomes include new renal replacement therapy receipt, persistent renal dysfunction, and incidence of stage 2 or higher acute kidney injury. This ongoing pragmatic trial will provide the largest and most comprehensive comparison to date of clinical outcomes with saline versus balanced crystalloids among critically ill adults. For logistical reasons, SMART was prospectively registered separately for the medical ICU (SMART-MED; ClinicalTrials.gov identifier: NCT02444988 ; registered on 11 May 2015; date of first patient enrollment: 1 June 2015) and the nonmedical ICUs (SMART-SURG; ClinicalTrials.gov identifier: NCT02547779 ; registered on 9 September 2015; date of first patient enrollment: 1 October 2015).

Temperature effect on high salinity depuration of Vibrio vulnificus and V. parahaemolyticus from the Eastern oyster (Crassostrea virginica).

PubMed

Larsen, A M; Rikard, F S; Walton, W C; Arias, C R

2015-01-02

Vibrio vulnificus (Vv) and Vibrio parahaemolyticus (Vp) are opportunistic human pathogens naturally associated with the Eastern oyster Crassostrea virginica. The abundances of both pathogens in oysters are positively correlated with temperature, thus ingestion of raw oysters during the warm summer months is a risk factor for contracting illness from these bacteria. Current post-harvest processing (PHP) methods for elimination of these pathogens are expensive and kill the oyster, changing their organoleptic properties and making them less appealing to some consumers. High salinity has proven effective in reducing Vv numbers in the wild and our research aims at developing an indoor recirculating system to reduce pathogenic Vibrios while maintaining the taste and texture of live oysters. The goal of this study was to determine the influence of temperature on the efficacy of high salinity depuration. Vv was enumerated as most probable number (MPN) per gram of oyster tissue using the FDA-approved modified cellobiose polymyxin colistin (mCPC) protocol and with an alternative Vibrio specific media CHROMagar™ Vibrio (CaV). CaV was also used to quantify Vp. Oysters were held at 35 psu for 10 days at three temperatures: low (20°C), mid (22.5°C) and high (25°C). There was no difference in MPN/g of Vv between media; however more Vv isolates were obtained from mCPC than CaV. There was no significant effect of temperature on reduction of Vv or Vp throughout depuration but there was a tendency for low temperatures to be less effective than the higher ones. High salinity resulted in a significant decrease in Vv by day 3 and again by day 10, and a decrease in Vp by day 3. Oyster condition indices were maintained throughout depuration and mortality was low (4% across three trials). Overall these results support the use of mCPC for Vv enumeration and demonstrate the promise of high salinity depuration for PHP of the Eastern oyster. The trend for lower temperatures to be less effective is surprising and indicates a potential interaction between salinity and temperature that should be further investigated. Copyright © 2014. Published by Elsevier B.V.

An Upgrade on the Rabbit Model of Anthracycline-Induced Cardiomyopathy: Shorter Protocol, Reduced Mortality, and Higher Incidence of Overt Dilated Cardiomyopathy

PubMed Central

Talavera, Jesús; Fernández-Del-Palacio, María Josefa; García-Nicolás, Obdulio; Seva, Juan; Brooks, Gavin; Moraleda, Jose M.

2015-01-01

Current protocols of anthracycline-induced cardiomyopathy in rabbits present with high premature mortality and nephrotoxicity, thus rendering them unsuitable for studies requiring long-term functional evaluation of myocardial function (e.g., stem cell therapy). We compared two previously described protocols to an in-house developed protocol in three groups: Group DOX2 received doxorubicin 2 mg/kg/week (8 weeks); Group DAU3 received daunorubicin 3 mg/kg/week (10 weeks); and Group DAU4 received daunorubicin 4 mg/kg/week (6 weeks). A cohort of rabbits received saline (control). Results of blood tests, cardiac troponin I, echocardiography, and histopathology were analysed. Whilst DOX2 and DAU3 rabbits showed high premature mortality (50% and 33%, resp.), DAU4 rabbits showed 7.6% premature mortality. None of DOX2 rabbits developed overt dilated cardiomyopathy; 66% of DAU3 rabbits developed overt dilated cardiomyopathy and quickly progressed to severe congestive heart failure. Interestingly, 92% of DAU4 rabbits showed overt dilated cardiomyopathy and 67% developed congestive heart failure exhibiting stable disease. DOX2 and DAU3 rabbits showed alterations of renal function, with DAU3 also exhibiting hepatic function compromise. Thus, a shortened protocol of anthracycline-induced cardiomyopathy as in DAU4 group results in high incidence of overt dilated cardiomyopathy, which insidiously progressed to congestive heart failure, associated to reduced systemic compromise and very low premature mortality. PMID:26788502

Rabies neutralizing antibody response to different schedules of serum and vaccine inoculations in non-exposed persons

PubMed Central

Atanasiu, P.; Bahmanyar, M.; Baltazard, M.; Fox, J. P.; Habel, K.; Kaplan, M. M.; Kissling, R. E.; Komarov, A.; Koprowski, H.; Lépine, P.; Gallardo, F. Pérez; Schaeffer, M.

1957-01-01

Further studies were made on groups of adult humans, previously unexposed to rabies and with no history of rabies vaccination, who were inoculated with different schedules of phenolized inactivated vaccine given subcutaneously and high egg passage (HEP) Flury strain vaccine given intradermally, with and without inoculation of antirabies serum. Serum specimens of the inoculated individuals were studied for antibody up to the 60th day after the first inoculation of the vaccines and serum. Studies were also made on the effect of “booster” doses of HEP Flury strain vaccine given 6 months after preparatory inoculations. The results can be summarized as follows: 1. Fourteen daily inoculations of phenolized vaccine produced a superior antibody response to that obtained with 3 inoculations given 5 days apart. 2. Three intradermal inoculations of HEP Flury vaccine given 5 days apart gave a low level of antibody response, but these individuals responded efficiently by producing antibody to a “booster” dose of the same vaccine given 6 months later. 3. Administration of phenolized vaccine or of HEP Flury vaccine alone did not produce detectable antibody in most individuals until between the 10th and the 15th day after the first inoculation of the vaccine. 4. Passive antibody following inoculation of antirabies serum persisted in some individuals for as long as 42 days. Two inoculations of serum administered 5 days apart did not give levels of antibody higher than those obtained with one inoculation. 5. One inoculation of serum completely suppressed antibody response to 3 inoculations of Flury vaccine given intradermally 5 days apart, and also prevented the preparation of the individuals to respond to a later “booster” dose of this vaccine. 6. Three inoculation of phenolized vaccine given 5 days apart acted efficiently in producing antibody by the 60th day. However, the interfering action of one and two inoculations of serum was clearly defined in this schedule. 7. One inoculation of serum had no suppressive effect on the active antibody response to 14 daily doses of phenolized vaccine; two doses of serum given in the same combination definitely interfered with the production of an active antibody response. PMID:13511138

The effects of xylazine, detomidine, acepromazine and butorphanol on equine solid phase gastric emptying rate.

PubMed

Sutton, D G M; Preston, T; Christley, R M; Cohen, N D; Love, S; Roussel, A J

2002-07-01

The aim of this study was to measure the effects of specific commonly used sedative protocols on equine solid phase gastric emptying rate, using the 13C-octanoic acid breath test (13C-OABT). The gastric emptying of a standard 13C-labelled test meal was measured once weekly in 8 mature horses over two 4 week treatment periods. Each horse acted as its own control. In treatment Period 1, saline (2 ml i.v.), xylazine (0.5 mg/kg i.v.), detomidine (0.01 mg/kg i.v.) or detomidine/butorphanol combination (0.01/0.02 mg/kg i.v.) was administered in randomised order after ingestion of the test meal. During treatment Period 2, test meal consumption was followed by saline, xylazine (1.0 mg/kg i.v.), or detomidine (0.03 mg/kg i.v.) administration, or preceded by acepromazine (0.05 mg/kg i.m.) in randomised order. The 13C:12C ratio of sequential expiratory breath samples was determined by isotope ratio mass spectrometry, and used to measure the gastric half-emptying time, t 1/2, and duration of the lag phase, t lag, for each of the 64 tests. In treatment Period 1, detomidine/butorphanol prolonged both t 1/2 and t lag with respect to xylazine 0.5 mg/kg and the saline control (P < 0.05). In Period 2, detomidine 0.03 mg/kg delayed each parameter with respect to saline, acepromazine and xylazine 1.0 mg/kg (P < 0.001). Xylazine 1.0 mg/kg also lengthened t lag relative to the saline control (P = 0.0004), but did not cause a significant change in t 1/2. Comparison of treatment periods showed that the inhibitory effect of detomidine on gastric emptying rate was dose related (P<0.05). These findings may have clinical significance for case selection when these agents are used for purposes of sedation and/or analgesia.

The interactive effects of essential ions and salinity on the survival of Mysidopsis bahia in 96-H acute toxicity tests of effluents discharged to marine and estuarine receiving waters

DOE Office of Scientific and Technical Information (OSTI.GOV)

Douglas, W.S.; Horne, M.T.

1997-10-01

The importance of salinity in whole effluent toxicity tests using marine organisms has been acknowledged in most testing protocols. However, little if any attention has been given to the specific effects of alteration of the ionic composition of seawater solutions to the test organism. The presence of persistent toxicity in effluents with no apparent toxic agents prompted examination of the potential influence of essential ions on the survival of the opossum shrimp, Mysidopsis bahia, a common effluent toxicity indicator organism. Through stepwise additions of ionic salts to deionized water, the minimum complement of salts to maintain survival of M. bahiamore » during 96-h exposures was determined to be Ca, Mg, K, Br, Na, and Cl. The toxicity curves for Ca, Mg, K, and Br were then determined across test salinity ranging from 10 to 35 parts per thousand. These curves for Ca, Mg, and K revealed that there are significant negative effects on survival when the essential ions are present in either low or high concentrations relative to the levels in natural seawater. Although there were no statistically detectable effects of Br on organism survival over the concentration range tested (5--480 mg/L). Br toxicity at concentrations less than 5 mg/L and greater than 700 mg/L have been shown in other studies. In addition, the tolerance ranges for K, Ca, and Mg were shown to shift significantly with changes in salinity, with lower salinity causing an apparent decrease in tolerance to an excess of essential ions. Tests with toxic effluents from five industrial and municipal sources revealed that adjustment of the ionic balance prior to testing reduced or eliminated toxicity in four of the five whole effluents tested. Suggestions for integrating this information into biomonitoring programs and toxicity identification evaluations are presented.« less

Dextran or Saline Can Replace Contrast for Intravascular Optical Coherence Tomography in Lower Extremity Arteries.

PubMed

Kendrick, Daniel E; Allemang, Matthew T; Gosling, Andre F; Nagavalli, Anil; Kim, Ann H; Nishino, Setsu; Parikh, Sahil A; Bezerra, Hiram G; Kashyap, Vikram S

2016-10-01

To examine the hypothesis that alternative flush media could be used for lower extremity optical coherence tomography (OCT) imaging in long lesions that would normally require excessive use of contrast. The OPTical Imaging Measurement of Intravascular Solution Efficacy (OPTIMISE) trial was a single-center, prospective study (ClinicalTrials.gov identifier NCT01743872) that enrolled 23 patients (mean age 68±11 years; 14 men) undergoing endovascular intervention involving the superficial femoral artery. Four flush media (heparinized saline, dextran, carbon dioxide, and contrast) were used in succession in random order for each image pullback. Quality was defined as ≥270° visualization of vessel wall layers from each axial image. Mean proportions (± standard deviation) of image quality for each flush medium were assessed using 1-way analysis of variance and are reported with the 95% confidence intervals (CI). Four OCT catheters failed, leaving 19 patients who completed the OCT imaging protocol; from this cohort, 51 highest quality runs were selected for analysis. Average vessel diameter was 3.99±1.01 mm. OCT imaging allowed 10- to 15-μm resolution of the lumen border, with diminishing quality as vessel diameter increased. Plaque characterization revealed fibrotic lesions. Mean proportions of image quality were dextran 87.2%±12% (95% CI 0.81 to 0.94), heparinized saline 74.3%±24.8% (95% CI 0.66 to 0.93), contrast 70.1%±30.5% (95% CI 0.52 to 0.88), and carbon dioxide 10.0%±10.4% (95% CI 0.00 to 0.26). Dextran, saline, and contrast provided better quality than carbon dioxide (p<0.001). OCT is feasible in peripheral vessels <5 mm in diameter. Dextran or saline flush media can allow lesion characterization, avoiding iodinated contrast. Carbon dioxide is inadequate for peripheral OCT imaging. Axial imaging may aid in enhancing durability of peripheral endovascular interventions. © The Author(s) 2016.

Effect of thermo-tolerant actinomycetes inoculation on cellulose degradation and the formation of humic substances during composting.

PubMed

Zhao, Yi; Zhao, Yue; Zhang, Zhechao; Wei, Yuquan; Wang, Huan; Lu, Qian; Li, Yanjie; Wei, Zimin

2017-10-01

The inoculum containing four cellulolytic thermophilic actinomycetes was screened from compost samples, and was inoculated into co-composting during different inoculation phases. The effect of different inoculation phases on cellulose degradation, humic substances formation and the relationship between inoculation and physical-chemical parameters was determined. The results revealed that inoculation at different phases of composting improved cellulase activities, accelerated the degradation of cellulose, increased the content of humic substances and influenced the structure of actinomycetic community, but there were significant differences between different inoculation phases. Redundancy analysis showed that the different inoculation phases had different impacts on the relationship between exogenous actinobacteria and physical-chemical parameters. Therefore, based on the promoting effort of inoculation in thermophilic phase of composting for the formation of humic substances, we suggested an optimized inoculation strategy to increase the content of humic substances, alleviate CO 2 emission during composting. Copyright © 2017 Elsevier Ltd. All rights reserved.

Dry Transfer Inoculation of Low-Moisture Spices Containing Antimicrobial Compounds.

PubMed

Hildebrandt, Ian M; Hu, Chuxuan; Grasso-Kelley, Elizabeth M; Ye, Peiran; Anderson, Nathan M; Keller, Susanne E

2017-02-01

Inoculation of a food product for use in subsequent validation studies typically makes use of a high concentration cocktail of microorganisms suspended in aqueous media. However, this inoculation method may prove difficult particularly when the food product is a low-moisture food containing antimicrobial compounds, such as some dried spices. In this study, a dry transfer method for inoculation of clove powder, oregano leaves, ginger powder, and ground black pepper with a five-serovar cocktail of Salmonella was developed and compared with a traditional aqueous inoculation procedure. Spices were inoculated at three levels, 10, 8, and 6 log CFU/g, by using both an aqueous suspension of Salmonella and a dry transfer of Salmonella from previously inoculated silica beads. At the highest inoculation level, the dry transfer method resulted in a significantly higher microbial load (P < 0.05) for ground cloves and oregano, but not for ginger and ground black pepper. At the intermediate inoculation level, differences were apparent only for ginger and black pepper. Inoculation levels of 6 log CFU/g resulted in recoveries below detection limits for both methods of inoculation. Additional examination on the survival of Salmonella on silica beads after inoculation and in clove powder after dry transfer from silica beads showed linear rates of decline, with a rate of -0.011 log CFU/g/day for beads and -0.015 log CFU/g/day for clove powder. The results suggest that dry transfer of Salmonella via inoculated silica beads is a viable alternative when traditional aqueous inoculation is not feasible.

Is it possible to increase bioavailability but not environmental risk of PAHs in bioremediation?

PubMed

Ortega-Calvo, J J; Tejeda-Agredano, M C; Jimenez-Sanchez, C; Congiu, E; Sungthong, R; Niqui-Arroyo, J L; Cantos, M

2013-10-15

The current poor predictability of end points associated with the bioremediation of polycyclic aromatic hydrocarbons (PAHs) is a large limitation when evaluating its viability for treating contaminated soils and sediments. However, we have seen a wide range of innovations in recent years, such as an the improved use of surfactants, the chemotactic mobilization of bacterial inoculants, the selective biostimulation at pollutant interfaces, rhizoremediation and electrobioremediation, which increase the bioavailability of PAHs but do not necessarily increase the risk to the environment. The integration of these strategies into practical remediation protocols would be beneficial to the bioremediation industry, as well as improve the quality of the environment. Copyright © 2013 Elsevier B.V. All rights reserved.

256 Slice Multi-detector Computed Tomography Thoracic Aorta Computed Tomography Angiography: Improved Luminal Opacification Using a Patient-Specific Contrast Protocol and Caudocranial Scan Acquisition.

PubMed

Saade, Charbel; El-Merhi, Fadi; El-Achkar, Bassam; Kerek, Racha; Vogl, Thomas J; Maroun, Gilbert Georges; Jamjoom, Lamia; Al-Mohiy, Hussain; Naffaa, Lena

Caudocranial scan direction and contrast injection timing based on measured patient vessel dynamics can significantly improve arterial and aneurysmal opacification and reduce both contrast and radiation dose in the assessment of thoracic aortic aneurysms (TAA) using helical thoracic computed tomography angiography (CTA). To investigate opacification of the thoracic aorta and TAA using a caudocranial scan direction and a patient-specific contrast protocol. Thoracic aortic CTA was performed in 160 consecutive patients with suspected TAA using a 256-slice computed tomography scanner and a dual barrel contrast injector. Patients were subjected in equal numbers to one of two contrast protocols. Patient age and sex were equally distributed across both groups. Protocol A, the department's standard protocol, consisted of a craniocaudal scan direction with 100 mL of contrast, intravenously injected at a flow rate of 4.5 mL/s. Protocol B involved a caudocranial scan direction and a novel contrast formula based on patient cardiovascular dynamics, followed by 100 mL of saline at 4.5 mL/s. Each scan acquisition comprised of 120 kVp, 200 mA with modulation, temporal resolution 0.27 seconds, and pitch 0.889:1. The dose length product was measured between each protocol and data generated were compared using Mann-Whitney U nonparametric statistics. Receiver operating characteristic analysis, visual grading characteristic (VGC), and κ analyses were performed. Mean opacification in the thoracic aorta and aneurysm measured was 24 % and 55%, respectively. The mean contrast volume was significantly lower in protocol B (73 ± 10 mL) compared with A (100 ± 1 mL) (P<0.001). The contrast-to-noise ratio demonstrated significant differences between the protocols (protocol A, 18.2 ± 12.9; protocol B, 29.7 ± 0.61; P < 0.003). Mean effective dose in protocol B (2.6 ± 0.4 mSv) was reduced by 19% compared with A (3.2 ± 0.8 mSv) (P < 0.004). Aneurysmal detectability demonstrated significant increases by receiver operating characteristic and visual grading characteristic analysis for protocol B compared with A (P < 0.02), and reader agreement increased from poor to excellent. Significant increase in the visualization of TAAs following a caudocranial scan direction during helical thoracic CTA can be achieved using low-contrast volume based on patient-specific contrast formula.

Inducing Resistance to Conspiracy Theory Propaganda: Testing Inoculation and Metainoculation Strategies

ERIC Educational Resources Information Center

Banas, John A.; Miller, Gregory

2013-01-01

This investigation examined the boundaries of inoculation theory by examining how inoculation can be applied to conspiracy theory propaganda as well as inoculation itself (called metainoculation). A 3-phase experiment with 312 participants compared 3 main groups: no-treatment control, inoculation, and metainoculation. Research questions explored…

More on Inoculating Against Reactance to Persuasive Health Messages: The Paradox of Threat.

PubMed

Richards, Adam S; Banas, John A; Magid, Yoav

2017-07-01

This research examined the efficacy of inoculation as a strategy to mitigate psychological reactance based on the level of threat communicated in the forewarning and subsequent persuasive health appeal. Two 2 (inoculation) × 2 (freedom-threatening language) experiments were conducted. The first (N = 181) used elaborated inoculation designed to enhance the threat of impending reactance to a message advocating for responsible alcohol consumption. The second (N = 159) used limited inoculation designed to minimize the threat of impending reactance to a message advocating for responsible soft drink consumption. Results showed that elaborated inoculation increased reactance, whereas limited inoculation decreased reactance but only when the subsequent appeal used less freedom-threatening language. These findings suggest that inoculation has the potential to facilitate or buffer reactance depending on the level of threat communicated in inoculation forewarnings and in subsequent persuasive health appeals.

The Effect of Raffaelea quercus-mongolicae Inoculations on the Formation of Non-conductive Sapwood of Quercus mongolica.

PubMed

Torii, Masato; Matsuda, Yosuke; Seo, Sang Tae; Kim, Kyung Hee; Ito, Shin-Ichiro; Moon, Myung Jin; Kim, Seong Hwan; Yamada, Toshihiro

2014-06-01

In Korea, mass mortality of Quercus mongolica trees has become obvious since 2004. Raffaelea quercus-mongolicae is believed to be a causal fungus contributing the mortality. To evaluate the pathogenicity of the fungus to the trees, the fungus was multiple- and single-inoculated to the seedlings and twigs of the mature trees, respectively. In both the inoculations, the fungus was reisolated from more than 50% of inoculated twigs and seedlings. In the single inoculations, proportions of the transverse area of non-conductive sapwood at inoculation points and vertical lengths of discoloration expanded from the points were significantly different between the inoculation treatment and the control. In the multiple inoculations, no mortality was confirmed among the seedlings examined. These results showed that R. quercus-mongolicae can colonize sapwood, contribute to sapwood discoloration and disrupt sap flows around inoculation sites of Q. mongolica, although the pathogenicity of the fungus was not proven.

A new rabbit model of implant-related biofilm infection: development and evaluation

NASA Astrophysics Data System (ADS)

Chu, Cheng-Bing; Zeng, Hong; Shen, Ding-Xia; Wang, Hui; Wang, Ji-Fang; Cui, Fu-Zhai

2016-03-01

This study is to establish a rabbit model for human prosthetic joint infection and biofilm formation. Thirty-two healthy adult rabbits were randomly divided into four groups and implanted with stainless steel screws and ultra-high molecular weight polyethylene (UHMWPE) washers in the non-articular surface of the femoral lateral condyle of the right hind knees. The rabbit knee joints were inoculated with 1 mL saline containing 0, 102, 103, 104 CFU of Staphylococcus epidermidis ( S. epidermidis) isolated from the patient with total knee arthroplasty (TKA) infection, respectively. On the 14th postoperative day, the UHMWPE washers from the optimal 103 CFU group were further examined. The SEM examination showed a typical biofilm construction that circular S. epidermidis were embedded in a mucous-like matrix. In addition, the LCSM examination showed that the biofilm consisted of the polysaccharide stained bright green fluorescence and S. epidermidis radiating red fluorescence. Thus, we successfully create a rabbit model for prosthetic joint infection and biofilm formation, which should be valuable for biofilm studies.

Airway hyper-responsiveness to neurokinin A and bradykinin following Mycoplasma pneumoniae infection associated with reduced epithelial neutral endopeptidase.

PubMed

Tamaoki, J; Chiyotani, A; Tagaya, E; Araake, M; Nagai, A

1998-09-01

To determine whether mycoplasma infection produces airway hyper-responsiveness to tachykinins and bradykinin and, if so, to elucidate the role of neutral endopeptidase (NEP), isolated hamster tracheal segments were studied under isometric conditions in vitro. Nasal inoculation with Mycoplasma pneumoniae potentiated contractile responses to neurokinin A and bradykinin, causing a leftward shift of the dose-response curves to a lower concentration by 1 log unit for each agonist, whereas there was no response with acetylcholine. Pretreatment of tissues with the NEP inhibitor phosphoramidon augmented neurokinin A- and bradykinin-induced contractions in saline-treated control tissues, but did not further potentiate the responsiveness in M. pneumoniae-infected tissues. NEP activity in the tracheal epithelium, but not in epithelium-denuded tissues, was decreased in infected animals. These results suggest that M. pneumoniae infection causes airway bronchoconstrictor hyper-responsiveness to neurokinin A and bradykinin and that this effect may be associated with an inhibition of epithelial NEP activity.

Efficacy of contact lens disinfecting solutions against Staphylococcus aureus and Pseudomonas aeruginosa.

PubMed

Manuj, Kapur; Gunderson, Charlise; Troupe, John; Huber, Mary E

2006-07-01

To evaluate the disinfection properties of multipurpose contact lens disinfection solutions, based on the International Organization for Standardization (ISO) 14729 guidelines. ReNu with MoistureLoc Multi-Purpose Solution, OPTI-FREE Express with Aldox Multi-Purpose Solution, Betadine 5% sterile ophthalmic preparation solution (povidone iodine), and 0.9% normal saline solution were inoculated with strains of Staphylococcus aureus and Pseudomonas aeruginosa. Surviving bacteria were quantified at specified times. ReNu with MoistureLoc, OPTI-FREE Express, and 5% ophthalmic povidone iodine were effective in achieving a 5-log reduction in bacterial count. Additionally, all three products maintained their effectivity at 72 hours. However, ReNu with MoistureLoc and povidone iodine resulted in the greatest reduction in bacterial colonization. ReNu with MoistureLoc, OPTI-FREE Express, and 5% ophthalmic povidone iodine meet the ISO 14729 guidelines for standalone contact lens solutions. However, ReNu with MoistureLoc and 5% ophthalmic povidone iodine are most efficient in reducing and maintaining low bacterial count for a period of 72 hours.

In vivo laser-based imaging of the human fallopian tube for future cancer detection

NASA Astrophysics Data System (ADS)

Seibel, Eric J.; Melville, C. David; Johnston, Richard S.; Gong, Yuanzheng; Agnew, Kathy; Chiang, Seine; Swisher, Elizabeth M.

2015-03-01

Inherited mutations in BRCA1 and BRCA2 lead to 20-50% lifetime risk of ovarian, tubal, or peritoneal carcinoma. Clinical recommendations for women with these genetic mutations include the prophylactic removal of ovaries and fallopian tubes by age 40 after child-bearing. Recent findings suggest that many presumed ovarian or peritoneal carcinomas arise in fallopian tube epithelium. Although survival rate is <90% when ovarian cancer is detected early (Stage_I), 70% of women have advanced disease (Stage_III/IV) at presentation when survival is less than 30%. Over the years, effective early detection of ovarian cancer has remained elusive, possibly because screening techniques have mistakenly focused on the ovary as origin of ovarian carcinoma. Unlike ovaries, the fallopian tubes are amenable to direct visual imaging without invasive surgery, using access through the cervix. To develop future screening protocols, we investigated using our 1.2- mm diameter, forward-viewing, scanning fiber endoscope (SFE) to image luminal surfaces of the fallopian tube before laparoscopic surgical removal. Three anesthetized human subjects participated in our protocol development which eventually led to 70-80% of the length of fallopian tubes being imaged in scanning reflectance, using red (632nm), green (532nm), and blue (442nm) laser light. A hysteroscope with saline uterine distention was used to locate the tubal ostia. To facilitate passage of the SFE through the interstitial portion of the fallopian tube, an introducer catheter was inserted 1- cm through each ostia. During insertion, saline was flushed to reduce friction and provide clearer viewing. This is likely the first high-resolution intraluminal visualization of fallopian tubes.

The time-adjusted gradual replacement injection method enables better visualization of the right heart.

PubMed

Nakahara, Takehiro; Jinzaki, Masahiro; Niwamae, Nogiku; Saito, Yuuichirou; Arai, Masashi; Tsushima, Yoshito; Kuribayashi, Sachio; Kurabayashi, Masahiko

2014-01-01

Despite the improvement of cardiac CT, right heart visualization remains challenging. We herein describe a new method, called the time-adjusted gradual replacement injection protocol. The aim of this study was to compare this protocol with the split-bolus injection protocol. Fifty-two patients who had undergone dual-source cardiac CT were retrospectively recruited. Twenty-six patients were injected by using the split-bolus injection protocol, and 26 patients were injected by using the time-adjusted gradual replacement injection protocol. For this method, we injected contrast medium for 10 seconds at a flow rate of 0.07 × body weight mL/s, then gradually replaced the contrast material with saline until 2 seconds before finishing the scans. The CT attenuation was measured in 4 chambers, the aorta, and the coronary arteries. The visualization of the anatomic structures and the occurrence and severity of streak artifacts were scored for the cardiac structures in the heart. For the analyses, either Welch t-test or Student t-test was performed. In the right heart, the CT values and visualization scores were significantly higher in the time-adjusted replacement injection group than in the split-bolus injection group, whereas the artifact scores were comparable between the 2 groups. The CT values, visualization scores, and artifact scores of the left heart were not significantly different between the 2 groups. In this study, the time-adjusted gradual replacement injection protocol provided excellent attenuation for visualization of the right heart. This method may help to accurately evaluate the right cardiac anatomy and thereby identify any potential diseases. Copyright © 2014 Society of Cardiovascular Computed Tomography. Published by Elsevier Inc. All rights reserved.

Performing the Parryscope technique gently for office tubal patency assessment.

PubMed

Parry, J Preston; Riche, Daniel; Rushing, John; Linton, Brittany; Butler, Vicki; Lindheim, Steven R

2017-10-01

To demonstrate a novel approach to office tubal patency assessment through infusing air into saline during flexible office hysteroscopy. We also provide data addressing pain and patient experience relative to hysterosalpingography (HSG). Video presentation of clinical technique with supportive crossover data (Canadian Task Force classification II-1). Its University of Mississippi Medical Center Institutional Review Board protocol number is 2013-0230. Academic hospital. Women undergoing office hysteroscopy and ultrasound, with a subset also having HSG. Air infusion into saline during office hysteroscopy. The focus is on demonstrating how the technique appears and is performed, with supplemental Likert data addressing subjective pain and preference relative to HSG. When performed as described, this office technique has 98.3%-100% sensitivity to tubal occlusion and 83.7% specificity. The gentle technique is central to accurate outcomes, which is facilitated through use of a small-caliber (<3 mm), flexible hysteroscope and avoiding uterine overdistention. Patients are far more likely to report maximum discomfort with HSG. Among patients who also had HSG, 92% somewhat or strongly prefer hysteroscopic assessment. Also, 96% of patients reporting maximum discomfort with HSG had mild to no discomfort with the described technique. Air-infused saline at flexible office hysteroscopy can accurately, gently, and rapidly assess tubal patency. Coupled with strong patient preference for this technique over HSG, it is a promising option for evaluating fertility. NCT02005263. Published by Elsevier Inc.

Integral strategy for evaluation of fecal indicator performance in bird-influenced saline inland waters.

PubMed

Kirschner, Alexander K T; Zechmeister, Thomas C; Kavka, Gerhard G; Beiwl, Christian; Herzig, Alois; Mach, Robert L; Farnleitner, Andreas H

2004-12-01

Wild birds are an important nonpoint source of fecal contamination of surface waters, but their contribution to fecal pollution is mostly difficult to estimate. Thus, to evaluate the relation between feces production and input of fecal indicator bacteria (FIB) into aquatic environments by wild waterfowl, we introduced a new holistic approach for evaluating the performance of FIB in six shallow saline habitats. For this, we monitored bird abundance, fecal pellet production, and the abundance of FIB concomitantly with a set of environmental variables over a 9-month period. For estimating fecal pellet production, a new protocol of fecal pellet counting was introduced, which was called fecal taxation (FTX). We could show that, over the whole range of investigated habitats, bird abundance, FTX values, and FIB abundance were highly significantly correlated and could demonstrate the good applicability of the FTX as a meaningful surrogate parameter for recent bird abundances and fecal contamination by birds in shallow aquatic ecosystems. Presumptive enterococci (ENT) were an excellent surrogate parameter of recent fecal contamination in these saline environments for samples collected at biweekly to monthly sampling intervals while presumptive Escherichia coli and fecal coliforms (FC) were often undetectable. Significant negative correlations with salinity indicated that E. coli and FC survival was hampered by osmotic stress. Statistical analyses further revealed that fecal pollution-associated parameters represented one system component independent from other environmental variables and that, besides feces production, rainfall, total suspended solids (direct), and trophy (indirect) had significant positive effects on ENT concentrations. Our holistic approach of linking bird abundance, feces production, and FIB detection with environmental variables may serve as a powerful model for application to other aquatic ecosystems.

Integral Strategy for Evaluation of Fecal Indicator Performance in Bird-Influenced Saline Inland Waters

PubMed Central

Kirschner, Alexander K. T.; Zechmeister, Thomas C.; Kavka, Gerhard G.; Beiwl, Christian; Herzig, Alois; Mach, Robert L.; Farnleitner, Andreas H.

2004-01-01

Wild birds are an important nonpoint source of fecal contamination of surface waters, but their contribution to fecal pollution is mostly difficult to estimate. Thus, to evaluate the relation between feces production and input of fecal indicator bacteria (FIB) into aquatic environments by wild waterfowl, we introduced a new holistic approach for evaluating the performance of FIB in six shallow saline habitats. For this, we monitored bird abundance, fecal pellet production, and the abundance of FIB concomitantly with a set of environmental variables over a 9-month period. For estimating fecal pellet production, a new protocol of fecal pellet counting was introduced, which was called fecal taxation (FTX). We could show that, over the whole range of investigated habitats, bird abundance, FTX values, and FIB abundance were highly significantly correlated and could demonstrate the good applicability of the FTX as a meaningful surrogate parameter for recent bird abundances and fecal contamination by birds in shallow aquatic ecosystems. Presumptive enterococci (ENT) were an excellent surrogate parameter of recent fecal contamination in these saline environments for samples collected at biweekly to monthly sampling intervals while presumptive Escherichia coli and fecal coliforms (FC) were often undetectable. Significant negative correlations with salinity indicated that E. coli and FC survival was hampered by osmotic stress. Statistical analyses further revealed that fecal pollution-associated parameters represented one system component independent from other environmental variables and that, besides feces production, rainfall, total suspended solids (direct), and trophy (indirect) had significant positive effects on ENT concentrations. Our holistic approach of linking bird abundance, feces production, and FIB detection with environmental variables may serve as a powerful model for application to other aquatic ecosystems. PMID:15574941

Effect of prolonged LBNP and saline ingestion on plasma volume and orthostatic responses during bed rest

NASA Technical Reports Server (NTRS)

Fortney, Suzanne M.; Dussack, Larry; Rehbein, Tracy; Wood, Margie; Steinmann, Laura

1991-01-01

Orthostatic intolerance remains a significant problem following space flight despite frequent use of the saline fluid-loading countermeasure and volitional use of an anti-gravity suite during reentry and landing. The purpose of this project is to examine the plasma volume (PV), endocrine, and orthostatic responses of bedrested subjects following 2-hr and 4-hr treatments of lower body negative pressure (LBNP) and saline ingestion. Ten healthy men were randomly assigned into 2 groups. Group A underwent a 4-hr LBNP/saline treatment on best rest day 5 and the 2-hr treatment on day 11. Group B underwent the 2-hr treatment on day 6 and the 4-hr treatment on day 10. Blood volume was determined before and after bed rest using radiolabelling. Changes in PV between measurements were calculated from changes in hematocrit and estimated red cell volume. Urinary excretion of anti-diuretic hormone (ADH) and aldosterone (ALD) were measured each day during the study. Orthostatic responses were measured using a ramp LBNP protocol before bed rest, before each treatment, and 24 hours after each treatment. Both 2-hr and 4-hr treatments resulted in a restoration of PV to pre-bed rest levels which persisted at least 24 hours. This increase in PV was associated with significant increases in urinary excretion of ADH and ALD. Twenty-four hours after the 4-hr treatment, the heart rate and pulse pressure response to LBNP were significantly lower and stroke volumes during LBNP were increased. Twenty-four hours after the 2-hr treatment, there was no evidence of improvement in orthostatic responses. These results suggest that a countermeasure which simply restores PV during space flight may not be sufficient for restoring orthostatic responses.

Stable Expression of mtlD Gene Imparts Multiple Stress Tolerance in Finger Millet

PubMed Central

Hema, Ramanna; Vemanna, Ramu S.; Sreeramulu, Shivakumar; Reddy, Chandrasekhara P.; Senthil-Kumar, Muthappa; Udayakumar, Makarla

2014-01-01

Finger millet is susceptible to abiotic stresses, especially drought and salinity stress, in the field during seed germination and early stages of seedling development. Therefore developing stress tolerant finger millet plants combating drought, salinity and associated oxidative stress in these two growth stages is important. Cellular protection through osmotic adjustment and efficient free radical scavenging ability during abiotic stress are important components of stress tolerance mechanisms in plants. Mannitol, an osmolyte, is known to scavenge hydroxyl radicals generated during various abiotic stresses and thereby minimize stress damage in several plant species. In this study transgenic finger millet plants expressing the mannitol biosynthetic pathway gene from bacteria, mannitol-1-phosphate dehydrogenase (mtlD), were developed through Agrobacterium tumefaciens-mediated genetic transformation. mtlD gene integration in the putative transgenic plants was confirmed by Southern blot. Further, performance of transgenic finger millet under drought, salinity and oxidative stress was studied at plant level in T1 generation and in T1 and T2 generation seedlings. Results from these experiments showed that transgenic finger millet had better growth under drought and salinity stress compared to wild-type. At plant level, transgenic plants showed better osmotic adjustment and chlorophyll retention under drought stress compared to the wild-type. However, the overall increase in stress tolerance of transgenics for the three stresses, especially for oxidative stress, was only marginal compared to other mtlD gene expressing plant species reported in the literature. Moreover, the Agrobacterium-mediated genetic transformation protocol developed for finger millet in this study can be used to introduce diverse traits of agronomic importance in finger millet. PMID:24922513

The Effect of Raffaelea quercus-mongolicae Inoculations on the Formation of Non-conductive Sapwood of Quercus mongolica

PubMed Central

Matsuda, Yosuke; Seo, Sang Tae; Kim, Kyung Hee; Ito, Shin-ichiro; Moon, Myung Jin; Kim, Seong Hwan; Yamada, Toshihiro

2014-01-01

In Korea, mass mortality of Quercus mongolica trees has become obvious since 2004. Raffaelea quercus-mongolicae is believed to be a causal fungus contributing the mortality. To evaluate the pathogenicity of the fungus to the trees, the fungus was multiple- and single-inoculated to the seedlings and twigs of the mature trees, respectively. In both the inoculations, the fungus was reisolated from more than 50% of inoculated twigs and seedlings. In the single inoculations, proportions of the transverse area of non-conductive sapwood at inoculation points and vertical lengths of discoloration expanded from the points were significantly different between the inoculation treatment and the control. In the multiple inoculations, no mortality was confirmed among the seedlings examined. These results showed that R. quercus-mongolicae can colonize sapwood, contribute to sapwood discoloration and disrupt sap flows around inoculation sites of Q. mongolica, although the pathogenicity of the fungus was not proven. PMID:25071395

Aerobic Exercise Training Attenuates Tumor Growth and Reduces Insulin Secretion in Walker 256 Tumor-Bearing Rats

PubMed Central

Moreira, Veridiana Mota; da Silva Franco, Claudinéia Conationi; Prates, Kelly Valério; Gomes, Rodrigo Mello; de Moraes, Ana Maria Praxedes; Ribeiro, Tatiane Aparecida; Martins, Isabela Peixoto; Previate, Carina; Pavanello, Audrei; Matiusso, Camila Cristina Ianoni; Almeida, Douglas Lopes; Francisco, Flávio Andrade; Malta, Ananda; Tófolo, Laize Peron; da Silva Silveira, Sandra; Saavedra, Lucas Paulo Jacinto; Machado, Katia; da Silva, Paulo Henrique Olivieri; Fabrício, Gabriel S.; Palma-Rigo, Kesia; de Souza, Helenir Medri; de Fátima Silva, Flaviane; Biazi, Giuliana Regina; Pereira, Taís Susane; Vieira, Elaine; Miranda, Rosiane Aparecida; de Oliveira, Júlio Cezar; da Costa Lima, Luiz Delmar; Rinaldi, Wilson; Ravanelli, Maria Ida; de Freitas Mathias, Paulo Cezar

2018-01-01

Aerobic exercise training can improve insulin sensitivity in many tissues; however, the relationship among exercise, insulin, and cancer cell growth is unclear. We tested the hypothesis that aerobic exercise training begun during adolescence can attenuate Walker 256 tumor growth in adult rats and alter insulin secretion. Thirty-day-old male Wistar rats engaged in treadmill running for 8 weeks, 3 days/week, 44 min/day, at 55–65% VO2max until they were 90 days old (TC, Trained Control). An equivalently aged group was kept inactive during the same period (SC, Sedentary Control). Then, half the animals of the SC and TC groups were reserved as the control condition and the other half were inoculated with Walker 256 cancer cells, yielding two additional groups (Sedentary Walker and Trained Walker). Zero mortalities were observed in tumor-bearing rats. Body weight (BW), food intake, plasma glucose, insulin levels, and peripheral insulin sensitivity were analyzed before and after tumor cell inoculation. We also evaluated tumor growth, metastasis and cachexia. Isolated pancreatic islets secretory activity was analyzed. In addition, we evaluated mechanic sensibility. Our results showed improved physical performance according to the final workload and VO2max and reduced BW in trained rats at the end of the running protocol. Chronic adaptation to the aerobic exercise training decreased tumor weight, cachexia and metastasis and were associated with low glucose and insulin levels and high insulin sensitivity before and after tumor cell inoculation. Aerobic exercise started at young age also reduced pancreatic islet insulin content and insulin secretion in response to a glucose stimulus, without impairing islet morphology in trained rats. Walker 256 tumor-bearing sedentary rats also presented reduced pancreatic islet insulin content, without changing insulin secretion through isolated pancreatic islets. The mechanical sensitivity test indicated that aerobic exercise training did not cause injury or trigger inflammatory processes prior to tumor cell inoculation. Taken together, the current study suggests that aerobic exercise training applied during adolescence may mitigate tumor growth and related disorders in Walker 256 tumor-bearing adult rats. Improved insulin sensibility, lower glucose and insulin levels and/or reduced insulin secretion stimulated by glucose may be implicated in this tumor attenuation.

Aerobic Exercise Training Attenuates Tumor Growth and Reduces Insulin Secretion in Walker 256 Tumor-Bearing Rats.

PubMed

Moreira, Veridiana Mota; da Silva Franco, Claudinéia Conationi; Prates, Kelly Valério; Gomes, Rodrigo Mello; de Moraes, Ana Maria Praxedes; Ribeiro, Tatiane Aparecida; Martins, Isabela Peixoto; Previate, Carina; Pavanello, Audrei; Matiusso, Camila Cristina Ianoni; Almeida, Douglas Lopes; Francisco, Flávio Andrade; Malta, Ananda; Tófolo, Laize Peron; da Silva Silveira, Sandra; Saavedra, Lucas Paulo Jacinto; Machado, Katia; da Silva, Paulo Henrique Olivieri; Fabrício, Gabriel S; Palma-Rigo, Kesia; de Souza, Helenir Medri; de Fátima Silva, Flaviane; Biazi, Giuliana Regina; Pereira, Taís Susane; Vieira, Elaine; Miranda, Rosiane Aparecida; de Oliveira, Júlio Cezar; da Costa Lima, Luiz Delmar; Rinaldi, Wilson; Ravanelli, Maria Ida; de Freitas Mathias, Paulo Cezar

2018-01-01

Aerobic exercise training can improve insulin sensitivity in many tissues; however, the relationship among exercise, insulin, and cancer cell growth is unclear. We tested the hypothesis that aerobic exercise training begun during adolescence can attenuate Walker 256 tumor growth in adult rats and alter insulin secretion. Thirty-day-old male Wistar rats engaged in treadmill running for 8 weeks, 3 days/week, 44 min/day, at 55-65% VO 2max until they were 90 days old (TC, Trained Control). An equivalently aged group was kept inactive during the same period (SC, Sedentary Control). Then, half the animals of the SC and TC groups were reserved as the control condition and the other half were inoculated with Walker 256 cancer cells, yielding two additional groups (Sedentary Walker and Trained Walker). Zero mortalities were observed in tumor-bearing rats. Body weight (BW), food intake, plasma glucose, insulin levels, and peripheral insulin sensitivity were analyzed before and after tumor cell inoculation. We also evaluated tumor growth, metastasis and cachexia. Isolated pancreatic islets secretory activity was analyzed. In addition, we evaluated mechanic sensibility. Our results showed improved physical performance according to the final workload and VO 2max and reduced BW in trained rats at the end of the running protocol. Chronic adaptation to the aerobic exercise training decreased tumor weight, cachexia and metastasis and were associated with low glucose and insulin levels and high insulin sensitivity before and after tumor cell inoculation. Aerobic exercise started at young age also reduced pancreatic islet insulin content and insulin secretion in response to a glucose stimulus, without impairing islet morphology in trained rats. Walker 256 tumor-bearing sedentary rats also presented reduced pancreatic islet insulin content, without changing insulin secretion through isolated pancreatic islets. The mechanical sensitivity test indicated that aerobic exercise training did not cause injury or trigger inflammatory processes prior to tumor cell inoculation. Taken together, the current study suggests that aerobic exercise training applied during adolescence may mitigate tumor growth and related disorders in Walker 256 tumor-bearing adult rats. Improved insulin sensibility, lower glucose and insulin levels and/or reduced insulin secretion stimulated by glucose may be implicated in this tumor attenuation.

Consequences of inoculation with native arbuscular mycorrhizal fungi for root colonization and survival of Artemisia tridentata ssp. wyomingensis seedlings after transplanting.

PubMed

Davidson, Bill E; Novak, Stephen J; Serpe, Marcelo D

2016-08-01

In arid environments, the propagule density of arbuscular mycorrhizal fungi (AMF) may limit the extent of the plant-AMF symbiosis. Inoculation of seedlings with AMF could alleviate this problem, but the success of this practice largely depends on the ability of the inoculum to multiply and colonize the growing root system after transplanting. These phenomena were investigated in Artemisia tridentata ssp. wyomingensis (Wyoming big sagebrush) seedlings inoculated with native AMF. Seedlings were first grown in a greenhouse in soil without AMF (non-inoculated seedlings) or with AMF (inoculated seedlings). In spring and fall, 3-month-old seedlings were transplanted outdoors to 24-L pots containing soil from a sagebrush habitat (spring and fall mesocosm experiments) or to a recently burned sagebrush habitat (spring and fall field experiments). Five or 8 months after transplanting, colonization was about twofold higher in inoculated than non-inoculated seedlings, except for the spring field experiment. In the mesocosm experiments, inoculation increased survival during the summer by 24 % (p = 0.011). In the field experiments, increased AMF colonization was associated with increases in survival during cold and dry periods; 1 year after transplanting, survival of inoculated seedlings was 27 % higher than that of non-inoculated ones (p < 0.001). To investigate possible mechanisms by which AMF increased survival, we analyzed water use efficiency (WUE) based on foliar (13)C/(12)C isotope ratios (δ (13)C). A positive correlation between AMF colonization and δ (13)C values was observed in the spring mesocosm experiment. In contrast, inoculation did not affect the δ (13)C values of fall transplanted seedlings that were collected the subsequent spring. The effectiveness of AMF inoculation on enhancing colonization and reducing seedling mortality varied among the different experiments, but average effects were estimated by meta-analyses. Several months after transplanting, average AMF colonization was in proportion 84 % higher in inoculated than non-inoculated seedlings (p = 0.0042), while the average risk of seedling mortality was 42 % lower in inoculated than non-inoculated seedlings (p = 0.047). These results indicate that inoculation can increase AMF colonization over the background levels occurring in the soil, leading to higher rates of survival.

Diversity and persistence of ectomycorrhizal fungi and their effect on nursery-inoculated Pinus pinaster in a post-fire plantation in Northern Portugal.

PubMed

Franco, Albina R; Sousa, Nadine R; Ramos, Miguel A; Oliveira, Rui S; Castro, Paula M L

2014-11-01

Ectomycorrhizal fungi (ECMF) play an important role in forest ecosystems, often mitigating stress factors and increasing seedling performance. The aim of this study was to investigate the effects of a nursery inoculation on Pinus pinaster growth and on the fungal communities established when reforesting burned areas. Inoculated P. pinaster saplings showed 1.5-fold higher stem height than the non-inoculated controls after a 5 year growth period, suggesting that fungal inoculation could potentiate tree growth in the field. Ordination analysis revealed the presence of different ECMF communities on both plots. Among the nursery-inoculated fungi, Laccaria sp., Rhizopogon sp., Suillus bovinus and Pisolithus sp. were detected on inoculated Pinus saplings on both sampling periods, indicating that they persisted after field establishment. Other fungi were also detected in the inoculated plants. Phialocephala sp. was found on the first assessment, while Terfezia sp. was detected on both sampling periods. Laccaria sp. and Rhizopogon sp. were identified in the control saplings, belonging however to different species than those found in the inoculated plot. Inocybe sp., Thelephora sp. and Paxillus involutus were present on both sampling periods in the non-inoculated plots. The results suggest that ECMF inoculation at nursery stage can benefit plant growth after transplantation to a post-fire site and that the inoculated fungi can persist in the field. This approach has great potential as a biotechnological tool to aid in the reforestation of burned areas.

Growth and Yield Responses of Cowpea to Inoculation and Phosphorus Fertilization in Different Environments

PubMed Central

Kyei-Boahen, Stephen; Savala, Canon E. N.; Chikoye, David; Abaidoo, Robert

2017-01-01

Cowpea (Vigna unguiculata) is a major source of dietary protein and essential component of the cropping systems in semi-arid regions of Sub-Saharan Africa. However, yields are very low due to lack of improved cultivars, poor management practices, and limited inputs use. The objectives of this study were to assess the effects of rhizobia inoculant and P on nodulation, N accumulation and yield of two cowpea cultivars in Mozambique. Field study was conducted in three contrasting environments during the 2013/2014 and 2014/2015 seasons using randomized complete block design with four replications and four treatments. Treatments consisted of seed inoculation, application of 40 kg P2O5 ha-1, inoculation + P, and a non-inoculated control. The most probable number (MPN) technique was used to estimate the indigenous bradyrhizobia populations at the experimental sites. The rhizobia numbers at the sites varied from 5.27 × 102 to 1.07 × 103 cells g-1 soil. Inoculation increased nodule number by 34–76% and doubled nodule dry weight (78 to 160 mg plant-1). P application improved nodulation and interacted positively with the inoculant. Inoculation, P, and inoculant + P increased shoot dry weight, and shoot and grain N content across locations but increases in number of pods plant-1, seeds pod-1, and 100-seed weight were not consistent among treatments across locations. Shoot N content was consistently high for the inoculated plants and also for the inoculated + P fertilized plants, whereas the non-inoculated control plants had the lowest tissue N content. P uptake in shoot ranged from 1.72 to 3.77 g kg-1 and was higher for plants that received P fertilizer alone. Inoculation and P either alone or in combination consistently increased cowpea grain yield across locations with yields ranging from 1097 kg ha-1 for the non-inoculated control to 1674 kg ha-1 for the inoculant + P treatment. Grain protein concentration followed a similar trend as grain yield and ranged from 223 to 252 g kg-1 but a negative correlation between grain yield and protein concentration was observed. Inoculation increased net returns by $104–163 ha-1 over that for the control. The results demonstrate the potential of improving cowpea grain yield, quality and profitability using inoculant, although the cost-benefit for using P at the current fertilizer price is not attractive except when applied together with inoculant at low P site. PMID:28515729

Inoculating against reactance to persuasive health messages.

PubMed

Richards, Adam S; Banas, John A

2015-01-01

This investigation examined the possibility of decreasing psychological reactance to health campaigns through the use of inoculation messages. It was hypothesized that an inoculation message, which forewarned of the potential of subsequent reactance, would decrease participants' likelihood of reacting negatively to a freedom-threatening message aimed to reduce excessive alcohol consumption. Participants (N = 275) who were inoculated against potential reactance felt less threatened and experienced less reactance compared to those who did not read an inoculation message. Structural equation modeling showed that inoculation indirectly predicted lower intention to drink alcohol via the theorized mediated reactance process. This research suggests that it is possible to inoculate against self-generated cognitions that might otherwise lead toward negative health behaviors.

An efficient and high-throughput protocol for Agrobacterium-mediated transformation based on phosphomannose isomerase positive selection in Japonica rice (Oryza sativa L.).

PubMed

Duan, Yongbo; Zhai, Chenguang; Li, Hao; Li, Juan; Mei, Wenqian; Gui, Huaping; Ni, Dahu; Song, Fengshun; Li, Li; Zhang, Wanggen; Yang, Jianbo

2012-09-01

A number of Agrobacterium-mediated rice transformation systems have been developed and widely used in numerous laboratories and research institutes. However, those systems generally employ antibiotics like kanamycin and hygromycin, or herbicide as selectable agents, and are used for the small-scale experiments. To address high-throughput production of transgenic rice plants via Agrobacterium-mediated transformation, and to eliminate public concern on antibiotic markers, we developed a comprehensive efficient protocol, covering from explant preparation to the acquisition of low copy events by real-time PCR analysis before transplant to field, for high-throughput production of transgenic plants of Japonica rice varieties Wanjing97 and Nipponbare using Escherichia coli phosphomannose isomerase gene (pmi) as a selectable marker. The transformation frequencies (TF) of Wanjing97 and Nipponbare were achieved as high as 54.8 and 47.5%, respectively, in one round of selection of 7.5 or 12.5 g/L mannose appended with 5 g/L sucrose. High-throughput transformation from inoculation to transplant of low copy events was accomplished within 55-60 days. Moreover, the Taqman assay data from a large number of transformants showed 45.2% in Wanjing97 and 31.5% in Nipponbare as a low copy rate, and the transformants are fertile and follow the Mendelian segregation ratio. This protocol facilitates us to perform genome-wide functional annotation of the open reading frames and utilization of the agronomically important genes in rice under a reduced public concern on selectable markers. We describe a comprehensive protocol for large scale production of transgenic Japonica rice plants using non-antibiotic selectable agent, at simplified, cost- and labor-saving manners.

A new strategy for co-composting dairy manure with rice straw: Addition of different inocula at three stages of composting.

PubMed

Zhou, Cheng; Liu, Zhang; Huang, Zhao-Lin; Dong, Ming; Yu, Xiao-Long; Ning, Ping

2015-06-01

In considering the impact of inoculation time and the characteristics of composting material and inoculants on the usefulness of inoculation, a new composting strategy has been proposed and studied, in which three inocula were inoculated at three stages of composting process respectively: inoculum A (Thermoactinomyces sp. GF1 and GF2) was inoculated before fermentation to increase or maintain high temperature of pile, inoculum B (Coprinus cinerea and Coprinus comatus) was inoculated after thermophilic phase to promote degradation of lignin, and inoculum C (Trichoderma harzianum and Rhizopus oryzae) was inoculated after 30-day fermentation to promote degradation of cellulose. The results showed that the inoculations could significantly enhance the temperature of pile and the degradation of lignocelluloses. When inocula A, B, and C were inoculated into pile, temperature increased from 25°C to 65°C, from 33°C to 39°C and from 33°C to 38°C respectively and 35% lignin and 43% cellulose had been degraded in inoculated pile compared to the degradation of 15% lignin and 25% cellulose in control pile. As a result, the C/N ratio dropped more rapidly degraded in the inoculated pile (reached 20 after 33-day fermentation) than that in the control pile (reached 21.7 after 45-day fermentation). In addition, the volume loss in inoculated pile (76.5%) was higher than that in control pile (53.2%). The study, therefore, indicated that inoculating proper microorganisms at appropriate time improved the composting process and our new composting strategy would be propitious to the co-composting dairy manure with rice straw. Copyright © 2015 Elsevier Ltd. All rights reserved.

[Quality of commercial inoculants for soybean crop in Argentina: concentration of viable rhizobia and presence of contaminants].

PubMed

Benintende, S

2010-01-01

In view of the inoculant production technology available, quality control is a necessary tool to improve soybean inoculants commercialized in Argentina. In 1988, the Facultad de Ciencias Agropecuarias de la Universidad Nacional de Entre Ríos (Argentina) created a quality control service for soybean crop inoculants to offer to farmers. The aim of this study was to evaluate the quality of soybean crop inoculants for seven cropping seasons and to contrast these results with those from previous investigations conducted in our country. This work was developed using 128 inoculant samples from 30 different trade names. The analyzed variables were: inoculant label information, number of viable rhizobia and presence of contaminants. Twenty per cent of the labels showed defects that did not comply with the Argentine legislation. The detected problems in inoculant labels were related to lot numbers or the expiry date, which lacked, was easy to remove or not visible. Eighty seven per cent of the analyzed inoculants were formulated in liquid carriers. Seventy six per cent of the samples had a number of rhizobia above 10(8) CFU/g or ml, the minimum quantity required by the legislation. Thirty per cent of the analyzed inoculants had contaminants and their presence was related to low rhizobia counts, as shown in a correspondence analysis. The relationship between liquid inoculants and the absence of contaminants was expressed. It can be concluded from the comparison of results found in this investigation with those in previous works published on Argentinean inoculants, that inoculant quality has been improved, although the situation is far from ideal. Adequate manufacturing and commercialization controls are necessary to ensure product quality.

Enhanced stability of superparamagnetic iron oxide nanoparticles in biological media using a pH adjusted-BSA adsorption protocol

NASA Astrophysics Data System (ADS)

Yu, Si-Ming; Laromaine, Anna; Roig, Anna

2014-07-01

Superparamagnetic iron oxide nanoparticles (SPIONs) are widely used for biological applications due to their unique properties compared to their bulk counterparts, simplified SPIONs stabilization protocols applicable for a wide spectra of biological media remains a challenging issue. In this work, SPIONs with different surface coatings, tetramethylammonium hydroxide-coated SPIONs (T-SPIONs), and citrate-coated SPIONs (C-SPIONs) were synthesized by a facile, rapid and cost effective microwave-assisted method. C-SPIONs show robust stability in biological media of phosphate buffered saline and Roswell Park Memorial Institute Medium, while destabilize in DMEM. T-SPIONs were found to aggregate rapidly and significantly in all tested media. Then, a modified pH adjusted-BSA adsorption protocol and an addition of excess trisodium citrate dihydrate (Na3Cit) were used to enhance their stability in the media. The BSA adsorption protocol showed great efficiency in stabilizing the dispersed state of both SPIONs in the tested media, while the addition of excess Na3Cit showed limited effect, and it was only applicable for C-SPIONs. The formed BSA layer on SPIONs could be imaged by negative staining TEM, and revealed by Cryo-TEM, FTIR, DLS, and the zeta potential measurements. Results indicated that BSA forms a monolayer of a thickness of about 3 ± 1 nm and BSA interacts with C-SPIONs and T-SPIONs through their coating, rather than by replacing them. This synthetic method and stabilization protocol offer a general methodology to obtain SPIONs with a variety of surfactants, stable in different biological media in few minutes.

An optimised protocol for molecular identification of Eimeria from chickens☆

PubMed Central

Kumar, Saroj; Garg, Rajat; Moftah, Abdalgader; Clark, Emily L.; Macdonald, Sarah E.; Chaudhry, Abdul S.; Sparagano, Olivier; Banerjee, Partha S.; Kundu, Krishnendu; Tomley, Fiona M.; Blake, Damer P.

2014-01-01

Molecular approaches supporting identification of Eimeria parasites infecting chickens have been available for more than 20 years, although they have largely failed to replace traditional measures such as microscopy and pathology. Limitations of microscopy-led diagnostics, including a requirement for specialist parasitological expertise and low sample throughput, are yet to be outweighed by the difficulties associated with accessing genomic DNA from environmental Eimeria samples. A key step towards the use of Eimeria species-specific PCR as a sensitive and reproducible discriminatory tool for use in the field is the production of a standardised protocol that includes sample collection and DNA template preparation, as well as primer selection from the numerous PCR assays now published. Such a protocol will facilitate development of valuable epidemiological datasets which may be easily compared between studies and laboratories. The outcome of an optimisation process undertaken in laboratories in India and the UK is described here, identifying four steps. First, samples were collected into a 2% (w/v) potassium dichromate solution. Second, oocysts were enriched by flotation in saturated saline. Third, genomic DNA was extracted using a QIAamp DNA Stool mini kit protocol including a mechanical homogenisation step. Finally, nested PCR was carried out using previously published primers targeting the internal transcribed spacer region 1 (ITS-1). Alternative methods tested included sample processing in the presence of faecal material, DNA extraction using a traditional phenol/chloroform protocol, the use of SCAR multiplex PCR (one tube and two tube versions) and speciation using the morphometric tool COCCIMORPH for the first time with field samples. PMID:24138724

Evaluation and single-laboratory verification of a proposed modification to the U.S. Food and Drug Administration method for detection and identification of Campylobacter jejuni or Campylobacter coli from raw silo milk.

PubMed

Gharst, Greg; Bark, Don H; Newkirk, Robert; Guillen, Lacey; Wang, Qian; Abeyta, Carlos

2013-01-01

The current U.S. Food and Drug Administration (FDA) methodology for detection of Campylobacter, a leading source for foodborne illness, is outdated. The purpose of this study, therefore, was to improve and update the cultural and identification methods found in the FDA/Bacteriological Analytical Manual (BAM). Raw silo milk samples containing typical and atypical strains of Campylobacter jejuni and Campylobacter coli at different levels (5 CFU/25 g, 50 CFU/25 g, and 125 CFU/25 g) were analyzed. Valid results were obtained from 240 test portions. Six inoculated (at the levels described above) and two uninoculated samples were sent to a participating laboratory to mimic a "real-world" scenario. These combined data indicated that the use of sheep blood in combination with enrichment is not necessary. R & F Campylobacter jejuni/Campylobacter coli Chromogenic Plating Medium is significantly (P < 0.05) more sensitive for detection of C. jejuni or C. coli at low inoculation levels than the modified Cefoperazone Charcoal Deoxycholate Agar used in the BAM. The quantitative PCR method described demonstrated rapid confirmation and identification of C. jejuni or C. coli. It reduced the time to isolate C. jejuni or C. coli, and increased the sensitivity compared to the current BAM protocol.

Spinoculation Enhances HBV Infection in NTCP-Reconstituted Hepatocytes.

PubMed

Yan, Ran; Zhang, Yongmei; Cai, Dawei; Liu, Yuanjie; Cuconati, Andrea; Guo, Haitao

2015-01-01

Hepatitis B virus (HBV) infection and its sequelae remain a major public health burden, but both HBV basic research and the development of antiviral therapeutics have been hindered by the lack of an efficient in vitro infection system. Recently, sodium taurocholate cotransporting polypeptide (NTCP) has been identified as the HBV receptor. We herein report that we established a NTCP-complemented HepG2 cell line (HepG2-NTCP12) that supports HBV infection, albeit at a low infectivity level following the reported infection procedures. In our attempts to optimize the infection conditions, we found that the centrifugation of HepG2-NTCP12 cells during HBV inoculation (termed "spinoculation") significantly enhanced the virus infectivity. Moreover, the infection level gradually increased with accelerated speed of spinoculation up to 1,000g tested. However, the enhancement of HBV infection was not significantly dependent upon the duration of centrifugation. Furthermore, covalently closed circular (ccc) DNA was detected in infected cells under optimized infection condition by conventional Southern blot, suggesting a successful establishment of HBV infection after spinoculation. Finally, the parental HepG2 cells remained uninfected under HBV spinoculation, and HBV entry inhibitors targeting NTCP blocked HBV infection when cells were spinoculated, suggesting the authentic virus entry mechanism is unaltered under centrifugal inoculation. Our data suggest that spinoculation could serve as a standard protocol for enhancing the efficiency of HBV infection in vitro.

Spinoculation Enhances HBV Infection in NTCP-Reconstituted Hepatocytes

PubMed Central

Yan, Ran; Zhang, Yongmei; Cai, Dawei; Liu, Yuanjie; Cuconati, Andrea; Guo, Haitao

2015-01-01

Hepatitis B virus (HBV) infection and its sequelae remain a major public health burden, but both HBV basic research and the development of antiviral therapeutics have been hindered by the lack of an efficient in vitro infection system. Recently, sodium taurocholate cotransporting polypeptide (NTCP) has been identified as the HBV receptor. We herein report that we established a NTCP-complemented HepG2 cell line (HepG2-NTCP12) that supports HBV infection, albeit at a low infectivity level following the reported infection procedures. In our attempts to optimize the infection conditions, we found that the centrifugation of HepG2-NTCP12 cells during HBV inoculation (termed “spinoculation”) significantly enhanced the virus infectivity. Moreover, the infection level gradually increased with accelerated speed of spinoculation up to 1,000g tested. However, the enhancement of HBV infection was not significantly dependent upon the duration of centrifugation. Furthermore, covalently closed circular (ccc) DNA was detected in infected cells under optimized infection condition by conventional Southern blot, suggesting a successful establishment of HBV infection after spinoculation. Finally, the parental HepG2 cells remained uninfected under HBV spinoculation, and HBV entry inhibitors targeting NTCP blocked HBV infection when cells were spinoculated, suggesting the authentic virus entry mechanism is unaltered under centrifugal inoculation. Our data suggest that spinoculation could serve as a standard protocol for enhancing the efficiency of HBV infection in vitro. PMID:26070202

Field monitoring of plant-growth-promoting rhizobacteria by colony immunoblotting.

PubMed

Krishnen, Ganisan; Kecskés, Mihály L; Rose, Michael T; Geelan-Small, Peter; Amprayn, Khanok-on; Pereg, Lily; Kennedy, Ivan R

2011-11-01

Inoculant plant-growth-promoting bacteria are emerging as an important component of sustainable agriculture. There is a need to develop inexpensive methods for enumerating these organisms after their application in the field, to better understand their survival and impacts on yields. Immunoblotting is one potential method to measure viable cells, but the high cost of the conventionally used nylon membranes makes this method prohibitive. In this study, less expensive alternative materials such as filter papers, glossy photo papers, and transparencies for the purpose of colony immunoblotting were evaluated and the best substance was chosen for further studies. Whatman filter paper No. 541 combined with a 0.01 mol·L(-1) H(2)SO(4) rinsing step gave similar results to nylon membranes but <20% of the overall cost of the original colony immunoblotting assay. The application of the modified immunoblot method was tested on nonsterile clay soil samples that were spiked with high numbers (>10(7) CFU·g(-1)) of the plant-growth-promoting bacteria Pseudomonas fluorescens , Azospirillum brasilense , or Rhizobium leguminosarum . The modified protocol allowed the identification and recovery of over 50% of the inoculated cells of all three strains, amidst a background of the native soil microflora. Subsequently, the survival of P. fluorescens was successfully monitored for several months after application to field-grown rice at Jerilderie, New South Wales, Australia, thus validating the procedure.

The synergistic effect of combined immunization with a DNA vaccine and chimeric yellow fever/dengue virus leads to strong protection against dengue.

PubMed

Azevedo, Adriana S; Gonçalves, Antônio J S; Archer, Marcia; Freire, Marcos S; Galler, Ricardo; Alves, Ada M B

2013-01-01

The dengue envelope glycoprotein (E) is the major component of virion surface and its ectodomain is composed of domains I, II and III. This protein is the main target for the development of a dengue vaccine with induction of neutralizing antibodies. In the present work, we tested two different vaccination strategies, with combined immunizations in a prime/booster regimen or simultaneous inoculation with a DNA vaccine (pE1D2) and a chimeric yellow fever/dengue 2 virus (YF17D-D2). The pE1D2 DNA vaccine encodes the ectodomain of the envelope DENV2 protein fused to t-PA signal peptide, while the YF17D-D2 was constructed by replacing the prM and E genes from the 17D yellow fever vaccine virus by those from DENV2. Balb/c mice were inoculated with these two vaccines by different prime/booster or simultaneous immunization protocols and most of them induced a synergistic effect on the elicited immune response, mainly in neutralizing antibody production. Furthermore, combined immunization remarkably increased protection against a lethal dose of DENV2, when compared to each vaccine administered alone. Results also revealed that immunization with the DNA vaccine, regardless of the combination with the chimeric virus, induced a robust cell immune response, with production of IFN-γ by CD8+ T lymphocytes.

[Assessment of two DNA extraction methods to amplify the pneumolysin gene (PLY) from blood culture samples of Streptococcus pneumoniae].

PubMed

Hernández, Carolina; Durán, Claudia; Ulloa, María Teresa; Prado, Valeria

2004-05-01

Streptococcus pneumoniae is a common etiologic agent of invasive respiratory infections among children under 5 years of age and older adults. Isolation rates of S. pneumoniae by traditional culture techniques are low. To study the sensitivity and specificity of two different DNA extraction methods to amplify the ply gene, applied to three different types of blood culture broths, experimentally inoculated with S. pneumoniae. DNA was extracted from the cultures using an organic method or a technique that consists in dilution, washing with NaOH and concentration of the sample. This was followed by PCR amplification of a 355 pb fragment of the pneumolysin gene (ply). The organic DNA extraction method inhibited the PCR reaction at all concentrations studied (0.6 to 10(6) colony forming units/mL). Using the NaOH extraction, ply gene amplification was positive in all three blood culture broths, but only at concentrations of 10(3) colony forming units/mL, or higher. Using the same DNA extraction method, PCR was negative when the broths were inoculated with seven other related bacterial species, which results in a 100% specificity. Detection of S. pneumoniae by amplification of ply gene from blood cultures using the protocol of NaOH for DNA extraction is specific and provides results in a short lapse. However, the diagnostic sensitivity is not optimal, which limits its clinical use.

Mice chronically infected with chimeric HIV resist peripheral and brain superinfection: a model of protective immunity to HIV.

PubMed

Kelschenbach, Jennifer L; Saini, Manisha; Hadas, Eran; Gu, Chao-Jiang; Chao, Wei; Bentsman, Galina; Hong, Jessie P; Hanke, Tomas; Sharer, Leroy R; Potash, Mary Jane; Volsky, David J

2012-06-01

Infection by some viruses induces immunity to reinfection, providing a means to identify protective epitopes. To investigate resistance to reinfection in an animal model of HIV disease and its control, we employed infection of mice with chimeric HIV, EcoHIV. When immunocompetent mice were infected by intraperitoneal (IP) injection of EcoHIV, they resisted subsequent secondary infection by IP injection, consistent with a systemic antiviral immune response. To investigate the potential role of these responses in restricting neurotropic HIV infection, we established a protocol for efficient EcoHIV expression in the brain following intracranial (IC) inoculation of virus. When mice were inoculated by IP injection and secondarily by IC injection, they also controlled EcoHIV replication in the brain. To investigate their role in EcoHIV antiviral responses, CD8+ T lymphocytes were isolated from spleens of EcoHIV infected and uninfected mice and adoptively transferred to isogenic recipients. Recipients of EcoHIV primed CD8+ cells resisted subsequent EcoHIV infection compared to recipients of cells from uninfected donors. CD8+ spleen cells from EcoHIV-infected mice also mounted modest but significant interferon-γ responses to two HIV Gag peptide pools. These findings suggest EcoHIV-infected mice may serve as a useful system to investigate the induction of anti-HIV protective immunity for eventual translation to human beings.

Development and optimization of an in vitro cultivation protocol allows for isolation of Borrelia miyamotoi from patients with hard tick-borne relapsing fever.

PubMed

Koetsveld, J; Kolyasnikova, N M; Wagemakers, A; Toporkova, M G; Sarksyan, D S; Oei, A; Platonov, A E; Hovius, J W

2017-07-01

Borrelia miyamotoi has been shown to infect humans in Eurasia and North America causing hard tick-borne relapsing fever (HTBRF). In vitro cultivation of B. miyamotoi was described recently; but clinical isolation of relapsing fever Borrelia is cumbersome. Our aim was to develop a straightforward protocol enabling B. miyamotoi isolation directly from the blood of patients. Modified Kelly-Pettenkorfer (MKP-F) medium, with or without anticoagulants, or blood from healthy human volunteers, was spiked with B. miyamotoi spirochaetes in vitro. Subsequently, either media or plasma was used for cultivation directly, or after an additional centrifugation step. This isolation protocol was tested in a clinical setting on patients suspected of HTBRF. Dipotassium-EDTA, trisodium citrate and lithium heparin inhibited growth of B. miyamotoi at concentrations ≥250 μg/mL, 2.5 mM and 1 IU/mL, respectively. However, when plasma originating from human blood containing B. miyamotoi spirochaetes was subjected to an additional centrifugation step at 8000 g, suspended and inoculated into fresh MKP-F media, positive cultures were observed within 2 weeks. Of importance, this straightforward protocol allowed for isolation of B. miyamotoi from six out of nine patients with confirmed HTBRF. Direct culture from K 2 -EDTA, trisodium citrate and lithium heparin plasma containing B. miyamotoi is hampered due to anticoagulants. Using a simple centrifugation protocol we were able to circumvent this detrimental effect, allowing for the first clinical isolation of B. miyamotoi. This will be of value for future research on the pathogenesis, genetics, diagnosis, therapy and epidemiology of HTBRF and other tick-borne relapsing fevers. Copyright © 2017 European Society of Clinical Microbiology and Infectious Diseases. Published by Elsevier Ltd. All rights reserved.

Strain-specific quantification of root colonization by plant growth promoting rhizobacteria Bacillus firmus I-1582 and Bacillus amyloliquefaciens QST713 in non-sterile soil and field conditions.

PubMed

Mendis, Hajeewaka C; Thomas, Varghese P; Schwientek, Patrick; Salamzade, Rauf; Chien, Jung-Ting; Waidyarathne, Pramuditha; Kloepper, Joseph; De La Fuente, Leonardo

2018-01-01

Bacillus amyloliquefaciens QST713 and B. firmus I-1582 are bacterial strains which are used as active ingredients of commercially-available soil application and seed treatment products Serenade® and VOTiVO®, respectively. These bacteria colonize plant roots promoting plant growth and offering protection against pathogens/pests. The objective of this study was to develop a qPCR protocol to quantitate the dynamics of root colonization by these two strains under field conditions. Primers and TaqMan® probes were designed based on genome comparisons of the two strains with publicly-available and unpublished bacterial genomes of the same species. An optimized qPCR protocol was developed to quantify bacterial colonization of corn roots after seed treatment. Treated corn seeds were planted in non-sterile soil in the greenhouse and grown for 28 days. Specific detection of bacteria was quantified weekly, and showed stable colonization between ~104-105 CFU/g during the experimental period for both bacteria, and the protocol detected as low as 103 CFU/g bacteria on roots. In a separate experiment, streptomycin-resistant QST713 and rifampicin-resistant I-1582 strains were used to compare dilution-plating on TSA with the newly developed qPCR method. Results also indicated that the presence of natural microflora and another inoculated strain does not affect root colonization of either one of these strains. The same qPCR protocol was used to quantitate root colonization by QST713 and I-1582 in two corn and two soybean varieties grown in the field. Both bacteria were quantitated up to two weeks after seeds were planted in the field and there were no significant differences in root colonization in either bacteria strain among varieties. Results presented here confirm that the developed qPCR protocol can be successfully used to understand dynamics of root colonization by these bacteria in plants growing in growth chamber, greenhouse and the field.

Intrauterine inoculation of minipigs with Chlamydia trachomatis during diestrus establishes a longer lasting infection compared to vaginal inoculation during estrus.

PubMed

Lorenzen, Emma; Follmann, Frank; Secher, Jan O; Goericke-Pesch, Sandra; Hansen, Mette S; Zakariassen, Hannah; Olsen, Anja W; Andersen, Peter; Jungersen, Gregers; Agerholm, Jørgen S

2017-06-01

Advanced animal models, such as minipigs, are needed for the development of a globally requested human Chlamydia vaccine. Previous studies have shown that vaginal inoculation of sexually mature Göttingen minipigs with Chlamydia trachomatis resulted in an infection lasting only 3-5 days. The aim of this study was to evaluate the effect of targeting the upper porcine genital tract by transcervical and transabdominal intrauterine inoculation, compared to previously performed vaginal inoculation. Furthermore, we investigated the effect of the hormonal cycle, estrus vs. diestrus, on the establishment of a C. trachomatis infection in the minipig. Targeting the upper genital tract (transcervical inoculation) resulted in a longer lasting infection (at least 7 days) compared to vaginal inoculation (3-5 days). When comparing intrauterine inoculation during estrus and diestrus, inoculation during diestrus resulted in a longer lasting infection (at least 10 days) compared to estrus (3-5 days). Furthermore, we found a significant C. trachomatis specific IFN-γ response in pigs inoculated during estrus correlating with the accelerated clearance of infection in these pigs. These findings suggest that for implementation of an optimal model of C. trachomatis in minipigs, inoculation should bypass the cervix and preferable be performed during diestrus. Copyright © 2017 The Author(s). Published by Elsevier Masson SAS.. All rights reserved.

Evaluation the virulence of Mycobacterium bovis isolated from milk samples through histopathological study in laboratory animals.

PubMed

Al-Saqur, I M; Al-Thwani, A N; Al-Attar, I M; Al-Mashhadani, M S

2016-12-01

Mycobacterium bovis has a broad host range, and it is the principal agent responsible for tuberculosis (TB) in bovine, domestic and wild mammals. M. bovis also infects human, causing zoonotic TB through ingestion, inhalation and, less frequently by contact with mucous membranes and broken skin. Zoonotic TB was formerly an endemic disease, usually transmitted to man by consumption of raw cow's milk. It is indistinguishable clinically or pathologically from TB caused by M. tuberculosis. The aims of this study were, to isolate and identified M. bovis from raw milk samples by different methods, and evaluate the virulence of M. bovis in laboratory animals (Rabbit). To conduct the study, ninety three cow's milk samples were collected from farms around Baghdad governorate. The decontamination of milk samples was firstly carried out, then samples were subjected to routine tests which include, direct smear for Ziehl Neelsen acid fast stain, culture, each sample was cultured on Lowenstein Jensen media with Sodium pyruvite (All cultures incubated on 37°C for 4-10weeks with continuous observation), and biochemical testes as Nitrate reduction test, Niacin paper strip test and pyrazinamidase test, were employed to diagnose and identified the bacteria. Beside molecular assay was used to confirm the identification of the isolates by Polymerase Chain Reaction (PCR) using specific primers for M. bovis. The virulence of these isolates were investigated through inoculate it in group of laboratory animals consist of 8 rabbit in addition to other group of 4 animals as control (inoculate with Phosphate Buffer Saline). The animals were scarified after 6weeks of inoculation, post- mortem examination was carried out, smears were taken from lesions, and tissue samples were collected from lymph nodes and different organs. The results revealed five isolates of M. bovis in direct smear by acid fast Ziehl-Neelsen stain, while eight isolates observed by culture, the colonies appeared with characteristic feature of cream color, rough, and with irregular edge. The molecular assay using PCR technique confirmed the diagnosis of eight positive isolates in smears and culture. The virulence of these isolates were investigated through the pathological effects appeared in inoculated rabbit which showed lesions scattered mainly in lymph nodes and different organs as lung, liver, spleen and kidney when compared with control group which were naive. Beside the infiltration of mononuclear cells in the internal organs particularly in the lungs. The result of histopathological examination clarified the virulence of M. bovis isolates, and its impact on tissue and organs of the rabbit. Our study conclude the presence of M. bovis isolates in milk in high percentage pause important source of tuberculosis infection for human being. Copyright © 2016.

Evaluation of protective efficacy of Spirulina platensis in Balb/C mice with candidiasis.

PubMed

Soltani, M; Khosravi, A-R; Asadi, F; Shokri, H

2012-12-01

This study was aimed at evaluating the immunostimulatory effect of Spirulina platensis in prophylaxis of Balb/C mice with systemic candidiasis. In first experiment, 40 mice were divided into four groups, ten mice per each group, for cytokines assay. Animals received a dose of 800mg/kg of S. platensis for 4days and then were intravenously inoculated with 1×10(6) Candida albicans. Control groups received 0.2mL and 0.1mL normal saline for prophylaxis and inoculation, respectively. Five mice from each group were euthanized after 24hours and 72hours and the serum levels of IFN-γ and TNF-α were measured by Enzyme-linked immunosorbent assay (ELISA) method. In second experiment, two mice groups with systemic candidiasis, 11 mice per each group, were included to evaluate the survival rate. Animals were monitored for 30days and the kidneys, liver, lungs and spleen were analyzed for fungal invasion. The results indicated that the Spirulina-treated mice produced more IFN-g and TNF-α level than their control groups. This infected group showed that the mean survival time (28.86±2.7) was significantly (P<0.05) higher than control group (13.9±3.34). They also exhibited that fungal clearance in selected organs at death time represents significant differences between spleen and liver (P<0.05). Prophylaxis with S. platensis had synergistic effect through producing cytokines such as TNF-α and IFN-γ. Our results provide important information for the potential application of S. platensis in the treatment and resistance of Balb/C mice with systemic candidiasis. Copyright © 2012 Elsevier Masson SAS. All rights reserved.

Characterization of phenotypes of immune cells and cytokines associated with chronic exposure to Premolis semirufa caterpillar bristles extract.

PubMed

Villas-Boas, Isadora Maria; Gonçalves-de-Andrade, Rute Maria; Squaiella-Baptistão, Carla Cristina; Sant'Anna, Osvaldo Augusto; Tambourgi, Denise V

2013-01-01

The Brazilian moth Premolis semirufa (Walker, 1856), usually called pararama, is a parasite of the rubber Hevea genus. Contact with the bristles causes symptoms of acute inflammation. A chronic inflammatory reaction frequently occurs in individuals after multiple contacts, and this reaction is characterised by articular synovial membrane thickening with joint deformities, common characteristics of chronic synovitis. Extract from the bristles has been shown to induce an intense inflammatory response in a murine model, and this reaction was characterised by the presence of neutrophils in the paw tissues of injected mice and a strong, specific antibody response. There is not yet an effective treatment for incidents involving contact with pararama. In this study, we evaluated the phenotype of the immunological response and cytokine production in BALB/c mice subcutaneously injected in the footpad with P. semirufa bristle extract or sterile saline (control) seven times at 15 day intervals. An analysis of cells from the draining lymph node by flow cytometry showed that the absolute numbers of TCD4, TCD8 and B lymphocytes, as well as the expression of activation molecules, were higher in the extract-treated group. Furthermore, immunohistochemistry and immunofluorescence analyses showed a mixed inflammatory infiltrate composed of neutrophils and macrophages at the inoculation site. In addition, an analysis of paw cytokines showed elevated levels of IL-6, IL-12, IL-10, IL-17 and IL-23 after the 7(th) inoculation. In conclusion, these data provide evidence of pro-inflammatory changes in the phenotypes of immune cells and cytokine production in animals subjected to injections with an extract from Premolis semirufa bristles, which may explain the intense and prolonged inflammatory response that characterises this disorder.

Glutamine supplementation improves intestinal barrier function in a weaned piglet model of Escherichia coli infection.

PubMed

Ewaschuk, Julia B; Murdoch, Gordon K; Johnson, Ian R; Madsen, Karen L; Field, Catherine J

2011-09-01

The weaning period is associated with an increased prevalence of gastrointestinal infection in many species. Glutamine (Gln) has been shown to improve intestinal barrier function and immune function in both in vivo and in vitro models. The objective of the present study was to determine the effect of dietary Gln supplementation on intestinal barrier function and intestinal cytokines in a model of Escherichia coli infection. We randomised 21-d-old piglets (n 20) to nutritionally complete isonitrogenous diets with or without Gln (4·4 %, w/w) for 2 weeks. Intestinal loops were isolated from anaesthetised pigs and inoculated with either saline or one of the two E. coli (K88AC or K88 wild-type)-containing solutions. Intestinal tissue was studied for permeability, cytokine expression, fluid secretion and tight-junction protein expression. Animals receiving Gln supplementation had decreased potential difference (PD) and short-circuit current (I(sc)) in E. coli-inoculated intestinal loops (PD 0·628 (SEM 0·151) mV; I(sc) 13·0 (SEM 3·07) μA/cm(2)) compared with control-fed animals (PD 1·36 (SEM 0·227) mV; I(sc) 22·4 (SEM 2·24) μA/cm(2)). Intestinal tissue from control, but not from Gln-supplemented, animals responded to E. coli with a significant increase in mucosal cytokine mRNA (IL-1β, IL-6, transforming growth factor-β and IL-10). Tight-junction protein expression (claudin-1 and occludin) was reduced with exposure to E. coli in control-fed animals and was not influenced in Gln-supplemented piglets. Gln supplementation may be useful in reducing the severity of weaning-related gastrointestinal infections, by reducing the mucosal cytokine response and altering intestinal barrier function.

Gibberellins Producing Endophytic Fungus Porostereum spadiceum AGH786 Rescues Growth of Salt Affected Soybean

PubMed Central

Hamayun, Muhammad; Hussain, Anwar; Khan, Sumera A.; Kim, Ho-Youn; Khan, Abdul L.; Waqas, Muhammad; Irshad, Muhammad; Iqbal, Amjad; Rehman, Gauhar; Jan, Samin; Lee, In-Jung

2017-01-01

In the pursuit of sustainable agriculture through environment and human health friendly practices, we evaluated the potential of a novel gibberellins (GAs) producing basidiomycetous endophytic fungus Porostereum spadiceum AGH786, for alleviating salt stress and promoting health benefits of soybean. Soybean seedlings exposed to different levels of NaCl stress (70 and 140 mM) under greenhouse conditions, were inoculated with the AGH786 strain. Levels of phytohormones including GAs, JA and ABA, and isoflavones were compared in control and the inoculated seedlings to understand the mechanism through which the stress is alleviated. Gibberellins producing endophytic fungi have been vital for promoting plant growth under normal and stress conditions. We report P. spadiceum AGH786 as the ever first GAs producing basidiomycetous fungus capable of producing six types of GAs. In comparison to the so for most efficient GAs producing Gibberella fujikuroi, AGH786 produced significantly higher amount of the bioactive GA3. Salt-stressed phenotype of soybean seedlings was characterized by low content of GAs and high amount of ABA and JA with reduced shoot length, biomass, leaf area, chlorophyll contents, and rate of photosynthesis. Mitigation of salt stress by AGH786 was always accompanied by high GAs, and low ABA and JA, suggesting that this endophytic fungus reduces the effect of salinity by modulating endogenous phytohormones of the seedlings. Additionally, this strain also enhanced the endogenous level of two isoflavones including daidzen and genistein in soybean seedlings under normal as well as salt stress conditions as compared to their respective controls. P. spadiceum AGH786 boosted the NaCl stress tolerance and growth in soybean, by modulating seedlings endogenous phytohormones and isoflavones suggesting a valuable contribution of this potent fungal biofertilizer in sustainable agriculture in salt affected soils. PMID:28473818

Antibacterial Efficacy of Super-Oxidized Water on Enterococcus faecalis Biofilms in Root Canal

PubMed Central

Zan, Recai; Alacam, Tayfun; Hubbezoglu, Ihsan; Tunc, Tutku; Sumer, Zeynep; Alici, Oguzhan

2016-01-01

Background The success of endodontic treatment depends on a few crucial factors. One of these factors is the complete chemomechanic preparation of root canal against various bacteria. In particular, the effect of resistant bacteria may cause intense pain with flare-up and formation of periapical lesions. Therefore, the strong effect of irrigants plays an important role in terms of the complete elimination of these bacteria to achieve long-term successful treatment. Objectives The aim of this study was to investigate the antibacterial effects of super-oxidized water (SPO) in root canals infected with Enterococcus faecalis biofilms. Methods One hundred twenty single-root, premolar teeth were selected. Initially, the teeth were prepared and then disinfected. E. faecalis were inoculated and kept at 37°C for 24 hours in the root canals. The re-inoculation procedure was repeated on the first, fourth, seventh, and tenth days. The infected root canals were divided into one negative (saline) and one positive (sodium hypochlorite) control group and four experimental groups (super-oxidized water: 1, 2, 3, or 5 minutes) (n = 20). Paper points were placed in the root canals to control and evaluate the biofilm formation. Biofilms were counted on blood agar plates, and data was evaluated and statistically analyzed using one-way ANOVA and Tukey’s test. Results Although sodium hypochlorite (NaOCl) showed no statistically significant difference when compared with three and five minutes of SPO irrigation (P > 0.05), NaOCl showed statistically significant differences among all other groups (P < 0.05). Conclusions Super-oxidized water indicated a remarkable and similar bactericidal effect to that of traditional NaOCl against E. faecalis biofilms. In terms of successful endodontic treatment approaches, super-oxidized water may be used as an effective irrigation solution in clinics. PMID:27800142

Resistance exercise attenuates skeletal muscle oxidative stress, systemic pro-inflammatory state, and cachexia in Walker-256 tumor-bearing rats.

PubMed

Padilha, Camila Souza; Borges, Fernando Henrique; Costa Mendes da Silva, Lilian Eslaine; Frajacomo, Fernando Tadeu Trevisan; Jordao, Alceu Afonso; Duarte, José Alberto; Cecchini, Rubens; Guarnier, Flávia Alessandra; Deminice, Rafael

2017-09-01

The aim of this study was to investigate the effects of resistance exercise training (RET) on oxidative stress, systemic inflammatory markers, and muscle wasting in Walker-256 tumor-bearing rats. Male (Wistar) rats were divided into 4 groups: sedentary controls (n = 9), tumor-bearing (n = 9), exercised (n = 9), and tumor-bearing exercised (n = 10). Exercised and tumor-bearing exercised rats were exposed to resistance exercise of climbing a ladder apparatus with weights tied to their tails for 6 weeks. The physical activity of control and tumor-bearing rats was confined to the space of the cage. After this period, tumor-bearing and tumor-bearing exercised animals were inoculated subcutaneously with Walker-256 tumor cells (11.0 × 10 7 cells in 0.5 mL of phosphate-buffered saline) while control and exercised rats were injected with vehicle. Following inoculation, rats maintained resistance exercise training (exercised and tumor-bearing exercised) or sedentary behavior (control and tumor-bearing) for 12 more days, after which they were euthanized. Results showed muscle wasting in the tumor-bearing group, with body weight loss, increased systemic leukocytes, and inflammatory interleukins as well as muscular oxidative stress and reduced mTOR signaling. In contrast, RET in the tumor-bearing exercised group was able to mitigate the reduced body weight and muscle wasting with the attenuation of muscle oxidative stress and systemic inflammatory markers. RET also prevented loss of muscle strength associated with tumor development. RET, however, did not prevent the muscle proteolysis signaling via FBXO32 gene messenger RNA expression in the tumor-bearing group. In conclusion, RET performed prior tumor implantation prevents cachexia development by attenuating tumor-induced systemic pro-inflammatory condition with muscle oxidative stress and muscle damage.

Pock forming ability of fowl pox virus isolated from layer chicken and its adaptation in chicken embryo fibroblast cell culture

PubMed Central

Gilhare, Varsha Rani; Hirpurkar, S. D.; Kumar, Ashish; Naik, Surendra Kumar; Sahu, Tarini

2015-01-01

Aim: The objective of the present study was to examine pock forming ability of field strain and vaccine strain of fowl pox virus (FPV) in chorioallantoic membrane (CAM) of embryonated chicken eggs and its adaptation in chicken embryo fibroblast (CEF) cell culture. Materials and Methods: Dry scabs were collected from 25 affected birds in glycerin-saline and preserved at 4°C until processed. Virus was isolated in 10-day-old embryonated chicken eggs by dropped CAM method. The identity of the virus is confirmed by clinical findings of affected birds, pock morphology and histopathology of infected CAM. In addition one field isolate and vaccine strain of FPV was adapted to CEF cell culture. CEF cell culture was prepared from 9-day-old embryonated chicken eggs. Result: Clinical symptoms observed in affected birds include pox lesion on comb, wattle, eyelids and legs, no internal lesions were observed. All field isolates produced similar findings in CAM. Pocks produced by field isolates ranged from 3 mm to 5 mm at the third passage while initial passages edematous thickening and necrosis of CAM was observed. Pocks formed by lyophilized strain were ranges from 0.5 mm to 2.5 mm in diameter scattered all over the membrane at the first passage. Intra-cytoplasmic inclusion bodies are found on histopathology of CAM. At third passage level, the CEF inoculated with FPV showed characteristic cytopathic effect (CPE) included aggregation of cells, syncytia and plaque formation. Conclusion: FPV field isolates and vaccine strain produced distinct pock lesions on CAMs. Infected CAM showed intracytoplasmic inclusion bodies. The CEF inoculated with FPV field isolate as well as a vaccine strain showed characteristic CPE at third passage level. PMID:27047081

Pock forming ability of fowl pox virus isolated from layer chicken and its adaptation in chicken embryo fibroblast cell culture.

PubMed

Gilhare, Varsha Rani; Hirpurkar, S D; Kumar, Ashish; Naik, Surendra Kumar; Sahu, Tarini

2015-03-01

The objective of the present study was to examine pock forming ability of field strain and vaccine strain of fowl pox virus (FPV) in chorioallantoic membrane (CAM) of embryonated chicken eggs and its adaptation in chicken embryo fibroblast (CEF) cell culture. Dry scabs were collected from 25 affected birds in glycerin-saline and preserved at 4°C until processed. Virus was isolated in 10-day-old embryonated chicken eggs by dropped CAM method. The identity of the virus is confirmed by clinical findings of affected birds, pock morphology and histopathology of infected CAM. In addition one field isolate and vaccine strain of FPV was adapted to CEF cell culture. CEF cell culture was prepared from 9-day-old embryonated chicken eggs. Clinical symptoms observed in affected birds include pox lesion on comb, wattle, eyelids and legs, no internal lesions were observed. All field isolates produced similar findings in CAM. Pocks produced by field isolates ranged from 3 mm to 5 mm at the third passage while initial passages edematous thickening and necrosis of CAM was observed. Pocks formed by lyophilized strain were ranges from 0.5 mm to 2.5 mm in diameter scattered all over the membrane at the first passage. Intra-cytoplasmic inclusion bodies are found on histopathology of CAM. At third passage level, the CEF inoculated with FPV showed characteristic cytopathic effect (CPE) included aggregation of cells, syncytia and plaque formation. FPV field isolates and vaccine strain produced distinct pock lesions on CAMs. Infected CAM showed intracytoplasmic inclusion bodies. The CEF inoculated with FPV field isolate as well as a vaccine strain showed characteristic CPE at third passage level.

Inefficacy of marketed contact lens disinfection solutions against keratitis-causing Acanthamoeba castellanii belonging to the T4 genotype.

PubMed

Lakhundi, Sahreena; Khan, Naveed Ahmed; Siddiqui, Ruqaiyyah

2014-06-01

The aim of this study was to assess the anti-amoebic effects of marketed contact lens disinfecting solutions. Using amoebistatic, amoebicidal, and cysticidal assays, nine different contact lens solutions were tested including: ReNu MultiPlus, DuraPlus, Ultimate Plus, OptiFree Replenish, OptiFree Express, Kontex Clean, Kontex Normal, Kontex Multisol extra+, Kontex Soak. In vitro growth inhibition (amoebistatic) assays were performed by incubating Acanthamoeba castellanii with aforementioned contact lens disinfection solutions as per manufacturer's instructions in the growth medium for up to 48h at 30°C. To determine amoebicidal and cysticidal effects, amoebae were incubated with contact lens solutions in phosphate buffered saline for 24h and viability was determined by haemocytometer counting as well as re-inoculating them in the growth medium. For controls, solutions were tested against bacterial corneal pathogen, Pseudomonas aeruginosa, as well as amoebae were incubated with the solvent alone. Of the nine contact lens solutions tested, none of them showed potent amoebicidal effects. Only DuraPlus and OptiFree Replenish exhibited trophozoite lysis of 85.3% and 73.7% respectively. In contrast, all contact lens disinfection solutions except Kontex Clean, Kontex Normal, Kontex Multisol extra+, tested showed amoebistatic effects. Importantly, none of the contact lens disinfection solutions exhibited cysticidal effects using qualitative assays, i.e., cysts treated with aforementioned solutions re-emerged as viable amoebae upon inoculation in the growth medium. However, more than 3-log reduction was observed when ReNu MultiPlus, DuraPlus and OptiFree Express were tested against P. aeruginosa which is in accordance with the ISO Stand-Alone Primary acceptance criteria. These findings are of great concern for contact lens users. Copyright © 2014 Elsevier Inc. All rights reserved.

Alteration in gene expression profile and oncogenicity of esophageal squamous cell carcinoma by RIZ1 upregulation.

PubMed

Dong, Shang-Wen; Li, Dong; Xu, Cong; Sun, Pei; Wang, Yuan-Guo; Zhang, Peng

2013-10-07

To investigate the effect of retinoblastoma protein-interacting zinc finger gene 1 (RIZ1) upregulation in gene expression profile and oncogenicity of human esophageal squamous cell carcinoma (ESCC) cell line TE13. TE13 cells were transfected with pcDNA3.1(+)/RIZ1 and pcDNA3.1(+). Changes in gene expression profile were screened and the microarray results were confirmed by reverse transcription-polymerase chain reaction (RT-PCR). Nude mice were inoculated with TE13 cells to establish ESCC xenografts. After two weeks, the inoculated mice were randomly divided into three groups. Tumors were injected with normal saline, transfection reagent pcDNA3.1(+) and transfection reagent pcDNA3.1(+)/RIZ1, respectively. Tumor development was quantified, and changes in gene expression of RIZ1 transfected tumors were detected by RT-PCR and Western blotting. DNA microarray data showed that RIZ1 transfection induced widespread changes in gene expression profile of cell line TE13, with 960 genes upregulated and 1163 downregulated. Treatment of tumor xenografts with RIZ1 recombinant plasmid significantly inhibited tumor growth, decreased tumor size, and increased expression of RIZ1 mRNA compared to control groups. The changes in gene expression profile were also observed in vivo after RIZ1 transfection. Most of the differentially expressed genes were associated with cell development, supervision of viral replication, lymphocyte costimulatory and immune system development in esophageal cells. RIZ1 gene may be involved in multiple cancer pathways, such as cytokine receptor interaction and transforming growth factor beta signaling. The development and progression of esophageal cancer are related to the inactivation of RIZ1. Virus infection may also be an important factor.

Quantification of Azospirillum brasilense FP2 Bacteria in Wheat Roots by Strain-Specific Quantitative PCR.

PubMed

Stets, Maria Isabel; Alqueres, Sylvia Maria Campbell; Souza, Emanuel Maltempi; Pedrosa, Fábio de Oliveira; Schmid, Michael; Hartmann, Anton; Cruz, Leonardo Magalhães

2015-10-01

Azospirillum is a rhizobacterial genus containing plant growth-promoting species associated with different crops worldwide. Azospirillum brasilense strains exhibit a growth-promoting effect by means of phytohormone production and possibly by N2 fixation. However, one of the most important factors for achieving an increase in crop yield by plant growth-promoting rhizobacteria is the survival of the inoculant in the rhizosphere, which is not always achieved. The objective of this study was to develop quantitative PCR protocols for the strain-specific quantification of A. brasilense FP2. A novel approach was applied to identify strain-specific DNA sequences based on a comparison of the genomic sequences within the same species. The draft genome sequences of A. brasilense FP2 and Sp245 were aligned, and FP2-specific regions were filtered and checked for other possible matches in public databases. Strain-specific regions were then selected to design and evaluate strain-specific primer pairs. The primer pairs AzoR2.1, AzoR2.2, AzoR5.1, AzoR5.2, and AzoR5.3 were specific for the A. brasilense FP2 strain. These primer pairs were used to monitor quantitatively the population of A. brasilense in wheat roots under sterile and nonsterile growth conditions. In addition, coinoculations with other plant growth-promoting bacteria in wheat were performed under nonsterile conditions. The results showed that A. brasilense FP2 inoculated into wheat roots is highly competitive and achieves high cell numbers (∼10(7) CFU/g [fresh weight] of root) in the rhizosphere even under nonsterile conditions and when coinoculated with other rhizobacteria, maintaining the population at rather stable levels for at least up to 13 days after inoculation. The strategy used here can be applied to other organisms whose genome sequences are available. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Quantification of Azospirillum brasilense FP2 Bacteria in Wheat Roots by Strain-Specific Quantitative PCR

PubMed Central

Stets, Maria Isabel; Alqueres, Sylvia Maria Campbell; Souza, Emanuel Maltempi; Pedrosa, Fábio de Oliveira; Schmid, Michael

2015-01-01

Azospirillum is a rhizobacterial genus containing plant growth-promoting species associated with different crops worldwide. Azospirillum brasilense strains exhibit a growth-promoting effect by means of phytohormone production and possibly by N2 fixation. However, one of the most important factors for achieving an increase in crop yield by plant growth-promoting rhizobacteria is the survival of the inoculant in the rhizosphere, which is not always achieved. The objective of this study was to develop quantitative PCR protocols for the strain-specific quantification of A. brasilense FP2. A novel approach was applied to identify strain-specific DNA sequences based on a comparison of the genomic sequences within the same species. The draft genome sequences of A. brasilense FP2 and Sp245 were aligned, and FP2-specific regions were filtered and checked for other possible matches in public databases. Strain-specific regions were then selected to design and evaluate strain-specific primer pairs. The primer pairs AzoR2.1, AzoR2.2, AzoR5.1, AzoR5.2, and AzoR5.3 were specific for the A. brasilense FP2 strain. These primer pairs were used to monitor quantitatively the population of A. brasilense in wheat roots under sterile and nonsterile growth conditions. In addition, coinoculations with other plant growth-promoting bacteria in wheat were performed under nonsterile conditions. The results showed that A. brasilense FP2 inoculated into wheat roots is highly competitive and achieves high cell numbers (∼107 CFU/g [fresh weight] of root) in the rhizosphere even under nonsterile conditions and when coinoculated with other rhizobacteria, maintaining the population at rather stable levels for at least up to 13 days after inoculation. The strategy used here can be applied to other organisms whose genome sequences are available. PMID:26187960

Heat Melt Compaction as an Effective Treatment for Eliminating Microorganisms from Solid Waste

NASA Technical Reports Server (NTRS)

Hummerick, Mary P.; Strayer, Richard; McCoy, LaShelle; Richard, Jeffrey; Ruby, Anna; Wheeler, Raymond

2012-01-01

One of the technologies being tested at Ames Research Center as part of the logistics and repurposing project is heat melt compaction (HMC) of solid waste to reduce volume, remove water and render a biologically stable and safe product. Studies at Kennedy Space Center have focused on the efficacy of the heat melt compaction process for killing microorganisms in waste and specific compacter operation protocols, i.e., time and temperature, required to achieve a sterile, stable product. The work reported here includes a controlled study to examine the survival and potential re-growth of specific microorganisms over a 6-month period of storage after heating and compaction. Before heating and compaction, ersatz solid wastes were inoculated with Bacillus amyloliquefaciens and Rhodotorula mucilaginosa, previously isolated from recovered space shuttle mission food and packaging waste. Compacted HMC tiles were sampled for microbiological analysis at time points between 0 and 180 days of storage in a controlled environment chamber. In addition, biological indicator strips containing spores of Bacillus atrophaeus and Ceo bacillus stearothermophilus were imbedded in trash to assess the efficacy of the HMC process to achieve sterilization. Analysis of several tiles compacted at 180 C for times of 40 minutes to over 2 hours detected organisms in all tile samples with the exception of one exposed to 180 C for approximately 2 hours. Neither of the inoculated organisms was recovered, and the biological indicator strips were negative for growth in all tiles indicating at least local sterilization of tile areas. The findings suggest that minimum time/temperature combination is required for complete sterilization. Microbial analysis of tiles processed at lower temperatures from 130 C-150 C at varying times will be discussed, as well as analysis of the bacteria and fungi present on the compactor hardware as a result of exposure to the waste and the surrounding environment. The two organisms inoculated into the waste were among those isolated and identified from the HMC surfaces indicating the possibility of cross contamination.

Novel Production Protocol for Small-scale Manufacture of Probiotic Fermented Foods

PubMed Central

Westerik, Nieke; Wacoo, Alex Paul; Sybesma, Wilbert; Kort, Remco

2016-01-01

A novel dried bacterial consortium of Lactobacillus rhamnosus yoba 2012 and Streptococcus thermophilus C106 is cultured in 1 L of milk. This fresh starter can be used for the production of fermented milk and other fermented foods either at home or at small-scale in rural settings. For the fresh starter, 1 L of milk is pasteurized in a pan that fits into a larger pan containing water, placed on a source of heat. In this water bath, the milk is heated and incubated at 85 °C for 30 min. Thereafter, the milk is cooled down to 45 °C, transferred to a vacuum flask, inoculated with the dried bacteria and left for at least 16 hr between 30 °C and 45 °C. For the purpose of frequent home production, the fresh starter is frozen into ice cubes, which can be used for the production of small volumes of up to 2 L of fermented milk. For the purpose of small-scale production in resource-poor countries, pasteurization of up to 100 L of milk is conducted in milk cans that are placed in a large sauce pan filled with water and heated on a fire at 85 °C for 30 min, and subsequently cooled to 45 °C. Next, the 100 L batch is inoculated with the 1 L freshly prepared starter mentioned before. To assure an effective fermentation at a temperature between 30 and 45 °C, the milk can is covered with a blanket for 12 hr. For the production of non-dairy fermented foods, the fresh starter is left in a cheese cloth for 12 hr, and the drained-off whey can be subsequently used for the inoculation of a wide range of food raw materials, including vegetables and cereal-based foods. PMID:27684196

Novel Production Protocol for Small-scale Manufacture of Probiotic Fermented Foods.

PubMed

Westerik, Nieke; Wacoo, Alex Paul; Sybesma, Wilbert; Kort, Remco

2016-09-10

A novel dried bacterial consortium of Lactobacillus rhamnosus yoba 2012 and Streptococcus thermophilus C106 is cultured in 1 L of milk. This fresh starter can be used for the production of fermented milk and other fermented foods either at home or at small-scale in rural settings. For the fresh starter, 1 L of milk is pasteurized in a pan that fits into a larger pan containing water, placed on a source of heat. In this water bath, the milk is heated and incubated at 85 °C for 30 min. Thereafter, the milk is cooled down to 45 °C, transferred to a vacuum flask, inoculated with the dried bacteria and left for at least 16 hr between 30 °C and 45 °C. For the purpose of frequent home production, the fresh starter is frozen into ice cubes, which can be used for the production of small volumes of up to 2 L of fermented milk. For the purpose of small-scale production in resource-poor countries, pasteurization of up to 100 L of milk is conducted in milk cans that are placed in a large sauce pan filled with water and heated on a fire at 85 °C for 30 min, and subsequently cooled to 45 °C. Next, the 100 L batch is inoculated with the 1 L freshly prepared starter mentioned before. To assure an effective fermentation at a temperature between 30 and 45 °C, the milk can is covered with a blanket for 12 hr. For the production of non-dairy fermented foods, the fresh starter is left in a cheese cloth for 12 hr, and the drained-off whey can be subsequently used for the inoculation of a wide range of food raw materials, including vegetables and cereal-based foods.

Validation of a modification to Performance-Tested Method 010403: microwell DNA hybridization assay for detection of Listeria spp. in selected foods and selected environmental surfaces.

PubMed

Alles, Susan; Peng, Linda X; Mozola, Mark A

2009-01-01

A modification to Performance-Tested Method 010403, GeneQuence Listeria Test (DNAH method), is described. The modified method uses a new media formulation, LESS enrichment broth, in single-step enrichment protocols for both foods and environmental sponge and swab samples. Food samples are enriched for 27-30 h at 30 degrees C, and environmental samples for 24-48 h at 30 degrees C. Implementation of these abbreviated enrichment procedures allows test results to be obtained on a next-day basis. In testing of 14 food types in internal comparative studies with inoculated samples, there were statistically significant differences in method performance between the DNAH method and reference culture procedures for only 2 foods (pasteurized crab meat and lettuce) at the 27 h enrichment time point and for only a single food (pasteurized crab meat) in one trial at the 30 h enrichment time point. Independent laboratory testing with 3 foods showed statistical equivalence between the methods for all foods, and results support the findings of the internal trials. Overall, considering both internal and independent laboratory trials, sensitivity of the DNAH method relative to the reference culture procedures was 90.5%. Results of testing 5 environmental surfaces inoculated with various strains of Listeria spp. showed that the DNAH method was more productive than the reference U.S. Department of Agriculture-Food Safety and Inspection Service (USDA-FSIS) culture procedure for 3 surfaces (stainless steel, plastic, and cast iron), whereas results were statistically equivalent to the reference method for the other 2 surfaces (ceramic tile and sealed concrete). An independent laboratory trial with ceramic tile inoculated with L. monocytogenes confirmed the effectiveness of the DNAH method at the 24 h time point. Overall, sensitivity of the DNAH method at 24 h relative to that of the USDA-FSIS method was 152%. The DNAH method exhibited extremely high specificity, with only 1% false-positive reactions overall.

Nontuberculous mycobacterial infection of the musculoskeletal system in immunocompetent hosts.

PubMed

Gundavda, Manit K; Patil, Hitendra G; Agashe, Vikas M; Soman, Rajeev; Rodriques, Camilla; Deshpande, Ramesh B

2017-01-01

Nontuberculous mycobacteria (NTM) were considered saprophytic organisms for many years but now are recognized as human pathogens. Although humans are routinely exposed to NTM, the rate of clinical infection is low. Such infections usually occur in the elderly and in patients who are immunocompromised. However, there has been an increasing incidence in recent years of infections in immunocompetent hosts. NTM infections in immunocompetent individuals are secondary to direct inoculation either contamination from surgical procedures or penetrating injuries rather than hematogenous dissemination. Clinically and on histopathology, musculoskeletal infections caused by NTM resemble those caused by Mycobacterium tuberculosis but are mostly resistant to routine antituberculosis medicines. Six cases of NTM infection in immunocompetent hosts presenting to the department from 2004 to 2015 were included in study. Of which two cases (one patella and one humerus) of infection were following an open wound due to trauma while two cases (one hip and one shoulder) of infection were by inoculation following an intraarticular injection for arthrogram of the joint, one case was infection following arthroscopy of knee joint and one case (calcaneum) was infection following local injection for the treatment of plantar fasciitis. All patients underwent inaging and tissue diagnosis with samples being sent for culture, staining, and histopathology. Clinical suspicion of NTM inoculation led to the correct diagnosis (four cases with culture positive and two cases with histopathological diagnosis). There treatment protocol for extrapulmonary NTM infection was radical surgical debridement and medical management based on drug sensitivity testing in culture positive cases. At a mean follow up of 3 years (range1-9 years) all patients had total remission and excellent results. Whenever a case of chronic granulomatous infection is encountered that does not respond to standard anti-tuberculous treatment, with a history of open trauma, surgical intervention, or injection as shown in this study, a possible NTM infection should be considered and managed appropriately.

The airway inflammation induced by nasal inoculation of PM2.5 and the treatment of bacterial lysates in rats.

PubMed

Shen, Yang; Zhang, Zhi-Hai; Hu, Di; Ke, Xia; Gu, Zheng; Zou, Qi-Yuan; Hu, Guo-Hua; Song, Shang-Hua; Kang, Hou-Yong; Hong, Su-Ling

2018-06-29

Particulate matter (PM) is one of the most important environmental issues in China. This study aimed to explore the correlation between PM2.5 and airway inflammation in healthy rats. The PM2.5 group was given an intranasal instillation of PM2.5 suspension on 15 consecutive days, and each received oral saline from day 16 to 90. The BV intervention group was treated as the PM2.5 exposure group, except that BV instead of saline was given daily. A histopathologic examination was performed to evaluate the airway inflammation. The prevalence and function of Th1/Th2/Treg/Th17 cells were detected by flow cytometry and ELISA. The expression of AhR was detected by western blot and real-time PCR. We found that epithelial damage and increased infiltration of inflammatory cell were present in the airways after PM2.5 exposure; there was an immune imbalance of Th cells in the PM2.5 group; the expression of AhR was increased in the airways after PM2.5 exposure. In the PM2.5 + BV group, we demonstrated alleviated immune imbalance and reduced inflammatory cell infiltration in the airways. Our study showed that exposure to PM2.5 induced airway inflammation. The imbalance of Th1/Th2/Treg/Th17 in PM2.5-induced airway inflammation might be associated with activation of the AhR pathway. Oral BV reduces PM2.5-induced airway inflammation and regulates systemic immune responses in rats.

Success rates of intrauterine inoculations of layers via the vagina.

PubMed

Landman, W J M; Matthijs, M G R; van Eck, J H H

2013-02-01

Intrauterine inoculation of layer hens has been documented previously in the literature; however, its efficiency has only been assessed on a very small scale. Attempts were therefore made to inoculate 14 experimental groups each consisting of 10 commercial table egg-producing hens intrauterinely with methylene blue. The effect of four variables-oviposition (natural, hormonally induced or not), position of the hen at inoculation (vertical/horizontal), technique to access the uterus (three methods) and inoculation device (four types)-on the success rate of intrauterine inoculation of layers of different breed and age was studied. Immediately after inoculation, hens were euthanized and the presence of methylene blue and perforations in the oviduct were examined. Successful inoculation rates ranged from 0/10 to 10/10. The 100% success rate was obtained in birds without an egg in the uterus, which were restrained in a horizontal position and inoculated with a hard catheter after exposure of the uterovaginal junction. The second best score was obtained in hens inoculated shortly after natural oviposition, maintained in a vertical position and using a corkscrew-shaped saliva ejector after exposure of the vaginal orifice (7/10). In all other groups the success rate was 5/10 or less. Vaginal perforation occurred in none to eight birds per group. It is concluded that intrauterine inoculation of laying hens is only reliable in birds without an egg in the uterus after exposure of the uterovaginal junction. All other methods proved unreliable and often caused vaginal perforations. However, intravaginal inoculation can be performed with 100% accuracy providing the vaginal orifice is exposed.

Effect of controlled inoculation with specific mycorrhizal fungi from the urban environment on growth and physiology of containerized shade tree species growing under different water regimes.

PubMed

Fini, Alessio; Frangi, Piero; Amoroso, Gabriele; Piatti, Riccardo; Faoro, Marco; Bellasio, Chandra; Ferrini, Francesco

2011-11-01

The aim of this work was to evaluate the effects of selected mycorrhiza obtained in the urban environment on growth, leaf gas exchange, and drought tolerance of containerized plants growing in the nursery. Two-year-old uniform Acer campestre L., Tilia cordata Mill., and Quercus robur L. were inoculated with a mixture of infected roots and mycelium of selected arbuscular (maple, linden) and/or ectomycorrhiza (linden, oak) fungi and grown in well-watered or water shortage conditions. Plant biomass and leaf area were measured 1 and 2 years after inoculation. Leaf gas exchange, chlorophyll fluorescence, and water relations were measured during the first and second growing seasons after inoculation. Our data suggest that the mycelium-based inoculum used in this experiment was able to colonize the roots of the tree species growing in the nursery. Plant biomass was affected by water shortage, but not by inoculation. Leaf area was affected by water regime and, in oak and linden, by inoculation. Leaf gas exchange was affected by inoculation and water stress. V(cmax) and J(max) were increased by inoculation and decreased by water shortage in all species. F(v)/F(m) was also generally higher in inoculated plants than in control. Changes in PSII photochemistry and photosynthesis may be related to the capacity of inoculated plants to maintain less negative leaf water potential under drought conditions. The overall data suggest that inoculated plants were better able to maintain physiological activity during water stress in comparison to non-inoculated plants.

Micropropagation of Asparagus by in vitro shoot culture.

PubMed

Stajner, Nataša

2013-01-01

Asparagus officinalis is most extensively studied species within the genus Asparagus, which is well known as garden asparagus. This species is dioecious with unisexual flowers, which means that generative propagation gives roughly equal number of male and female plants. Male plants are high yielders and preferred commercially over female plants. Tissue culture techniques could efficiently promote vegetative propagation of male plants and pave the way for efficient plant breeding.This chapter describes an efficient micropropagation protocol for developing rapid growing in vitro Asparagus shoot cultures. The source of explants, inoculation, and shoot proliferation, followed by shoot propagation, rooting, and acclimatization is described. The optimal medium for Asparagus micropropagation described in this chapter is composed of MS macro- and microelements and a combination of auxins and cytokinins. Plant growth regulators NAA, kinetin, and BA were used in various concentrations. Three different media representing the whole micropropagation protocol of Asparagus are described; medium for shoot initiation, medium for shoot multiplication, and medium for root formation. By in vitro propagation of Asparagus, root initiation is difficult, but can be promoted by adding growth retardant ancymidol which also greatly promotes shoot development and suppresses callus formation.

Generation of Neural Progenitor Spheres from Human Pluripotent Stem Cells in a Suspension Bioreactor.

PubMed

Yan, Yuanwei; Song, Liqing; Tsai, Ang-Chen; Ma, Teng; Li, Yan

2016-01-01

Conventional two-dimensional (2-D) culture systems cannot provide large numbers of human pluripotent stem cells (hPSCs) and their derivatives that are demanded for commercial and clinical applications in in vitro drug screening, disease modeling, and potentially cell therapy. The technologies that support three-dimensional (3-D) suspension culture, such as a stirred bioreactor, are generally considered as promising approaches to produce the required cells. Recently, suspension bioreactors have also been used to generate mini-brain-like structure from hPSCs for disease modeling, showing the important role of bioreactor in stem cell culture. This chapter describes a detailed culture protocol for neural commitment of hPSCs into neural progenitor cell (NPC) spheres using a spinner bioreactor. The basic steps to prepare hPSCs for bioreactor inoculation are illustrated from cell thawing to cell propagation. The method for generating NPCs from hPSCs in the spinner bioreactor along with the static control is then described. The protocol in this study can be applied to the generation of NPCs from hPSCs for further neural subtype specification, 3-D neural tissue development, or potential preclinical studies or clinical applications in neurological diseases.

Effect of Inoculation of Acacia senegal mature trees with Mycorrhiza and Rhizobia on soil properties and microbial community structure

NASA Astrophysics Data System (ADS)

Assigbetsé, K.; Ciss, I.; Bakhoum, N.; Dieng, L.

2012-04-01

Inoculation of legume plants with symbiotic microorganisms is widely used to improve their development and productivity. The objective of this study was to investigate the effect of inoculation of Acacia senegal mature trees with rhizobium (Sinorhizobium) and arbuscular mycorrhizal fungus (G. mosseae, G. fasciculatum, G. intraradices) either singly or in combination, on soil properties, activity and the genetic structure of soil microbial communities. The experiment set up in Southern Senegal consisted of 4 randomized blocks of A. senegal mature trees with 4 treatments including inoculated trees with Rhizobium (R), mycorrhizal fungus (M) and with Rhizobium+mycorhizal fungus (RM) and non-inoculated control (CON). Soil were sampled 2 years after the inoculation. Soil pH, C and N and available P contents were measured. The microbial abundance and activity were measured in terms of microbial biomass C (MBC) and basal soil respiration. The community structure of the total bacterial, diazotrophic and denitrifying communities was assessed by denaturing gradient gel electrophoresis of 16S rDNA, nifH and nirK genes respectively. Inoculations with symbiont under field conditions have increased soil pH. The C and N contents were enhanced in the dual-inoculated treatments (RM). The mycorrhized treatment have displayed the lowest available P contents while RM and R treatments exhibited higher contents rates. The microbial biomass C rates were higher in treatments co-inoculated with AM fungi and Rhizobium than in those inoculated singly with AM fungi or Rhizobium strains. The basal soil respiration were positively correlated to MBC, and the highest rates were found in the co-inoculated treatments. Fingerprints of 16S rDNA gene exhibited similar patterns between inoculated treatments and the control showing that the inoculation of mature trees have not impacted the total bacterial community structure. In contrast, the inoculated treatments have displayed individually different diazotrophic and denitrifying communities fingerprints, indicating that the inoculation with microsymbionts have modified the genetic structure of the two functional communities in soil. Further, the diazotrophic community richness was reduced over the control indicating the impact of the addition of symbionts on the free-living N2-fixing bacterial (nifH) diversity. This study shows that inoculation of A. senegal mature trees with rhizobium and arbuscular mycorrhizal fungus has enhanced soil biofunctioning and modified the genetic structure of microbial community involved in N-cycling. Combined inoculation of AM fungi and Rhizobium have improved these effects on chemical characteristics, microbial community abundance and activity demonstrating synergism between the two microsymbionts.

An evaluation of varying protocols for high-level disinfection of flexible fiberoptic laryngoscopes.

PubMed

Liming, Bryan; Funnell, Ian; Jones, Anthony; Demons, Samandra; Marshall, Kathryn; Harsha, Wayne

2014-11-01

The use of flexible fiberoptic laryngoscopes (FFLs) is ubiquitous in otolaryngology practices. As with any medical device, there exists a small risk for transmission of pathogenic microorganisms between patients, necessitating high-level decontamination between uses. Most of the literature to date has studied channeled scopes such as those used in esophagogastroduodenoscopy and colonoscopy. A recent study of nonchanneled flexible laryngoscopes suggested that current high-level decontamination practices in use at some institutions, including ours, may be overly aggressive. We sought to evaluate and compare the efficacy of varying techniques of high-level disinfection of FFLs. FFLs were used in routine clinical encounters and then disinfected with a variety of techniques. The FFLs were then cultured for bacteria and fungi, and the rates of positive cultures were compared between the techniques and the controls. In this study, we took FFLs following use in routine clinical practice and disinfected them using one of eight decontamination protocols. We compared the bacterial and fungal culture results to positive and negative controls. We demonstrated that each of the eight cleaning protocols was statistically efficacious at removing bacterial contamination. Our results for fungal cultures did not reach statistical significance. Using in vitro inoculation of FFLs, this study demonstrated that quicker and more cost-effective practices are equally efficacious to more time-consuming and expensive techniques with regard to bacterial contamination of FFLs. NA © 2014 The American Laryngological, Rhinological and Otological Society, Inc.

Potential biodefense model applications for portable chlorine dioxide gas production.

PubMed

Stubblefield, Jeannie M; Newsome, Anthony L

2015-01-01

Development of decontamination methods and strategies to address potential infectious disease outbreaks and bioterrorism events are pertinent to this nation's biodefense strategies and general biosecurity. Chlorine dioxide (ClO2) gas has a history of use as a decontamination agent in response to an act of bioterrorism. However, the more widespread use of ClO2 gas to meet current and unforeseen decontamination needs has been hampered because the gas is too unstable for shipment and must be prepared at the application site. Newer technology allows for easy, onsite gas generation without the need for dedicated equipment, electricity, water, or personnel with advanced training. In a laboratory model system, 2 unique applications (personal protective equipment [PPE] and animal skin) were investigated in the context of potential development of decontamination protocols. Such protocols could serve to reduce human exposure to bacteria in a decontamination response effort. Chlorine dioxide gas was capable of reducing (2-7 logs of vegetative and spore-forming bacteria), and in some instances eliminating, culturable bacteria from difficult to clean areas on PPE facepieces. The gas was effective in eliminating naturally occurring bacteria on animal skin and also on skin inoculated with Bacillus spores. The culturable bacteria, including Bacillus spores, were eliminated in a time- and dose-dependent manner. Results of these studies suggested portable, easily used ClO2 gas generation systems have excellent potential for protocol development to contribute to biodefense strategies and decontamination responses to infectious disease outbreaks or other biothreat events.

Inoculating wheat (Triticum aestivum L.) with the endophytic bacterium Serratia sp. PW7 to reduce pyrene contamination.

PubMed

Zhu, Xuezhu; Wang, Wanqing; Sun, Kai; Lin, Xianghao; Li, Shuang; Waigi, Michael Gatheru; Ling, Wanting

2017-08-03

This research was conducted to find an optimal inoculation way for a pyrene-degrading endophytic Serratia sp. PW7 to colonize wheat for reducing pyrene contamination. Three inoculation ways, which are soaking seeds in inocula (TS), dipping roots of seedlings in inocula (TR), and spraying inocula on leaves of seedlings (TL), were used in this study. Inoculated seedlings and noninoculated seedlings (CK) were, respectively, cultivated in Hoagland solutions supplemented with pyrene in a growth chamber. The results showed that strain PW7 successfully colonized the inoculated seedlings in high numbers, and significantly promoted the growth of seedlings (TS and TR). More importantly, strain PW7 reduced pyrene levels in the seedlings and the Hoagland solutions. Compared to the noninoculated seedlings, the pyrene contents of the inoculated seedlings were decreased by 35.7-86.3% in the shoots and by 26.8-60.1% in the roots after 8-day cultivation. By comparing the efficiencies of decreasing pyrene residues, it can be concluded that TR was an optimal inoculation way for endophytic strains to colonize the inoculated plants and to reduce the pyrene contamination. Our findings provide an optimized inoculation way to reduce organic contamination in crops by inoculating plants with functional endophytic bacteria.

Role of the major antigenic membrane protein in phytoplasma transmission by two insect vector species.

PubMed

Rashidi, Mahnaz; Galetto, Luciana; Bosco, Domenico; Bulgarelli, Andrea; Vallino, Marta; Veratti, Flavio; Marzachì, Cristina

2015-09-30

Phytoplasmas are bacterial plant pathogens (class Mollicutes), transmitted by phloem feeding leafhoppers, planthoppers and psyllids in a persistent/propagative manner. Transmission of phytoplasmas is under the control of behavioral, environmental and geographical factors, but molecular interactions between membrane proteins of phytoplasma and vectors may also be involved. The aim of the work was to provide experimental evidence that in vivo interaction between phytoplasma antigenic membrane protein (Amp) and vector proteins has a role in the transmission process. In doing so, we also investigated the topology of the interaction at the gut epithelium and at the salivary glands, the two barriers encountered by the phytoplasma during vector colonization. Experiments were performed on the 'Candidatus Phytoplasma asteris' chrysanthemum yellows strain (CYP), and the two leafhopper vectors Macrosteles quadripunctulatus Kirschbaum and Euscelidius variegatus Kirschbaum. To specifically address the interaction of CYP Amp at the gut epithelium barrier, insects were artificially fed with media containing either the recombinant phytoplasma protein Amp, or the antibody (A416) or both, and transmission, acquisition and inoculation efficiencies were measured. An abdominal microinjection protocol was employed to specifically address the interaction of CYP Amp at the salivary gland barrier. Phytoplasma suspension was added with Amp or A416 or both, injected into healthy E. variegatus adults and then infection and inoculation efficiencies were measured. An internalization assay was developed, consisting of dissected salivary glands from healthy E. variegatus exposed to phytoplasma suspension alone or together with A416 antibody. The organs were then either observed in confocal microscopy or subjected to DNA extraction and phytoplasma quantification by qPCR, to visualize and quantify possible differences among treatments in localization/presence/number of CYP cells. Artificial feeding and abdominal microinjection protocols were developed to address the two barriers separately. The in vivo interactions between Amp of 'Candidatus Phytoplasma asteris' Chrysanthemum yellows strain (CYP) and vector proteins were studied by evaluating their effects on phytoplasma transmission by Euscelidius variegatus and Macrosteles quadripunctulatus leafhoppers. An internalization assay was developed, consisting of dissected salivary glands from healthy E. variegatus exposed to phytoplasma suspension alone or together with anti-Amp antibody. To visualize possible differences among treatments in localization/presence of CYP cells, the organs were observed in confocal microscopy. Pre-feeding of E. variegatus and M. quadripunctulatus on anti-Amp antibody resulted in a significant decrease of acquisition efficiencies in both species. Inoculation efficiency of microinjected E. variegatus with CYP suspension and anti-Amp antibody was significantly reduced compared to that of the control with phytoplasma suspension only. The possibility that this was due to reduced infection efficiency or antibody-mediated inhibition of phytoplasma multiplication was ruled out. These results provided the first indirect proof of the role of Amp in the transmission process. Protocols were developed to assess the in vivo role of the phytoplasma native major antigenic membrane protein in two phases of the vector transmission process: movement through the midgut epithelium and colonization of the salivary glands. These methods will be useful also to characterize other phytoplasma-vector combinations. Results indicated for the first time that native CYP Amp is involved in vivo in specific crossing of the gut epithelium and salivary gland colonization during early phases of vector infection.

Standardization and validation of real time PCR assays for the diagnosis of histoplasmosis using three molecular targets in an animal model

PubMed Central

López, Luisa F.; Muñoz, César O.; Cáceres, Diego H.; Tobón, Ángela M.; Loparev, Vladimir; Clay, Oliver; Chiller, Tom; Litvintseva, Anastasia; Gade, Lalitha; González, Ángel

2017-01-01

Histoplasmosis is considered one of the most important endemic and systemic mycoses worldwide. Until now few molecular techniques have been developed for its diagnosis. The aim of this study was to develop and evaluate three real time PCR (qPCR) protocols for different protein-coding genes (100-kDa, H and M antigens) using an animal model. Fresh and formalin-fixed and paraffin-embedded (FFPE) lung tissues from BALB/c mice inoculated i.n. with 2.5x106 Histoplasma capsulatum yeast or PBS were obtained at 1, 2, 3, 4, 8, 12 and 16 weeks post-infection. A collection of DNA from cultures representing different clades of H. capsulatum (30 strains) and other medically relevant pathogens (36 strains of related fungi and Mycobacterium tuberculosis) were used to analyze sensitivity and specificity. Analytical sensitivity and specificity were 100% when DNAs from the different strains were tested. The highest fungal burden occurred at first week post-infection and complete fungal clearance was observed after the third week; similar results were obtained when the presence of H. capsulatum yeast cells was demonstrated in histopathological analysis. In the first week post-infection, all fresh and FFPE lung tissues from H. capsulatum-infected animals were positive for the qPCR protocols tested except for the M antigen protocol, which gave variable results when fresh lung tissue samples were analyzed. In the second week, all qPCR protocols showed variable results for both fresh and FFPE tissues. Samples from the infected mice at the remaining times post-infection and uninfected mice (controls) were negative for all protocols. Good agreement was observed between CFUs, histopathological analysis and qPCR results for the 100-kDa and H antigen protocols. We successfully standardized and validated three qPCR assays for detecting H. capsulatum DNA in fresh and FFPE tissues, and conclude that the 100-kDa and H antigen molecular assays are promising tests for diagnosing this mycosis. PMID:29287097

Effects of vesicular-arbuscular mycorrhizal (VAM) fungi on the seedling growth of three Pistacia species.

PubMed

Caglar, S; Akgun, A

2006-07-01

The experiment was undertaken to test the efficiency of inoculation of vesicular-arbuscular mycorrhizal (VAM) fungi on the seedling growth of three Pistacia species used as rootstocks. The stratified Pistacia seeds were inoculated with VAM fungi. The highest rate of inoculated roots was 96.7% in P. khinjuck seedlings with G. clarum and G. etunicatum, 83.3% in P. vera seedlings with G. caledonium and 73.3% in P. terebinthus seedlings with G. caledonium. Mycorrhizal inoculations improved seedling height only in P. terebinthus. Certain mycorrhizal inoculations increased the leaf N, but not P and K contents. Seedlings inoculated with G. caledonium had higher reducing sugar contents. It was concluded that pre-inoculated Pistacia seedlings could have a better growth in the harsh field conditions.

Comparison of Inoculation with the InoqulA and WASP Automated Systems with Manual Inoculation

PubMed Central

Croxatto, Antony; Dijkstra, Klaas; Prod'hom, Guy

2015-01-01

The quality of sample inoculation is critical for achieving an optimal yield of discrete colonies in both monomicrobial and polymicrobial samples to perform identification and antibiotic susceptibility testing. Consequently, we compared the performance between the InoqulA (BD Kiestra), the WASP (Copan), and manual inoculation methods. Defined mono- and polymicrobial samples of 4 bacterial species and cloudy urine specimens were inoculated on chromogenic agar by the InoqulA, the WASP, and manual methods. Images taken with ImagA (BD Kiestra) were analyzed with the VisionLab version 3.43 image analysis software to assess the quality of growth and to prevent subjective interpretation of the data. A 3- to 10-fold higher yield of discrete colonies was observed following automated inoculation with both the InoqulA and WASP systems than that with manual inoculation. The difference in performance between automated and manual inoculation was mainly observed at concentrations of >106 bacteria/ml. Inoculation with the InoqulA system allowed us to obtain significantly more discrete colonies than the WASP system at concentrations of >107 bacteria/ml. However, the level of difference observed was bacterial species dependent. Discrete colonies of bacteria present in 100- to 1,000-fold lower concentrations than the most concentrated populations in defined polymicrobial samples were not reproducibly recovered, even with the automated systems. The analysis of cloudy urine specimens showed that InoqulA inoculation provided a statistically significantly higher number of discrete colonies than that with WASP and manual inoculation. Consequently, the automated InoqulA inoculation greatly decreased the requirement for bacterial subculture and thus resulted in a significant reduction in the time to results, laboratory workload, and laboratory costs. PMID:25972424

Restoration of motor function after operative reconstruction of the acutely transected spinal cord in the canine model.

PubMed

Liu, Zehan; Ren, Shuai; Fu, Kuang; Wu, Qiong; Wu, Jun; Hou, Liting; Pan, Hong; Sun, Linlin; Zhang, Jian; Wang, Bingjian; Miao, Qing; Sun, Guiyin; Bonicalzi, Vincenzo; Canavero, Sergio; Ren, Xiaoping

2018-05-01

Cephalosomatic anastomosis or what has been called a "head transplantation" requires full reconnection of the respective transected ends of the spinal cords. The GEMINI spinal cord fusion protocol has been developed for this reason. Here, we report the first randomized, controlled study of the GEMINI protocol in large animals. We conducted a randomized, controlled study of a complete transection of the spinal cord at the level of T10 in dogs at Harbin Medical University, Harbin, China. These dogs were followed for up to 8 weeks postoperatively by assessments of recovery of motor function, somato-sensory evoked potentials, and diffusion tensor imaging using magnetic resonance imaging. A total of 12 dogs were subjected to operative exposure of the dorsal aspect of the spinal cord after laminectomy and longitudinal durotomy followed by a very sharp, controlled, full-thickness, complete transection of the spinal cord at T10. The fusogen, polyethylene glycol, was applied topically to the site of the spinal cord transection in 7 of 12 dogs; 0.9% NaCl saline was applied to the site of transection in the remaining 5 control dogs. Dogs were selected randomly to receive polyethylene glycol or saline. All polyethylene glycol-treated dogs reacquired a substantial amount of motor function versus none in controls over these first 2 months as assessed on the 20-point (0-19), canine, Basso-Beattie-Bresnahan rating scale (P<.006). Somatosensory evoked potentials confirmed restoration of electrical conduction cranially across the site of spinal cord transection which improved over time. Diffusion tensor imaging, a magnetic resonance permutation that assesses the integrity of nerve fibers and cells, showed restitution of the transected spinal cord with polyethylene glycol treatment (at-injury level difference: P<.02). A sharply and fully transected spinal cord at the level of T10 can be reconstructed with restoration of many aspects of electrical continuity in large animals following the GEMINI spinal cord fusion protocol, with objective evidence of motor recovery and of electrical continuity across the site of transection, opening the way to the first cephalosomatic anastomosis. (Surgery 2017;160:XXX-XXX.). Copyright © 2017. Published by Elsevier Inc.

Inoculation of the nonlegume Capsicum annuum (L.) with Rhizobium strains. 1. Effect on bioactive compounds, antioxidant activity, and fruit ripeness.

PubMed

Silva, Luís R; Azevedo, Jessica; Pereira, Maria J; Carro, Lorena; Velazquez, Encarna; Peix, Alvaro; Valentão, Patrícia; Andrade, Paula B

2014-01-22

Pepper (Capsicum annuum L.) is an economically important agricultural crop and an excellent dietary source of natural colors and antioxidant compounds. The levels of these compounds can vary according to agricultural practices, like inoculation with plant growth-promoting rhizobacteria. In this work we evaluated for the first time the effect of the inoculation of two Rhizobium strains on C. annuum metabolites and bioactivity. The results revealed a decrease of organic acids and no effect on phenolics and capsaicinoids of leaves from inoculated plants. In the fruits from inoculated plants organic acids and phenolic compounds decreased, showing that fruits from inoculated plants present a higher ripeness stage than those from uninoculated ones. In general, the inoculation with Rhizobium did not improve the antioxidant activity of pepper fruits and leaves. Considering the positive effect on fruit ripening, the inoculation of C. annuum with Rhizobium is a beneficious agricultural practice for this nonlegume.

The start-up of denitrifying phosphorus removal system by using nitrite as electron acceptor

NASA Astrophysics Data System (ADS)

Li, W.; Liu, J.; Sun, H. Z.; Fu, J. X.; Gao, Y.; Sun, J.

2017-08-01

The inoculation of short-cut denitrifying polyphosphate-accumulating organisms (DPAOs) mainly included two-phase inoculation and three-phase inoculation. The short-cut denitrifying phosphorus removal bacteria were quickly inoculated by sequencing batch reactor (SBR) to treatment domestic wastewater. The results showed that the average effluent concentration of TP was 0.85 mg/L after 132 cycles by 44 days in two-phase inoculation. The removal rates of NO2 --N, TP and COD were 94.73%, 95.47% and 89.96% after 126 cycles by 42 days in three-phase inoculation, and the effluent concentrations were separately 1.31 mg/L, 0.45 mg/L and 17.07 mg/L, which reached the first A class requirement of Urban sewage treatment plant pollutant discharge standard. It was indicated that the efficiency of three-phase inoculation was higher. Anoxic phosphorus uptake was influenced seriously by anaerobic residual carbon, and it was the difference of the two inoculations.

Microbial-enhanced Selenium and Iron Biofortification of Wheat (Triticum aestivum L.)--Applications in Phytoremediation and Biofortification.

PubMed

Yasin, Muhammad; El-Mehdawi, Ali Farag; Anwar, Aneela; Pilon-Smits, Elizabeth A H; Faisal, Muhammad

2015-01-01

Selenium (Se) is an essential trace element for humans and other mammals. Most dietary Se is derived from crops. To develop a Se biofortification strategy for wheat, the effect of selenate fertilization and bacterial inoculation on Se uptake and plant growth was investigated. YAM2, a bacterium with 99% similarity to Bacillus pichinotyi, showed many plant growth promoting characteristics. Inoculation with YAM2 enhanced wheat growth, both in the presence and absence of selenate: YAM2-inoculated plants showed significantly higher dry weight, shoot length and spike length compared to un-inoculated plants. Selenate also stimulated wheat growth; Un-inoculated Se-treated plants showed a significantly higher dry weight and shoot length compared to control plants without Se. Bacterial inoculation significantly enhanced Se concentration in wheat kernels (167%) and stems (252%), as well as iron (Fe) levels in kernels (70%) and stems (147%), compared to un-inoculated plants. Inoculated Se-treated plants showed a significant increase in acid phosphatase activity, which may have contributed to the enhanced growth. In conclusion; Inoculation with Bacillus sp. YAM2 is a promising Se biofortification strategy for wheat and potentially other crops.

Extinction of conditioned opiate withdrawal in rats in a two-chambered place conditioning apparatus

PubMed Central

Myers, Karyn M.; Bechtholt-Gompf, Anita J.; Coleman, Brian R.; Carlezon, William A.

2016-01-01

Conditioned opiate withdrawal contributes to relapse in addicts and can be studied in rats using the opiate withdrawal-induced conditioned place aversion (OW-CPA) paradigm. Attenuation of conditioned withdrawal through extinction may be beneficial in the treatment of addiction. Here we describe a protocol for studying OW-CPA extinction using a two-chambered place conditioning apparatus. Rats are made dependent on morphine through subcutaneous implantation of morphine pellets and then trained to acquire OW-CPA through pairings of one chamber with naloxone-precipitated withdrawal and the other chamber with saline. Extinction training consists of re-exposures to both chambers in the absence of precipitated withdrawal. Rats tested following the completion of training show a decline in avoidance of the formerly naloxone-paired chamber with increasing numbers of extinction training sessions. The protocol takes a minimum of seven days; the exact duration varies with the amount of extinction training, which is determined by the goals of the experiment. PMID:22362157

Nasal irrigation: From empiricism to evidence-based medicine. A review.

PubMed

Bastier, P-L; Lechot, A; Bordenave, L; Durand, M; de Gabory, L

2015-11-01

Nasal irrigation plays a non-negligible role in the treatment of numerous sinonasal pathologies and postoperative care. There is, however, a wide variety of protocols. The present review of the evidence-based literature sought objective arguments for optimization and efficacy. It emerged that large-volume low-pressure nasal douche optimizes the distribution and cleansing power of the irrigation solution in the nasal cavity. Ionic composition and pH also influence mucociliary clearance and epithelium trophicity. Seawater is less rich in sodium ions and richer in bicarbonates, potassium, calcium and magnesium than is isotonic normal saline, while alkaline pH and elevated calcium concentration optimized ciliary motility in vitro. Bicarbonates reduce secretion viscosity. Potassium and magnesium promote healing and limit local inflammation. These results show that the efficacy of nasal irrigation is multifactorial. Large-volume low-pressure nasal irrigation using undiluted seawater seems, in the present state of knowledge, to be the most effective protocol. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

Surgical treatment of ulcers caused by extravasation of cytotoxic drugs.

PubMed

D'Andrea, Francesco; Onesti, Maria Giuseppina; Nicoletti, Giovanni Francesco; Grella, Elisa; Renzi, Luca Francesco; Spalvieri, Cristina; Scuderi, Nicolò

2004-01-01

Despite preventive measures, the extravasation of cytotoxic drugs still occurs in 0.6% to 6% of cases. The aetiology is thought to be that tissue necrosis develops into a chronic ulcer, which causes problems if the harmful action of the drug is not blocked. From 1988-2002 at the Department of Plastic Surgery of Rome University "La Sapienza", 240 patients presented with extravasation of cytotoxic drugs; all had been treated with an original conservative protocol first described in 1994, based on the repeated local infiltration of a large quantity of saline solution (90-540 ml) into the area of extravasation. We considered only cases with actively necrotic lesions. Eleven of the 240 patients (5%) had ulcers ranging from small ulcers to extensive areas of tissue necrosis. Of the 11 patients, eight had already had ulcers, while the remaining three were those in whom our conservative protocol had not prevented necrosis. They were all operated on and given grafts, local flaps, reverse radial flaps, and free flaps.

Paleo-reconstruction: Using multiple biomarker parameters

NASA Astrophysics Data System (ADS)

Chen, Zhengzheng

Advanced technologies have played essential roles in the development of molecular organic geochemistry. In this thesis, we have developed several new techniques and explored their applications, alone and with previous techniques, to paleo-reconstruction. First, we developed a protocol to separate biomarker fractions for accurate measurement of compound-specific isotope analysis. This protocol involves combination of zeolite adduction and HPLC separation. Second, an integrated study of traditional biomarker parameters, diamondoids and compound-specific biomarker isotopes, differentiated oil groups from Saudi Arabia. Specifically, Cretaceous reservoired oils were divided into three groups and the Jurassic reservoired oils were divided into two groups. Third, biomarker acids provide an alternative way to characterize biodegradation. Oils from San Joaquin Valley, U.S.A. and oils from Mediterranean display drastically different acid profiles. These differences in biomarker acids probably reflect different processes of biodegradation. Fourth, by analyzing biomarker distributions in the organic-rich rocks recording the onset of Late Ordovician extinction, we propose that changes in salinity associated with eustatic sea-level fall, contributed at least locally to the extinction of graptolite species.

Soil C and N statuses determine the effect of maize inoculation by plant growth-promoting rhizobacteria on nitrifying and denitrifying communities.

PubMed

Florio, Alessandro; Pommier, Thomas; Gervaix, Jonathan; Bérard, Annette; Le Roux, Xavier

2017-08-21

Maize inoculation by Azospirillum stimulates root growth, along with soil nitrogen (N) uptake and root carbon (C) exudation, thus increasing N use efficiency. However, inoculation effects on soil N-cycling microbial communities have been overlooked. We hypothesized that inoculation would (i) increase roots-nitrifiers competition for ammonium, and thus decrease nitrifier abundance; and (ii) increase roots-denitrifiers competition for nitrate and C supply to denitrifiers by root exudation, and thus limit or benefit denitrifiers depending on the resource (N or C) mostly limiting these microorganisms. We quantified (de)nitrifiers abundance and activity in the rhizosphere of inoculated and non-inoculated maize on 4 sites over 2 years, and ancillary soil variables. Inoculation effects on nitrification and nitrifiers (AOA, AOB) were not consistent between the three sampling dates. Inoculation influenced denitrifiers abundance (nirK, nirS) differently among sites. In sites with high C limitation for denitrifiers (i.e. limitation of denitrification by C > 66%), inoculation increased nirS-denitrifier abundance (up to 56%) and gross N 2 O production (up to 84%), likely due to increased root C exudation. Conversely, in sites with low C limitation (<47%), inoculation decreased nirS-denitrifier abundance (down to -23%) and gross N 2 O production (down to -18%) likely due to an increased roots-denitrifiers competition for nitrate.

Evaluation of the efficacy of ProHeart® 6 (moxidectin) against a resistant isolate of Dirofilaria immitis (JYD-34) in dogs.

PubMed

Bowman, Dwight D; McTier, Tom L; Adams, Eric L; Mahabir, Sean P; Login, Joyce A; Bidgood, Tara; Woods, Debra J

2017-11-09

In a previous study, it was demonstrated that ProHeart ® 6 (PH6) (moxidectin, Zoetis) provided only about 20% efficacy in a small six-dog study against a macrocyclic lactone -resistant Dirofilaria immitis isolate (Jd2009-2) when dogs were inoculated with infective third-stage larvae (L3) at the end of the dosing period (ie, 180 days post treatment). The objective of the current study was to determine the prophylactic efficacy of a moxidectin sustained-release formulation (PH6) against a confirmed macrocyclic lactone-resistant isolate of D. immitis (JYD-34) in dogs when administered by subcutaneous injection at the labeled dose of 0.17 mg/kg 2 days before L3 inoculation. This was intended to model the scenario where dogs become infected with resistant heartworms at the end of the PH6 treatment period (ie, 6 months post treatment) when dogs would routinely be given another injection under normal field use. Twelve purpose-bred Beagle dogs (six males and six females) were selected and randomly allocated to two groups, untreated controls and PH6-treated dogs in groups of six each. The dogs were ≥8 months old at the start of the study, and using blood samples collected on Day -7 were shown to be negative for adult heartworm antigen and microfilariae. On Day 0, the dogs in the untreated control group were administered saline subcutaneously by injection, and the dogs in the treated group were administered PH6 according to label instructions. On Day 2, each dog was inoculated in the inguinal area with 50 L3 of D. immitis. The dogs were necropsied on Day 150 (148 days post infection), and the worms were collected and counted. All of the six control dogs were infected and harbored a range of 21 to 37 worms (geometric mean, 25.4; 10.9 males and 13.9 females). Only one of the six PH6 dogs was found to be infected, harboring a single male worm. Efficacy was 99.5% (geometric mean). ProHeart ® 6 was highly effective in preventing the development of heartworms in dogs challenged with a confirmed macrocyclic lactone-resistant heartworm isolate (JYD-34) 2 days prior to treatment.

Repeated rat-forced swim test: reducing the number of animals to evaluate gradual effects of antidepressants.

PubMed

Mezadri, T J; Batista, G M; Portes, A C; Marino-Neto, J; Lino-de-Oliveira, C

2011-02-15

The forced swim test (FST) is a pre-clinical test to short and long term treatment with antidepressant drugs (ADT), which requires between-subject designs. Herein a modified protocol of the FST using within-subject design (repeated rat-FST) was evaluated. Male Wistar rats were submitted to 15 min of swimming (Day 1: pretest) followed by three subsequent 5 min-swimming tests one week apart (Day 2: test, Day 7: retest 1, Day 14: retest 2). To determine the temporal and factorial characteristics of the variables scored in the repeated rat-FST, the protocol was carried out in untreated animals (E1). To validate the method, daily injections of Fluoxetine (FLX, 2.5mg/kg, i.p.) or saline were given over a 2-week period (E2). Tests and retests have been videotaped for further register of the latency, frequency and duration of behaviors. Over retesting the latency to immobility decreased whereas duration of immobility tended to increase. Factorial analysis revealed that the test, the retest 1 as well as the retest 2 have variables suitable to detection of antidepressant-like effects of ADT. Compared to saline, FLX chronically administrated reduced duration of immobility whereas increased duration of swimming in retest 2. The data suggest that repeated rat-FST detected the gradual increase in the efficacy of low doses of FLX over time. Therefore, repeated rat-FST seemed suitable to detect short and long term effects of selective serotonin reuptake inhibitors, or other ADT, thus reducing the number of animals used in the screenings of this type of compounds. © 2010 Elsevier B.V. All rights reserved.

Efficacy of Intravenous Mannitol in the Management of Orbital Compartment Syndrome: A Nonhuman Primate Model.

PubMed

Johnson, Davin; Winterborn, Andrew; Kratky, Vladimir

2016-01-01

To report the efficacy of intravenous mannitol in the treatment of orbital compartment syndrome. An experimental study was conducted on 4 nonhuman primates (8 orbits). Orbital compartment syndrome was simulated by injecting autologous blood into both orbits of each nonhuman primate until a pressure of 80 mm Hg was reached (time 0). After 10 minutes, nonhuman primates were randomized to receive an infusion of either mannitol or saline, given over 15 minutes. Five minutes after the infusion was complete, lateral canthotomy and cantholysis was performed on both orbits in isolated steps every 5 minutes. During the study protocol, orbital and intraocular pressures were recorded every 5 minutes, with a final set of measurements at 60 minutes. The primary outcome measures were the mean change in pressure from time 0 to 60 minutes, as well as the mean change in pressure during the infusion period. There was no statistically significant difference in the mean changes in orbital or intraocular pressure from time 0 to 60 minutes of the protocol. However, during the infusion period there was significantly greater decrease in both orbital and intraocular pressure in the mannitol compared with saline group (-34.0 vs. -9.3 mm Hg for orbital pressure [p = 0.03]; -34.8 vs. -9.7 mm Hg for intraocular pressure [p = 0.04]). While the definitive treatment of orbital compartment syndrome is lateral canthotomy and cantholysis, mannitol results in a rapid and clinically meaningful drop in orbital and intraocular pressure. The authors believe that their data support the routine use of mannitol in orbital compartment syndrome, especially when there is a delay in timely surgical management.

Umbilical cord mesenchyme stem cell local intramuscular injection for treatment of uterine niche: Protocol for a prospective, randomized, double-blinded, placebo-controlled clinical trial.

PubMed

Fan, Dazhi; Wu, Shuzhen; Ye, Shaoxin; Wang, Wen; Guo, Xiaoling; Liu, Zhengping

2017-11-01

Uterine niche is defined as a triangular anechoic structure at the site of the scar or a gap in the myometrium at the site of a previous caesarean section. The main clinical manifestations are postmenstrual spotting and intrauterine infection, which may seriously affect the daily life of nonpregnant women. Trials have shown an excellent safety and efficacy for the potential of mesenchymal stem cells (MSCs) as a therapeutic option for scar reconstruction. Therefore, this study is designed to investigate the safety and efficacy of using MSCs in the treatment for the uterine niche. This phase II clinical trial is a single-center, prospective, randomized, double-blind, placebo-controlled with 2 arms. One hundred twenty primiparous participants will be randomly (1:1 ratio) assigned to receive direct intramuscular injection of MSCs (a dose of 1*10 cells in 1 mL of 0.9% saline) (MSCs group) or an identical-appearing 1 mL of 0.9% saline (placebo-controlled group) near the uterine incision. The primary outcome of this trial is to evaluate the proportion of participants at 6 months who is found uterine niche in the uterus by transvaginal utrasonography. Adverse events will be documented in a case report form. The study will be conducted at the Department of Obstetric of Southern Medical University Affiliated Maternal & Child Health Hospital of Foshan. This trial is the first investigation of the potential for therapeutic use of MSCs for the management of uterine niche after cesarean delivery. This protocol will help to determine the efficacy and safety of MSCs treatment in uterine niche and bridge the gap with regards to the current preclinical and clinical evidence. NCT02968459 (Clinical Trials.gov: http://clinicaltrials.gov/).

Effects of Rapid Intravenous Rehydration in Children With Mild-to-Moderate Dehydration.

PubMed

Janet, Sophie; Molina, Juan Carlos; Marañón, Rafael; García-Ros, Marta

2015-08-01

New guidelines for "rapid or ultrarapid" intravenous rehydration are being developed in different emergency departments. These new guidelines propose a faster administration of fluids and electrolytes than in traditional protocols. However, there is still insufficient evidence to establish a standard protocol. Our objective was to determine the effects of an outpatient rapid intravenous rehydration regimen based on the administration of 0.9% saline + 2.5% dextrose, at a rate of 20 mL/kg per hour for 2 hours, in children with mild-to-moderate isonatremic dehydration resulting from acute gastroenteritis. We performed a 2-institution, prospective, observational, descriptive study. Eighty-three patients were included in the study. All patients underwent a first evaluation, including physical examination, laboratory tests, and assessment of clinical degree of dehydration. After this initial evaluation, all children received our intravenous rehydration regimen. A second evaluation including the same items as in the first one was made after in all the children. Intravenous rehydration was successful in 69 patients (83.1%). It failed in 14 patients (16.8%), who required hospitalization because of persistent vomiting in 9 patients and poor general appearance in 5 patients. After intravenous rehydration, we observed a statistically significant decrease in the levels of ketonemia and uremia and in the Gorelick scale score. However, no significant changes were observed in sodium, chloride, potassium, and osmolarity values. We conclude that, in children with mild-to-moderate dehydration, the administration of 20 mL/kg per hour for 2 hours of 0.9% saline solution + 2.5% glucose improved clinical scores and may be used as an alternative and safe way for intravenous rehydration.

Changes in rumen bacterial community composition following feeding of silage inoculated with a commercial silage inoculant

USDA-ARS?s Scientific Manuscript database

Some silage inoculants yield an increase in milk production without increasing fiber digestibility, possibly through altering the rumen microflora. We hypothesized that silage treated with a commercial inoculant (Lactobacillus plantarum, LP) would improve milk production and would alter rumen bacter...

Comparison of bend angle measurements in fresh cryopreserved cartilage specimens after electromechanical reshaping

NASA Astrophysics Data System (ADS)

Karimi, Koohyar; Protsenko, Dimitry; Wu, Edward C.; Foulad, Allen; Manuel, Cyrus T.; Lim, Amanda; Wong, Brian J. F.

2010-02-01

Cryopreservation of cartilage has been investigated for decades and is currently an established protocol. However, the reliability and applicability of cartilage cryopreservation for the use in electromechanical reshaping (EMR) has not been studied exclusively. A system to cryopreserve large numbers of tissue specimens provides a steady source of cartilage of similar quality for experimentation at later dates. This will reduce error that may arise from different cartilage stock, and has the potential to maximize efficiency under time constraints. Our study utilizes a unique methodology to cryopreserve septal cartilage for use in EMR studies. Rabbit septal cartilage specimens were harvested and standardized to 20 x 8 x 1 mm, and placed in one of three solutions (normal saline, PBS, 10% DMSO in PBS) for four hours in a cold storage room at 4 degrees Celsius. Then, each cartilage specimen was vacuumed and sealed in an anti-frost plastic bag and stored in a freezer at -80 degrees Celsius for 1 to 3 weeks duration. EMR was performed using 2 and 6 volts for 2 minutes application time. Bend angle measurements of the cryopreserved cartilage specimens were compared to bend angles of fresh cartilage which underwent EMR using the same parameters. Results demonstrate that normal saline, phosphate buffered saline (PBS), and PBS with DMSO were effective in cryopreservation, and indicated no significant differences in bend angle measurements when compared to no cryopreservation. Our methodology to cryopreserve cartilage specimens provides a successful approach for use in conducting large-scale EMR studies.

Inhibition of angiotensin-converting enzyme increases oestradiol production in ewes submitted to oestrous synchronization protocol.

PubMed

Costa, A s; Junior, A S; Viana, G E N; Muratori, M C S; Reis, A M; Costa, A P R

2014-10-01

This study aimed at evaluating the effects of angiotensin-converting enzyme inhibitor (enalapril) and angiotensin II antagonist (valsartan) on the oestradiol and progesterone production in ewes submitted to oestrous synchronization protocol. The animals were weighed and randomly divided into three groups (n = 7). A pre-experiment conducted to verify the effectiveness and toxicity of enalapril (0.5 mg/kg LW) and valsartan (2.2 mg/kg LW) showed that, in the doses used, these drugs were effective in reducing blood pressure without producing toxic effects. In the experiment, all animals were subjected to oestrous synchronization protocol during 12 days. On D10, D11 and D12, animals received saline, enalapril or valsartan (same doses of the pre-experiment), according to the group randomly divided. The hormonal analysis showed an increase in oestradiol on the last day of the protocol (D12) in animals that received enalapril (p < 0.05), but not in other groups, without changing the concentration of progesterone in any of the treatments. It is concluded that valsartan and enalapril are safe and effective subcutaneously for use in sheep and that the angiotensin-converting enzyme (ACE) inhibition with enalapril leads to an increase in oestradiol production near ovulation without changing the concentration of progesterone. This shows that ACE inhibition may be a useful tool in reproductive biotechnologies involving induction and synchronization of oestrus and ovulation in sheep. © 2014 Blackwell Verlag GmbH.

Reliability of Vibrating Mesh Technology.

PubMed

Gowda, Ashwin A; Cuccia, Ann D; Smaldone, Gerald C

2017-01-01

For delivery of inhaled aerosols, vibrating mesh systems are more efficient than jet nebulizers are and do not require added gas flow. We assessed the reliability of a vibrating mesh nebulizer (Aerogen Solo, Aerogen Ltd, Galway Ireland) suitable for use in mechanical ventilation. An initial observational study was performed with 6 nebulizers to determine run time and efficiency using normal saline and distilled water. Nebulizers were run until cessation of aerosol production was noted, with residual volume and run time recorded. Three controllers were used to assess the impact of the controller on nebulizer function. Following the observational study, a more detailed experimental protocol was performed using 20 nebulizers. For this analysis, 2 controllers were used, and time to cessation of aerosol production was noted. Gravimetric techniques were used to measure residual volume. Total nebulization time and residual volume were recorded. Failure was defined as premature cessation of aerosol production represented by residual volume of > 10% of the nebulizer charge. In the initial observational protocol, an unexpected sporadic failure rate was noted of 25% in 55 experimental runs. In the experimental protocol, a failure rate was noted of 30% in 40 experimental runs. Failed runs in the experimental protocol exhibited a wide range of retained volume averaging ± SD 36 ± 21.3% compared with 3.2 ± 1.5% (P = .001) in successful runs. Small but significant differences existed in nebulization time between controllers. Aerogen Solo nebulization was often randomly interrupted with a wide range of retained volumes. Copyright © 2017 by Daedalus Enterprises.

Sodium sulphite inhibition of potato and cherry polyphenolics in nucleic acid extraction for virus detection by RT-PCR.

PubMed

Singh, R P; Nie, X; Singh, M; Coffin, R; Duplessis, P

2002-01-01

Phenolic compounds from plant tissues inhibit reverse transcription-polymerase chain reaction (RT-PCR). Multiple-step protocols using several additives to inhibit polyphenolic compounds during nucleic acid extraction are common, but time consuming and laborious. The current research highlights that the inclusion of 0.65 to 0.70% of sodium sulphite in the extraction buffer minimizes the pigmentation of nucleic acid extracts and improves the RT-PCR detection of Potato virus Y (PVY) and Potato leafroll virus (PLRV) in potato (Solanum tuberosum) tubers and Prune dwarf virus (PDV) and Prunus necrotic ringspot virus (PNRSV) in leaves and bark in the sweet cherry (Prunus avium) tree. Substituting sodium sulphite in the nucleic acid extraction buffer eliminated the use of proteinase K during extraction. Reagents phosphate buffered saline (PBS)-Tween 20 and polyvinylpyrrolidone (PVP) were also no longer required during RT or PCR phase. The resultant nucleic acid extracts were suitable for both duplex and multiplex RT-PCR. This simple and less expensive nucleic acid extraction protocol has proved very effective for potato cv. Russet Norkotah, which contains a high amount of polyphenolics. Comparing commercially available RNA extraction kits (Catrimox and RNeasy), the sodium sulphite based extraction protocol yielded two to three times higher amounts of RNA, while maintaining comparable virus detection by RT-PCR. The sodium sulphite based extraction protocol was equally effective in potato tubers, and in leaves and bark from the cherry tree.

Estuarine water quality in parks of the Northeast Coastal and Barrier Network: Development and early implementation of vital signs estuarine nutrient-enrichment monitoring, 2003-06

USGS Publications Warehouse

Kopp, Blaine S.; Nielsen, Martha; Glisic, Dejan; Neckles, Hilary A.

2009-01-01

This report documents results of pilot tests of a protocol for monitoring estuarine nutrient enrichment for the Vital Signs Monitoring Program of the National Park Service Northeast Coastal and Barrier Network. Data collected from four parks during protocol development in 2003-06 are presented: Gateway National Recreation Area, Colonial National Historic Park, Fire Island National Seashore, and Assateague Island National Seashore. The monitoring approach incorporates several spatial and temporal designs to address questions at a hierarchy of scales. Indicators of estuarine response to nutrient enrichment were sampled using a probability design within park estuaries during a late-summer index period. Monitoring variables consisted of dissolved-oxygen concentration, chlorophyll a concentration, water temperature, salinity, attenuation of downwelling photosynthetically available radiation (PAR), and turbidity. The statistical sampling design allowed the condition of unsampled locations to be inferred from the distribution of data from a set of randomly positioned "probability" stations. A subset of sampling stations was sampled repeatedly during the index period, and stations were not rerandomized in subsequent years. These "trend stations" allowed us to examine temporal variability within the index period, and to improve the sensitivity of the monitoring protocol to detecting change through time. Additionally, one index site in each park was equipped for continuous monitoring throughout the index period. Thus, the protocol includes elements of probabilistic and targeted spatial sampling, and the temporal intensity ranges from snapshot assessments to continuous monitoring.

Efficacy of methylene blue in an experimental model of calcium channel blocker-induced shock.

PubMed

Jang, David H; Donovan, Sean; Nelson, Lewis S; Bania, Theodore C; Hoffman, Robert S; Chu, Jason

2015-04-01

Calcium channel blocker poisonings account for a substantial number of reported deaths from cardiovascular drugs. Although supportive care is the mainstay of treatment, experimental therapies such as high-dose insulin-euglycemia and lipid emulsion have been studied in animal models and used in humans. In the most severe cases, even aggressive care is inadequate and deaths occur. In both experimental models and clinical cases of vasodilatory shock, methylene blue improves hemodynamic measures. It acts as a nitric oxide scavenger and inhibits guanylate cyclase that is responsible for the production of cyclic guanosine monophosphate (cGMP). Excessive cGMP production is associated with refractory vasodilatory shock in sepsis and anaphylaxis. The aim of this study is to determine the efficacy of methylene blue in an animal model of amlodipine-induced shock. Sprague-Dawley rats were anesthetized, ventilated, and instrumented for continuous blood pressure and pulse rate monitoring. The dose of amlodipine that produced death within 60 minutes was 17 mg/kg per hour (LD50). Rats were divided into 2 groups: amlodipine followed by methylene blue or amlodipine followed by normal saline solution, with 15 rats in each group. Rats received methylene blue at 2 mg/kg during 5 minutes or an equivalent amount of normal saline solution in 3 intervals from the start of the protocol: minutes 5, 30, and 60. The animals were observed for a total of 2 hours after the start of the protocol. Mortality risk and survival time were analyzed with Fisher's exact test and Kaplan-Meier survival analysis with the log rank test. Overall, 1 of 15 rats (7%) in the saline solution-treated group survived to 120 minutes compared with 5 of 15 (33%) in the methylene blue-treated group (difference -26%; 95% confidence interval [CI] -54% to 0.3%). The median survival time for the normal saline solution group was 42 minutes (95% CI 28.1 to 55.9 minutes); for the methylene blue group, 109 minutes (95% CI 93.9 to 124.1 minutes). Pulse rate and mean arterial pressure (MAP) differences between groups were analyzed until 60 minutes. Pulse rate was significantly higher in the methylene blue-treated group beginning 25 minutes after the start of the amlodipine infusion (95% CI 30 to 113 minutes) that was analyzed until 60 minutes. MAP was significantly higher in the methylene blue-treated group starting 25 minutes after the amlodipine infusion (95% CI 2 to 30 minutes) that was analyzed until 60 minutes. Methylene blue did not result in a significant difference in mortality risk. There was an increased pulse rate, MAP, and median survival time in the methylene blue group. Copyright © 2014 American College of Emergency Physicians. Published by Elsevier Inc. All rights reserved.

Laboratory evaluation of oil spill bioremediation products in salt and freshwater systems.

PubMed

Haines, John R; Kleiner, Eric J; McClellan, Kim A; Koran, Karen M; Holder, Edith L; King, Dennis W; Venosa, Albert D

2005-05-01

Ten oil spill bioremediation products were tested in the laboratory for their ability to enhance biodegradation of weathered Alaskan North Slope crude oil in both freshwater and saltwater media. The products included nutrients to stimulate inoculated microorganisms, nutrients plus an oil-degrading inoculum, nutrients plus compounds intended to stimulate oil-degrading activity, or other compounds intended to enhance microbial activity. The product tests were undertaken to evaluate significant modifications in the existing official United States Environmental Protection Agency (EPA) protocol used for qualifying commercial bioremediation agents for use in oil spills. The EPA protocol was modified to include defined formulas for the exposure waters (freshwater, saltwater), a positive control using a known inoculum and nutrients, two negative controls (one sterile, the other inoculated but nutrient-limited), and simplified oil chemical analysis. Three analysts conducted the product test independently in each type of exposure water in round-robin fashion. Statistical tests were performed on analyst variability, reproducibility, and repeatability, and the performance of the various products was quantified in both exposure media. Analysis of variance showed that the analyst error at each time-point was highly significant (P values ranged from 0.0001 to 0.008, depending on water type and oil fraction). In the saltwater tests, six products demonstrated various degrees of biodegradative activity against the alkane fraction of the crude oil and three degraded the aromatic hydrocarbons by >10%. In the freshwater tests, eight products caused >20% loss of alkane hydrocarbons, of which five degraded the alkanes by >50%. Only four products were able to degrade polycyclic aromatic hydrocarbons (PAHs) by >20%, one of which caused 88% removal. However, when the variability of the analysts was taken into consideration, only one of the ten products was found to yield significant percent removals of the PAH fraction and only in freshwater. Viable microorganism population analysis (most-probable-number method) was also performed on every sample by each operator to measure the changes in aromatic and alkane hydrocarbon-degrading organism numbers. In general, little evidence of significant growth of either alkane- or PAH-degraders occurred among any of the ten products in either the saltwater or freshwater testing.

Effect of post-inoculation relative humidity (RH) on peanut infection by Sclerotinia sclerotiorum

USDA-ARS?s Scientific Manuscript database

Stems of six-week-old plants of the cv Okrun (susceptible to Sclerotinia blight) were inoculated with S. sclerotiorum, isolated from pumpkin. Two post-inoculation humidity regimes of 100% RH were used. In the first RH regime, one inoculation chamber was kept open for the duration of experiment (DO...

Changes in ruminal bacterial community composition following feeding of alfalfa silage inoculated with a commercial silage inoculant

USDA-ARS?s Scientific Manuscript database

Some silage inoculants promote an increase in milk production, possibly through altering the rumen microflora. In this study, dairy cows fed alfalfa silage treated with the inoculant, Lactobacillus plantarum MTD/1 (LPS), were compared to cows fed untreated silage (Ctrl) with the objectives: 1) to de...

Seasonal trends in response to inoculation of coast live oak with Phytophthora ramorum

Treesearch

Richard S. Dodd; Daniel Hüberli; Tamar Y. Harnik; Brenda O' Dell; Matteo Garbelotto

2006-01-01

We developed a branch cutting inoculation method to provide a controlled system for studying variation in response to inoculation of coast live oak (Quercus agrifolia) with Phytophthora ramorum. This method has advantages over inoculations of trees in the field, in containing the inoculum and in allowing high levels of replication...

Effect of multi-stage inoculation on the bacterial and fungal community structure during organic municipal solid wastes composting.

PubMed

Xi, Beidou; He, Xiaosong; Dang, Qiuling; Yang, Tianxue; Li, Mingxiao; Wang, Xiaowei; Li, Dan; Tang, Jun

2015-11-01

In this study, PCR-DGGE method was applied to investigate the impact of multi-stage inoculation treatment on the community composition of bacterial and fungal during municipal solid wastes (MSW) composting process. The results showed that the high temperature period was extended by the multi-stage inoculation treatment, 1day longer than initial-stage inoculation treatment, and 5days longer than non-inoculation treatment. The temperature of the secondary fermentation increased to 51°C with multi-stage inoculation treatment. The multi-stage inoculation method improved the community diversity of bacteria and fungi that the diversity indexes reached the maximum on the 17days and 20days respectively, avoided the competition between inoculations and indigenous microbes, and enhanced the growth of dominant microorganisms. The DNA sequence indicated that various kinds of uncultured microorganisms with determined ratios were detected, which were dominant microbes during the whole fermentation process. These findings call for further researches of compost microbial cultivation technology. Copyright © 2015 Elsevier Ltd. All rights reserved.

Mineral Soils as Carriers for Rhizobium Inoculants

PubMed Central

Chao, W.-L.; Alexander, Martin

1984-01-01

Mineral soil-based inoculants of Rhizobium meliloti and Rhizobium phaseoli survived better at 4°C than at higher temperatures, but ca. 15% of the cells were viable at 37°C after 27 days. Soil-based inoculants of R. meliloti, R. phaseoli, Rhizobium japonicum, and a cowpea Rhizobium sp. applied to seeds of their host legumes also survived better at low temperatures, but the percent survival of such inoculants was higher than peat-based inoculants at 35°C. Survival of R. phaseoli, R. japonicum, and cowpea rhizobia was not markedly improved when the cells were suspended in sugar solutions before drying them in soil. Nodulation was abundant on Phaseolus vulgaris derived from seeds that had been coated with a soil-based inoculant and stored for 165 days at 25°C. The increase in yield and nitrogen content of Phaseolus angularis grown in the greenhouse was the same with soil-and peat-based inoculants. We suggest that certain mineral soils can be useful and readily available carriers for legume inoculants containing desiccation-resistant Rhizobium strains. PMID:16346460

Compatible bacterial mixture, tolerant to desiccation, improves maize plant growth.

PubMed

Molina-Romero, Dalia; Baez, Antonino; Quintero-Hernández, Verónica; Castañeda-Lucio, Miguel; Fuentes-Ramírez, Luis Ernesto; Bustillos-Cristales, María Del Rocío; Rodríguez-Andrade, Osvaldo; Morales-García, Yolanda Elizabeth; Munive, Antonio; Muñoz-Rojas, Jesús

2017-01-01

Plant growth-promoting rhizobacteria (PGPR) increase plant growth and crop productivity. The inoculation of plants with a bacterial mixture (consortium) apparently provides greater benefits to plant growth than inoculation with a single bacterial strain. In the present work, a bacterial consortium was formulated containing four compatible and desiccation-tolerant strains with potential as PGPR. The formulation had one moderately (Pseudomonas putida KT2440) and three highly desiccation-tolerant (Sphingomonas sp. OF178, Azospirillum brasilense Sp7 and Acinetobacter sp. EMM02) strains. The four bacterial strains were able to adhere to seeds and colonize the rhizosphere of plants when applied in both mono-inoculation and multi-inoculation treatments, showing that they can also coexist without antagonistic effects in association with plants. The effects of the bacterial consortium on the growth of blue maize were evaluated. Seeds inoculated with either individual bacterial strains or the bacterial consortium were subjected to two experimental conditions before sowing: normal hydration or desiccation. In general, inoculation with the bacterial consortium increased the shoot and root dry weight, plant height and plant diameter compared to the non-inoculated control or mono-inoculation treatments. The bacterial consortium formulated in this work had greater benefits for blue maize plants even when the inoculated seeds underwent desiccation stress before germination, making this formulation attractive for future field applications.

Compatible bacterial mixture, tolerant to desiccation, improves maize plant growth

PubMed Central

Molina-Romero, Dalia; Baez, Antonino; Quintero-Hernández, Verónica; Castañeda-Lucio, Miguel; Fuentes-Ramírez, Luis Ernesto; Bustillos-Cristales, María del Rocío; Rodríguez-Andrade, Osvaldo; Morales-García, Yolanda Elizabeth; Munive, Antonio

2017-01-01

Plant growth-promoting rhizobacteria (PGPR) increase plant growth and crop productivity. The inoculation of plants with a bacterial mixture (consortium) apparently provides greater benefits to plant growth than inoculation with a single bacterial strain. In the present work, a bacterial consortium was formulated containing four compatible and desiccation-tolerant strains with potential as PGPR. The formulation had one moderately (Pseudomonas putida KT2440) and three highly desiccation-tolerant (Sphingomonas sp. OF178, Azospirillum brasilense Sp7 and Acinetobacter sp. EMM02) strains. The four bacterial strains were able to adhere to seeds and colonize the rhizosphere of plants when applied in both mono-inoculation and multi-inoculation treatments, showing that they can also coexist without antagonistic effects in association with plants. The effects of the bacterial consortium on the growth of blue maize were evaluated. Seeds inoculated with either individual bacterial strains or the bacterial consortium were subjected to two experimental conditions before sowing: normal hydration or desiccation. In general, inoculation with the bacterial consortium increased the shoot and root dry weight, plant height and plant diameter compared to the non-inoculated control or mono-inoculation treatments. The bacterial consortium formulated in this work had greater benefits for blue maize plants even when the inoculated seeds underwent desiccation stress before germination, making this formulation attractive for future field applications. PMID:29117218

Effects of arbuscular mycorrhizal fungi inoculation on carbon and nitrogen distribution and grain yield and nutritional quality in rice (Oryza sativa L.).

PubMed

Zhang, Xue; Wang, Li; Ma, Fang; Yang, Jixian; Su, Meng

2017-07-01

The importance of arbuscular mycorrhizal fungi (AMF) for nutrient uptake and growth in rice has been widely recognized. However, little is known about the distribution of carbon (C) and nitrogen (N) in rice under AMF inoculation, which can affect grain yield and quality. This study was conducted to investigate the distribution of C and N within rice plants under AMF inoculation and the effects on grain yield and quality. AMF inoculation significantly increased N accumulation and distribution in vegetative tissues at tillering, and N translocation into seeds from heading to maturity. Consequently, AMF inoculation more strongly impacted the distribution of N than that of C in seeds, with significantly reduced C:N ratios and increased protein content (by 7.4%). Additionally, AMF inoculation significantly increased grain yield by 28.2% through increasing the grain:straw ratio by 18.4%. In addition, the roots of inoculated rice exhibited greater change in C distribution, with significantly higher C concentrations, C accumulations, and C:N ratios at tillering and maturity. AMF inoculation affected the distribution of N in seeds and C in roots. As such, AMF inoculation may be a potential method for improving grain yield and quality. © 2016 Society of Chemical Industry. © 2016 Society of Chemical Industry.

[Effect of Glomus versiforme and Trichoderma harzianum on growth and quality of Salvia miltiorrhiza].

PubMed

Wang, Xue; Chen, Mei-Lan; Yang, Guang; Li, Xiao-Ming; Li, Peng-Ying; Chen, Min

2014-05-01

The present study aimed to investigate the effect of Glomus versiforme and Trichodema harzianum on the growth and quality of Salvia miltiorrhiza continuous cropping under field conditions. The field plot experiment was conducted, these active components in the plant were analyzed by HPLC, the root diseases incidence rate of S. miltiorrhiza determined by observation and counting, and relative parameters were measured. The data was statistically processed. The result showed that inoculation of G. versiforme and combined inoculation of G. versiforme with T. harzianum significantly decreased the root diseases incidence rate of S. miltiorrhiza, and combined inoculation of G. versiforme with T. harzianum was better than other treatments. All treatments improved accumulation of active ingredients in root. Inoculation of G. versiforme and combined inoculation of G. versiforme with T. harzianum significantly increased the content of salvianolic acid B and cryptotanshinone of root (P < 0.05), Inoculation of G. versiforme, T. harzianum and combined inoculation of G. versiforme with T. harzianum significantly enhanced the content of tanshinone I and tanshinone II(A) of the root (P < 0.05). It may conclude that inoculation of G. versiforme and combined inoculation of G. versiforme with T. harzianum can effectively reduce the root diseases incidence of continuous cropping S. miltiorrhiza, and improve the quality of S. miltiorrhiza.

Cost-effectiveness of wound management in France: pressure ulcers and venous leg ulcers.

PubMed

Meaume, S; Gemmen, E

2002-06-01

This study set out to define realistic protocols of care for the treatment of chronic venous leg ulcers and pressure ulcers in France and, by developing cost-effectiveness models, to compare the different protocols of care for the two ulcer groups, enabling a calculation of direct medical costs per ulcer healed in a typical French health insurance plan. Clinical outcomes and some treatment patterns were obtained from published literature. Validations of different treatment patterns were developed using an expert consensus panel similar to the Delphi approach. Costs were calculated based on national averages and estimates from the UK and Germany. The models were used to measure costs per healed ulcer over a 12-week period. For both the pressure ulcer and venous leg ulcer models, three protocols of care were identified. For pressure ulcers and venous leg ulcers, the hydrocolloid DuoDERM (ConvaTec, also known as Granuflex in the UK and Varihesive in Germany) was most cost-effective in France. The combination of published data and expert consensus opinion is a valid technique, and in this case suggests that treating pressure ulcers and venous leg ulcers with hydrocolloid dressings is more cost-effective than treating them with saline gauze, in spite of the lower unit cost of the latter.

Non-anthocyanin polyphenolic transformation by native yeast and bacteria co-inoculation strategy during vinification.

PubMed

Devi, Apramita; Archana, Kodira Muthanna; Bhavya, Panikuttria Kuttappa; Anu-Appaiah, Konerira Aiyappaa

2018-02-01

Co-inoculation has been adapted by many wine-producing countries because it enhances the success of malolactic fermentation and reduces the fermentation cost, as well as time. However, wine phenolics have been sparsely highlighted during co-inoculation, even though polyphenols are an important parameter affecting wine colour, astringency and aroma. In the present study, we investigated the impact of co-inoculation on non-anthocyanin polyphenol profile for two different grape varieties. Co-inoculation of native yeast strain (AAV2) along with Oenococcus oeni was adapted for Cabernet Sauvignon and Shiraz wine. It was observed that the co-inoculation had minimal yet significant impact on the phenolic composition of wines for both the grape varieties. Color loss, as well as fruity aroma development, was observed in co-inoculated wines. The wines were on a par with the commercial wine, as well as wines without malolactic fermentation, in terms of phenolic compounds and overall organoleptic acceptance. Principal component analysis and hierarchical cluster analysis further suggested that the varietal influence on phenolic composition was dominating compared to inoculation strategies. Among the varieties, the inoculation strategies have significantly influenced the Cabernet wines compared to Shiraz wines. The results of the present study demonstrate that the phenolic compounds are not drastically affected by metabolic activities of malolactic bacteria during co-inoculation and, hence, are equally suitable for wine fermentation. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Topical Administration Is a Promising Inoculating Route versus Intramuscular Inoculation for the Nanoparticle-Carried DNA Vaccine to Prevent Corneal Infections.

PubMed

Hu, Kai; Malla, Tejsu; Zhai, Yujia; Dong, Lili; Tang, Ru

2015-01-01

To evaluate the comparative effect of topical versus intramuscular administration of nanoparticle-carried DNA vaccine in preventing corneal herpes simplex virus type 1 (HSV-1) infection. Nanoparticle [polyethylenimine (PEI)-Fe3O4]-carried DNA vaccine (PEI-Fe3O4-pRSC-gD-IL-21) or DNA vaccine (pRSC-gD-IL-21) alone were topically versus intramuscularly inoculated into one eye each of mice on days 0, 14 and 28. Three weeks after the final immunization, the specific immune responses and clinical degrees of primary herpes simplex keratitis were evaluated. Topical inoculation of nanoparticle-carried DNA vaccine induced mice to generate similar levels of specific HSV-1-neutralizing antibody, IFN-γ and IL-4 in serum and specific killing (cytotoxicity) and proliferative activities of the splenic lymphocytes, but a significantly higher level of secretory IgA in tears compared to those of intramuscular inoculation. More importantly, the mice inoculated topically showed a significantly decreased herpes simplex keratitis severity than the mice inoculated intramuscularly after HSV-1 challenge on the corneas of the mice. Topical inoculation of nanoparticle-carried DNA vaccine elicits a stronger specific local immune response and more effectively inhibits herpes simplex keratitis as compared to intramuscular inoculation in an HSV-1 ocular challenge mouse model. Thus, topical administration may be a promising inoculating route for the nanoparticle-carried DNA vaccine to prevent corneal infections. © 2015 S. Karger AG, Basel.

Plant growth-promoting rhizobacteria allow reduced application rates of chemical fertilizers.

PubMed

Adesemoye, A O; Torbert, H A; Kloepper, J W

2009-11-01

The search for microorganisms that improve soil fertility and enhance plant nutrition has continued to attract attention due to the increasing cost of fertilizers and some of their negative environmental impacts. The objectives of this greenhouse study with tomato were to determine (1) if reduced rates of inorganic fertilizer coupled with microbial inoculants will produce plant growth, yield, and nutrient uptake levels equivalent to those with full rates of the fertilizer and (2) the minimum level to which fertilizer could be reduced when inoculants were used. The microbial inoculants used in the study were a mixture of plant growth-promoting rhizobacteria (PGPR) strains Bacillus amyloliquefaciens IN937a and Bacillus pumilus T4, a formulated PGPR product, and the arbuscular mycorrhiza fungus (AMF), Glomus intraradices. Results showed that supplementing 75% of the recommended fertilizer rate with inoculants produced plant growth, yield, and nutrient (nitrogen and phosphorus) uptake that were statistically equivalent to the full fertilizer rate without inoculants. When inoculants were used with rates of fertilizer below 75% of the recommended rate, the beneficial effects were usually not consistent; however, inoculation with the mixture of PGPR and AMF at 70% fertility consistently produced the same yield as the full fertility rate without inoculants. Without inoculants, use of fertilizer rates lower than the recommended resulted in significantly less plant growth, yield, and nutrient uptake or inconsistent impacts. The results suggest that PGPR-based inoculants can be used and should be further evaluated as components of integrated nutrient management strategies.

Lewis and Fischer 344 rats as a model for genetic differences in spatial learning and memory: Cocaine effects.

PubMed

Fole, Alberto; Miguéns, Miguel; Morales, Lidia; González-Martín, Carmen; Ambrosio, Emilio; Del Olmo, Nuria

2017-06-02

Lewis (LEW) and Fischer 344 (F344) rats are considered a model of genetic vulnerability to drug addiction. We previously showed important differences in spatial learning and memory between them, but in contrast with previous experiments demonstrating cocaine-induced enhanced learning in Morris water maze (MWM) highly demanding tasks, the eight-arm radial maze (RAM) performance was not modified either in LEW or F344 rats after chronic cocaine treatment. In the present work, chronically cocaine-treated LEW and F344 adult rats have been evaluated in learning and memory performance using the Y-maze, two RAM protocols that differ in difficulty, and a reversal protocol that tests cognitive flexibility. After one of the RAM protocols, we quantified dendritic spine density in hippocampal CA1 neurons and compared it to animals treated with cocaine but not submitted to RAM. LEW cocaine treated rats showed a better performance in the Y maze than their saline counterparts, an effect that was not evident in the F344 strain. F344 rats significantly took more time to learn the RAM task and made a greater number of errors than LEW animals in both protocols tested, whereas cocaine treatment induced deleterious effects in learning and memory in the highly difficult protocol. Moreover, hippocampal spine density was cocaine-modulated in LEW animals whereas no effects were found in F344 rats. We propose that differences in addictive-like behavior between LEW and F344 rats could be related to differences in hippocampal learning and memory processes that could be on the basis of individual vulnerability to cocaine addiction. Copyright © 2017 Elsevier Inc. All rights reserved.

A survey-based study on the protocols for therapeutic hypothermia in cardiac arrest patients in Korea: focusing on the differences between level 1 and 2 centers.

PubMed

Oh, Tae Gwan; Cha, Won Chul; Jo, Ik Joon; Kang, Mun Ju; Lee, Dong Woo

2015-12-01

We aimed to summarize the therapeutic hypothermia (TH) protocols used in emergency departments (EDs) in Korea and to investigate the differences between level 1 and 2 centers. The chief residents from 56 EDs were given a structured survey containing questions on the indications for TH, methods for TH induction, maintaining, and finalizing TH treatments. The participants were divided into 2 groups based on their work place (level 1 vs. level 2 centers). We received 36 responses to the survey. The majority of the participants (94.4%) reported that they routinely used TH. An average of 5.9 (standard deviation, 3.4) and 3.3 (standard deviation, 2.9) TH procedures were performed monthly in level 1 and 2 centers, respectively (P=0.01). The majority of level 1 and 2 centers (80.0% and 73.1%, respectively) had written TH protocols. Rectal (50.0%) and esophageal probes (38.9%) were most commonly used to monitor the patients' body temperatures. Midazolam (80.6%) and remifentanyl (47.2%) were the most commonly used sedatives. For TH induction, external cooling devices (77.8%) and cold saline infusion (66.1%) were predominant. Between level 1 and 2 centers, only the number of TH, the usage of remifentanyl, and application of external cooling device showed significant differences (P<0.05). Our study summarizes the TH protocols used in 36 EDs. The majority of study participants performed TH using a written protocol. We observed small number of differences in TH induction and maintenance methods between level 1 and 2 centers.

Effects of gasoline engine emissions on preexisting allergic airway responses in mice.

PubMed

Day, Kimberly C; Reed, Matthew D; McDonald, Jacob D; Seilkop, Steven K; Barrett, Edward G

2008-10-01

Gasoline-powered vehicle emissions contribute significantly to ambient air pollution. We hypothesized that exposure to gasoline engine emissions (GEE) may exacerbate preexisting allergic airway responses. Male BALB/c mice were sensitized by injection with ovalbumin (OVA) and then received a 10-min aerosolized OVA challenge. Parallel groups were sham-sensitized with saline. Mice were exposed 6 h/day to air (control, C) or GEE containing particulate matter (PM) at low (L), medium (M), or high (H) concentrations, or to the H level with PM removed by filtration (high-filtered, HF). Immediately after GEE exposure mice received another 10-min aerosol OVA challenge (pre-OVA protocol). In a second (post-OVA) protocol, mice were similarly sensitized but only challenged to OVA before air or GEE exposure. Measurements of airway hyperresponsiveness (AHR), bronchoalveolar lavage (BAL), and blood collection were performed approximately 24 h after the last exposure. In both protocols, M, H, and HF GEE exposure significantly decreased BAL neutrophils from nonsensitized mice but had no significant effect on BAL cells from OVA-sensitized mice. In the pre-OVA protocol, GEE exposure increased OVA-specific IgG(1) but had no effect on BAL interleukin (IL)-2, IL-4, IL-13, or interferon (IFN)-gamma in OVA-sensitized mice. Nonsensitized GEE-exposed mice had increased OVA-specific IgG(2a), IgE, and IL-2, but decreased total IgE. In the post-OVA protocol, GEE exposure reduced BAL IL-4, IL-5, and IFN-gamma in nonsensitized mice but had no effect on sensitized mice. These results suggest acute exposure to the gas-vapor phase of GEE suppressed inflammatory cells and cytokines from nonsensitized mice but did not substantially exacerbate allergic responses.

An optimised protocol for molecular identification of Eimeria from chickens.

PubMed

Kumar, Saroj; Garg, Rajat; Moftah, Abdalgader; Clark, Emily L; Macdonald, Sarah E; Chaudhry, Abdul S; Sparagano, Olivier; Banerjee, Partha S; Kundu, Krishnendu; Tomley, Fiona M; Blake, Damer P

2014-01-17

Molecular approaches supporting identification of Eimeria parasites infecting chickens have been available for more than 20 years, although they have largely failed to replace traditional measures such as microscopy and pathology. Limitations of microscopy-led diagnostics, including a requirement for specialist parasitological expertise and low sample throughput, are yet to be outweighed by the difficulties associated with accessing genomic DNA from environmental Eimeria samples. A key step towards the use of Eimeria species-specific PCR as a sensitive and reproducible discriminatory tool for use in the field is the production of a standardised protocol that includes sample collection and DNA template preparation, as well as primer selection from the numerous PCR assays now published. Such a protocol will facilitate development of valuable epidemiological datasets which may be easily compared between studies and laboratories. The outcome of an optimisation process undertaken in laboratories in India and the UK is described here, identifying four steps. First, samples were collected into a 2% (w/v) potassium dichromate solution. Second, oocysts were enriched by flotation in saturated saline. Third, genomic DNA was extracted using a QIAamp DNA Stool mini kit protocol including a mechanical homogenisation step. Finally, nested PCR was carried out using previously published primers targeting the internal transcribed spacer region 1 (ITS-1). Alternative methods tested included sample processing in the presence of faecal material, DNA extraction using a traditional phenol/chloroform protocol, the use of SCAR multiplex PCR (one tube and two tube versions) and speciation using the morphometric tool COCCIMORPH for the first time with field samples. Copyright © 2013 Dirk Vulpius The Authors. Published by Elsevier B.V. All rights reserved.

Co-Inoculation with Rhizobia and AMF Inhibited Soybean Red Crown Rot: From Field Study to Plant Defense-Related Gene Expression Analysis

PubMed Central

Gao, Xiang; Lu, Xing; Wu, Man; Zhang, Haiyan; Pan, Ruqian; Tian, Jiang; Li, Shuxian; Liao, Hong

2012-01-01

Background Soybean red crown rot is a major soil-borne disease all over the world, which severely affects soybean production. Efficient and sustainable methods are strongly desired to control the soil-borne diseases. Principal Findings We firstly investigated the disease incidence and index of soybean red crown rot under different phosphorus (P) additions in field and found that the natural inoculation of rhizobia and arbuscular mycorrhizal fungi (AMF) could affect soybean red crown rot, particularly without P addition. Further studies in sand culture experiments showed that inoculation with rhizobia or AMF significantly decreased severity and incidence of soybean red crown rot, especially for co-inoculation with rhizobia and AMF at low P. The root colony forming unit (CFU) decreased over 50% when inoculated by rhizobia and/or AMF at low P. However, P addition only enhanced CFU when inoculated with AMF. Furthermore, root exudates of soybean inoculated with rhizobia and/or AMF significantly inhibited pathogen growth and reproduction. Quantitative RT-PCR results indicated that the transcripts of the most tested pathogen defense-related (PR) genes in roots were significantly increased by rhizobium and/or AMF inoculation. Among them, PR2, PR3, PR4 and PR10 reached the highest level with co-inoculation of rhizobium and AMF. Conclusions Our results indicated that inoculation with rhizobia and AMF could directly inhibit pathogen growth and reproduction, and activate the plant overall defense system through increasing PR gene expressions. Combined with optimal P fertilization, inoculation with rhizobia and AMF could be considered as an efficient method to control soybean red crown rot in acid soils. PMID:22442737

Co-inoculation of Lolium perenne with Funneliformis mosseae and the dark septate endophyte Cadophora sp. in a trace element-polluted soil.

PubMed

Berthelot, Charlotte; Blaudez, Damien; Beguiristain, Thierry; Chalot, Michel; Leyval, Corinne

2018-04-01

The presence of dark septate endophytes (DSEs) or arbuscular mycorrhizal fungi (AMF) in plant roots and their effects on plant fitness have been extensively described. However, little is known about their interactions when they are simultaneously colonizing a plant root, especially in trace element (TE)-polluted soils. We therefore investigated the effects of Cadophora sp. and Funneliformis mosseae on ryegrass (Lolium perenne) growth and element uptake in a Cd/Zn/Pb-polluted soil. The experiment included four treatments, i.e., inoculation with Cadophora sp., inoculation with F. mosseae, co-inoculation with Cadophora sp. and F. mosseae, and no inoculation. Ryegrass biomass and shoot Na, P, K, and Mg concentrations significantly increased following AMF inoculation as compared to non-inoculated controls. Similarly, DSE inoculation increased shoot Na concentration, whereas dual inoculation significantly decreased shoot Cd concentration. Moreover, oxidative stress determined by ryegrass leaf malondialdehyde concentration was alleviated both in the AMF and dual inoculation treatments. We used quantitative PCR and microscope observations to quantify colonization rates. They demonstrated that DSEs had no effect on AMF colonization, while AMF colonization slightly decreased DSE frequency. We also monitored fluorescein diacetate (FDA) hydrolysis and alkaline phosphatase (AP) activity in the rhizosphere soils. FDA hydrolysis remained unchanged in the three inoculated treatments, but AMF colonization increased AP activity and P mobility in the soil whereas DSE colonization did not alter AP activity. In this experiment, we unveiled the interactions between two ecologically important fungal groups likely to occur in roots which involved a decrease of oxidative stress and Cd accumulation in shoots. These results open promising perspectives on the fungal-based phytomanagement of TE-contaminated sites by the production of uncontaminated and marketable plant biomass.

Chronic Bee Paralysis Virus and Nosema ceranae Experimental Co-Infection of Winter Honey Bee Workers (Apis mellifera L.)

PubMed Central

Toplak, Ivan; Jamnikar Ciglenečki, Urška; Aronstein, Katherine; Gregorc, Aleš

2013-01-01

Chronic bee paralysis virus (CBPV) is an important viral disease of adult bees which induces significant losses in honey bee colonies. Despite comprehensive research, only limited data is available from experimental infection for this virus. In the present study winter worker bees were experimentally infected in three different experiments. Bees were first inoculated per os (p/o) or per cuticle (p/c) with CBPV field strain M92/2010 in order to evaluate the virus replication in individual bees. In addition, potential synergistic effects of co-infection with CBPV and Nosema ceranae (N. ceranae) on bees were investigated. In total 558 individual bees were inoculated in small cages and data were analyzed using quantitative real time RT-PCR (RT-qPCR). Our results revealed successful replication of CBPV after p/o inoculation, while it was less effective when bees were inoculated p/c. Dead bees harbored about 1,000 times higher copy numbers of the virus than live bees. Co-infection of workers with CBPV and N. ceranae using either method of virus inoculation (p/c or p/o) showed increased replication ability for CBPV. In the third experiment the effect of inoculation on bee mortality was evaluated. The highest level of bee mortality was observed in a group of bees inoculated with CBPV p/o, followed by a group of workers simultaneously inoculated with CBPV and N. ceranae p/o, followed by the group inoculated with CBPV p/c and the group with only N. ceranae p/o. The experimental infection with CBPV showed important differences after p/o or p/c inoculation in winter bees, while simultaneous infection with CBPV and N. ceranae suggesting a synergistic effect after inoculation. PMID:24056674

Nitrogen fertilizer enhances growth and nutrient uptake of Medicago sativa inoculated with Glomus tortuosum grown in Cd-contaminated acidic soil.

PubMed

Liu, Mohan; Sun, Jian; Li, Yang; Xiao, Yan

2017-01-01

This study aimed to explore whether nitrogen availability could influence mycorrhizal function and their associations with host plants in Cd-contaminated acidic soils or not. A greenhouse pot experiment was conducted to assess the effects of mycorrhizal inoculation (non-mycorrhizal inoculation (NM), Glomus aggregatum (Ga), G. tortuosum (Gt) and G. versiforme (Gv)) and inorganic N amendment on the growth, nutrient and Cd uptake of Medicago sativa grown in Cd-contaminated acidic soils (10 mg Cd kg -1 soil). AMF inoculations significantly increased the shoot and total biomass and decreased the shoot Cd concentration in comparison to plants uninoculated. N addition increased markedly concentration and content of N and decreased those of P in plants at all inoculation treatments. Shoot K, Na and Mg concentration in plants inoculated with Ga and Gv were decreased by N addition, whereas shoot K, Na, Ca and Mg concentration in plants inoculated with Gt were not negatively affected. It was observed that N addition only increased mycorrhizal colonization, shoot biomass, shoot K, Ca and Mg content of plants inoculated with Gt. Irrespective of N addition, plants with Gt inoculation got the maximum shoot and root P concentration and content, as well as P/Cd concentration molar ratio among all inoculation treatment. Neither AMF nor N fertilizer contributed to the decrease of soil exchangeable Cd and increase of soil pH. These results suggested that N fertilizer only elevated plant performance of alfalfa with Gt inoculation grown in acidic soil, by diluting Cd concentration and alleviating of nutrient deficiency, especially P. Copyright © 2016 Elsevier Ltd. All rights reserved.