Trans-shell infection by pathogenic micro-organisms reduces the shelf life of non-incubated bird's eggs: a constraint on the onset of incubation?

PubMed

Cook, Mark I; Beissinger, Steven R; Toranzos, Gary A; Rodriguez, Roberto A; Arendt, Wayne J

2003-11-07

Many birds initiate incubation before clutch completion, which results in asynchronous hatching. The ensuing within-brood size disparity often places later-hatched nestlings at a developmental disadvantage, but the functional significance of the timing of the onset of incubation is poorly understood. Early incubation may serve to maintain the viability of early-laid eggs, which declines over time owing to the putative effects of ambient temperature. An unexplored risk to egg viability is trans-shell infection by micro-organisms. We experimentally investigated the rate and magnitude of microbial trans-shell infection of the egg, and the relative effects of ambient temperature and micro-organisms on hatching success. We show that infection of egg contents is prevalent and occurs within the time required to lay a clutch. The probability of infection depends on the climatic conditions, the exposure period and the phylogenetic composition of the eggshell microbiota. We also demonstrate that microbial infection and ambient temperature act independently to reduce egg viability considerably. Our results suggest that these two factors could affect the onset of avian incubation in a wide range of environments.

Trans-shell infection by pathogenic micro-organisms reduces the shelf life of non-incubated bird's eggs: a constraint on the onset of incubation?

PubMed Central

Cook, Mark I; Beissinger, Steven R; Toranzos, Gary A; Rodriguez, Roberto A; Arendt, Wayne J

2003-01-01

Many birds initiate incubation before clutch completion, which results in asynchronous hatching. The ensuing within-brood size disparity often places later-hatched nestlings at a developmental disadvantage, but the functional significance of the timing of the onset of incubation is poorly understood. Early incubation may serve to maintain the viability of early-laid eggs, which declines over time owing to the putative effects of ambient temperature. An unexplored risk to egg viability is trans-shell infection by micro-organisms. We experimentally investigated the rate and magnitude of microbial trans-shell infection of the egg, and the relative effects of ambient temperature and micro-organisms on hatching success. We show that infection of egg contents is prevalent and occurs within the time required to lay a clutch. The probability of infection depends on the climatic conditions, the exposure period and the phylogenetic composition of the eggshell microbiota. We also demonstrate that microbial infection and ambient temperature act independently to reduce egg viability considerably. Our results suggest that these two factors could affect the onset of avian incubation in a wide range of environments. PMID:14613609

Flow cytometry and cell sorting of heterogeneous microbial populations: the importance of single-cell analyses.

PubMed Central

Davey, H M; Kell, D B

1996-01-01

The most fundamental questions such as whether a cell is alive, in the sense of being able to divide or to form a colony, may sometimes be very hard to answer, since even axenic microbial cultures are extremely heterogeneous. Analyses that seek to correlate such things as viability, which is a property of an individual cell, with macroscopic measurements of culture variables such as ATP content, respiratory activity, and so on, must inevitably fail. It is therefore necessary to make physiological measurements on individual cells. Flow cytometry is such a technique, which allows one to analyze cells rapidly and individually and permits the quantitative analysis of microbial heterogeneity. It therefore offers many advantages over conventional measurements for both routine and more exploratory analyses of microbial properties. While the technique has been widely applied to the study of mammalian cells, is use in microbiology has until recently been much more limited, largely because of the smaller size of microbes and the consequently smaller optical signals obtainable from them. Since these technical barriers no longer hold, flow cytometry with appropriate stains has been used for the rapid discrimination and identification of microbial cells, for the rapid assessment of viability and of the heterogeneous distributions of a wealth of other more detailed physiological properties, for the analysis of antimicrobial drug-cell interactions, and for the isolation of high-yielding strains of biotechnological interest. Flow cytometric analyses provide an abundance of multivariate data, and special methods have been devised to exploit these. Ongoing advances mean that modern flow cytometers may now be used by nonspecialists to effect a renaissance in our understanding of microbial heterogeneity. PMID:8987359

Microbial viability in preparations packaged for single use.

PubMed

Obayashi, Akiko; Oie, Shigeharu; Kamiya, Akira

2003-05-01

We evaluated microbial viability in preparations packaged for single use only which mandate that residual solution be discarded such as albumin and globulin preparations as blood products, preparations containing albumin (such as urokinase and interferon), fat emulsions, and a preparation containing fat emulsions (propofol). In most preparations, Serratia marcescens and Burkholderia cepacia proliferated rapidly at 30 degrees C. However, in globulin preparations containing 1-2.25% glycine to prevent protein degradation (Gamma-Venin P, Venilon-I, Globulin Injection, and Ahlbulin), no growth of S. marcescens and B. cepacia was detected over 24 h at 30 degrees C. For globulin preparations containing 1-2.25% glycine, the injunction to "Discard residual solution after the package has been used" in the package inserts can be revised to "It is possible to use residual solution within 24 h after the package has been used with storage in a cool place."

Biodiesel production by various oleaginous microorganisms from organic wastes.

PubMed

Cho, Hyun Uk; Park, Jong Moon

2018-05-01

Biodiesel is a biodegradable and renewable fuel. A large amount of research has considered microbial oil production using oleaginous microorganisms, but the commercialization of microbial lipids produced in this way remains uncertain due to the high cost of feedstock or low lipid yield. Microbial lipids can be typically produced by microalgae, yeasts, and bacteria; the lipid yields of these microorganisms can be improved by using sufficient concentrations of organic carbon sources. Therefore, combining low-cost organic compounds contained in organic wastes with cultivation of oleaginous microorganisms can be a promising approach to obtain commercial viability. However, to achieve effective bioconversion of low-cost substrates to microbial lipids, the characteristics of each microorganism and each substrate should be considered simultaneously. This article discusses recent approaches to developing cost-effective microbial lipid production processes that use various oleaginous microorganisms and organic wastes. Copyright © 2018 Elsevier Ltd. All rights reserved.

The Philippine National Collection of Microorganisms (PNCM): Repository of microbial diversity of the country

NASA Astrophysics Data System (ADS)

Monsalud, R. G.; Magbanua, F. O.; Parungao, M. P.; Banaay, C. G. B.; Bayer, M. H. D.; Yap, J. K.; Tapay, L. M.

2002-04-01

The prime function of the Philippine National Collection of Microorganisms (PNCM), being the national repository of microbial strains, is to collect and preserve strains for their continued viability and availability for future use. To date, a total of 2144 strains of bacteria (1357), yeasts (250), filamentous, fungi (377), algae (14), and strains still to be identified (146) are maintained at the PNCM. These are preserved and maintained using various methods which include modified liquid drying (lyophilization), ultra-low temperature (-70°C) storage in 10% glycerol, storage in sterile soil, distilled water and overlaying with mineral oil. Periodic viability testing is done to assess the stability of these preserved cultures under storage. Aside from preservation and maintenance of cultures, the PNCM is also involved in several research activities. One of these is the isolation, characterization and identification of some Vibrio isolates from the Philippines. Details on this particular study is presented in this report.

Viability of commercial cucumber fermentation without nitrogen or air purging

USDA-ARS?s Scientific Manuscript database

Bloater defect in cucumber fermentation refers to the formation of gas pockets in the seed cavity or endocarp as the result of carbon dioxide production. Bloater damage is known to cause economic losses for the pickling industry. Microbial activity during fermentation, tissue respiration within the ...

ACETOGENIC AND SULPHATE-REDUCING BACTERIA INHABITING THE RHIZOPLANE AND DEEP CORTEX CELLS OF THE SEAGRASS HALODULE WRIGHTII

EPA Science Inventory

Recent declines in sea grass distribution underscore the importance of understanding microbial community structure-function relationships in sea grass rhizosphere that might affect the viability of these plants. Phospholipid fatty acid analyses showed that sulfate-reducing bacter...

Node position influences viability and contamination in hazelnut shoot

USDA-ARS?s Scientific Manuscript database

Initiation of shoot cultures is difficult in many woody plants due to internal microbial contaminants and general lack of juvenility in material from the source plants. Hazelnuts (Corylus avellana L.) are generally difficult to initiate into culture for these same reasons. This study was designed to...

Microbial response to space environment, part B

NASA Technical Reports Server (NTRS)

Taylor, G. R.; Chassay, C. E.; Ellis, W. L.; Foster, B. G.; Volz, P. A.; Spizizen, J.; Buecker, H.; Wrenn, R. T.; Simmonds, R. C.; Long, R. A.

1972-01-01

The performance of the microbial response to space environment experiment is considered excellent by all investigators. For most microbial systems, only preliminary survival data are available at this time. None of the available data indicate space flight-mediated changes in cell viability or recovery. One quite important observation has been made at this early date, however. The eggs produced after mice had been infected with N. dubius larvae demonstrated a significant decrease in hatchability when compared to identical ground controls. Except for the fact that the Apollo 16 flight larvae had been on board the command module, treatment of the flown larvae and ground control larvae was the same; neither had been exposed to UV irradiation. The significance and implications of this finding are currently being studied.

Investigating the effect of aqueous extracts of basil and savory on antioxidant activity, microbial and sensory properties of probiotic yogurt.

PubMed

Mosiyani, Zohreh Ghaleh; Pourahmad, Rezvan; Eshaghi, Mohammad Reza

2017-01-01

The low viability of probiotics causes the short shelf life of fermented products. Therefore compounds which prolong the viability of probiotic bacteria can increase or at least maintain the health- benefiting properties of these products. On the other hand, the addition of antioxidants is one of the methods to increase the shelf life of food products which has recently become more prevalent. In this respect, herbal extracts which are a good source of antioxidants can be appropriate alternative. The aim of this study was  to evaluate the effect of adding basil and savory extracts on antioxidant activity, and on the microbial and organoleptic characteristics of probiotic yogurt. The effect of adding basil extract (8% and 10%) and savory extract (6% and 8%) separately to low fat yogurt (1.5% fat) containing Lactobacillus paracasei subsp. paracasei was investigated. The samples were stored at 4°C. The viability of Lactobacillus paracasei subsp. paracasei, antioxidant activ- ity and sensory properties of probiotic yogurt were evaluated on the 1st, 7th, 14th and 21st days. Basil and savory extracts significantly increased the viability of probiotic bacteria (p < 0.05). Dur- ing storage, probiotic counts markedly decreased (p < 0.05) in comparison to the control sample. The addi- tion of herbal extracts significantly increased antioxidant activity, but this activity decreased during storage (p < 0.05). The scores for taste, odor, color and overall acceptance decreased as herbal extracts increased, but there was no significant difference between the test samples and control sample in terms of the texture score (p > 0.05). During storage, there was no significant difference between the organoleptic scores of the samples (p > 0.05), but the taste score did increase significantly (p < 0.05). It can be concluded that adding herbal extracts had a positive effect on the viability of probiotics and antioxidant activity of probiotic yogurt.

Synergistic effect of polyaniline coverage and surface microstructure on the inhibition of Pseudomonas aeruginosa biofilm formation.

PubMed

Gallarato, L A; Mulko, L E; Dardanelli, M S; Barbero, C A; Acevedo, D F; Yslas, E I

2017-02-01

Biofilm Formation is a survival strategy for microorganisms to adapt to their environment. Microbial cells in biofilm become tolerant and resistant to antibiotics and immune responses, increasing the difficulties for the clinical treatment of microbial infections. The surface chemistry and the micro/nano-topography of solid interfaces play a major role in mediating microorganism activity and adhesion. The effect of the surface chemical composition and topography on the adhesion and viability of Pseudomonas aeruginosa was studied. Polymeric (polyethylene terephthalate) surfaces were covered with a conducting polymer (polyaniline, PANI) film by in-situ polymerization and microstructured by Direct Laser Interference Patterning (DLIP). The viability of Pseudomonas aeruginosa on the different surfaces was investigated. The physicochemical properties of the surfaces were characterized by water contact angle measurements, scanning electron microscopy and atomic force microscopy. Bacterial biofilms were imaged by atomic force and scanning electron microscopies. The bacterial viability decreased on PANI compared with the substrate (polyethylene terephthalate) and it decreased even more upon micro-structuring the PANI films. In addition, the biofilm reduction could be improved using polymers with different chemical composition and/or the same polymer with different topographies. Both methods presented diminish the bacterial attachment and biofilm formation. These findings present a high impact related to materials for biomedical engineer applications regarding medical devices, as prostheses or catheters. Copyright © 2016 Elsevier B.V. All rights reserved.

Magnetic core/shell nanoparticle thin films deposited by MAPLE: Investigation by chemical, morphological and in vitro biological assays

NASA Astrophysics Data System (ADS)

Cristescu, R.; Popescu, C.; Socol, G.; Iordache, I.; Mihailescu, I. N.; Mihaiescu, D. E.; Grumezescu, A. M.; Balan, A.; Stamatin, I.; Chifiriuc, C.; Bleotu, C.; Saviuc, C.; Popa, M.; Chrisey, D. B.

2012-09-01

We report on thin film deposition of nanostructured Fe3O4/oleic acid/ceftriaxone and Fe3O4/oleic acid/cefepime nanoparticles (core/shell/adsorption-shell) were fabricated by matrix assisted pulsed laser evaporation (MAPLE) onto inert substrates. The thin films were characterized by profilometry, Fourier transform infrared spectroscopy, atomic force microscopy, and investigated by in vitro biological assays. The biological properties tested included the investigation of the microbial viability and the microbial adherence to the glass coverslip nanoparticle film, using Gram-negative and Gram-positive bacterial strains with known antibiotic susceptibility behavior, the microbial adherence to the HeLa cells monolayer grown on the nanoparticle pellicle, and the cytotoxicity on eukaryotic cells. The proposed system, based on MAPLE, could be used for the development of novel anti-microbial materials or strategies for fighting pathogenic biofilms frequently implicated in the etiology of biofilm associated chronic infections.

Possible overestimation of surface disinfection efficiency by assessment methods based on liquid sampling procedures as demonstrated by in situ quantification of spore viability.

PubMed

Grand, I; Bellon-Fontaine, M-N; Herry, J-M; Hilaire, D; Moriconi, F-X; Naïtali, M

2011-09-01

The standard test methods used to assess the efficiency of a disinfectant applied to surfaces are often based on counting the microbial survivors sampled in a liquid, but total cell removal from surfaces is seldom achieved. One might therefore wonder whether evaluations of microbial survivors in liquid-sampled cells are representative of the levels of survivors in whole populations. The present study was thus designed to determine the "damaged/undamaged" status induced by a peracetic acid disinfection for Bacillus atrophaeus spores deposited on glass coupons directly on this substrate and to compare it to the status of spores collected in liquid by a sampling procedure. The method utilized to assess the viability of both surface-associated and liquid-sampled spores included fluorescence labeling with a combination of Syto 61 and Chemchrome V6 dyes and quantifications by analyzing the images acquired by confocal laser scanning microscopy. The principal result of the study was that the viability of spores sampled in the liquid was found to be poorer than that of surface-associated spores. For example, after 2 min of peracetic acid disinfection, less than 17% ± 5% of viable cells were detected among liquid-sampled cells compared to 79% ± 5% or 47% ± 4%, respectively, when the viability was evaluated on the surface after or without the sampling procedure. Moreover, assessments of the survivors collected in the liquid phase, evaluated using the microscopic method and standard plate counts, were well correlated. Evaluations based on the determination of survivors among the liquid-sampled cells can thus overestimate the efficiency of surface disinfection procedures.

Possible Overestimation of Surface Disinfection Efficiency by Assessment Methods Based on Liquid Sampling Procedures as Demonstrated by In Situ Quantification of Spore Viability ▿

PubMed Central

Grand, I.; Bellon-Fontaine, M.-N.; Herry, J.-M.; Hilaire, D.; Moriconi, F.-X.; Naïtali, M.

2011-01-01

The standard test methods used to assess the efficiency of a disinfectant applied to surfaces are often based on counting the microbial survivors sampled in a liquid, but total cell removal from surfaces is seldom achieved. One might therefore wonder whether evaluations of microbial survivors in liquid-sampled cells are representative of the levels of survivors in whole populations. The present study was thus designed to determine the “damaged/undamaged” status induced by a peracetic acid disinfection for Bacillus atrophaeus spores deposited on glass coupons directly on this substrate and to compare it to the status of spores collected in liquid by a sampling procedure. The method utilized to assess the viability of both surface-associated and liquid-sampled spores included fluorescence labeling with a combination of Syto 61 and Chemchrome V6 dyes and quantifications by analyzing the images acquired by confocal laser scanning microscopy. The principal result of the study was that the viability of spores sampled in the liquid was found to be poorer than that of surface-associated spores. For example, after 2 min of peracetic acid disinfection, less than 17% ± 5% of viable cells were detected among liquid-sampled cells compared to 79% ± 5% or 47% ± 4%, respectively, when the viability was evaluated on the surface after or without the sampling procedure. Moreover, assessments of the survivors collected in the liquid phase, evaluated using the microscopic method and standard plate counts, were well correlated. Evaluations based on the determination of survivors among the liquid-sampled cells can thus overestimate the efficiency of surface disinfection procedures. PMID:21742922

The possible interplanetary transfer of microbes: assessing the viability of Deinococcus spp. under the ISS Environmental conditions for performing exposure experiments of microbes in the Tanpopo mission.

PubMed

Kawaguchi, Yuko; Yang, Yinjie; Kawashiri, Narutoshi; Shiraishi, Keisuke; Takasu, Masako; Narumi, Issay; Satoh, Katsuya; Hashimoto, Hirofumi; Nakagawa, Kazumichi; Tanigawa, Yoshiaki; Momoki, Yoh-Hei; Tanabe, Maiko; Sugino, Tomohiro; Takahashi, Yuta; Shimizu, Yasuyuki; Yoshida, Satoshi; Kobayashi, Kensei; Yokobori, Shin-Ichi; Yamagishi, Akihiko

2013-10-01

To investigate the possible interplanetary transfer of life, numerous exposure experiments have been carried out on various microbes in space since the 1960s. In the Tanpopo mission, we have proposed to carry out experiments on capture and space exposure of microbes at the Exposure Facility of the Japanese Experimental Module of the International Space Station (ISS). Microbial candidates for the exposure experiments in space include Deinococcus spp.: Deinococcus radiodurans, D. aerius and D. aetherius. In this paper, we have examined the survivability of Deinococcus spp. under the environmental conditions in ISS in orbit (i.e., long exposure to heavy-ion beams, temperature cycles, vacuum and UV irradiation). A One-year dose of heavy-ion beam irradiation did not affect the viability of Deinococcus spp. within the detection limit. Vacuum (10(-1) Pa) also had little effect on the cell viability. Experiments to test the effects of changes in temperature from 80 °C to -80 °C in 90 min (± 80 °C/90 min cycle) or from 60 °C to -60 °C in 90 min (± 60 °C/90 min cycle) on cell viability revealed that the survival rate decreased severely by the ± 80 °C/90 min temperature cycle. Exposure of various thicknesses of deinococcal cell aggregates to UV radiation (172 nm and 254 nm, respectively) revealed that a few hundred micrometer thick aggregate of deinococcal cells would be able to withstand the solar UV radiation on ISS for 1 year. We concluded that aggregated deinococcal cells will survive the yearlong exposure experiments. We propose that microbial cells can aggregate as an ark for the interplanetary transfer of microbes, and we named it 'massapanspermia'.

Microbial activity at gigapascal pressures.

PubMed

Sharma, Anurag; Scott, James H; Cody, George D; Fogel, Marilyn L; Hazen, Robert M; Hemley, Russell J; Huntress, Wesley T

2002-02-22

We observed physiological and metabolic activity of Shewanella oneidensis strain MR1 and Escherichia coli strain MG1655 at pressures of 68 to 1680 megapascals (MPa) in diamond anvil cells. We measured biological formate oxidation at high pressures (68 to 1060 MPa). At pressures of 1200 to 1600 MPa, living bacteria resided in fluid inclusions in ice-VI crystals and continued to be viable upon subsequent release to ambient pressures (0.1 MPa). Evidence of microbial viability and activity at these extreme pressures expands by an order of magnitude the range of conditions representing the habitable zone in the solar system.

Microbial infection affects egg viability and incubation behavior in a tropical passerine.

Treesearch

Mark I. Cook; Steven R. Beissinger; Gary A. Toranzos; Roberto A. Arendt Rodriguez

2004-01-01

Many avian species initiate incubation before clutch completion, which causes eggs to hatch asynchronously. This influences brood competitive dynamics and often results in nestling mortality. The prevailing hypotheses contend that parents incubate early because asynchronous hatching provides fitness benefits to parents or surviving offspring. An alternative idea is...

Short communication: incorporation of inulin and transglutaminase in fermented goat milk containing probiotic bacteria.

PubMed

Mituniewicz-Małek, A; Ziarno, M; Dmytrów, I

2014-01-01

Goat milk is a good carrier for probiotic bacteria; however, it is difficult to produce fermented goat milk with a consistency comparable to that of fermented cow milks. It can be improved by the addition of functional stabilizers, such as inulin, or treatment with transglutaminase. The aim of this study was to determine the effect of cold storage of inulin and microbial transglutaminase on the viability of Lactobacillus acidophilus La-5 and Bifidobacterium animalis ssp. lactis Bb-12 in fermented goat milk. Microbiological analysis included the determination of the probiotic bacteria cell count in fermented milk samples, whereas physico-chemical analysis included the analysis of fat content, titratable acidity, and pH of raw, pasteurized, and fermented goat milk samples. No positive influence of inulin or microbial transglutaminase on the viability of probiotics in fermented goat's milk samples was observed. Nevertheless, the population of probiotics remained above 6 log cfu/g after 8 wk of storage at 5 °C. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Determination of water-soluble vitamins using a colorimetric microbial viability assay based on the reduction of water-soluble tetrazolium salts.

PubMed

Tsukatani, Tadayuki; Suenaga, Hikaru; Ishiyama, Munetaka; Ezoe, Takatoshi; Matsumoto, Kiyoshi

2011-07-15

A method for the determination of water-soluble vitamins using a colorimetric microbial viability assay based on the reduction of the tetrazolium salt {2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt (WST-8)} via 2-methyl-1,4-napthoquinone (NQ) was developed. Measurement conditions were optimized for the microbiological determination of water-soluble vitamins, such as vitamin B(6), biotin, folic acid, niacin, and pantothenic acid, using microorganisms that have a water-soluble vitamin requirement. A linear relationship between absorbance and water-soluble vitamin concentration was obtained. The proposed method was applied to determine the concentration of vitamin B(6) in various foodstuffs. There was good agreement between vitamin B(6) concentrations determined after 24h using the WST-8 colorimetric method and those obtained after 48h using a conventional method. The results suggest that the WST-8 colorimetric assay is a useful method for the rapid determination of water-soluble vitamins in a 96-well microtiter plate. Copyright © 2011 Elsevier Ltd. All rights reserved.

Cytometric methods for measuring bacteria in water: advantages, pitfalls and applications.

PubMed

Hammes, Frederik; Egli, Thomas

2010-06-01

Rapid detection of microbial cells is a challenge in microbiology, particularly when complex indigenous communities or subpopulations varying in viability, activity and physiological state are investigated. Flow cytometry (FCM) has developed during the last 30 years into a multidisciplinary technique for analysing bacteria. When used correctly, FCM can provide a broad range of information at the single-cell level, including (but not limited to) total counts, size measurements, nucleic acid content, cell viability and activity, and detection of specific bacterial groups or species. The main advantage of FCM is that it is fast and easy to perform. It is a robust technique, which is adaptable to different types of samples and methods, and has much potential for automation. Hence, numerous FCM applications have emerged in industrial biotechnology, food and pharmaceutical quality control, routine monitoring of drinking water and wastewater systems, and microbial ecological research in soils and natural aquatic habitats. This review focuses on the information that can be gained from the analysis of bacteria in water, highlighting some of the main advantages, pitfalls and applications.

Optimization of a Viability PCR Method for the Detection of Listeria monocytogenes in Food Samples.

PubMed

Agustí, Gemma; Fittipaldi, Mariana; Codony, Francesc

2018-06-01

Rapid detection of Listeria and other microbial pathogens in food is an essential part of quality control and it is critical for ensuring the safety of consumers. Culture-based methods for detecting foodborne pathogens are time-consuming, laborious and cannot detect viable but non-culturable microorganism, whereas viability PCR methodology provides quick results; it is able to detect viable but non-culturable cells, and allows for easier handling of large amount of samples. Although the most critical point to use viability PCR technique is achieving the complete exclusion of dead cell amplification signals, many improvements are being introduced to overcome this. In the present work, the yield of dead cell DNA neutralization was enhanced by incorporating two new sample treatment strategies: tube change combined with a double light treatment. This procedure was successfully tested using artificially contaminated food samples, showing improved neutralization of dead cell DNA.

Biomass viability: An experimental study and the development of an empirical mathematical model for submerged membrane bioreactor.

PubMed

Zuthi, M F R; Ngo, H H; Guo, W S; Nghiem, L D; Hai, F I; Xia, S Q; Zhang, Z Q; Li, J X

2015-08-01

This study investigates the influence of key biomass parameters on specific oxygen uptake rate (SOUR) in a sponge submerged membrane bioreactor (SSMBR) to develop mathematical models of biomass viability. Extra-cellular polymeric substances (EPS) were considered as a lumped parameter of bound EPS (bEPS) and soluble microbial products (SMP). Statistical analyses of experimental results indicate that the bEPS, SMP, mixed liquor suspended solids and volatile suspended solids (MLSS and MLVSS) have functional relationships with SOUR and their relative influence on SOUR was in the order of EPS>bEPS>SMP>MLVSS/MLSS. Based on correlations among biomass parameters and SOUR, two independent empirical models of biomass viability were developed. The models were validated using results of the SSMBR. However, further validation of the models for different operating conditions is suggested. Copyright © 2015 Elsevier Ltd. All rights reserved.

Comparative effect of a stannous fluoride toothpaste and a sodium fluoride toothpaste on a multispecies biofilm.

PubMed

Cheng, Xingqun; Liu, Jinman; Li, Jiyao; Zhou, Xuedong; Wang, Lijiang; Liu, Jiquan; Xu, Xin

2017-02-01

This paper aimed to compare the mode of action of a stannous fluoride-containing toothpaste with a conventional sodium fluoride-containing toothpaste on anti-biofilm properties. A three-species biofilm model that consists of Streptococcus mutans, Streptococcus sanguinis and Porphyromonas gingivalis was established to compare the anti-biofilm properties of a stannous fluoride-containing toothpaste (CPH), a conventional sodium fluoride-containing toothpaste (CCP) and a negative control (PBS). The 48h biofilms were subjected to two-minute episodes of treatment with test agents twice a day for 5 consecutive days. Crystal violet staining and XTT assays were used to evaluate the biomass and viability of the treated biofilm. Live/dead staining and bacteria/extracellular polysaccharides (EPS) double-staining were used to visualize the biofilm structure and to quantify microbial/extracellular components of the treated biofilms. Species-specific fluorescent in situ hybridization and quantitative polymerase chain reaction (qPCR) were used to analyze microbial composition of the biofilms after treatment. The biomass and viability of the biofilms were significantly reduced after CPH toothpaste treatment. The inhibitory effect was further confirmed by the live/dead staining. The EPS amounts of the three-species biofilm were significantly reduced by CCP and CPH treatments, and CPH toothpaste demonstrated significant inhibition on EPS production. More importantly, CPH toothpaste significantly suppressed S. mutans and P. gingvalis, and enriched S. sanguinis in the three-species biofilm. In all experiments CPH had a significantly greater effect than CCP (p<0.05) and CCP had a greater effect than PBS (p<0.05). Stannous fluoride-containing toothpaste not only showed better inhibitory effect against oral microbial biofilm, but was also able to modulate microbial composition within multi-species biofilm compared with conventional sodium fluoride-containing toothpaste. Copyright © 2016 Elsevier Ltd. All rights reserved.

An in vitro antifungal efficacy of silver nanoparticles activated by diode laser to Candida albicans

NASA Astrophysics Data System (ADS)

Astuti, S. D.; Kharisma, D. H.; Kholimatussa'diah, S.; Zaidan, A. H.

2017-09-01

Microbial infectious diseases and increased resistance to antibiotics become urgent problems requiring immediate solutions. One promising alternative is the using of silver nanoparticles. The combination of the microbial inhibition characteristic of silver nanotechnology enhances the activity of antimicrobial effect. This study aims to determine effectiveness of antifungal silver nanoparticles with the activation of the diode laser on Candida albicans. The samples were culture of Candida albicans. Candida albicans cultures were incubated with silver nanoparticles (concentration 10-4 M) and treated with various exposure time of diode laser (15, 30, 45, 60, 75, 90)s. The suspension was planted on Sabouraud Dextrone Agar sterile media and incubated for 24 hours at temperature of 37oC. The number of colony-forming units per milliliter (CFU/ml) was determined after incubation. The results were log-transformed and analyzed by analysis of variance (ANOVA). In this analysis, P value ≤0.05 was considered to indicate a statistically significant difference. The result of this study showed the quantum yield of silver nanoparticles with diode laser 450 nm was 63,61%. Irradiating with diode laser 450 nm for 75 s resulted in the highest decreasing percentage of Candida albicans viability 65,03%. Irradiating with diode laser 450 nm 75 s with silver nanoparticles resulted in the higest decreasing percentage of Candida albicans viability 84,63%. Therefore, silver nanoparticles activated with diode laser irradiation of 450 nm resulted antifungal effect to Candida albicans viability.

Distinction of Gram-positive and -negative bacteria using a colorimetric microbial viability assay based on the reduction of water-soluble tetrazolium salts with a selection medium.

PubMed

Tsukatani, Tadayuki; Suenaga, Hikaru; Higuchi, Tomoko; Shiga, Masanobu; Noguchi, Katsuya; Matsumoto, Kiyoshi

2011-01-01

Bacteria are fundamentally divided into two groups: Gram-positive and Gram-negative. Although the Gram stain and other techniques can be used to differentiate these groups, some issues exist with traditional approaches. In this study, we developed a method for differentiating Gram-positive and -negative bacteria using a colorimetric microbial viability assay based on the reduction of the tetrazolium salt {2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium, monosodium salt} (WST-8) via 2-methyl-1,4-napthoquinone with a selection medium. We optimized the composition of the selection medium to allow the growth of Gram-negative bacteria while inhibiting the growth of Gram-positive bacteria. When the colorimetric viability assay was carried out in a selection medium containing 0.5µg/ml crystal violet, 5.0 µg/ml daptomycin, and 5.0µg/ml vancomycin, the reduction in WST-8 by Gram-positive bacteria was inhibited. On the other hand, Gram-negative bacteria produced WST-8-formazan in the selection medium. The proposed method was also applied to determine the Gram staining characteristics of bacteria isolated from various foodstuffs. There was good agreement between the results obtained using the present method and those obtained using a conventional staining method. These results suggest that the WST-8 colorimetric assay with selection medium is a useful technique for accurately differentiating Gram-positive and -negative bacteria.

Characterization of apple replant disease-associated microbial communities over multiple growth periods using next-generation sequencing

USDA-ARS?s Scientific Manuscript database

Replant disease in apple occurs as a result of incompletely understood and variable complexes of soil-borne pathogens that can build up over time in orchard soil. This disease limits economic viability of newly established orchards on replant sites and results in reduced productivity for the life of...

Connecting Water Quality With Air Quality Through Microbial Aerosols

NASA Astrophysics Data System (ADS)

Dueker, M. Elias

Aerosol production from surface waters results in the transfer of aquatic materials (including nutrients and bacteria) to air. These materials can then be transported by onshore winds to land, representing a biogeochemical connection between aquatic and terrestrial systems not normally considered. In urban waterfront environments, this transfer could result in emissions of pathogenic bacteria from contaminated waters. Despite the potential importance of this link, sources, near-shore deposition, identity and viability of microbial aerosols are largely uncharacterized. This dissertation focuses on the environmental and biological mechanisms that define this water-air connection, as a means to build our understanding of the biogeochemical, biogeographical, and public health implications of the transfer of surface water materials to the near-shore environment in both urban and non-urban environments. The effects of tidal height, wind speed and fog on coastal aerosols and microbial content were first quantified on a non-urban coast of Maine, USA. Culture-based, culture-independent, and molecular methods were used to simultaneously sample microbial aerosols while monitoring meteorological parameters. Aerosols at this site displayed clear marine influence and high concentrations of ecologically-relevant nutrients. Coarse aerosol concentrations significantly increased with tidal height, onshore wind speed, and fog presence. Tidal height and fog presence did not significantly influence total microbial aerosol concentrations, but did have a significant effect on culturable microbial aerosol fallout. Molecular analyses of the microbes settling out of near-shore aerosols provided further evidence of local ocean to terrestrial transport of microbes. Aerosol and surface ocean bacterial communities shared species and in general were dominated by organisms previously sampled in marine environments. Fog presence strengthened the microbial connection between water and land through air by increasing microbial aerosol settling rates and enhancing viability of aerosolized marine microbes. Using methods developed for the non-urban site, the role of local environment and winds in mediating water-air connections was further investigated in the urban environment. The local environment, including water surfaces, was an important source of microbial aerosols at urban sites. Large portions of the urban waterfront microbial aerosol communities were aquatic and, at a highly polluted Superfund waterfront, were closely related to bacteria previously described in environments contaminated with hydrocarbons, heavy metals, sewage and other industrial waste. Culturable urban aerosols and surface waters contained bacterial genera known to include human pathogens and asthma agents. High onshore winds strengthened this water-air connection by playing both a transport and production role. The microbial connection between water and air quality outlined by this dissertation highlights the need for information on the mechanisms that deliver surface water materials to terrestrial systems on a much larger scale. Moving from point measurements to landscape-level analyses will allow for the quantitative assessment of implications for this microbial water-air-land transfer in both urban and non-urban arenas.

Assessment of microbial viability in municipal sludge following ultrasound and microwave pretreatments and resulting impacts on the efficiency of anaerobic sludge digestion.

PubMed

Cella, Monica Angela; Akgul, Deniz; Eskicioglu, Cigdem

2016-03-01

A range of ultrasonication (US) and microwave irradiation (MW) sludge pretreatments were compared to determine the extent of cellular destruction in micro-organisms within secondary sludge and how this cellular destruction translated to anaerobic digestion (AD). Cellular lysis/inactivation was measured using two microbial viability assays, (1) Syto 16® Green and Sytox® Orange counter-assay to discern the integrity of cellular membranes and (2) a fluorescein diacetate assay to understand relative enzymatic activity. A range of MW intensities (2.17-6.48 kJ/g total solids or TS, coinciding temperatures of 60-160 °C) were selected for comparison via viability assays; a range of corresponding US intensities (2.37-27.71 kJ/g TS, coinciding sonication times of 10-60 min at different amplitudes) were also compared to this MW range. The MW pretreatment of thickened waste activated sludge (tWAS) caused fourfold to fivefold greater cell death than non-pretreated and US-pretreated tWAS. The greatest microbial destruction occurred at MW intensities greater than 2.62 kJ/g TS of sludge, after which increased energy input via MW did not appear to cause greater microbial death. In addition, the optimal MW pretreatment (80 °C, 2.62 kJ/g TS) and corresponding US pretreatment (10 min, 60 % amplitude, 2.37 kJ/g TS) were administered to the tWAS of a mixed sludge and fed to anaerobic digesters over sludge retention times (SRTs) of 20, 14, and 7 days to compare effects of feed pretreatment on AD efficiency. The digester utilizing MW-pretreated tWAS (80 °C, 2.62 kJ/g TS) had the greatest fecal coliform removal (73.4 and 69.8 % reduction, respectively), greatest solids removal (44.2 % TS reduction), and highest overall methane production (248.2 L CH4/kg volatile solids) at 14- and 7-day SRTs. However, despite the fourfold to fivefold increases in cell death upon pretreatment, improvements from the digester fed MW-pretreated sludge were marginal (i.e., increases in efficiency of less than 3-10 %) and likely due to a smaller proportion of cells (10-20 %) in the polymeric network and mixed sludge fed to digesters.

Review: Microbial Analysis in Dielectrophoretic Microfluidic Systems

PubMed Central

Fernandez, Renny E.; Rohani, Ali; Farmehini, Vahid; Swami, Nathan S.

2017-01-01

Infections caused by various known and emerging pathogenic microorganisms, including antibiotic-resistant strains, are a major threat to global health and well-being. This highlights the urgent need for detection systems for microbial identification, quantification and characterization towards assessing infections, prescribing therapies and understanding the dynamic cellular modifications. Current state-of-the-art microbial detection systems exhibit a trade-off between sensitivity and assay time, which could be alleviated by selective and label-free microbial capture onto the sensor surface from dilute samples. AC electrokinetic methods, such as dielectrophoresis, enable frequency-selective capture of viable microbial cells and spores due to polarization based on their distinguishing size, shape and sub-cellular compositional characteristics, for downstream coupling to various detection modalities. Following elucidation of the polarization mechanisms that distinguish bacterial cells from each other, as well as from mammalian cells, this review compares the microfluidic platforms for dielectrophoretic manipulation of microbials and their coupling to various detection modalities, including immuno-capture, impedance measurement, Raman spectroscopy and nucleic acid amplification methods, as well as for phenotypic assessment of microbial viability and antibiotic susceptibility. Based on the urgent need within point-of-care diagnostics towards reducing assay times and enhancing capture of the target organism, as well as the emerging interest in isolating intact microbials based on their phenotype and subcellular features, we envision widespread adoption of these label-free and selective electrokinetic techniques. PMID:28372723

Viability of fungal and actinomycetal spores after microwave radiation of building materials.

PubMed

Górny, Rafał L; Mainelis, Gediminas; Wlazło, Agnieszka; Niesler, Anna; Lis, Danuta O; Marzec, Stanisław; Siwińska, Ewa; Łudzeń-Izbińska, Beata; Harkawy, Aleksander; Kasznia-Kocot, Joanna

2007-01-01

The effects of microwave radiation on viability of fungal and actinomycetal spores growing on agar (medium optimal for growth) as well as on wooden panel and drywall (common building construction/finishing materials) were studied. All materials were incubated at high (97-99%) and low (32-33%) relative humidity to mimic "wet" and "dry" environmental conditions. Two microwave power densities (10 and 60 mW/cm2) and three times of exposure (5, 30, and 60 min) were tested to find the most effective parameters of radiation which could be applied to non-invasive reduction or cleaning of building materials from microbial contaminants. Additionally, a control of the surface temperature during the experiments allowed differentiation between thermal and microwave effect of such radiation. The results showed that the viability of studied microorganisms differed depending on their strains, growth conditions, power density of microwave radiation, time of exposure, and varied according to the applied combination of the two latter elements. The effect of radiation resulting in a decrease of spore viability on "wet" wooden panel and drywall was generally observed at 60 min exposure. Shorter exposure times decreased the viability of fungal spores only, while in actinomycetes colonizing the studied building materials, such radiation caused an opposite (supporting growth) effect.

Determination of Post-Culture Processing with Carbohydrates by MALDI-MS and TMS derivatization GC/MS.

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

Wunschel, David S.; Wahl, Karen L.; Melville, Angela M.

2011-09-30

Biological materials generally require stabilization to retain activity or viability in a dry form. A number of industrial products, such as vaccines, probiotics and biopesticides, have been produced as dry preparations. The same methods and materials used for stabilizing commercial microbial products may be applicable to preserving biothreat pathogens in a dry form.

Encapsulation method for maintaining biodecontamination activity

DOEpatents

Rogers, Robert D.; Hamilton, Melinda A.; Nelson, Lee O.; Benson, Jennifer; Green, Martin J.; Milner, Timothy N.

2002-01-01

A method for maintaining the viability and subsequent activity of microorganisms utilized in a variety of environments to promote biodecontamination of surfaces. One application involves the decontamination of concrete surfaces. Encapsulation of microbial influenced degradation (MID) microorganisms has shown that MID activity is effectively maintained under passive conditions, that is, without manual addition of moisture or nutrients, for an extended period of time.

Encapsulation method for maintaining biodecontamination activity

DOEpatents

Rogers, Robert D.; Hamilton, Melinda A.; Nelson, Lee O.; Benson, Jennifer; Green, Martin J.; Milner, Timothy N.

2006-04-11

A method for maintaining the viability and subsequent activity of microorganisms utilized in a variety of environments to promote biodecontamination of surfaces. One application involves the decontamination of concrete surfaces. Encapsulation of microbial influenced degradation (MID) microorganisms has shown that MID activity is effectively maintained under passive conditions, that is, without manual addition of moisture or nutrients, for an extended period of time.

Formation of resting cells by non-spore-forming microorganisms as a strategy of long-term survival in the environment

NASA Astrophysics Data System (ADS)

Mulyukin, Andrei L.; Soina, Vera S.; Demkina, Elena V.; Kozlova, Alla N.; Suzina, Natalia E.; Dmitriev, Vladimir V.; Duda, Vitalii I.; El'-Registan, Galina I.

2003-01-01

Non-spore-forming bacteria of the genera Micrococcus and Arthrobacter, including the isolates from permafrost sediments, were found to be able to form cystlike cells under special conditions. Cystlike cells maintained the viability during long-term storage (for up to several years), had undetectable respiratory activity and the elevated resistance to heating and other unfavorable conditions, possessed the specific fine structure and morphology, and were formed in the life cycles of the microorganism. These properties allow cystlike cells to be attributed to a new type of resting microbial forms. Furthermore, the distinctive feature of resting cystlike cells was their low P/S ratios and high Ca/K ratios in comparison to vegetative cells as shown by X-ray microanalysis. The experimentally obtained bacterial cystlike cells with thickened and laminated cell walls and altered texture of the cytoplasm were similar to the cells abundant in native microbial populations isolated from permafrost sediments and ancient soils of the Kolyma lowland (Siberia, Russia). Due to the inherent elevated resistance to adverse conditions and maintenance of viability for prolonged periods, resting cystlike cells are likely to ensure long-term survival of non-spore-forming bacteria in cold environments.

Survival of Bacillus pumilus spores for a prolonged period of time in real space conditions.

PubMed

Vaishampayan, Parag A; Rabbow, Elke; Horneck, Gerda; Venkateswaran, Kasthuri J

2012-05-01

To prevent forward contamination and maintain the scientific integrity of future life-detection missions, it is important to characterize and attempt to eliminate terrestrial microorganisms associated with exploratory spacecraft and landing vehicles. Among the organisms isolated from spacecraft-associated surfaces, spores of Bacillus pumilus SAFR-032 exhibited unusually high resistance to decontamination techniques such as UV radiation and peroxide treatment. Subsequently, B. pumilus SAFR-032 was flown to the International Space Station (ISS) and exposed to a variety of space conditions via the European Technology Exposure Facility (EuTEF). After 18 months of exposure in the EXPOSE facility of the European Space Agency (ESA) on EuTEF under dark space conditions, SAFR-032 spores showed 10-40% survivability, whereas a survival rate of 85-100% was observed when these spores were kept aboard the ISS under dark simulated martian atmospheric conditions. In contrast, when UV (>110 nm) was applied on SAFR-032 spores for the same time period and under the same conditions used in EXPOSE, a ∼7-log reduction in viability was observed. A parallel experiment was conducted on Earth with identical samples under simulated space conditions. Spores exposed to ground simulations showed less of a reduction in viability when compared with the "real space" exposed spores (∼3-log reduction in viability for "UV-Mars," and ∼4-log reduction in viability for "UV-Space"). A comparative proteomics analysis indicated that proteins conferring resistant traits (superoxide dismutase) were present in higher concentration in space-exposed spores when compared to controls. Also, the first-generation cells and spores derived from space-exposed samples exhibited elevated UVC resistance when compared with their ground control counterparts. The data generated are important for calculating the probability and mechanisms of microbial survival in space conditions and assessing microbial contaminants as risks for forward contamination and in situ life detection.

The immunomodulatory properties of probiotic microorganisms beyond their viability (ghost probiotics: proposal of paraprobiotic concept).

PubMed

Taverniti, Valentina; Guglielmetti, Simone

2011-08-01

The probiotic approach represents a potentially effective and mild alternative strategy for the prevention and treatment of either inflammatory or allergic diseases. Several studies have shown that different bacterial strains can exert their probiotic abilities by influencing the host's immune system, thereby modulating immune responses. However, the emerging concern regarding safety problems arising from the extensive use of live microbial cells is enhancing the interest in non-viable microorganisms or microbial cell extracts, as they could eliminate shelf-life problems and reduce the risks of microbial translocation and infection. The purpose of this review is to provide an overview of the scientific literature concerning studies in which dead microbial cells or crude microbial cell fractions have been used as health-promoting agents. Particular attention will be given to the modulation of host immune responses. Possible mechanisms determining the effect on the immune system will also be discussed. Finally, in the light of the FAO/WHO definition of probiotics, indicating that the word 'probiotic' should be restricted to products that contain live microorganisms, and considering the scientific evidence indicating that inactivated microbes can positively affect human health, we propose the new term 'paraprobiotic' to indicate the use of inactivated microbial cells or cell fractions to confer a health benefit to the consumer.

The effect of a toothpaste containing 2% zinc citrate and 0.3% Triclosan on bacterial viability and plaque growth in vivo compared to a toothpaste containing 0.3% Triclosan and 2% copolymer.

PubMed

Adams, S E; Theobald, A J; Jones, N M; Brading, M G; Cox, T F; Mendez, A; Chesters, D M; Gillam, D G; Hall, C; Holt, J

2003-12-01

To compare the antimicrobial efficacy and effect on plaque growth of a new silica-based fluoride toothpaste containing 2% zinc citrate/ 0.3% Triclosan with a silica-based fluoride toothpaste containing 0.3% Triclosan/2% copolymer. In Study 1, plaque was collected after one week's use of each toothpaste and assessed for bacterial viability, live/ dead ratio and microbial membrane integrity. In study 2, plaque was measured immediately and 18 hours after a single brushing with the specified toothpastes. The 2% zinc citrate/0.3% Triclosan formulation significantly reduced the total number of viable aerobic and anaerobic bacteria (p = 0.0223 and p = 0.0443 respectively) compared to the 0.3% Triclosan/2% copolymer formulation. Both toothpastes increased the bacterial membrane permeability significantly. However, the proportion of live bacteria for the 2% zinc citrate/0.3% Triclosan product was significantly reduced (p < 0.05). Study 2 showed significantly less plaque growth 18 hours after using the 2% zinc citrate/0.3% Triclosan toothpaste compared to the 0.3% Triclosan/2% copolymer toothpaste (p < 0.01). Regular use of a fluoride toothpaste containing 2% zinc citrate and 0.3% Triclosan, significantly reduced the viability of plaque bacteria compared to a fluoride toothpaste containing 0.3% Triclosan/ 2% copolymer 12 hours after brushing. In addition, a clinical plaque growth study confirmed that this anti-microbial efficacy leads to a significant reduction in plaque growth.

3-(1,3,4-Thiadiazole-2-yl)quinoline derivatives: synthesis, characterization and anti-microbial activity.

PubMed

Bhat, Abdul R; Tazeem; Azam, Amir; Choi, Inho; Athar, Fareeda

2011-07-01

A new series of thiadiazoles and intermediate thiosemicarbazones were synthesized from the chloroquinone molecule, with an aim to explore their effect on in vitro growth of microorganisms causing microbial infection. The chemical structures of the compound were elucidated by elemental analysis, FTIR, 1H and 13C NMR and ESI-MS spectral data. In vitro anti-microbial activity was performed against Staphylococcusaureus, Streptococcuspyogenes, Salmonellatyphimurium, and Escherichiacoli. The MIC was detected using the double dilution method. The results were compared by calculating percent inhibit area/μg of the compounds and the standard "amoxicillin". The selected compounds were tested for cytotoxic results using MTT assay H9c2 cardiac myoblasts cell line and the results showed that all the compounds offered remarkable >80% viability to a concentration of 200 μg/mL. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

The nanostructure of microbially-reduced graphene oxide fosters thick and highly-performing electrochemically-active biofilms

NASA Astrophysics Data System (ADS)

Virdis, Bernardino; Dennis, Paul G.

2017-07-01

Biofilms of electrochemically-active organisms are used in microbial electrochemical technologies (METs) to catalyze bioreactions otherwise not possible at bare electrodes. At present, however, achievable current outputs are still below levels considered sufficient for economic viability of large-scale METs implementations. Here, we report three-dimensional, self-aggregating biofilm composites comprising of microbial cells embedded with microbially-reduced graphene oxide (rGO) nanoparticles to form a thick macro-porous network with superior electrochemical properties. In the presence of metabolic substrate, these hybrid biofilms are capable of producing up to five times more catalytic current than the control biofilms. Cyclic voltammetry, linear sweep voltammetry, and electrochemical impedance spectroscopy, show that in spite of the increased thickness, the biofilms amended with GO display lower polarization/charge transfer resistance compared to the controls, which we ascribe to the incorporation of rGO into the biofilms, which (1) promotes fast electron transfer, yet conserving a macroporous structure that allows free diffusion of reactants and products, and (2) enhances the interfacial dynamics by allowing a higher load of microbial cells per electrode surface area. These results suggest an easy-to-apply and cost-effective method to produce high-performing electrochemically-active biofilms in situ.

Microbial contamination of water-soaked cotton gauze and its cause.

PubMed

Oie, S; Yoshida, H; Kamiya, A

2001-01-01

Seven in-use cotton gauze samples and three cotton balls soaked in sterile distilled water in canisters were investigated 7 days after they were prepared in hospital. All samples were contaminated with bacteria including 10(6) to 10(7) colony forming units/ml of Pseudomonas aeruginosa. In vitro viability tests using cotton gauze and cotton balls soaked in sterile distilled water revealed rapid proliferation of P. aeruginosa, Serratia marcescens and Candida albicans. Since the cotton gauze and the cotton balls were soaked in water containing nutrients, such as protein and glucose, these materials may be readily contaminated with bacteria including P. aeruginosa. Thus, when using cotton gauze and cotton balls containing water, microbial contamination should be expected.

Inhibition of Escherichia coli viability by external guide sequences complementary to two essential genes

PubMed Central

McKinney, Jeffrey; Guerrier-Takada, Cecilia; Wesolowski, Donna; Altman, Sidney

2001-01-01

Narrow spectrum antimicrobial activity has been designed to reduce the expression of two essential genes, one coding for the protein subunit of RNase P (C5 protein) and one for gyrase (gyrase A). In both cases, external guide sequences (EGS) have been designed to complex with either mRNA. Using the EGS technology, the level of microbial viability is reduced to less than 10% of the wild-type strain. The EGSs are additive when used together and depend on the number of nucleotides paired when attacking gyrase A mRNA. In the case of gyrase A, three nucleotides unpaired out of a 15-mer EGS still favor complete inhibition by the EGS but five unpaired nucleotides do not. PMID:11381134

Fate of Earth Microbes on Mars: UV Radiation Effects

NASA Technical Reports Server (NTRS)

Cockell, Charles

2000-01-01

A radiative transfer model is used to quantitatively investigate aspects of the martian ultraviolet radiation environment. Biological action spectra for DNA inactivation are used to estimate biologically effective irradiances for the martian surface under cloudless skies. Although the present-day martian UV flux is similar to early earth and thus may not be a limitation to life in the evolutionary context, it is a constraint to an unadapted biota and will rapidly kill spacecraft-borne microbes not covered by a martian dust layer. Here calculations for loss of microbial viability on the Pathfinder and Polar lander spacecraft are presented and the effects of martian dust on loss of viability are discussed. Details of the radiative transfer model are presented.

Fate of Earth Microbes on Mars -- UV Radiation Effects

NASA Technical Reports Server (NTRS)

Cockell, Charles

2000-01-01

A radiative transfer model is used to quantitatively investigate aspects of the martian ultraviolet radiation environment. Biological action spectra for DNA inactivation are used to estimate biologically effective irradiances for the martian surface under cloudless skies. Although the present-day martian UV flux is similar to early earth and thus may not be a limitation to life in the evolutionary context, it is a constraint to an unadapted biota and will rapidly kill spacecraft-borne microbes not covered by a martian dust layer. Here calculations for loss of microbial viability on the Pathfinder and Polar lander spacecraft are presented and the effects of martian dust on loss of viability are discussed. Details of the radiative transfer model are presented.

Viability loss of Escherichia coli cell populations in whey and corn meal snack treated at different temperatures with a twin screw extruder

USDA-ARS?s Scientific Manuscript database

Many studies on the development of new and/ or value added nutritional meals for the US consumer have been reported. However, information on the effect of treatment parameters on microbial safety of foods extruded below 100 deg C is limited. In this study, we investigated the effect of extrusion tre...

Inorganic polyphosphate in the microbial world. Emerging roles for a multifaceted biopolymer.

PubMed

Albi, Tomás; Serrano, Aurelio

2016-02-01

Inorganic polyphosphates (polyP) are linear polymers of tens to hundreds orthophosphate residues linked by phosphoanhydride bonds. These fairly abundant biopolymers occur in all extant forms of life, from prokaryotes to mammals, and could have played a relevant role in prebiotic evolution. Since the first identification of polyP deposits as metachromatic or volutin granules in yeasts in the nineteenth century, an increasing number of varied physiological functions have been reported. Due to their "high energy" bonds analogous to those in ATP and their properties as polyanions, polyP serve as microbial phosphagens for a variety of biochemical reactions, as a buffer against alkalis, as a storage of Ca(2+) and as a metal-chelating agent. In addition, recent studies have revealed polyP importance in signaling and regulatory processes, cell viability and proliferation, pathogen virulence, as a structural component and chemical chaperone, and as modulator of microbial stress response. This review summarizes the current status of knowledge and future perspectives of polyP functions and their related enzymes in the microbial world.

Enhanced performance of microbial fuel cell with in situ preparing dual graphene modified bioelectrode.

PubMed

Chen, Junfeng; Hu, Yongyou; Tan, Xiaojun; Zhang, Lihua; Huang, Wantang; Sun, Jian

2017-10-01

This study proposed a three-step method to prepare dual graphene modified bioelectrode (D-GM-BE) by in situ microbial-induced reduction of GO and polarity reversion in microbial fuel cell (MFC). Both graphene modified bioanode (GM-BA) and biocathode (GM-BC) were of 3D graphene/biofilm architectures; the viability and thickness of microbial biofilm decreased compared with control bioelectrode (C-BE). The coulombic efficiency (CE) of GM-BA was 2.1 times of the control bioanode (C-BA), which demonstrated higher rate of substrates oxidation; the relationship between peak current and scan rates data meant that GM-BC was of higher efficiency of catalyzing oxygen reduction than the control biocathode (C-BC). The maximum power density obtained in D-GM-BE MFC was 122.4±6.9mWm -2 , the interfacial charge transfer resistance of GM-BA and GM-BC were decreased by 79% and 75.7%. The excellent electrochemical performance of D-GM-BE MFC was attributed to the enhanced extracellular electron transfer (EET) process and catalyzing oxygen reduction. Copyright © 2017 Elsevier Ltd. All rights reserved.

Microbial stratification structure within cathodic biofilm of the microbial fuel cell using the freezing microtome method.

PubMed

Li, Xiao; Lu, Yaobin; Luo, Haiping; Liu, Guangli; Zhang, Renduo

2017-10-01

The aim of this study was to investigate the microbial stratification structure within cathodic biofilm of the microbial fuel cell (MFC) using the freezing microtome method. Experiments were conducted in a single-chamber air-cathode MFC with 0.8g/L maltodextrin as substrate for ∼30d operation. The maximum power density was 945±10mW/m 2 in the MFC. Maltodextrin resulted in the relative abundance of Candidatus Saccharibacteria of 37.0% in the anodic biofilm. Different bacterial communities were identified in different layers within the cathodic biofilm. The relative abundance of Enterococcus was 3.7%, 10.5%, and 1.6% in the top (100-150μm), middle (50-100μm), and bottom (0-50μm) layers, respectively. Higher bacterial viability was observed within the top and bottom layers of the cathodic biofilm. Understanding the stratification of bacterial community in cathodic biofilm should be important to control the cathodic biofilm in the MFC. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of cryopreservation and lyophilization on viability and growth of strict anaerobic human gut microbes.

PubMed

Bircher, Lea; Geirnaert, Annelies; Hammes, Frederik; Lacroix, Christophe; Schwab, Clarissa

2018-04-17

Strict anaerobic gut microbes have been suggested as 'next-generation probiotics' for treating several intestinal disorders. The development of preservation techniques is of major importance for therapeutic application. This study investigated cryopreservation (-80°C) and lyophilization survival and storage stability (4°C for 3 months) of the strict anaerobic gut microbes Bacteroides thetaiotaomicron, Faecalibacterium prausnitzii, Roseburia intestinalis, Anaerostipes caccae, Eubacterium hallii and Blautia obeum. To improve preservation survival, protectants sucrose and inulin (both 5% w/v) were added for lyophilization and were also combined with glycerol (15% v/v) for cryopreservation. Bacterial fitness, evaluated by maximum growth rate and lag phase, viability and membrane integrity were determined using a standardized growth assay and by flow cytometry as markers for preservation resistance. Lyophilization was more detrimental to viability and fitness than cryopreservation, but led to better storage stability. Adding sucrose and inulin enhanced viability and the proportion of intact cells during lyophilization of all strains. Viability of protectant-free B. thetaiotaomicron, A. caccae and F. prausnitzii was above 50% after cryopreservation and storage and increased to above 80% if protectants were present. The addition of glycerol, sucrose and inulin strongly enhanced the viability of B. obeum, E. hallii and R. intestinalis from 0.03-2% in protectant-free cultures to 11-37%. This is the first study that quantitatively compared the effect of cryopreservation and lyophilization and the addition of selected protectants on viability and fitness of six strict anaerobic gut microbes. Our results suggest that efficiency of protectants is process- and species-specific. © 2018 The Authors. Microbial Biotechnology published by John Wiley & Sons Ltd and Society for Applied Microbiology.

A new microtitre plate screening method for evaluating the viability of aerobic respiring bacteria in high surface biofilms.

PubMed

Pérez, L M; Alvarez, B L; Codony, F; Fittipaldi, M; Adrados, B; Peñuela, G; Morató, J

2010-09-01

It is difficult to determine the effects of bactericidal compounds against bacteria in a biofilm because classical procedures for determining cell viability require several working days, multiple complicated steps and are frequently only applicable to cells in suspension. We attempt to develop a compact, inexpensive and versatile system to measure directly the extent of biofilm formation from water systems and to determine the viability of respiring bacteria in high surface biofilms. It has been reported that the reduction of tetrazolium sodium salts, such as XTT (sodium 3,3'-[1-[(phenylamino)carbonyl]-3,4-tetrazolium]Bis(4-methoxy)-6-nitro)benzene sulfonic acid hydrate), during active bacterial metabolism can be incorporated into a colorimetric method for quantifying cell viability. XTT is reduced to a soluble formazan compound during bacterial aerobic metabolism such that the amount of formazan generated is proportional to the bacterial biomass. We show here, for the first time, that this colorimetric approach can be used to determine the metabolic activity of adherent aerobic bacteria in a biofilm as a measure of cell viability. This technique has been used to estimate viability and proliferation of bacteria in suspension, but this is the first application to microbial communities in a real undisturbed biofilm. This simple new system can be used to evaluate the complex biofilm community without separating the bacteria from their support. Thus, the results obtained by this practice may be more representative of the circumstances in a natural system, opening the possibility to multiple potential applications.

Species-specific viability analysis of Pseudomonas aeruginosa, Burkholderia cepacia and Staphylococcus aureus in mixed culture by flow cytometry

PubMed Central

2014-01-01

Background Bacterial species coexist commonly in mixed communities, for instance those occurring in microbial infections of humans. Interspecies effects contribute to alterations in composition of communities with respect to species and thus, to the course and severity of infection. Therefore, knowledge concerning growth and viability of single species in medically-relevant mixed communities is of high interest to resolve complexity of interspecies dynamics and to support development of treatment strategies. In this study, a flow cytometric method was established to assess the species-specific viability in defined three-species mixed cultures. The method enables the characterization of viability of Pseudomonas aeruginosa, Burkholderia cepacia and Staphylococcus aureus, which are relevant to lung infections of Cystic Fibrosis (CF) patients. The method combines fluorescence detection by antibody and lectin labeling with viability fluorescence staining using SYBR®Green I and propidium iodide. In addition, species-specific cell enumeration analysis using quantitative terminal restriction fragment length polymorphisms (qT-RFLP) was used to monitor the growth dynamics. Finally, to investigate the impact of substrate availability on growth and viability, concentrations of main substrates and metabolites released were determined. Results For each species, the time course of growth and viability during mixed culture cultivations was obtained by using qT-RFLP analysis in combination with flow cytometry. Comparison between mixed and pure cultures revealed for every species differences in growth properties, e.g. enhanced growth of P. aeruginosa in mixed culture. Differences were also observed for B. cepacia and S. aureus in the time course of viability, e.g. an early and drastic reduction of viability of S. aureus in mixed culture. Overall, P. aeruginosa clearly dominated the mixed culture with regard to obtained cell concentrations. Conclusions In combination with qT-RFLP analysis, the methods enabled monitoring of species-specific cell concentrations and viability during co-cultivation of theses strains. Experimental findings suggest that the predominance of P. aeruginosa over B. cepacia and S. aureus in mixed culture under the chosen cultivation conditions is promoted by more efficient substrate consumption of P. aeruginosa, and antagonistic interspecies effects induced by P. aeruginosa. PMID:24606608

Effects of inorganic nanoparticles on viability and catabolic activities of Agrobacterium sp. PH-08 during biodegradation of dibenzofuran.

PubMed

Le, Thao Thanh; Murugesan, Kumarasamy; Kim, Eun-Ju; Chang, Yoon-Seok

2014-09-01

This study investigated the cytotoxicity, genotoxicity, and growth inhibition effects of four different inorganic nanoparticles (NPs) such as aluminum (nAl), iron (nFe), nickel (nNi), and zinc (nZn) on a dibenzofuran (DF) degrading bacterium Agrobacterium sp. PH-08. NP (0-1,000 mg L(-1)) -treated bacterial cells were assessed for cytotoxicity, genotoxicity, growth and biodegradation activities at biochemical and molecular levels. In an aqueous system, the bacterial cells treated with nAl, nZn and nNi at 500 mg L(-1) showed significant reduction in cell viability (30-93.6 %, p < 0.05), while nFe had no significant inhibition on bacterial cell viability. In the presence of nAl, nZn and nNi, the cells exhibited elevated levels of reactive oxygen species (ROS), DNA damage and cell death. Furthermore, NP exposure showed significant (p < 0.05) impairment in DF and catechol biodegradation activities. The reduction in DF biodegradation was ranged about 71.7-91.6 % with single NPs treatments while reached up to 96.3 % with a mixture of NPs. Molecular and biochemical investigations also clearly revealed that NP exposure drastically affected the catechol-2,3-dioxygenase activities and its gene (c23o) expression. However, no significant inhibition was observed in nFe treatment. The bacterial extracellular polymeric materials and by-products from DF degradation can be assumed as key factors in diminishing the toxic effects of NPs, especially for nFe. This study clearly demonstrates the impact of single and mixed NPs on the microbial catabolism of xenobiotic-degrading bacteria at biochemical and molecular levels. This is the first study on estimating the impact of mixed NPs on microbial biodegradation.

Effect of impaired twitching motility and biofilm dispersion on performance of Pseudomonas aeruginosa-powered microbial fuel cells.

PubMed

Shreeram, Devesh D; Panmanee, Warunya; McDaniel, Cameron T; Daniel, Susan; Schaefer, Dale W; Hassett, Daniel J

2018-02-01

Pseudomonas aeruginosa is a metabolically voracious bacterium that is easily manipulated genetically. We have previously shown that the organism is also highly electrogenic in microbial fuel cells (MFCs). Polarization studies were performed in MFCs with wild-type strain PAO1 and three mutant strains (pilT, bdlA and pilT bdlA). The pilT mutant was hyperpiliated, while the bdlA mutant was suppressed in biofilm dispersion chemotaxis. The double pilT bdlA mutant was expected to have properties of both mutations. Polarization data indicate that the pilT mutant showed 5.0- and 3.2-fold increases in peak power compared to the wild type and the pilT bdlA mutant, respectively. The performance of the bdlA mutant was surprisingly the lowest, while the pilT bdlA electrogenic performance fell between the pilT mutant and wild-type bacteria. Measurements of biofilm thickness and bacterial viability showed equal viability among the different strains. The thickness of the bdlA mutant, however, was twice that of wild-type strain PAO1. This observation implicates the presence of dead or dormant bacteria in the bdlA mutant MFCs, which increases biofilm internal resistance as confirmed by electrochemical measurements.

Growth and Survival of Perchlorate-Reducing Bacteria in Media Containing Elevated Perchlorate Concentrations and UV-C Conditions

NASA Technical Reports Server (NTRS)

Bywaters, K. F.; Mckay, C. P.; Quinn, R. C.

2017-01-01

Introduction: The identification of perchlorate (ClO4(-)) on Mars has led to the possibility that complete redox couples are available for microbial metabolism in contemporary surface environments. Perchlorate-reducing bacteria (PRB) utilize ClO4(-) and chlorate (ClO3(-)) as terminal electron acceptors due to the high reduction potential. Additionally, ClO4(-) salts have been suggested as a possible source of brines on Mars and spectral evidence indicates that the hydration of ClO4(-) salts in the regolith of Martian is linked to the surface recurring slope lineae (RSL). For these reasons PRB may serve as analog organisms for possible life on Mars. However, there is very little information on the viability of PRB in aqueous environments that contain high levels of perchlorate Microorganisms on or near the surface of Mars, such as in the RSL, would potentially be exposed to high-salinity and high ultraviolet radiation environments. Under these extreme conditions, microorganisms must possess mechanisms for maintaining continued high genome fidelity. To assess possible microbial viability in contemporary Mars analog environments we are investigating the tolerance of two PRB strains in aqueous conditions under high UV-C conditions and high ClO4(-) concentrations.

An investigation into the preservation of microbial cell banks for α-amylase production during 5 l fed-batch Bacillus licheniformis fermentations.

PubMed

Hancocks, Nichola H; Thomas, Colin R; Stocks, Stuart M; Hewitt, Christopher J

2010-10-01

Fluorescent staining techniques were used for a systematic examination of methods used to cryopreserve microbial cell banks. The aim of cryopreservation here is to ensure subsequent reproducible fermentation performance rather than just post thaw viability. Bacillus licheniformis cell physiology post-thaw is dependent on the cryopreservant (either Tween 80, glycerol or dimethyl sulphoxide) and whilst this had a profound effect on the length of the lag phase, during subsequent 5 l fed-batch fermentations, it had little effect on maximum specific growth rate, final biomass concentration or α-amylase activity. Tween 80 not only protected the cells during freezing but also helped them recover post-thaw resulting in shorter process times.

Lead Toxicity to the Performance, Viability, And Community Composition of Activated Sludge Microorganisms

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

Yuan, L; Zhi, W; Liu, YS

Lead (Pb) is a prominent toxic metal in natural and engineered systems. Current knowledge on Pb toxicity to the activated sludge has been limited to short-term (<= 24 h) toxicity. The effect of extended Pb exposure on process performance, bacterial viability, and community compositions remains unknown. We quantified the 24-h and 7-day Pb toxicity to chemical oxygen demand (COD) and NH3-N removal, bacterial viability, and community compositions using lab-scale experiments. Our results showed that 7-day toxicity was significantly higher than the short-term 24-h toxicity. Ammonia-oxidizing bacteria were more susceptible than the heterotrophs to Pb toxicity. The specific oxygen uptake ratemore » responded quickly to Pb addition and could serve as a rapid indicator for detecting Pb pollutions. Microbial viability decreased linearly with the amount of added Pb at extended exposure. The bacterial community diversity was markedly reduced with elevated Pb concentrations. Surface analysis suggested that the adsorbed form of Pb could have contributed to its toxicity along with the dissolved form. Our study provides for the first time a systematic investigation of the effect of extended exposure of Pb on the performance and microbiology of aerobic treatment processes, and it indicates that long-term Pb toxicity has been underappreciated by previous studies.« less

Long-term performance and characterization of microbial desalination cells in treating domestic wastewater.

PubMed

Luo, Haiping; Xu, Pei; Ren, Zhiyong

2012-09-01

Microbial desalination cell represents a new technology for simultaneous wastewater treatment, water desalination, and energy production. This study characterized the long-term performance of MDC during wastewater treatment and identified the key factors that caused performance decline. The 8-month operation shows that MDC performance decreased over time, as indicated by a 47% decline in current density, a 46% drop in Columbic efficiency, and a 27% decrease in desalination efficiency. Advanced electrochemical, microscopy, and spectroscopy analyses all confirmed biofouling on the anion exchange membrane, which increased system resistance and reduced ionic transfer and energy conversion efficiency. Minor chemical scaling was found on the cation exchange membrane surface. Microbial communities became less diverse at the end of operation, and Proteobacteria spp. was dominant on both anode and AEM fouling layer surface. These results provide insights into the viability of long-term MDC operation on reactor performance and direct system development through membrane optimization. Copyright © 2012 Elsevier Ltd. All rights reserved.

Sensory evaluation of locally-grown fruit purees and inulin fibre on probiotic yogurt in Mwanza, Tanzania and the Microbial Analysis of Probiotic Yogurt Fortified with Moringa oleifera.

PubMed

Hekmat, Sharareh; Morgan, Kathryn; Soltani, Mohammad; Gough, Robert

2015-03-01

The purpose of this study was to establish new food products that increase the nutritional value and health benefits of the probiotic yogurt currently used in the Western Heads East (WHE) Project in Mwanza, Tanzania. The probiotic yogurt has established health benefits, and product development through fortification must not adversely affect the acceptability of yogurt or the viability of the probiotics. Both sensory testing and microbial analysis testing were conducted. The products tested were yogurt fortified with locally-grown fruit purees with inulin and Moringa oleifera. The results of the sensory evaluation showed that all yogurts were not rated significantly different from the control, except for appearance. The avocado puree without inulin rated significantly lower in all categories. The microbial analysis showed that Moringa oleifera did not negatively affect the growth of Lactobacillus rhamnosus GR-1 in MRS, milk or yogurt, although a significant decrease was found after 5 weeks of storage at 4 (o)C.

Evidence of pathogenic microbes in the International Space Station drinking water: reason for concern?

NASA Technical Reports Server (NTRS)

La Duc, Myron T.; Sumner, Randall; Pierson, Duane; Venkat, Parth; Venkateswaran, Kasthuri

2004-01-01

Molecular analyses were carried out on four preflight and six postflight International Space Station (ISS)-associated potable water samples at various stages of purification, storage, and transport, to ascertain their associated microbial diversities and overall microbial burdens. Following DNA extraction, PCR amplification, and molecular cloning procedures, rDNA sequences closely related to pathogenic species of Acidovorax, Afipia, Brevundimonas, Propionibacterium, Serratia, and others were recovered in varying abundance. Retrieval of sequences arising from the iodine (biocide)-reducing Delftia acidovorans in postflight waters is also of concern. Total microbial burdens of ISS potable waters were derived from data generated by an ATP-based enumeration procedure, with results ranging from 0 to 4.9 x 10(4) cells/ml. Regardless of innate biases in sample collection and analysis, such circumstantial evidence for the presence of viable, intact pathogenic cells should not be taken lightly. Implementation of new cultivation approaches and/or viability-based assays are requisite to confirm such an occurrence.

Deciphering the Counterplay of Aspergillus fumigatus Infection and Host Inflammation by Evolutionary Games on Graphs

NASA Astrophysics Data System (ADS)

Pollmächer, Johannes; Timme, Sandra; Schuster, Stefan; Brakhage, Axel A.; Zipfel, Peter F.; Figge, Marc Thilo

2016-06-01

Microbial invaders are ubiquitously present and pose the constant risk of infections that are opposed by various defence mechanisms of the human immune system. A tight regulation of the immune response ensures clearance of microbial invaders and concomitantly limits host damage that is crucial for host viability. To investigate the counterplay of infection and inflammation, we simulated the invasion of the human-pathogenic fungus Aspergillus fumigatus in lung alveoli by evolutionary games on graphs. The layered structure of the innate immune system is represented by a sequence of games in the virtual model. We show that the inflammatory cascade of the immune response is essential for microbial clearance and that the inflammation level correlates with the infection-dose. At low infection-doses, corresponding to daily inhalation of conidia, the resident alveolar macrophages may be sufficient to clear infections, however, at higher infection-doses their primary task shifts towards recruitment of neutrophils to infection sites.

Deciphering the Counterplay of Aspergillus fumigatus Infection and Host Inflammation by Evolutionary Games on Graphs

PubMed Central

Pollmächer, Johannes; Timme, Sandra; Schuster, Stefan; Brakhage, Axel A.; Zipfel, Peter F.; Figge, Marc Thilo

2016-01-01

Microbial invaders are ubiquitously present and pose the constant risk of infections that are opposed by various defence mechanisms of the human immune system. A tight regulation of the immune response ensures clearance of microbial invaders and concomitantly limits host damage that is crucial for host viability. To investigate the counterplay of infection and inflammation, we simulated the invasion of the human-pathogenic fungus Aspergillus fumigatus in lung alveoli by evolutionary games on graphs. The layered structure of the innate immune system is represented by a sequence of games in the virtual model. We show that the inflammatory cascade of the immune response is essential for microbial clearance and that the inflammation level correlates with the infection-dose. At low infection-doses, corresponding to daily inhalation of conidia, the resident alveolar macrophages may be sufficient to clear infections, however, at higher infection-doses their primary task shifts towards recruitment of neutrophils to infection sites. PMID:27291424

Molecular improvements in microbial α-amylases for enhanced stability and catalytic efficiency.

PubMed

Sindhu, Raveendran; Binod, Parameswaran; Madhavan, Aravind; Beevi, Ummalyma Sabeela; Mathew, Anil Kuruvilla; Abraham, Amith; Pandey, Ashok; Kumar, Vinod

2017-12-01

α-Amylases is one of the most important industrial enzyme which contributes to 25% of the industrial enzyme market. Though it is produced by plant, animals and microbial source, those from microbial source seems to have potential applications due to their stability and economic viability. However a large number of α-amylases from different sources have been detailed in the literature, only few numbers of them could withstand the harsh industrial conditions. Thermo-stability, pH tolerance, calcium independency and oxidant stability and starch hydrolyzing efficiency are the crucial qualities for α-amylase in starch based industries. Microbes can be genetically modified and fine tuning can be done for the production of enzymes with desired characteristics for specific applications. This review focuses on the native and recombinant α-amylases from microorganisms, their heterologous production and the recent molecular strategies which help to improve the properties of this industrial enzyme. Copyright © 2017 Elsevier Ltd. All rights reserved.

Biology of the Coarse Aerosol Mode: Insights Into Urban Aerosol Ecology

NASA Astrophysics Data System (ADS)

Dueker, E.; O'Mullan, G. D.; Montero, A.

2015-12-01

Microbial aerosols have been understudied, despite implications for climate studies, public health, and biogeochemical cycling. Because viable bacterial aerosols are often associated with coarse aerosol particles, our limited understanding of the coarse aerosol mode further impedes our ability to develop models of viable bacterial aerosol production, transport, and fate in the outdoor environment, particularly in crowded urban centers. To address this knowledge gap, we studied aerosol particle biology and size distributions in a broad range of urban and rural settings. Our previously published findings suggest a link between microbial viability and local production of coarse aerosols from waterways, waste treatment facilities, and terrestrial systems in urban and rural environments. Both in coastal Maine and in New York Harbor, coarse aerosols and viable bacterial aerosols increased with increasing wind speeds above 4 m s-1, a dynamic that was observed over time scales ranging from minutes to hours. At a New York City superfund-designated waterway regularly contaminated with raw sewage, aeration remediation efforts resulted in significant increases of coarse aerosols and bacterial aerosols above that waterway. Our current research indicates that bacterial communities in aerosols at this superfund site have a greater similarity to bacterial communities in the contaminated waterway with wind speeds above 4 m s-1. Size-fractionated sampling of viable microbial aerosols along the urban waterfront has also revealed significant shifts in bacterial aerosols, and specifically bacteria associated with coarse aerosols, when wind direction changes from onshore to offshore. This research highlights the key connections between bacterial aerosol viability and the coarse aerosol fraction, which is important in assessments of production, transport, and fate of bacterial contamination in the urban environment.

Batch-batch stable microbial community in the traditional fermentation process of huyumei broad bean pastes.

PubMed

Zhu, Linjiang; Fan, Zihao; Kuai, Hui; Li, Qi

2017-09-01

During natural fermentation processes, a characteristic microbial community structure (MCS) is naturally formed, and it is interesting to know about its batch-batch stability. This issue was explored in a traditional semi-solid-state fermentation process of huyumei, a Chinese broad bean paste product. The results showed that this MCS mainly contained four aerobic Bacillus species (8 log CFU per g), including B. subtilis, B. amyloliquefaciens, B. methylotrophicus, and B. tequilensis, and the facultative anaerobe B. cereus with a low concentration (4 log CFU per g), besides a very small amount of the yeast Zygosaccharomyces rouxii (2 log CFU per g). The dynamic change of the MCS in the brine fermentation process showed that the abundance of dominant species varied within a small range, and in the beginning of process the growth of lactic acid bacteria was inhibited and Staphylococcus spp. lost its viability. Also, the MCS and its dynamic change were proved to be highly reproducible among seven batches of fermentation. Therefore, the MCS naturally and stably forms between different batches of the traditional semi-solid-state fermentation of huyumei. Revealing microbial community structure and its batch-batch stability is helpful for understanding the mechanisms of community formation and flavour production in a traditional fermentation. This issue in a traditional semi-solid-state fermentation of huyumei broad bean paste was firstly explored. This fermentation process was revealed to be dominated by a high concentration of four aerobic species of Bacillus, a low concentration of B. cereus and a small amount of Zygosaccharomyces rouxii. Lactic acid bacteria and Staphylococcus spp. lost its viability at the beginning of fermentation. Such the community structure was proved to be highly reproducible among seven batches. © 2017 The Society for Applied Microbiology.

Effects of superheated steam on Geobacillus stearothermophilus spore viability.

PubMed

Head, D S; Cenkowski, S; Holley, R; Blank, G

2008-04-01

To examine the effect of processing with superheated steam (SS) on Geobacillus stearothermophilus ATCC 10149 spores. Two inoculum levels of spores of G. stearothermophilus were mixed with sterile sand and exposed to SS at 105-175 degrees C. The decimal reduction time (D-value) and the thermal resistance constant (z-value) were calculated. The effect of cooling of spores between periods of exposure to SS was also examined. A mean z-value of 25.4 degrees C was calculated for both inoculum levels for SS processing temperatures between 130 degrees C and 175 degrees C. Spore response to SS treatment depends on inoculum size. SS treatment may be effective for reduction in viability of thermally resistant bacterial spores provided treatments are separated by intermittent cooling periods. There is a need for technologies that require short thermal processing times to eliminate bacterial spores in foods. The SS processing technique has the potential to reduce microbial load and to modify food texture with less energy in comparison to commonly used hot air treatment. This work provides information on the effect of SS processing parameters on the viability of G. stearothermophilus spores.

Probiotic viability and storage stability of yogurts and fermented milks prepared with several mixtures of lactic acid bacteria.

PubMed

Mani-López, E; Palou, E; López-Malo, A

2014-05-01

Currently, the food industry wants to expand the range of probiotic yogurts but each probiotic bacteria offers different and specific health benefits. Little information exists on the influence of probiotic strains on physicochemical properties and sensory characteristics of yogurts and fermented milks. Six probiotic yogurts or fermented milks and 1 control yogurt were prepared, and we evaluated several physicochemical properties (pH, titratable acidity, texture, color, and syneresis), microbial viability of starter cultures (Lactobacillus delbrueckii ssp. bulgaricus and Streptococcus thermophilus) and probiotics (Lactobacillus acidophilus, Lactobacillus casei, and Lactobacillus reuteri) during fermentation and storage (35 d at 5°C), as well as sensory preference among them. Decreases in pH (0.17 to 0.50 units) and increases in titratable acidity (0.09 to 0.29%) were observed during storage. Only the yogurt with S. thermophilus, L. delbrueckii ssp. bulgaricus, and L. reuteri differed in firmness. No differences in adhesiveness were determined among the tested yogurts, fermented milks, and the control. Syneresis was in the range of 45 to 58%. No changes in color during storage were observed and no color differences were detected among the evaluated fermented milk products. Counts of S. thermophilus decreased from 1.8 to 3.5 log during storage. Counts of L. delbrueckii ssp. bulgaricus also decreased in probiotic yogurts and varied from 30 to 50% of initial population. Probiotic bacteria also lost viability throughout storage, although the 3 probiotic fermented milks maintained counts ≥ 10(7)cfu/mL for 3 wk. Probiotic bacteria had variable viability in yogurts, maintaining counts of L. acidophilus ≥ 10(7) cfu/mL for 35 d, of L. casei for 7d, and of L. reuteri for 14 d. We found no significant sensory preference among the 6 probiotic yogurts and fermented milks or the control. However, the yogurt and fermented milk made with L. casei were better accepted. This study presents relevant information on physicochemical, sensory, and microbial properties of probiotic yogurts and fermented milks, which could guide the dairy industry in developing new probiotic products. Copyright © 2014 American Dairy Science Association. Published by Elsevier Inc. All rights reserved.

Quality challenges associated with microbial-based cleaning products from the Industry Perspective.

PubMed

Teasdale, Steve M; Kademi, Ali

2018-06-01

Microbial-based cleaning products (MBCPs) continue to gain popularity in the market as environmentally friendly cleaners. The majority of these products contain spores of various Bacillus species. Although the microorganisms used in MBCPs are subject to regulation in Canada under the Canadian Environmental Protection Act, the products themselves are not. Unlike other types of microbial products such as probiotics and biopesticides, the use, manufacture and quality parameters of MBCPs in Canada and other countries are poorly defined and not specifically subject to any required standards. Due to their complexity and nature, these products feature unique quality challenges. We noted the existing MBCPs we analyzed vary vastly in quality; external microbial contaminants, viability of the spores and the biocompatibility of the ingredients are issues that greatly affect product quality. A proper taxonomic identification of the bacterial species used also seems to be a major challenge for a number of manufacturers. A good understanding of the mechanisms governing these quality challenges and the adoption of good practices for the cultivation, harvesting, formulation, and manufacture of these types of products are essential for achieving high-quality performance standards. Copyright © 2017 Elsevier Ltd. All rights reserved.

Effect of multiwalled carbon nanotubes on UASB microbial consortium.

PubMed

Yadav, Tushar; Mungray, Alka A; Mungray, Arvind K

2016-03-01

The continuous rise in production and applications of carbon nanotubes (CNTs) has grown a concern about their fate and toxicity in the environment. After use, these nanomaterials pass through sewage and accumulate in wastewater treatment plants. Since, such plants rely on biological degradation of wastes; their activity may decrease due to the presence of CNTs. This study investigated the effect of multiwalled carbon nanotubes (MWCNTs) on upflow anaerobic sludge blanket (UASB) microbial activity. The toxic effect on microbial viability, extracellular polymeric substances (EPS), volatile fatty acids (VFA), and biogas generation was determined. The reduction in a colony-forming unit (CFU) was 29 and 58 % in 1 and 100 mg/L test samples, respectively, as compared to control. The volatile fatty acids and biogas production was also found reduced. The scanning electron microscopy (SEM) and fluorescent microscopy images confirmed that the MWCNT mediated microbial cell damage. This damage caused the increase in EPS carbohydrate, protein, and DNA concentration. Fourier transform infrared (FTIR) spectroscopy results supported the alterations in sludge EPS due to MWCNT. Our observations offer a new insight to understand the nanotoxic effect of MWCNTs on UASB microflora in a complex environment system.

Biological activity of the non-microbial fraction of kefir: antagonism against intestinal pathogens.

PubMed

Iraporda, Carolina; Abatemarco Júnior, Mário; Neumann, Elisabeth; Nunes, Álvaro Cantini; Nicoli, Jacques R; Abraham, Analía G; Garrote, Graciela L

2017-08-01

Kefir is a fermented milk obtained by the activity of kefir grains which are composed of lactic and acetic acid bacteria, and yeasts. Many beneficial health effects have been associated with kefir consumption such as stimulation of the immune system and inhibition of pathogenic microorganisms. The biological activity of kefir may be attributed to the presence of a complex microbiota as well as the microbial metabolites that are released during fermentation. The aim of this work was to characterise the non-microbial fraction of kefir and to study its antagonism against Escherichia coli, Salmonella spp. and Bacillus cereus. During milk fermentation there was a production of organic acids, mainly lactic and acetic acid, with a consequent decrease in pH and lactose content. The non-microbial fraction of kefir added to nutrient broth at concentrations above 75% v/v induced a complete inhibition of pathogenic growth that could be ascribed to the presence of un-dissociated lactic acid. In vitro assays using an intestinal epithelial cell model indicated that pre-incubation of cells with the non-microbial fraction of kefir did not modify the association/invasion of Salmonella whereas pre-incubation of Salmonella with this fraction under conditions that did not affect their viability significantly decreased the pathogen's ability to invade epithelial cells. Lactate exerted a protective effect against Salmonella in a mouse model, demonstrating the relevance of metabolites present in the non-microbial fraction of kefir produced during milk fermentation.

Sodium ascorbate kills Candida albicans in vitro via iron-catalyzed Fenton reaction: importance of oxygenation and metabolism

PubMed Central

Avci, Pinar; Freire, Fernanda; Banvolgyi, Andras; Mylonakis, Eleftherios; Wikonkal, Norbert M; Hamblin, Michael R

2016-01-01

Aim: Ascorbate can inhibit growth and even decrease viability of various microbial species including Candida albicans. However the optimum conditions and the mechanism of action are unclear. Materials/methodology: Candida albicans shaken for 90 min in a buffered solution of ascorbate (90 mM) gave a 5-log reduction of cell viability, while there was no killing without shaking, in growth media with different carbon sources or at 4°C. Killing was inhibited by the iron chelator 2,2′-bipyridyl. Hydroxyphenyl fluorescein probe showed the intracellular generation of hydroxyl radicals. Results/conclusion: Ascorbate-mediated killing of C. albicans depends on oxygenation and metabolism, involves iron-catalyzed generation of hydroxyl radicals via Fenton reaction and depletion of intracellular NADH. Ascorbate could serve as a component of a topical antifungal therapy. PMID:27855492

The model of fungal population dynamics affected by nystatin

NASA Astrophysics Data System (ADS)

Voychuk, Sergei I.; Gromozova, Elena N.; Sadovskiy, Mikhail G.

Fungal diseases are acute problems of the up-to-day medicine. Significant increase of resistance of microorganisms to the medically used antibiotics and a lack of new effective drugs follows in a growth of dosage of existing chemicals to solve the problem. Quite often such approach results in side effects on humans. Detailed study of fungi-antibiotic dynamics can identify new mechanisms and bring new ideas to overcome the microbial resistance with a lower dosage of antibiotics. In this study, the dynamics of the microbial population under antibiotic treatment was investigated. The effects of nystatin on the population of Saccharomyces cerevisiae yeasts were used as a model system. Nystatin effects were investigated both in liquid and solid media by viability tests. Dependence of nystatin action on osmotic gradient was evaluated in NaCl solutions. Influences of glucose and yeast extract were additionally analyzed. A "stepwise" pattern of the cell death caused by nystatin was the most intriguing. This pattern manifested in periodical changes of the stages of cell death against stages of resistance to the antibiotic. The mathematical model was proposed to describe cell-antibiotic interactions and nystatin viability effects in the liquid medium. The model implies that antibiotic ability to cause a cells death is significantly affected by the intracellular compounds, which came out of cells after their osmotic barriers were damaged

Anti-cancer Effect of Luminacin, a Marine Microbial Extract, in Head and Neck Squamous Cell Carcinoma Progression via Autophagic Cell Death.

PubMed

Shin, Yoo Seob; Cha, Hyun Young; Lee, Bok-Soon; Kang, Sung Un; Hwang, Hye Sook; Kwon, Hak Cheol; Kim, Chul-Ho; Choi, Eun Chang

2016-04-01

The purpose of this study is to determine whether luminacin, a marine microbial extract from the Streptomyces species, has anti-tumor effects on head and neck squamous cell carcinoma (HNSCC) cell lines via autophagic cell death. Inhibition of cell survival and increased cell death was measured using cell viability, colony forming, and apoptosis assays. Migration and invasion abilities of head and cancer cells were evaluated using wound healing, scattering, and invasion assays. Changes in the signal pathway related to autophagic cell death were investigated. Drug toxicity of luminacin was examined in in vitro HaCaT cells and an in vivo zebrafish model. Luminacin showed potent cytotoxicity in HNSCC cells in cell viability, colony forming, and fluorescence-activated cell sorting analysis. In vitro migration and invasion of HNSCC cells were attenuated by luminacin treatment. Combined with Beclin-1 and LC3B, Luminacin induced autophagic cell death in head and neck cancer cells. In addition, in a zebrafish model and human keratinocyte cell line used for toxicity testing, luminacin treatment with a cytotoxic concentration to HNSCC cells did not cause toxicity. Taken together, these results demonstrate that luminacin induces the inhibition of growth and cancer progression via autophagic cell death in HNSCC cell lines, indicating a possible alternative chemotherapeutic approach for treatment of HNSCC.

Viability and resistance of lactobacilli isolated from cocoa fermentation to simulated gastrointestinal digestive steps in soy yogurt.

PubMed

Saito, V S T; Dos Santos, T F; Vinderola, C G; Romano, C; Nicoli, J R; Araújo, L S; Costa, M M; Andrioli, J L; Uetanabaro, A P T

2014-02-01

To study the potential probiotic characteristics such as decrease of pH, microbial viability, and tolerance to simulated digestive steps of fermented soy beverage ("soy yogurt") produced with lactobacilli isolated from cocoa fermentation (Lactobacillus fermentum TcUESC01 and Lactobacillus plantarum TcUESC02) during fermentation and refrigerated storage. The sensory acceptance of the yogurts was also tested. Samples of soy yogurt produced with L. fermentum TcUESC01 or L. plantarum TcUESC02 were collected during fermentation (0, 4, 8, and 12 h) and refrigerated storage (1, 9, 18, and 27 d), and submitted to pH and bacterial viability determinations. Tolerance to simulated digestion steps was done with refrigerated storage samples at 9 °C. Simulated digestion was performed in 3 successive steps: exposure to pepsin-HCl solution, bile shock, and simulated small intestinal juice. During storage, a decrease in pH and lactobacillus viability was observed. L. fermentum TcUESC01 showed to be more resistant than L. plantarum TcUESC02 to simulated gastrointestinal digestion. All soy yogurts showed acceptable hedonic scores (greater than 5 in a 9-point hedonic scale ranging from "like extremely" to "dislike extremely") in sensory evaluation for flavor, aroma, color, consistency, and overall impression. L. plantarum TcUESC02 and, especially, L. fermentum TcUESC01 showed potential probiotic characteristics when considering pH, cell viability, and tolerance to simulated digestive steps and did not affect the sensory characteristics when supplemented to soy yogurt during storage. © 2014 Institute of Food Technologists®

Molecular characterization of microbial population dynamics during sildenafil citrate degradation.

PubMed

De Felice, Bruna; Argenziano, Carolina; Guida, Marco; Trifuoggi, Marco; Russo, Francesca; Condorelli, Valerio; Inglese, Mafalda

2009-02-01

Little is known about pharmaceutical and personal care products pollutants (PPCPs), but there is a growing interest in how they might impact the environment and microbial communities. The widespread use of Viagra (sildenafil citrate) has attracted great attention because of the high usage rate, the unpredictable disposal and the unknown potential effects on wildlife and the environment. Until now information regarding the impact of Viagra on microbial community in water environment has not been reported. In this research, for the first time, the genetic profile of the microbial community, developing in a Viagra polluted water environment, was evaluated by means of the 16S and 18S rRNA genes, for bacteria and fungi, respectively, amplified by polymerase chain reaction (PCR) and separated using the denaturing gradient gel electrophoresis (DGGE) technique. The DGGE results revealed a complex microbial community structure with most of the population persisting throughout the experimental period. DNA sequences from bands observed in the different denaturing gradient gel electrophoresis profiles exhibited the highest degree of identity to uncultured bacteria and fungi found previously mainly in polluted environmental and treating bioreactors. Biotransformation ability of sildenafil citrate by the microbial pool was studied and the capability of these microorganisms to detoxify a polluted water ecosystem was assessed. The bacterial and fungal population was able to degrade sildenafil citrate entirely. Additionally, assays conducted on Daphnia magna, algal growth inhibition assay and cell viability determination on HepG2 human cells showed that biotransformation products obtained from the bacterial growth was not toxic. The higher removal efficiency for sildenafil citrate and the lack of toxicity by the biotransformation products obtained showed that the microbial community identified here represented a composite population that might have biotechnological relevance to retrieve sildenafil citrate contaminated sites.

An Evaluation of Subsurface Microbial Activity Conditional to Subsurface Temperature, Porosity, and Permeability at North American Carbon Sequestration Sites

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

Wilson, B.; Mordensky, S.; Verba, Circe

Several nations, including the United States, recognize global climate change as a force transforming the global ecosphere. Carbon dioxide (CO 2) is a greenhouse gas that contributes to the evolving climate. Reduction of atmospheric CO 2 levels is a goal for many nations and carbon sequestration which traps CO 2 in the Earth’s subsurface is one method to reduce atmospheric CO 2 levels. Among the variables that must be considered in developing this technology to a national scale is microbial activity. Microbial activity or biomass can change rock permeability, alter artificial seals around boreholes, and play a key role inmore » biogeochemistry and accordingly may determine how CO 2 is sequestered underground. Certain physical parameters of a reservoir found in literature (e.g., temperature, porosity, and permeability) may indicate whether a reservoir can host microbial communities. In order to estimate which subsurface formations may host microbes, this report examines the subsurface temperature, porosity, and permeability of underground rock formations that have high potential to be targeted for CO 2 sequestration. Of the 268 North American wellbore locations from the National Carbon Sequestration Database (NATCARB; National Energy and Technology Laboratory, 2015) and 35 sites from Nelson and Kibler (2003), 96 sequestration sites contain temperature data. Of these 96 sites, 36 sites have temperatures that would be favorable for microbial survival, 48 sites have mixed conditions for supporting microbial populations, and 11 sites would appear to be unfavorable to support microbial populations. Future studies of microbe viability would benefit from a larger database with more formation parameters (e.g. mineralogy, structure, and groundwater chemistry), which would help to increase understanding of where CO 2 sequestration could be most efficiently implemented.« less

Resistance of Terrestrial Microbial Communities to Impack of Physical Conditinos of Subsurface Layers of Martian Regolith

NASA Astrophysics Data System (ADS)

Cheptsov, V. S.; Vorobyova, E. A.

2017-05-01

Currently, astrobiology is focused on Mars as one of the most perspective objects in the Solar System to search for microbial life. It was assumed that the putative biosphere of Mars could be cryopreserved and had been stored for billions of years in anabiotic state like microbial communities of Arctic and Antarctic permafrost deposits have been preserved till now for millions of years. In this case microbial cells should be not able to repair the damages or these processes have to be significantly depressed, and the main factor causing cell's death should be ionizing radiation. In a series of experiments we simulated the effects of combination of physical factors known as characteristics of the Martian regolith (and close to the space environment) on the natural microbial communities inhabiting xerophytic harsh habitats with extreme temperature conditions: polar permafrost and desert soils. The aim of the study was to examine the cumulative effect of factors (gamma radiation, low temperature, low pressure) to assess the possibility of metabolic reactions, and to find limits of the viability of natural microbial communities after exposure to the given conditions. It was found that microbial biomarkers could be reliably detected in soil samples after radiation dose accumulation up to 1 MGy (not further investigated) in combination with exposure to low temperature and low pressure. Resistance to extremely high doses of radiation in simulated conditions proves that if there was an Earth-like biosphere on the early Mars microorganisms could survive in the surface or subsurface layers of the Martian regolith for more than tens of millions of years after climate change. The study gives also some new grounds for the approval of transfer of viable microorganisms in space.

Novel method for enumeration of viable Lactobacillus plantarum WCFS1 cells after single-droplet drying.

PubMed

Perdana, Jimmy; Bereschenko, Ludmila; Roghair, Mark; Fox, Martijn B; Boom, Remko M; Kleerebezem, Michiel; Schutyser, Maarten A I

2012-11-01

Survival of probiotic bacteria during drying is not trivial. Survival percentages are very specific for each probiotic strain and can be improved by careful selection of drying conditions and proper drying carrier formulation. An experimental approach is presented, comprising a single-droplet drying method and a subsequent novel screening methodology, to assess the microbial viability within single particles. The drying method involves the drying of a single droplet deposited on a flat, hydrophobic surface under well-defined drying conditions and carrier formulations. Semidried or dried particles were subjected to rehydration, fluorescence staining, and live/dead enumeration using fluorescence microscopy. The novel screening methodology provided accurate survival percentages in line with conventional plating enumeration and was evaluated in single-droplet drying experiments with Lactobacillus plantarum WCFS1 as a model probiotic strain. Parameters such as bulk air temperatures and the carrier matrices (glucose, trehalose, and maltodextrin DE 6) were varied. Following the experimental approach, the influence on the viability as a function of the drying history could be monitored. Finally, the applicability of the novel viability assessment was demonstrated for samples obtained from drying experiments at a larger scale.

Novel Method for Enumeration of Viable Lactobacillus plantarum WCFS1 Cells after Single-Droplet Drying

PubMed Central

Perdana, Jimmy; Bereschenko, Ludmila; Roghair, Mark; Fox, Martijn B.; Boom, Remko M.; Kleerebezem, Michiel

2012-01-01

Survival of probiotic bacteria during drying is not trivial. Survival percentages are very specific for each probiotic strain and can be improved by careful selection of drying conditions and proper drying carrier formulation. An experimental approach is presented, comprising a single-droplet drying method and a subsequent novel screening methodology, to assess the microbial viability within single particles. The drying method involves the drying of a single droplet deposited on a flat, hydrophobic surface under well-defined drying conditions and carrier formulations. Semidried or dried particles were subjected to rehydration, fluorescence staining, and live/dead enumeration using fluorescence microscopy. The novel screening methodology provided accurate survival percentages in line with conventional plating enumeration and was evaluated in single-droplet drying experiments with Lactobacillus plantarum WCFS1 as a model probiotic strain. Parameters such as bulk air temperatures and the carrier matrices (glucose, trehalose, and maltodextrin DE 6) were varied. Following the experimental approach, the influence on the viability as a function of the drying history could be monitored. Finally, the applicability of the novel viability assessment was demonstrated for samples obtained from drying experiments at a larger scale. PMID:22983965

Evaluation of a potentially probiotic non-dairy beverage developed with honey and kefir grains: Fermentation kinetics and storage study.

PubMed

Fiorda, Fernanda A; de Melo Pereira, Gilberto V; Thomaz-Soccol, Vanete; Rakshit, Sudip K; Soccol, Carlos R

2016-12-01

The aim of this work was to study the fermentation process of honey with kefir grains through a comprehensive understanding of its rheological properties, probiotic cell viability, instrumental color parameters and kinetic aspects in a batch bioreactor and during storage. The results showed that kefir grains were well adapted to bioreactor conditions, reaching high levels of cell viability (over 10 6 CFU mL -1 for total yeast and bacteria), phenolic compounds content (190 GAE/100 g) and acidification after 24 h of fermentation at 30 ℃. Colorimetric analysis showed that lightness (L*) and redness (a*) remained constant, while yellowness intensities (b*) decreased during fermentation time. After 35 days of storage, honey kefir beverage maintained its chemical characteristics and microbial viability as required to be classified as a probiotic product. The Ostwald-de-Waele (R 2  ≥ 0.98) and Herschel-Bulkley (R 2  ≥ 0.99) models can be used to predict the behavior of honey kefir beverage. The parameters analyzed in this study should be taken into account for industrial production of this novel non-dairy beverage. © The Author(s) 2016.

Sensory Evaluation of Locally-grown Fruit Purees and Inulin Fibre on Probiotic Yogurt in Mwanza, Tanzania and the Microbial Analysis of Probiotic Yogurt Fortified with Moringa oleifera

PubMed Central

Morgan, Kathryn; Soltani, Mohammad; Gough, Robert

2015-01-01

ABSTRACT The purpose of this study was to establish new food products that increase the nutritional value and health benefits of the probiotic yogurt currently used in the Western Heads East (WHE) Project in Mwanza, Tanzania. The probiotic yogurt has established health benefits, and product development through fortification must not adversely affect the acceptability of yogurt or the viability of the probiotics. Both sensory testing and microbial analysis testing were conducted. The products tested were yogurt fortified with locally-grown fruit purees with inulin and Moringa oleifera. The results of the sensory evaluation showed that all yogurts were not rated significantly different from the control, except for appearance. The avocado puree without inulin rated significantly lower in all categories. The microbial analysis showed that Moringa oleifera did not negatively affect the growth of Lactobacillus rhamnosus GR-1 in MRS, milk or yogurt, although a significant decrease was found after 5 weeks of storage at 4 oC. PMID:25995722

Observation to Theory in Deep Subsurface Microbiology Research: Can We Piece It Together?

NASA Astrophysics Data System (ADS)

Colwell, F. S.; Thurber, A. R.

2016-12-01

Three decades of observations of microbes in deep environments have led to startling discoveries of life in the subsurface. Now, a few theoretical frameworks exist that help to define Stygian life. Temperature, redox gradients, productivity (e.g., in the overlying ocean), and microbial power requirements are thought to determine the distribution of microbes in the subsurface. Still, we struggle to comprehend the spatial and temporal spectra of Earth processes that define how deep microbe communities survive. Stommel diagrams, originally used to guide oceanographic sampling, may be useful in depicting the subsurface where microbial communities are impacted by co-occurring spatial and temporal phenomena that range across exponential scales. Spatially, the geological settings that influence the activity and distribution of microbes range from individual molecules or minerals all the way up to the planetary-scale where geological formations, occupying up to 105 km3, dictate the bio- and functional geography of microbial communities. Temporally, life in the subsurface may respond in time units familiar to humans (e.g., seconds to days) or to events that unfold over hundred millennial time periods. While surface community dynamics are underpinned by solar and lunar cycles, these cycles only fractionally dictate survival underground where phenomena like tectonic activity, isostatic rebound, and radioactive decay are plausible drivers of microbial life. Geological or planetary processes that occur on thousand or million year cycles could be uniquely important to microbial viability in the subsurface. Such an approach aims at a holistic comprehension of the interaction of Earth system dynamics with microbial ecology.

Modification of the surfaces of medical devices to prevent microbial adhesion and biofilm formation.

PubMed

Desrousseaux, C; Sautou, V; Descamps, S; Traoré, O

2013-10-01

The development of devices with surfaces that have an effect against microbial adhesion or viability is a promising approach to the prevention of device-related infections. To review the strategies used to design devices with surfaces able to limit microbial adhesion and/or growth. A PubMed search of the published literature. One strategy is to design medical devices with a biocidal agent. Biocides can be incorporated into the materials or coated or covalently bonded, resulting either in release of the biocide or in contact killing without release of the biocide. The use of biocides in medical devices is debated because of the risk of bacterial resistance and potential toxicity. Another strategy is to modify the chemical or physical surface properties of the materials to prevent microbial adhesion, a complex phenomenon that also depends directly on microbial biological structure and the environment. Anti-adhesive chemical surface modifications mostly target the hydrophobicity features of the materials. Topographical modifications are focused on roughness and nanostructures, whose size and spatial organization are controlled. The most effective physical parameters to reduce bacterial adhesion remain to be determined and could depend on shape and other bacterial characteristics. A prevention strategy based on reducing microbial attachment rather than on releasing a biocide is promising. Evidence of the clinical efficacy of these surface-modified devices is lacking. Additional studies are needed to determine which physical features have the greatest potential for reducing adhesion and to assess the usefulness of antimicrobial coatings other than antibiotics. Copyright © 2013 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.

Back to the kitchen: food-grade agar is a low-cost alternative to bacteriological agar.

PubMed

Petrovski, Steve; Tillett, Daniel

2012-10-15

Food-grade agar can be used as a low-cost substitute for bacteriological agar in the preparation of solid microbial media. No difference was observed in the colony morphology, growth rate, or viability of bacteria grown on solid media prepared using food-grade agar as compared with using bacteriological-grade agar. This simple tip can reduce the cost of the most common solid media by 80% or more. Copyright © 2012 Elsevier Inc. All rights reserved.

A Comparison of the Microbial Production and Combustion Characteristics of Three Alcohol Biofuels: Ethanol, 1-Butanol, and 1-Octanol.

PubMed

Kremer, Florian; Blank, Lars M; Jones, Patrik R; Akhtar, M Kalim

2015-01-01

Over the last decade, microbes have been engineered for the manufacture of a variety of biofuels. Saturated linear-chain alcohols have great potential as transport biofuels. Their hydrocarbon backbones, as well as oxygenated content, confer combustive properties that make it suitable for use in internal combustion engines. Herein, we compared the microbial production and combustion characteristics of ethanol, 1-butanol, and 1-octanol. In terms of productivity and efficiency, current microbial platforms favor the production of ethanol. From a combustion standpoint, the most suitable fuel for spark-ignition engines would be ethanol, while for compression-ignition engines it would be 1-octanol. However, any general conclusions drawn at this stage regarding the most superior biofuel would be premature, as there are still many areas that need to be addressed, such as large-scale purification and pipeline compatibility. So far, the difficulties in developing and optimizing microbial platforms for fuel production, particularly for newer fuel candidates, stem from our poor understanding of the myriad biological factors underpinning them. A great deal of attention therefore needs to be given to the fundamental mechanisms that govern biological processes. Additionally, research needs to be undertaken across a wide range of disciplines to overcome issues of sustainability and commercial viability.

A Comparison of the Microbial Production and Combustion Characteristics of Three Alcohol Biofuels: Ethanol, 1-Butanol, and 1-Octanol

PubMed Central

Kremer, Florian; Blank, Lars M.; Jones, Patrik R.; Akhtar, M. Kalim

2015-01-01

Over the last decade, microbes have been engineered for the manufacture of a variety of biofuels. Saturated linear-chain alcohols have great potential as transport biofuels. Their hydrocarbon backbones, as well as oxygenated content, confer combustive properties that make it suitable for use in internal combustion engines. Herein, we compared the microbial production and combustion characteristics of ethanol, 1-butanol, and 1-octanol. In terms of productivity and efficiency, current microbial platforms favor the production of ethanol. From a combustion standpoint, the most suitable fuel for spark-ignition engines would be ethanol, while for compression-ignition engines it would be 1-octanol. However, any general conclusions drawn at this stage regarding the most superior biofuel would be premature, as there are still many areas that need to be addressed, such as large-scale purification and pipeline compatibility. So far, the difficulties in developing and optimizing microbial platforms for fuel production, particularly for newer fuel candidates, stem from our poor understanding of the myriad biological factors underpinning them. A great deal of attention therefore needs to be given to the fundamental mechanisms that govern biological processes. Additionally, research needs to be undertaken across a wide range of disciplines to overcome issues of sustainability and commercial viability. PMID:26301219

Marine sources influence fog bioaerosol composition in Namibia and Maine

NASA Astrophysics Data System (ADS)

Evans, S. E.; Dueker, E.; Logan, J. R. V.; Weathers, K. C.

2017-12-01

Organic aerosol particles act as condensation nuclei for fogs and clouds (CCN) and are main determinants of fog evolution, chemical processing, and overall aerosol-fog-cloud interactions. Recent work has confirmed the presence of marine bioaerosols, but little is known about their sources, transport, taxonomic diversity or viability. The few studies that have characterized bioaerosols in fog have been limited to culture-based approaches that capture only a fraction of microbial diversity. We characterized fungal and bacterial communities in the fog in two iconic fog systems, the Coast of Maine (USA) and the Namib Desert (Namibia). The biology of fog in both systems was diverse and distinct, by geography, from dry aerosols, and from local sources. The local environment had a dominant influence on fog in both the Namib and Maine; in particular, the biology of fog in Maine, which was collected near the coast, was more similar to microbial communities from the ocean surface. In both systems, differences between pre- and post-fog aerosol communities suggest that fog events can significantly alter microbial aerosol diversity and composition. This insight into the microbial composition of fog indicates that its origin and frequency has the potential to influence the number and diversity of microorganisms that settle in a given environment, and the composition of microbial aerosol communities in ambient or clear conditions. Here we suggest that fog microbes can possess specific traits that enhance nucleation, altering the transport and deposition of marine- and soil-derived organic matter in terrestrial systems.

Fetal Tissue Procurement for Karyotype Analysis: Clinician or Pathologist - Which is Better?

PubMed

Conant, Joanna L; Tang, Mary E; Waters, Brenda L

2016-01-01

Chromosomal abnormalities are detected in up to 13% of stillbirths and over 20% of those with developmental anomalies. These estimates may be low since up to 50% of samples fail to achieve a result due to microbial overgrowth or nonviability. Tissue for cytogenetics can be procured at bedside by the clinician or by the pathologist in the laboratory. With clinical collection, tissue is placed into culture media immediately, increasing chances of growth. However, collection competes for attention with other activities, which may result in microbial overgrowth or selection of maternal rather than fetal tissue. Laboratory procurement occurs in a controlled environment using sterile technique, but delay in collection may decrease viability. Our goal was to determine which collection method yields better results. We reviewed cases from 2007-2013 that had two samples submitted for cytogenetics, one from the clinician and one from the pathologist. Specimen source, delivery, collection, and culture setup times, harvest date, cell growth, microbial overgrowth, maternal contamination and final result were obtained from medical records and cytogenetic culture sheets. There was no difference in growth rate, maternal cell contamination, or reporting time between clinician- and pathologist-procured samples despite delay in collection time for laboratory samples. Clinical samples had more microbial overgrowth. Compared to samples collected at bedside, samples collected in the laboratory had a lower rate of microbial contamination with similar growth and maternal cell contamination rates, despite prolonged time to collection. Collecting samples both at bedside and in the laboratory is unnecessary.

State transitions and physicochemical aspects of cryoprotection and stabilization in freeze-drying of Lactobacillus rhamnosus GG (LGG).

PubMed

Pehkonen, K S; Roos, Y H; Miao, S; Ross, R P; Stanton, C

2008-06-01

The frozen and dehydrated state transitions of lactose and trehalose were determined and studied as factors affecting the stability of probiotic bacteria to understand physicochemical aspects of protection against freezing and dehydration of probiotic cultures. Lactobacillus rhamnosus GG was frozen (-22 or -43 degrees C), freeze-dried and stored under controlled water vapour pressure (0%, 11%, 23% and 33% relative vapour pressure) conditions. Lactose, trehalose and their mixture (1 : 1) were used as protective media. These systems were confirmed to exhibit relatively similar state transition and water plasticization behaviour in freeze-concentrated and dehydrated states as determined by differential scanning calorimetry. Ice formation and dehydrated materials were studied using cold-stage microscopy and scanning electron microscopy. Trehalose and lactose-trehalose gave the most effective protection of cell viability as observed from colony forming units after freezing, dehydration and storage. Enhanced cell viability was observed when the freezing temperature was -43 degrees C. State transitions of protective media affect ice formation and cell viability in freeze-drying and storage. Formation of a maximally freeze-concentrated matrix with entrapped microbial cells is essential in freezing prior to freeze-drying. Freeze-drying must retain a solid amorphous state of protectant matrices. Freeze-dried matrices contain cells entrapped in the protective matrices in the freezing process. The retention of viability during storage seems to be controlled by water plasticization of the protectant matrix and possibly interactions of water with the dehydrated cells. Highest cell viability was obtained in glassy protective media. This study shows that physicochemical properties of protective media affect the stability of dehydrated cultures. Trehalose and lactose may be used in combination, which is particularly important for the stabilization of probiotic bacteria in dairy systems.

Blue light (470 nm) effectively inhibits bacterial and fungal growth.

PubMed

De Lucca, A J; Carter-Wientjes, C; Williams, K A; Bhatnagar, D

2012-12-01

Blue light (470 nm) LED antimicrobial properties were studied alone against bacteria and with or without the food grade photosensitizer, erythrosine (ERY) against filamentous fungi. Leuconostoc mesenteroides (LM), Bacillus atrophaeus (BA) or Pseudomonas aeruginosa (PA) aliquots were exposed on nutrient agar plates to Array 1 (AR1, 0·2 mW cm(-2)) or Array 2 (AR2, 80 mW cm(-2)), which emitted impure or pure blue light (0-300 J cm(-2)), respectively. Inoculated control (room light only) plates were incubated (48 h) and colonies enumerated. The antifungal properties of blue light combined with ERY (11·4 and 22·8 μmol l(-1)) on Penicillium digitatum (PD) and Fusarium graminearum (FG) conidia were determined. Conidial controls consisted of: no light, room light-treated conidia and ERY plus room light. Light-treated (ERY + blue light) conidial samples were exposed only to AR2 (0-100 J cm(-2)), aliquots spread on potato dextrose agar plates, incubated (48 h, 30°C) and colonies counted. Blue light alone significantly reduced bacterial and FG viability. Combined with ERY, it significantly reduced PD viability. Blue light is lethal to bacteria and filamentous fungi although effectiveness is dependent on light purity, energy levels and microbial genus. Light from two arrays of different blue LEDs significantly reduced bacterial (Leuconostoc mesenteroides, Bacillus atrophaeus and Pseudomonas aeruginosa) viabilities. Significant in vitro viability loss was observed for the filamentous fungi, Penicillium digitatum and Fusarium graminearum when exposed to pure blue light only plus a photosensitizer. F. graminearum viability was significantly reduced by blue light alone. Results suggest that (i) the amount of significant loss in bacterial viability observed for blue light that is pure or with traces of other wavelengths is genus dependent and (ii) depending on fungal genera, pure blue light is fungicidal with or without a photosensitizer. © 2012 The Society for Applied Microbiology.

Microbial Community Structure of an Alluvial Aquifer Treated to Encourage Microbial Induced Calcite Precipitation

NASA Astrophysics Data System (ADS)

Ohan, J.; Saneiyan, S.; Lee, J.; Ntarlagiannis, D.; Burns, S.; Colwell, F. S.

2017-12-01

An oligotrophic aquifer in the Colorado River floodplain (Rifle, CO) was treated with molasses and urea to encourage microbial induced calcite precipitation (MICP). This would stabilize the soil mass by reducing porosity and strengthening the mineral fabric. Over the course of a 15-day treatment period, microbial biomass was collected from monitoring well groundwater for DNA extraction and sequencing. Bromide, a conservative tracer, was co-injected and subsequently detected in downgradient wells, confirming effective nutrient delivery. Conductivity increased during the injection regime and an overall decrease in pH was observed. Groundwater chemistry showed a marked increase in ammonia, suggesting urea hydrolysis - a process catalyzed by the enzyme urease - the primary enzyme implicated in MICP. Additionally, soluble iron was detected, suggesting a general increase in microbial activity; possibly as iron-reducing bacteria changed insoluble ferric oxide to soluble ferrous hydroxide in the anoxic aquifer. DNA sequencing of the 16S rRNA gene confirmed the presence of iron reducing bacteria, including Shewanella and Desulfuromonadales. Generally, a decrease in microbial community diversity was observed when pre-injection community taxa were compared with post-injection community taxa. Phyla indicative of anoxic aquifers were represented in accordance with previous literature at the Rifle site. Linear discriminant analysis showed significant differences in representative phyla over the course of the injection series. Geophysical monitoring of the site further suggested changes that could be due to MICP. Induced polarization increased the phase shift in the primary treated area, in agreement with laboratory experiments. Cross-hole seismic testing confirmed that the shear wave velocities increased in the treated soil mass, implying the soil matrix became more stable. Future investigations will help elucidate the viability and efficacy of MICP treatment in changing microbial community structure, and the functional attributes associated with community change, so that physical strengthening of soils by microbial action can be accomplished.

Effects of the antimicrobial peptide protegrine 1 on sperm viability and bacterial load of boar seminal doses.

PubMed

Sancho, S; Briz, M; Yeste, M; Bonet, S; Bussalleu, E

2017-10-01

The presence of bacteria adversely affects boar sperm quality of seminal doses intended for artificial insemination. Currently, the most common measure to prevent bacteriospermia is the addition of antibiotics in semen extenders; however, mounting evidence shows that microbial resistance exists. A promising alternative to replace antibiotics are antimicrobial peptides. In this study, the effects of the antimicrobial peptide protegrine 1 (PG1) on the sperm viability and bacterial load of boar seminal doses were evaluated. Three different concentrations of PG1 (2.5, 25 and 100 μg/ml) were tested over a storing period of 10 days at 17°C. Sperm viability was analysed by fluorescence microscopy (SYBR14/propidium iodide), and bacterial load was assessed by plating 100 μl of each sample in Luria-Bertani medium and incubated at 37°C for 72 hr under aerobic conditions. Protegrine 1 was effective in controlling the bacterial load in all the assessed concentrations (p < .05), reaching the lowest values at the highest concentrations of the antimicrobial peptide. Nevertheless, sperm viability was significantly (p < .05) reduced by all tested concentrations of this peptide, the most cytotoxic effects being observed at the highest PG1 concentrations. Despite these results, the use of PG1 as an alternative to antibiotics cannot be totally discarded, as further studies using the truncated form of this peptide are needed. © 2017 Blackwell Verlag GmbH.

EMA-qPCR to monitor the efficiency of a closed-coupled solar pasteurization system in reducing Legionella contamination of roof-harvested rainwater.

PubMed

Reyneke, B; Dobrowsky, P H; Ndlovu, T; Khan, S; Khan, W

2016-05-15

Solar pasteurization is effective in reducing the level of indicator organisms in stored rainwater to within drinking water standards. However, Legionella spp. were detected at temperatures exceeding the recommended pasteurization temperatures using polymerase chain reaction assays. The aim of the current study was thus to apply EMA quantitative polymerase chain reaction (EMA-qPCR) to determine whether the Legionella spp. detected were intact cells and therefore possibly viable at pasteurization temperatures >70°C. The BacTiter-Glo™ Microbial Cell Viability Assay was also used to detect the presence of ATP in the tested samples, as ATP indicates the presence of metabolically active cells. Chemical analysis also indicated that all anions and cations were within the respective drinking water guidelines, with the exception of iron (mean: 186.76 μg/L) and aluminium (mean: 188.13 μg/L), which were detected in the pasteurized tank water samples at levels exceeding recommended guidelines. The BacTiter-Glo™ Microbial Cell Viability Assay indicated the presence of viable cells for all pasteurized temperatures tested, with the percentage of ATP (in the form of relative light units) decreasing with increasing temperature [70-79°C (96.7%); 80- 89°C (99.2%); 90-95°C (99.7%)]. EMA-qPCR then indicated that while solar pasteurization significantly reduced (p<0.05) the genomic copy numbers of intact Legionella cells in the pasteurized tank water (~99%), no significant difference (p>0.05) in the mean copy numbers was detected with an increase in the pasteurization temperature, with 6 × 10(3) genomic copies/mL DNA sample obtained at 95°C. As intact Legionella cells were detected in the pasteurized tank water samples, quantitative microbial risk assessment studies need to be conducted to determine the potential health risk associated with using the water for domestic purposes. Copyright © 2016 Elsevier B.V. All rights reserved.

Cryostored autologous skull bone for cranioplasty? A study on cranial bone flaps' viability and microbial contamination after deep-frozen storage at -80°C.

PubMed

Chan, David Yuen Chung; Mok, Yi Tan; Lam, Ping Kuen; Tong, Cindy See Wai; Ng, Stephanie Chi Ping; Sun, Tin Fung David; Poon, Wai Sang

2017-08-01

Craniectomy is a life-saving procedure. Subsequent cranioplasty with autologous skull bone has a bone resorption rate from 4% to 22.8% and an infection rate from 3.3% to 26%. There are concerns with their viability and the potential microbial contamination as they were explanted for a long period of time. Eighteen cranial bone flaps stored at Prince of Wales Hospital Skull Bone Bank during the period from June 2011 to March 2016 were identified. Ethics approval was obtained. Bone chips and deep bone swabs were collected for osteoblast culture and microbial culture. Skull Bone Bank was kept at -80°C under strict aseptic technique during the study period. The storage period ranged from 4months to 55months. For the osteoblast culture, all eighteen bone flaps had no viable osteoblast growth. For the bacterial culture, five had positive bacteria growth (27.8%). Three were Pasteurella multocida and two were Methicillin-resistant Staphylococcus aureus. The mean duration of storage of the infected bone flap was 32.9months (±15.1months) versus 19.9months (±17.9months) of those bone flaps with no bacterial growth (p=0.1716). The mean size of the infected versus non-infected bone flaps was 117.7cm 2 (±44.96cm 2 ) versus 76.8cm 2 (±50.24cm 2 ) respectively (p=0.1318). Although in this study statistical significance was not reached, it was postulated that infected bone flaps tended to be larger in size and had a longer duration of storage. In conclusion, cryostored skull bone flaps beyond four months showed no viable osteoblasts. Bacterial contamination rate of bone flaps was 27.8% in this study. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Prokaryotic Abundance and Activity in Permafrost of the Northern Victoria Land and Upper Victoria Valley (Antarctica).

PubMed

La Ferla, Rosabruna; Azzaro, Maurizio; Michaud, Luigi; Caruso, Gabriella; Lo Giudice, Angelina; Paranhos, Rodolfo; Cabral, Anderson S; Conte, Antonella; Cosenza, Alessandro; Maimone, Giovanna; Papale, Maria; Rappazzo, Alessandro Ciro; Guglielmin, Mauro

2017-08-01

Victoria Land permafrost harbours a potentially large pool of cold-affected microorganisms whose metabolic potential still remains underestimated. Three cores (BC-1, BC-2 and BC-3) drilled at different depths in Boulder Clay (Northern Victoria Land) and one sample (DY) collected from a core in the Dry Valleys (Upper Victoria Valley) were analysed to assess the prokaryotic abundance, viability, physiological profiles and potential metabolic rates. The cores drilled at Boulder Clay were a template of different ecological conditions (different temperature regime, ice content, exchanges with atmosphere and with liquid water) in the same small basin while the Dry Valleys site was very similar to BC-2 conditions but with a complete different geological history and ground ice type. Image analysis was adopted to determine cell abundance, size and shape as well as to quantify the potential viable and respiring cells by live/dead and 5-cyano-2,3-ditolyl-tetrazolium chloride staining, respectively. Subpopulation recognition by apparent nucleic acid contents was obtained by flow cytometry. Moreover, the physiological profiles at community level by Biolog-Ecoplate™ as well as the ectoenzymatic potential rates on proteinaceous (leucine-aminopeptidase) and glucidic (ß-glucosidase) organic matter and on organic phosphates (alkaline-phosphatase) by fluorogenic substrates were tested. The adopted methodological approach gave useful information regarding viability and metabolic performances of microbial community in permafrost. The occurrence of a multifaceted prokaryotic community in the Victoria Land permafrost and a large number of potentially viable and respiring cells (in the order of 10 4 -10 5 ) were recognised. Subpopulations with a different apparent DNA content within the different samples were observed. The physiological profiles stressed various potential metabolic pathways among the samples and intense utilisation rates of polymeric carbon compounds and carbohydrates, mainly in deep samples. The measured enzymatic activity rates suggested the potential capability of the microbial community to decompose proteins and polysaccharides. The microbial community seems to be appropriate to contribute to biogeochemical cycling in this extreme environment.

Utilization of subsurface microbial electrochemical systems to elucidate the mechanisms of competition between methanogenesis and microbial iron(III)/humic acid reduction in Arctic peat soils

NASA Astrophysics Data System (ADS)

Friedman, E. S.; Miller, K.; Lipson, D.; Angenent, L. T.

2012-12-01

High-latitude peat soils are a major carbon reservoir, and there is growing concern that previously dormant carbon from this reservoir could be released to the atmosphere as a result of continued climate change. Microbial processes, such as methanogenesis and carbon dioxide production via iron(III) or humic acid reduction, are at the heart of the carbon cycle in Arctic peat soils [1]. A deeper understanding of the factors governing microbial dominance in these soils is crucial for predicting the effects of continued climate change. In previous years, we have demonstrated the viability of a potentiostatically-controlled subsurface microbial electrochemical system-based biosensor that measures microbial respiration via exocellular electron transfer [2]. This system utilizes a graphite working electrode poised at 0.1 V NHE to mimic ferric iron and humic acid compounds. Microbes that would normally utilize these compounds as electron acceptors donate electrons to the electrode instead. The resulting current is a measure of microbial respiration with the electrode and is recorded with respect to time. Here, we examine the mechanistic relationship between methanogenesis and iron(III)- or humic acid-reduction by using these same microbial-three electrode systems to provide an inexhaustible source of alternate electron acceptor to microbes in these soils. Chamber-based carbon dioxide and methane fluxes were measured from soil collars with and without microbial three-electrode systems over a period of four weeks. In addition, in some collars we simulated increased fermentation by applying acetate treatments to understand possible effects of continued climate change on microbial processes in these carbon-rich soils. The results from this work aim to increase our fundamental understanding of competition between electron acceptors, and will provide valuable data for climate modeling scenarios. 1. Lipson, D.A., et al., Reduction of iron (III) and humic substances plays a major role in anaerobic respiration in an Arctic peat soil. Journal of Geophysical Research-Biogeosciences, 2010. 115. 2. Friedman, E.S., et al., A cost-effective and field-ready potentiostat that poises subsurface electrodes to monitor bacterial respiration. Biosensors and Bioelectronics, 2012. 32(1): p. 309-313.

Nest sanitation through defecation: antifungal properties of wood cockroach feces.

PubMed

Rosengaus, Rebeca B; Mead, Kerry; Du Comb, William S; Benson, Ryan W; Godoy, Veronica G

2013-11-01

The wood cockroach Cryptocercus punctulatus nests as family units inside decayed wood, a substrate known for its high microbial load. We tested the hypothesis that defecation within their nests, a common occurrence in this species, reduces the probability of fungal development. Conidia of the entomopathogenic fungus, Metarhizium anisopliae, were incubated with crushed feces and subsequently plated on potato dextrose agar. Relative to controls, the viability of fungal conidia was significantly reduced following incubation with feces and was negatively correlated with incubation time. Although the cockroach's hindgut contained abundant β-1,3-glucanase activity, its feces had no detectable enzymatic function. Hence, these enzymes are unlikely the source of the fungistasis. Instead, the antifungal compound(s) of the feces involved heat-sensitive factor(s) of potential microbial origin. When feces were boiled or when they were subjected to ultraviolet radiation and subsequently incubated with conidia, viability was "rescued" and germination rates were similar to those of controls. Filtration experiments indicate that the fungistatic activity of feces results from chemical interference. Because Cryptocercidae cockroaches have been considered appropriate models to make inferences about the factors fostering the evolution of termite sociality, we suggest that nesting in microbe-rich environments likely selected for the coupling of intranest defecation and feces fungistasis in the common ancestor of wood cockroaches and termites. This might in turn have served as a preadaptation that prevented mycosis as these phylogenetically related taxa diverged and evolved respectively into subsocial and eusocial organizations.

Nest sanitation through defecation: antifungal properties of wood cockroach feces

NASA Astrophysics Data System (ADS)

Rosengaus, Rebeca B.; Mead, Kerry; Du Comb, William S.; Benson, Ryan W.; Godoy, Veronica G.

2013-11-01

The wood cockroach Cryptocercus punctulatus nests as family units inside decayed wood, a substrate known for its high microbial load. We tested the hypothesis that defecation within their nests, a common occurrence in this species, reduces the probability of fungal development. Conidia of the entomopathogenic fungus, Metarhizium anisopliae, were incubated with crushed feces and subsequently plated on potato dextrose agar. Relative to controls, the viability of fungal conidia was significantly reduced following incubation with feces and was negatively correlated with incubation time. Although the cockroach's hindgut contained abundant β-1,3-glucanase activity, its feces had no detectable enzymatic function. Hence, these enzymes are unlikely the source of the fungistasis. Instead, the antifungal compound(s) of the feces involved heat-sensitive factor(s) of potential microbial origin. When feces were boiled or when they were subjected to ultraviolet radiation and subsequently incubated with conidia, viability was "rescued" and germination rates were similar to those of controls. Filtration experiments indicate that the fungistatic activity of feces results from chemical interference. Because Cryptocercidae cockroaches have been considered appropriate models to make inferences about the factors fostering the evolution of termite sociality, we suggest that nesting in microbe-rich environments likely selected for the coupling of intranest defecation and feces fungistasis in the common ancestor of wood cockroaches and termites. This might in turn have served as a preadaptation that prevented mycosis as these phylogenetically related taxa diverged and evolved respectively into subsocial and eusocial organizations.

Microbial mito-pathogens: fact or fiction?

PubMed

Bongaerts, Ger P A; van den Heuvel, Lambert P

2008-01-01

Mitochondria are bacteria-like semi-autonomous intracellular organelles that function as the powerhouses of eukaryotic cells. Inactivation or destruction of these organelles may have far-reaching consequences regarding the viability of the cells and thus of tissues, organs and finally even the body. Since mitochondria resemble (degenerated) bacteria, we have extrapolated from both cytological and microbiological facts the existence of various (kinds of) mitochondrion-specific microbial pathogens, i.e., pathogenic micro-organisms that may damage or destroy the mitochondria from within. These mito-pathogens may include mitoviruses, mitoviroids and mitobacteria. Although these mito-pathogens have not yet been demonstrated in humans, their theoretical degenerative effect regarding energy production from energy-rich substrates, such as carbohydrates and fats, might explain diseases that have not yet been understood, such as prion diseases and post-traumatic muscle dystrophy. Therefore, these kinds of micro-organisms should be kept in mind.

Control of adaptive immunity by the innate immune system.

PubMed

Iwasaki, Akiko; Medzhitov, Ruslan

2015-04-01

Microbial infections are recognized by the innate immune system both to elicit immediate defense and to generate long-lasting adaptive immunity. To detect and respond to vastly different groups of pathogens, the innate immune system uses several recognition systems that rely on sensing common structural and functional features associated with different classes of microorganisms. These recognition systems determine microbial location, viability, replication and pathogenicity. Detection of these features by recognition pathways of the innate immune system is translated into different classes of effector responses though specialized populations of dendritic cells. Multiple mechanisms for the induction of immune responses are variations on a common design principle wherein the cells that sense infections produce one set of cytokines to induce lymphocytes to produce another set of cytokines, which in turn activate effector responses. Here we discuss these emerging principles of innate control of adaptive immunity.

Enhanced microbial coalbed methane generation: A review of research, commercial activity, and remaining challenges

USGS Publications Warehouse

Ritter, Daniel J.; Vinson, David S.; Barnhart, Elliott P.; Akob, Denise M.; Fields, Matthew W.; Cunningham, Al B.; Orem, William H.; McIntosh, Jennifer C.

2015-01-01

Coalbed methane (CBM) makes up a significant portion of the world’s natural gas resources. The discovery that approximately 20% of natural gas is microbial in origin has led to interest in microbially enhanced CBM (MECoM), which involves stimulating microorganisms to produce additional CBM from existing production wells. This paper reviews current laboratory and field research on understanding processes and reservoir conditions which are essential for microbial CBM generation, the progress of efforts to stimulate microbial methane generation in coal beds, and key remaining knowledge gaps. Research has been primarily focused on identifying microbial communities present in areas of CBM generation and attempting to determine their function, in-situ reservoir conditions that are most favorable for microbial CBM generation, and geochemical indicators of metabolic pathways of methanogenesis (i.e., acetoclastic or hydrogenotrophic methanogenesis). Meanwhile, researchers at universities, government agencies, and companies have focused on four primary MECoM strategies: 1) microbial stimulation (i.e., addition of nutrients to stimulate native microbes); 2) microbial augmentation (i.e., addition of microbes not native to or abundant in the reservoir of interest); 3) physically increasing microbial access to coal and distribution of amendments; and 4) chemically increasing the bioavailability of coal organics. Most companies interested in MECoM have pursued microbial stimulation: Luca Technologies, Inc., successfully completed a pilot scale field test of their stimulation strategy, while two others, Ciris Energy and Next Fuel, Inc., have undertaken smaller scale field tests. Several key knowledge gaps remain that need to be addressed before MECoM strategies can be implemented commercially. Little is known about the bacterial community responsible for coal biodegradation and how these microorganisms may be stimulated to enhance microbial methanogenesis. In addition, research is needed to understand what fraction of coal is available for biodegradation, and methods need to be developed to determine the extent of in-situ coal biodegradation by MECoM processes for monitoring changes to coal quality. Questions also remain about how well field-scale pilot tests will scale to commercial production, how often amendments will need to be added to maintain new methane generation, and how well MECoM strategies transfer between coal basins with different formation water geochemistries and coal ranks. Addressing these knowledge gaps will be key in determining the feasibility and commercial viability of MECoM technology.

Microbial astronauts: assembling microbial communities for advanced life support systems.

PubMed

Roberts, M S; Garland, J L; Mills, A L

2004-02-01

Extension of human habitation into space requires that humans carry with them many of the microorganisms with which they coexist on Earth. The ubiquity of microorganisms in close association with all living things and biogeochemical processes on Earth predicates that they must also play a critical role in maintaining the viability of human life in space. Even though bacterial populations exist as locally adapted ecotypes, the abundance of individuals in microbial species is so large that dispersal is unlikely to be limited by geographical barriers on Earth (i.e., for most environments "everything is everywhere" given enough time). This will not be true for microbial communities in space where local species richness will be relatively low because of sterilization protocols prior to launch and physical barriers between Earth and spacecraft after launch. Although community diversity will be sufficient to sustain ecosystem function at the onset, richness and evenness may decline over time such that biological systems either lose functional potential (e.g., bioreactors may fail to reduce BOD or nitrogen load) or become susceptible to invasion by human-associated microorganisms (pathogens) over time. Research at the John F. Kennedy Space Center has evaluated fundamental properties of microbial diversity and community assembly in prototype bioregenerative systems for NASA Advanced Life Support. Successional trends related to increased niche specialization, including an apparent increase in the proportion of nonculturable types of organisms, have been consistently observed. In addition, the stability of the microbial communities, as defined by their resistance to invasion by human-associated microorganisms, has been correlated to their diversity. Overall, these results reflect the significant challenges ahead for the assembly of stable, functional communities using gnotobiotic approaches, and the need to better define the basic biological principles that define ecosystem processes in the space environment. Copyright 2004 Springer-Verlag

Microbial astronauts: assembling microbial communities for advanced life support systems

NASA Technical Reports Server (NTRS)

Roberts, M. S.; Garland, J. L.; Mills, A. L.

2004-01-01

Extension of human habitation into space requires that humans carry with them many of the microorganisms with which they coexist on Earth. The ubiquity of microorganisms in close association with all living things and biogeochemical processes on Earth predicates that they must also play a critical role in maintaining the viability of human life in space. Even though bacterial populations exist as locally adapted ecotypes, the abundance of individuals in microbial species is so large that dispersal is unlikely to be limited by geographical barriers on Earth (i.e., for most environments "everything is everywhere" given enough time). This will not be true for microbial communities in space where local species richness will be relatively low because of sterilization protocols prior to launch and physical barriers between Earth and spacecraft after launch. Although community diversity will be sufficient to sustain ecosystem function at the onset, richness and evenness may decline over time such that biological systems either lose functional potential (e.g., bioreactors may fail to reduce BOD or nitrogen load) or become susceptible to invasion by human-associated microorganisms (pathogens) over time. Research at the John F. Kennedy Space Center has evaluated fundamental properties of microbial diversity and community assembly in prototype bioregenerative systems for NASA Advanced Life Support. Successional trends related to increased niche specialization, including an apparent increase in the proportion of nonculturable types of organisms, have been consistently observed. In addition, the stability of the microbial communities, as defined by their resistance to invasion by human-associated microorganisms, has been correlated to their diversity. Overall, these results reflect the significant challenges ahead for the assembly of stable, functional communities using gnotobiotic approaches, and the need to better define the basic biological principles that define ecosystem processes in the space environment. Copyright 2004 Springer-Verlag.

Current density reversibly alters metabolic spatial structure of exoelectrogenic anode biofilms

NASA Astrophysics Data System (ADS)

Sun, Dan; Cheng, Shaoan; Zhang, Fang; Logan, Bruce E.

2017-07-01

Understanding how current densities affect electrogenic biofilm activity is important for wastewater treatment as current densities can substantially decrease at COD concentrations greater than those suitable for discharge to the environment. We examined the biofilm's response, in terms of viability and enzymatic activity, to different current densities using microbial electrolysis cells with a lower (0.7 V) or higher (0.9 V) added voltage to alter current production. Viability was assessed using florescent dyes, with dead cells identified on the basis of dye penetration due to a compromised cell outer-membrane (red), and live cells (intact membrane) fluorescing green. Biofilms operated with 0.7 V produced 2.4 ± 0.2 A m-2, and had an inactive layer near the electrode and a viable layer at the biofilm-solution interface. The lack of cell activity near the electrode surface was confirmed by using an additional dye that fluoresces only with enzymatic activity. Adding 0.9 V increased the current by 61%, and resulted in a single, more homogeneous and active biofilm layer. Switching biofilms between these two voltages produced outcomes associated with the new current rather than the previous biofilm conditions. These findings suggest that maintaining higher current densities will be needed to ensure long-term viability electrogenic biofilms.

Evaluation of the suitability and performance of cassava waste (peel) extracts in a microbial fuel cell for supplementary and sustainable energy production.

PubMed

Adekunle, Ademola; Raghavan, Vijaya

2017-01-01

In a number of energy-poor nations, peel from cassava processing represents one of the most abundant sources of lignocellulosic biomass. This peel is mostly discarded indiscriminately and eventually constitutes a problem to the environment. However, energy can be extracted from this peel in a microbial fuel cell. In this study, the viability of cassava peel extract as a substrate in a single-chamber air cathode microbial fuel cell is demonstrated, and optimum performance conditions are explored. The effects of different pretreatments on the extract are also discussed in the context of observed changes in the internal resistances, conductivity and Coulombic efficiencies. At the best conditions examined, the extract from cassava peel fermented for 168 h and adjusted to a pH of 7.63 attained a peak voltage of 687 mV ± 21 mV, a power density of 155 mW m -3 of reactor volume and a Coulombic efficiency of 11 %. Although this energy is limited to direct use, systems exist that can effectively harvest and boost the energy to levels sufficient for supplementary energy usage in cassava producing regions.

Therapeutic Application of Synbiotics, a Fusion of Probiotics and Prebiotics, and Biogenics as a New Concept for Oral Candida Infections: A Mini Review

PubMed Central

Ohshima, Tomoko; Kojima, Yukako; Seneviratne, Chaminda J.; Maeda, Nobuko

2016-01-01

Candida is a major human fungal pathogen causing infectious conditions predominantly in the elderly and immunocompromised hosts. Although Candida resides as a member of the oral indigenous microbiota in symbiosis, some circumstances may cause microbial imbalance leading to dysbiosis and resultant oral candidiasis. Therefore, oral microbial symbiosis that suppresses the overgrowth of Candida is important for a healthy oral ecosystem. In this regard, probiotics, prebiotics, and synbiotics can be considered a potential therapeutic and preventive strategy against oral candidiasis. Prebiotics have a direct effect on microbial growth as they stimulate the growth of beneficial bacteria and suppress the growth of pathogens. Probiotics render a local protective effect against pathogens and a systemic indirect effect on immunological amelioration. Synbiotics are fusion products of prebiotics and probiotics. This mini review discusses the potential use and associated limitations of probiotics, prebiotics, and synbiotics for the prevention and treatment of oral candidiasis. We will also introduce biogenics, a recent concept derived from the work on probiotics. Biogenics advocates the use of beneficial bioactive substances produced by probiotic bacteria, whose activities are independent from the viability of probiotic bacteria in human bodies. PMID:26834728

Microorganisms under high pressure--adaptation, growth and biotechnological potential.

PubMed

Mota, Maria J; Lopes, Rita P; Delgadillo, Ivonne; Saraiva, Jorge A

2013-12-01

Hydrostatic pressure is a well-known physical parameter which is now considered an important variable of life, since organisms have the ability to adapt to pressure changes, by the development of resistance against this variable. In the past decades a huge interest in high hydrostatic pressure (HHP) technology is increasingly emerging among food and biosciences researchers. Microbial specific stress responses to HHP are currently being investigated, through the evaluation of pressure effects on biomolecules, cell structure, metabolic behavior, growth and viability. The knowledge development in this field allows a better comprehension of pressure resistance mechanisms acquired at sub-lethal pressures. In addition, new applications of HHP could arise from these studies, particularly in what concerns to biotechnology. For instance, the modulation of microbial metabolic pathways, as a response to different pressure conditions, may lead to the production of novel compounds with potential biotechnological and industrial applications. Considering pressure as an extreme life condition, this review intends to present the main findings so far reported in the scientific literature, focusing on microorganisms with the ability to withstand and to grow in high pressure conditions, whether they have innated or acquired resistance, and show the potential of the application of HHP technology for microbial biotechnology. © 2013.

Sol-gel immobilization as a suitable technique for enhancement of α-amylase activity of Aspergillus oryzae PP.

PubMed

Evstatieva, Yana; Yordanova, Mariya; Chernev, Georgi; Ruseva, Yanislava; Nikolova, Dilyana

2014-07-04

Bioencapsulation of microbial cells in silica-based matrices has proved to be a good strategy to enhance the biosynthetic capabilities and viability of bioproducers. In the present study, mycelium and pellet cultures of strain Aspergillus oryzae PP were successfully immobilized in sol-gel hybrid matrices composed of tetraethylorthosilicate as an inorganic precursor, 5% (w/v) starch and 10 or 15% (w/v) polyethylene oxide, or 10% (w/v) calcium alginate as organic compounds. Biosynthetic activity of immobilized cultures was investigated by batch and fed-batch cultivation and the obtained results of 3042.04 IU cm -3 were comparable with the enzyme activity of the free cell culture. Immobilized cultures retained their viability and biosynthetic capabilities up to the 744th h during fed-batch fermentation processes. Consequently, sol-gel encapsulation in hybrid matrices could be considered as a promising technique for immobilization of Aspergillus oryzae PP in order to increase the α-amylase production.

Effect of gamma irradiation on hyperthermal composting microorganisms for feasible application in space

NASA Astrophysics Data System (ADS)

Yoon, Minchul; Choi, Jong-il; Yamashita, Masamichi

2013-05-01

The composting system is the most efficient method for processing organic waste in space; however, the composting activity of microorganisms can be altered by cosmic rays. In this study, the effect of ionizing irradiation on composting bacteria was investigated. Sequence analyses of amplified 16S rRNA, 18S rRNA, and amoA genes were used to identify hyperthermal composting microorganisms. The viability of microorganisms in compost soil after gamma irradiation was directly determined using LIVE/DEAD BacLight viability kit. The dominant bacterial genera were Weissella cibaria and Leuconostoc sp., and the fungal genera were Metschnikowia bicuspidata and Pichia guilliermondii. Gamma irradiation up to a dose of 10 kGy did not significantly alter the microbial population. Furthermore, amylase and cellulase activities were maintained after high-dose gamma irradiation. Our results show that hyperthermal microorganisms can be used to recycle agricultural and fermented material in space stations and other human-inhabiting facilities on the Moon, Mars, and other planets.

Feasibility study of the sterilization of pigskin used as wound dressings by neutral electrolyzed water.

PubMed

Ge, Liangpeng; Zhang, Xiaochun; Cao, Chuan; Gu, Zhaobin; Liu, Zuohua; Liu, Lubin; Lin, Baozhong

2012-06-01

Neutral electrolyzed water (NEW) is considered to be a high-level biodegradable disinfectant with sporicidal, bactericidal, and virucidal activity. It has also been reported to accelerate wound healing; thus, it is particularly attractive for the elimination or minimization of the microbial population of skin grafts to be used as wound dressings. Pigskins were sterilized with different concentrations of NEW and with different methods. The feasibility of pigskin sterilization by NEW was evaluated through microbiological analyses, viability assays, histologic assessments, contact cytotoxicity assays, and extract cytotoxicity assays. NEW has strong bactericidal effects on pigskin microorganisms, does not change skin graft histologic properties, and has no cytotoxicity; however, skin viability was significantly reduced after NEW treatment. Although NEW treatment is a very safe and effective method for nonviable pigskin dressing sterilization, to obtain a complete sterilization of pigskin grafts, available chlorine concentration of NEW as well as sterilization time and methods should be optimized. Copyright © 2012 by Lippincott Williams & Wilkins.

Microbial Electrochemistry and its Application to Energy and Environmental Issues

NASA Astrophysics Data System (ADS)

Hastings, Jason Thomas

Microbial electrochemistry forms the basis of a wide range of topics from microbial fuel cells to fermentation of carbon food sources. The ability to harness microbial electron transfer processes can lead to a greener and cleaner future. This study focuses on microbial electron transfer for liquid fuel production, novel electrode materials, subsurface environments and removal of unwanted byproducts. In the first chapter, exocellular electron transfer through direct contact utilizing passive electrodes for the enhancement of bio-fuel production was tested. Through the application of microbial growth in a 2-cell apparatus on an electrode surface ethanol production was enhanced by 22.7% over traditional fermentation. Ethanol production efficiencies of close to 95% were achieved in a fraction of the time required by traditional fermentation. Also, in this chapter, the effect of exogenous electron shuttles, electrode material selection and resistance was investigated. Power generation was observed using the 2-cell passive electrode system. An encapsulation method, which would also utilize exocellular transfer of electrons through direct contact, was hypothesized for the suspension of viable cells in a conductive polymer substrate. This conductive polymer substrate could have applications in bio-fuel production. Carbon black was added to a polymer solution to test electrospun polymer conductivity and cell viability. Polymer morphology and cell viability were imaged using electron and optical microscopy. Through proper encapsulation, higher fuel production efficiencies would be achievable. Electron transfer through endogenous exocellular protein shuttles was observed in this study. Secretion of a soluble redox active exocellular protein by Clostridium sp. have been shown utilizing a 2-cell apparatus. Cyclic voltammetry and gel electrophoresis were used to show the presence of the protein. The exocellular protein is capable of reducing ferrous iron in a membrane separated chamber. In experiments where the redox active protein was allowed to pass through the permeable membrane, iron dissolution was 14-fold greater than experiments where the protein was held to one chamber by a non-permeable membrane. Confirmation of a redox active protein could reshape or understanding of subsurface redox processes. The final topic in this study discusses electron transfer within the cell for production of fermentation products. Glycerol, which is an unwanted side-product of biodiesel transesterfication, is utilized as a carbon source for fermentation. Bacterial samples harvested from Galena Creek soil (NGC) are shown in this study to be efficient consumers of glycerol. NGC microbe was characterized through 16s rDNA genetic sequencing and determined to belong to genus Clostridium. Clostridium NGC was able to consume glycerol at 29.7gpl within 72hrs grown in a media containing 50gpl glycerol. All observed fermentation metabolites were characterized and quantified through an HPLC. Glycerol consumption rates and metabolite production rates were observed using varying media recipes. This study has found that NGC has higher selectivity for low weight acids at lower yeast extract concentration and higher selectivity for larger acids and alcohols at higher yeast extract concentrations.

Cleanroom Maintenance Significantly Reduces Abundance but Not Diversity of Indoor Microbiomes.

PubMed

Mahnert, Alexander; Vaishampayan, Parag; Probst, Alexander J; Auerbach, Anna; Moissl-Eichinger, Christine; Venkateswaran, Kasthuri; Berg, Gabriele

2015-01-01

Cleanrooms have been considered microbially-reduced environments and are used to protect human health and industrial product assembly. However, recent analyses have deciphered a rather broad diversity of microbes in cleanrooms, whose origin as well as physiological status has not been fully understood. Here, we examined the input of intact microbial cells from a surrounding built environment into a spacecraft assembly cleanroom by applying a molecular viability assay based on propidium monoazide (PMA). The controlled cleanroom (CCR) was characterized by ~6.2*103 16S rRNA gene copies of intact bacterial cells per m2 floor surface, which only represented 1% of the total community that could be captured via molecular assays without viability marker. This was in contrast to the uncontrolled adjoining facility (UAF) that had 12 times more living bacteria. Regarding diversity measures retrieved from 16S rRNA Illumina-tag analyzes, we observed, however, only a minor drop in the cleanroom facility allowing the conclusion that the number but not the diversity of microbes is strongly affected by cleaning procedures. Network analyses allowed tracking a substantial input of living microbes to the cleanroom and a potential enrichment of survival specialists like bacterial spore formers and archaeal halophiles and mesophiles. Moreover, the cleanroom harbored a unique community including 11 exclusive genera, e.g., Haloferax and Sporosarcina, which are herein suggested as indicators of cleanroom environments. In sum, our findings provide evidence that archaea are alive in cleanrooms and that cleaning efforts and cleanroom maintenance substantially decrease the number but not the diversity of indoor microbiomes.

Cleanroom Maintenance Significantly Reduces Abundance but Not Diversity of Indoor Microbiomes

PubMed Central

Mahnert, Alexander; Vaishampayan, Parag; Probst, Alexander J.; Auerbach, Anna; Moissl-Eichinger, Christine; Venkateswaran, Kasthuri; Berg, Gabriele

2015-01-01

Cleanrooms have been considered microbially-reduced environments and are used to protect human health and industrial product assembly. However, recent analyses have deciphered a rather broad diversity of microbes in cleanrooms, whose origin as well as physiological status has not been fully understood. Here, we examined the input of intact microbial cells from a surrounding built environment into a spacecraft assembly cleanroom by applying a molecular viability assay based on propidium monoazide (PMA). The controlled cleanroom (CCR) was characterized by ~6.2*103 16S rRNA gene copies of intact bacterial cells per m2 floor surface, which only represented 1% of the total community that could be captured via molecular assays without viability marker. This was in contrast to the uncontrolled adjoining facility (UAF) that had 12 times more living bacteria. Regarding diversity measures retrieved from 16S rRNA Illumina-tag analyzes, we observed, however, only a minor drop in the cleanroom facility allowing the conclusion that the number but not the diversity of microbes is strongly affected by cleaning procedures. Network analyses allowed tracking a substantial input of living microbes to the cleanroom and a potential enrichment of survival specialists like bacterial spore formers and archaeal halophiles and mesophiles. Moreover, the cleanroom harbored a unique community including 11 exclusive genera, e.g., Haloferax and Sporosarcina, which are herein suggested as indicators of cleanroom environments. In sum, our findings provide evidence that archaea are alive in cleanrooms and that cleaning efforts and cleanroom maintenance substantially decrease the number but not the diversity of indoor microbiomes. PMID:26273838

Design and evaluation of a novel subatmospheric pressure bioreactor for the preconditioning of tissue-engineered vascular constructs.

PubMed

Coakley, Daniel N; Shaikh, Faisal M; O'Sullivan, Kathleen; Kavanagh, Eamon G; Grace, Pierce A; McGloughlin, Tim M

2016-02-01

The pre-conditioning of tissue-engineered vascular scaffolds with mechanical stimuli is being recognised as an essential step in producing a functional vascular construct. In this study we design and evaluate a novel bioreactor, which exerts a mechanical strain on developing vascular scaffolds via subatmospheric pressure. We design and construct a bioreactor, which exerts subatmospheric pressure via a vacuum assisted closure unit. Vascular scaffolds seeded with human umbilical endothelial cells were evaluated for structural integrity, microbial contamination, cellular viability, von Willebrand factor (VWF) production, cell proliferation and morphology under a range of subatmospheric pressures (75-200mmHg). The bioreactor produced sustained subatmospheric pressures, which exerted a mechanical strain on the vascular scaffold. No microbial contamination was found during the study. The structural integrity of the vascular construct was maintained. There was no difference in cellular viability between control or subatmospheric pressure groups (p = 0.817). Cells continued to produce VWF under a range of subatmospheric pressures. Cells subjected to subatmospheric pressures of 125mmHg and 200mmHg exhibited higher levels of growth than cells in atmospheric pressure at 24 (p≤0.016) and 48 hour (p≤0.001). Negative pressure affected cellular morphology, which were more organised, elongated and expanded when exposed to subatmospheric pressure. We have constructed and validated a novel subatmospheric bioreactor. The bioreactor maintained a continuous subatmospheric pressure to the vascular scaffolds in a stable, sterile and constant environment. The bioreactor exerted a strain on the vascular sheets, which was shown to alter cellular morphology and enhance cellular proliferation.

Microbial Interactions within a Cheese Microbial Community▿ †

PubMed Central

Mounier, Jérôme; Monnet, Christophe; Vallaeys, Tatiana; Arditi, Roger; Sarthou, Anne-Sophie; Hélias, Arnaud; Irlinger, Françoise

2008-01-01

The interactions that occur during the ripening of smear cheeses are not well understood. Yeast-yeast interactions and yeast-bacterium interactions were investigated within a microbial community composed of three yeasts and six bacteria found in cheese. The growth dynamics of this community was precisely described during the ripening of a model cheese, and the Lotka-Volterra model was used to evaluate species interactions. Subsequently, the effects on ecosystem functioning of yeast omissions in the microbial community were evaluated. It was found both in the Lotka-Volterra model and in the omission study that negative interactions occurred between yeasts. Yarrowia lipolytica inhibited mycelial expansion of Geotrichum candidum, whereas Y. lipolytica and G. candidum inhibited Debaryomyces hansenii cell viability during the stationary phase. However, the mechanisms involved in these interactions remain unclear. It was also shown that yeast-bacterium interactions played a significant role in the establishment of this multispecies ecosystem on the cheese surface. Yeasts were key species in bacterial development, but their influences on the bacteria differed. It appeared that the growth of Arthrobacter arilaitensis or Hafnia alvei relied less on a specific yeast function because these species dominated the bacterial flora, regardless of which yeasts were present in the ecosystem. For other bacteria, such as Leucobacter sp. or Brevibacterium aurantiacum, growth relied on a specific yeast, i.e., G. candidum. Furthermore, B. aurantiacum, Corynebacterium casei, and Staphylococcus xylosus showed reduced colonization capacities in comparison with the other bacteria in this model cheese. Bacterium-bacterium interactions could not be clearly identified. PMID:17981942

Antimicrobial drugs encapsulated in fibrin nanoparticles for treating microbial infested wounds.

PubMed

Alphonsa, B Maria; Sudheesh Kumar, P T; Praveen, G; Biswas, Raja; Chennazhi, K P; Jayakumar, R

2014-05-01

In vitro evaluation of antibacterial and antifungal drugs encapsulated fibrin nanoparticles to prove their potential prospect of using these nanocomponent for effective treatment of microbial infested wounds. Surfactant-free oil-in-water emulsification-diffusion method was adopted to encapsulate 1 mg/ml each of antimicrobial drugs (Ciprofloxacin and Fluconazole) in 4 ml of aqueous fibrinogen suspension and subsequent thrombin mediated cross linking to synthesize drug loaded fibrin nanoparticles. Ciprofloxacin loaded fibrin nanoparticles (CFNPs) showed size range of 253 ± 6 nm whereas that of Fluconazole loaded fibrin nanoparticles (FFNPs) was 260 ± 10 nm. Physico chemical characterizations revealed the firm integration of antimicrobial drugs within fibrin nanoparticles. Drug release studies performed at physiological pH 7.4 showed a release of 16% ciprofloxacin and 8% of fluconazole while as the release of ciprofloxacin at alkaline pH 8.5, was 48% and that of fluconazole was 37%. The antimicrobial activity evaluations of both drug loaded systems independently showed good antibacterial activity against Escherichia coli (E.coli), Staphylococcus aureus (S. aureus) and antifungal activity against Candida albicans (C. albicans). The in vitro toxicity of the prepared drug loaded nanoparticles were further analyzed using Human dermal fibroblast cells (HDF) and showed adequate cell viability. The efficacies of both CFNPs and FFNPs for sustained delivery of encapsulated anti microbial drugs were evaluated in vitro suggesting its potential use for treating microbial infested wounds (diabetic foot ulcer).

A protocol for assessing the biotreatability of hydrocarbon contaminated exploration and production site soils

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

Tezak, J.; Miller, J.A.; Lawrence, A.W.

1995-12-01

It is estimated that there are over 260,000 natural gas production wells in the continental United States. Production or reserve pits exist which ma require remediation depending on several conditions such as: the manner in which they were initially closed; whether or not they were lined; and the local climate, soil type, and depth to groundwater. As part of the Gas Research Institute (GRI) research program on exploration and production (E&P) site remediation, a treatability Protocol is being developed to facilitate the rapid assessment of the amenability of the contaminated soils to remediation by biological processes. This paper describes themore » treatability protocol and the results of a series of treatability tests on a spectrum of hydrocarbon contaminated E&P soils collected from various operating locations throughout the United States. The soils are subjected to physical and chemical characterization prior to treatability testing. Potential biotoxic characteristics of the soils are determined by a respirometry screening technique. Presuming that the soils are not toxic to aerobic soil microorganisms, 20 percent by weight aqueous slurries of the soils are prepared and subjected to continuous batch aeration for a six week period. Conditions favorable to microbial growth are maintained in the reactors by monitoring and augmentation is needed of pH, microbial nutrients and oxygen for microbial respiration. The extent of microbial degradation of the contaminant hydrocarbons is monitored by periodic measurement of total petroleum hydrocarbons (TPH), oil and grease, and individual hydrocarbon compounds as determined by gas chromatography. Microbial plate counts are prepared to document the biological viability of the treatment process. The factors influencing the amenability of these soils to bioremediation as determined from the test results are discussed.« less

Deep-Sea Trench Microbiology Down to 10.9 Kilometers Below the Surface

NASA Astrophysics Data System (ADS)

Bartlett, D. H.

2012-12-01

Deep-sea trenches, extending to more than 10.9 km below the sea surface, are among the most remote and infrequently sampled habitats. As a result a global perspective of microbial diversity and adaptation is lacking in these extreme settings. I will present the results of studies of deep-sea trench microbes collected in the Puerto Rico Trench (PRT), Tonga Trench, New Britain Trench and Mariana Trench. The samples collected include sediment, seawater and animals in baited traps. The analyses to be described include microbial community activity and viability measurements as a function of hydrostatic pressure, microbial culturing at high pressure under various physiological conditions, phylogenetics and metagenome and single-cell genome characterizations. Most of the results to date stem from samples recovered from the PRT. The deep-sea PRT Trench microbes have more in common at the species level with other deep-sea microbial communities previously characterized in the Pacific Ocean and the Mediterranean Sea than with the microbial populations above them in shallow waters. They also harbor larger genomes with more genes assigned to signal transduction, transcription, replication, recombination and repair and inorganic ion transport. The overrepresented transporters in the PRT metagenome include di- and tri-carboxylate transporters that correspond to the prevailing catabolic processes such as butanoate, glyoxylate and dicarboxylate metabolism. A surprisingly high abundance of sulfatases for the degradation of sulfated polysaccharides were also present in the PRT. But, perhaps the most dramatic adaptational feature of the PRT microbes is heavy metal resistance, as reflected in the high numbers of metal efflux systems present. Single-cell genomics approaches have proven particularly useful for placing PRT metagenomic data into context.

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.

Water Recovery System Design to Accommodate Dormant Periods for Manned Missions

NASA Technical Reports Server (NTRS)

Tabb, David; Carter, Layne

2015-01-01

Future manned missions beyond lower Earth orbit may include intermittent periods of extended dormancy. Under the NASA Advanced Exploration System (AES) project, NASA personnel evaluated the viability of the ISS Water Recovery System (WRS) to support such a mission. The mission requirement includes the capability for life support systems to support crew activity, followed by a dormant period of up to one year, and subsequently for the life support systems to come back online for additional crewed missions. Dormancy could be a critical issue due to concerns with microbial growth or chemical degradation that might prevent water systems from operating properly when the crewed mission began. As such, it is critical that the water systems be designed to accommodate this dormant period. This paper details the results of this evaluation, which include identification of dormancy issues, results of testing performed to assess microbial stability of pretreated urine during dormancy periods, and concepts for updating to the WRS architecture and operational concepts that will enable the ISS WRS to support the dormancy requirement.

Apple replant disease: role of microbial ecology in cause and control.

PubMed

Mazzola, Mark; Manici, Luisa M

2012-01-01

Replant disease of apple is common to all major apple growing regions of the world. Difficulties in defining disease etiology, which can be exacerbated by abiotic factors, have limited progress toward developing alternatives to soil fumigation for disease control. However, the preponderance of data derived from studies of orchard soil biology employing multidisciplinary approaches has defined a complex of pathogens/parasites as causal agents of the disease. Approaches to manipulate microbial resources endemic to the orchard soil system have been proposed to induce a state of general soil suppressiveness to replant disease. Such a long-term strategy may benefit the existing orchard through extending the period of economic viability and reduce overall disease pressure to which young trees are exposed during establishment of successive plantings on the site. Alternatively, more near-term methods have been devised to achieve specific quantitative and qualitative changes in soil biology during the period of orchard renovation that may lead to effective disease suppression.

Microbial Removals by a Novel Biofilter Water Treatment System

PubMed Central

Wendt, Christopher; Ives, Rebecca; Hoyt, Anne L.; Conrad, Ken E.; Longstaff, Stephanie; Kuennen, Roy W.; Rose, Joan B.

2015-01-01

Two point-of-use drinking water treatment systems designed using a carbon filter and foam material as a possible alternative to traditional biosand systems were evaluated for removal of bacteria, protozoa, and viruses. Two configurations were tested: the foam material was positioned vertically around the carbon filter in the sleeve unit or horizontally in the disk unit. The filtration systems were challenged with Cryptosporidium parvum, Raoultella terrigena, and bacteriophages P22 and MS2 before and after biofilm development to determine average log reduction (ALR) for each organism and the role of the biofilm. There was no significant difference in performance between the two designs, and both designs showed significant levels of removal (at least 4 log10 reduction in viruses, 6 log10 for protozoa, and 8 log10 for bacteria). Removal levels meet or exceeded Environmental Protection Agency (EPA) standards for microbial purifiers. Exploratory test results suggested that mature biofilm formation contributed 1–2 log10 reductions. Future work is recommended to determine field viability. PMID:25758649

Storage and qualification of viable intact human amniotic graft and technology transfer to a tissue bank.

PubMed

Laurent, Romain; Nallet, Aurélie; Obert, Laurent; Nicod, Laurence; Gindraux, Florelle

2014-06-01

Human amniotic membrane (hAM) is known to have good potential to help the regeneration of tissue. It has been used for over 100 years in many medical disciplines because of its properties, namely a scaffold containing stem cells and growth factors, with low immunogenicity and anti-microbial, anti-inflammatory, anti-fibrotic and analgesic properties. In order to use this "boosted membrane" as an advanced therapeutic medicinal product for bone repair, we aimed to observe the influence of tissue culture and/or cryopreservation on cell viability and tissue structure, and secondly, to adapt to a tissue bank, identify easy processes to store hAM containing viable cells and to verify the quality of the graft before its release for use. To this end, we tested different published culture or cryopreservation storage conditions and cell viability assays. Tissue structure was evaluated by Giemsa staining and was compared to histological analysis. Preliminary results show no dramatic decrease in cell viability in cultured hAM as compared to cryopreserved hAM, but tissue structure alterations were observed with both storage conditions. Histological and immunohistochemical data highlight that tissue damage was associated with significantly modified protein expression, which could lead to a possible loss of differentiation potential. Finally, we report that trypan blue and Giemsa staining could constitute controls that are "materially and easily transferable" to a tissue bank.

A Novel Double Subculture Method and Its Theory for the Enumeration of Injured Cells in Stressed Microbial Population.

PubMed

Tsuchido, Tetsuaki

2017-01-01

　A novel double subculture method, termed DiVSaL (Differential Viabilities between Solid and Liquid media) method, for the enumeration of injured cell population of a microorganism, which occurs after some sublethal to lethal treatment, was proposed. In this method injured cells were enumerated as the differential value between viabilities determined with two different techniques, the conventional plate counting using a solid agar medium and the growth delay analysis using a liquid medium. In the former technique, the viable cell number is obtained as colony forming unit (CFU) formed on an agar medium where sublethally injured cells are as much rescued as possible. In the latter technique, on the other hand," the integrated viability" defined by Takano and Tsuchido (1982) is introduced and is calculated from the growth delay of a stressed population, referred to unstressed one. For the growth delay analysis, in this paper, not only the original theoretical model, where the specific growth rate (and therefore the defined G 10 value) does not change after the exposure to a stress treatment, but also a novel modified theory, where the parameter changes, is proposed. On the theoretical background, this DiVSaL method as a double subculture method can be used to enumerate the injured cells without selection by addition of some inhibitor or by nutritional shortage.

Short-term differential adaptation to anaerobic stress via genomic mutations by Escherichia coli strains K-12 and B lacking alcohol dehydrogenase.

PubMed

Kim, Hyun Ju; Jeong, Haeyoung; Hwang, Seungwoo; Lee, Moo-Seung; Lee, Yong-Jik; Lee, Dong-Woo; Lee, Sang Jun

2014-01-01

Microbial adaptations often occur via genomic mutations under adverse environmental conditions. This study used Escherichia coli ΔadhE cells as a model system to investigate adaptation to anaerobic conditions, which we then compared with the adaptive mechanisms of two closely related E. coli strains, K-12 and B. In contrast to K-12 ΔadhE cells, the E. coli B ΔadhE cells exhibited significantly delayed adaptive growth under anaerobic conditions. Adaptation by the K-12 and B strains mainly employed anaerobic lactate fermentation to restore cellular growth. Several mutations were identified in the pta or pflB genes of adapted K-12 cells, but mostly in the pta gene of the B strains. However, the types of mutation in the adapted K-12 and B strains were similar. Cellular viability was affected directly by severe redox imbalance in B ΔadhE cells, which also impaired their ability to adapt to anaerobic conditions. This study demonstrates that closely related microorganisms may undergo different adaptations under the same set of adverse conditions, which might be associated with the specific metabolic characteristics of each strain. This study provides new insights into short-term microbial adaptation to stressful conditions, which may reflect dynamic microbial population changes in nature.

TOF-SIMS imaging of chlorhexidine-digluconate transport in frozen hydrated biofilms of the fungus Candida albicans

NASA Astrophysics Data System (ADS)

Tyler, Bonnie J.; Rangaranjan, Srinath; Möller, Jörg; Beumer, Andre'; Arlinghaus, Heinrich F.

2006-07-01

The diffusion of the anti-microbial chlorhexidine digluconate (CHG) has been studied in C. albicans biofilms by time-of-flight secondary-ion mass spectrometry (TOF-SIMS). C. albicans has been shown to become resistant to common anti-microbial agents, including CHG, when growing as a biofilm. Mass transport resistance within biofilms has commonly been suggested as a resistance mechanism, but measurement of transport for most anti-microbial agents in biofilms has proven extremely difficult because of the heterogeneity of the biofilms and the difficulty in detecting these agents within an intact biofilm. In this study, TOF-SIMS has been used to study the transport of CHG and glucose in a frozen hydrated biofilm. The TOF-SIMS images reveal a progression of CHG from the top of the biofilm to its base with time. Images suggest that there are channels within the biofilm and show preferential binding of CHG to cellular components of the biofilm. Additionally, both living and dead cells can be identified in the TOF-SIMS images by the sequestration of K + and the presence of cell markers. This study demonstrates that TOF-SIMS has the unique potential to simultaneously observe the presence of an antimicrobial agent, concentration of nutrients, and the viability of the cell population.

Anti-microbial properties of histone H2A from skin secretions of rainbow trout, Oncorhynchus mykiss.

PubMed Central

Fernandes, Jorge M O; Kemp, Graham D; Molle, M Gerard; Smith, Valerie J

2002-01-01

Skin exudates of rainbow trout contain a potent 13.6 kDa anti-microbial protein which, from partial internal amino acid sequencing, peptide mass fingerprinting, matrix-associated laser desorption/ionization MS and amino acid analysis, seems to be histone H2A, acetylated at the N-terminus. The protein, purified to homogeneity by ion-exchange and reversed-phase chromatography, exhibits powerful anti-bacterial activity against Gram-positive bacteria, with minimal inhibitory concentrations in the submicromolar range. Kinetic analysis revealed that at a concentration of 0.3 microM all test bacteria lose viability after 30 min incubation. Weaker activity is also displayed against the yeast Saccharomyces cerevisiae. The protein is salt-sensitive and has no haemolytic activity towards trout erythrocytes at concentrations below 0.3 microM. Reconstitution of the protein in a planar lipid bilayer strongly disturbs the membrane but does not form stable ion channels, indicating that its anti-bacterial activity is probably not due to pore-forming properties. This is the first report to show that, in addition to its classical function in the cell, histone H2A has extremely strong anti-microbial properties and could therefore help contribute to protection against bacterial invasion. PMID:12164782

Microbial degradation of high impact polystyrene (HIPS), an e-plastic with decabromodiphenyl oxide and antimony trioxide.

PubMed

Sekhar, Vini C; Nampoothiri, K Madhavan; Mohan, Arya J; Nair, Nimisha R; Bhaskar, Thallada; Pandey, Ashok

2016-11-15

Accumulation of electronic waste has increased catastrophically and out of that various plastic resins constitute one of the leading thrown out materials in the electronic machinery. Enrichment medium, containing high impact polystyrene (HIPS) with decabromodiphenyl oxide and antimony trioxide as sole carbon source, was used to isolate microbial cultures. The viability of these cultures in the e-plastic containing mineral medium was further confirmed by triphenyl tetrazolium chloride (TTC) reduction test. Four cultures were identified by 16S rRNA sequencing as Enterobacter sp., Citrobacter sedlakii, Alcaligenes sp. and Brevundimonas diminuta. Biodegradation experiments were carried out in flask level and gelatin supplementation (0.1% w/v) along with HIPS had increased the degradation rate to a maximum of 12.4% (w/w) within 30days. This is the first report for this kind of material. The comparison of FTIR, NMR, and TGA analysis of original and degraded e-plastic films revealed structural changes under microbial treatment. Polystyrene degradation intermediates in the culture supernatant were also detected using HPLC analysis. The gravity of biodegradation was validated by morphological changes under scanning electron microscope. All isolates displayed depolymerase activity to substantiate enzymatic degradation of e-plastic. Copyright © 2016 Elsevier B.V. All rights reserved.

Blue light induced free radicals from riboflavin in degradation of crystal violet by microbial viability evaluation.

PubMed

Liang, Ji-Yuan; Yuann, Jeu-Ming P; Hsie, Zong-Jhe; Huang, Shiuh-Tsuen; Chen, Chiing-Chang

2017-09-01

Crystal violet (CV) is applied in daily use mainly as a commercial dye and antimicrobial agent. Waste water containing CV may affect aquatic ecosystems. Riboflavin, also known as vitamin B 2 , is non-toxic and an essential vitamin required for the functions of the human body. Riboflavin is photosensitive to UV and visible light in terms of generating reactive oxygen species. This study investigated the potential application of blue light on riboflavin, so as to come up with an effective way of degrading CV during its treatment. Photosensitivity of CV leading to degradation in the presence of riboflavin was investigated by light intensity, exposure time, and irradiation dosage. The degradation of CV during riboflavin photolysis treatment was studied by a UV/vis spectrometry and chromatography. The effects of CV degradation on microbial viability are relevant when considering the influences on the ecosystem. This study proved that riboflavin photochemical treatment with blue light degrades CV dye by ROS formation. The riboflavin photolysis-treated CV solution appeared to be transparent during conformational transformations of the CV that was rearranged by free radical species generated from riboflavin photolysis. After riboflavin photolysis, colony-forming units (CFUs) were determined for each CV solution. CFU preservation was 85.2% for the CV dissolved riboflavin solution treated with blue light irradiation at 2.0mW/cm 2 for 120min. Degradation of CV by riboflavin photochemical procedures can greatly reduce antimicrobial ability and serve as an environmental friendly waste water treatment method. Our results presented here concerning riboflavin photolysis in degradation of CV provide a novel technique, and a simple and safe practice for environmental decontamination processes. Copyright © 2017 Elsevier B.V. All rights reserved.

Successful Resolution of Recurrent Clostridium difficile Infection using Freeze-Dried, Encapsulated Fecal Microbiota; Pragmatic Cohort Study

PubMed Central

Staley, Christopher; Hamilton, Matthew J.; Vaughn, Byron P.; Graiziger, Carolyn T.; Newman, Krista M.; Kabage, Amanda J.; Sadowsky, Michael J.; Khoruts, Alexander

2017-01-01

OBJECTIVES Fecal microbiota transplantation (FMT) is increasingly being used for treatment of recurrent Clostridium difficile infection (R-CDI) that cannot be cured with antibiotics alone. In addition, FMT is being investigated for a variety of indications where restoration or restructuring of the gut microbial community is hypothesized to be beneficial. We sought to develop a stable, freeze-dried encapsulated preparation of standardized fecal microbiota that can be used for FMT with ease and convenience in clinical practice and research. METHODS We systematically developed a lyophilization protocol that preserved the viability of bacteria across the taxonomic spectrum found in fecal microbiota and yielded physicochemical properties that enabled consistent encapsulation. We also treated a cohort of R-CDI patients with a range of doses of encapsulated microbiota and analyzed the associated changes in the fecal microbiome of the recipients. RESULTS The optimized lyophilized preparation satisfied all our preset goals for physicochemical properties, encapsulation ease, stability at different temperatures, and microbiota viability in vitro and in vivo (germ-free mice). The capsule treatment was administered to 49 patients. Overall, 43/49 (88%) of patients achieved a clinical success, defined as no recurrence of CDI over 2 months. Analysis of the fecal microbiome demonstrated near normalization of the fecal microbial community by 1 month following FMT treatment. The simplest protocol using the lowest dose (2.1–2.5×1011 bacteria in 2–3 capsules) without any colon purgative performed equally well in terms of clinical outcomes and microbiota engraftment. CONCLUSIONS A single administration of encapsulated, freeze-dried fecal microbiota from a healthy donor was highly successful in treating antibiotic-refractory R-CDI syndrome. PMID:28195180

Elevated temperature alters carbon cycling in a model microbial community

NASA Astrophysics Data System (ADS)

Mosier, A.; Li, Z.; Thomas, B. C.; Hettich, R. L.; Pan, C.; Banfield, J. F.

2013-12-01

Earth's climate is regulated by biogeochemical carbon exchanges between the land, oceans and atmosphere that are chiefly driven by microorganisms. Microbial communities are therefore indispensible to the study of carbon cycling and its impacts on the global climate system. In spite of the critical role of microbial communities in carbon cycling processes, microbial activity is currently minimally represented or altogether absent from most Earth System Models. Method development and hypothesis-driven experimentation on tractable model ecosystems of reduced complexity, as presented here, are essential for building molecularly resolved, benchmarked carbon-climate models. Here, we use chemoautotropic acid mine drainage biofilms as a model community to determine how elevated temperature, a key parameter of global climate change, regulates the flow of carbon through microbial-based ecosystems. This study represents the first community proteomics analysis using tandem mass tags (TMT), which enable accurate, precise, and reproducible quantification of proteins. We compare protein expression levels of biofilms growing over a narrow temperature range expected to occur with predicted climate changes. We show that elevated temperature leads to up-regulation of proteins involved in amino acid metabolism and protein modification, and down-regulation of proteins involved in growth and reproduction. Closely related bacterial genotypes differ in their response to temperature: Elevated temperature represses carbon fixation by two Leptospirillum genotypes, whereas carbon fixation is significantly up-regulated at higher temperature by a third closely related genotypic group. Leptospirillum group III bacteria are more susceptible to viral stress at elevated temperature, which may lead to greater carbon turnover in the microbial food web through the release of viral lysate. Overall, this proteogenomics approach revealed the effects of climate change on carbon cycling pathways and other microbial activities. When scaled to more complex ecosystems and integrated into Earth System Models, this approach could significantly improve predictions of global carbon-climate feedbacks. Experiments such as these are a critical first step designed at understanding climate change impacts in order to better predict ecosystem adaptations, assess the viability of mitigation strategies, and inform relevant policy decisions.

Microbial whole‐cell arrays

PubMed Central

Elad, Tal; Lee, Jin Hyung; Belkin, Shimshon; Gu, Man Bock

2008-01-01

Summary The coming of age of whole‐cell biosensors, combined with the continuing advances in array technologies, has prepared the ground for the next step in the evolution of both disciplines – the whole‐cell array. In the present review, we highlight the state‐of‐the‐art in the different disciplines essential for a functional bacterial array. These include the genetic engineering of the biological components, their immobilization in different polymers, technologies for live cell deposition and patterning on different types of solid surfaces, and cellular viability maintenance. Also reviewed are the types of signals emitted by the reporter cell arrays, some of the transduction methodologies for reading these signals and the mathematical approaches proposed for their analysis. Finally, we review some of the potential applications for bacterial cell arrays, and list the future needs for their maturation: a richer arsenal of high‐performance reporter strains, better methodologies for their incorporation into hardware platforms, design of appropriate detection circuits, the continuing development of dedicated algorithms for multiplex signal analysis and – most importantly – enhanced long‐term maintenance of viability and activity on the fabricated biochips. PMID:21261831

Anaerobic Probiotics: The Key Microbes for Human Health.

PubMed

El Enshasy, Hesham; Malik, Khairuddin; Malek, Roslinda Abd; Othman, Nor Zalina; Elsayed, Elsayed Ahmed; Wadaan, Mohammad

Human gastrointestinal microbiota (HGIM) incorporate a large number of microbes from different species. Anaerobic bacteria are the dominant organisms in this microbial consortium and play a crucial role in human health. In addition to their functional role as the main source of many essential metabolites for human health, they are considered as biotherapeutic agents in the regulation of different human metabolites. They are also important in the prevention and in the treatment of different physical and mental diseases. Bifidobacteria are the dominant anaerobic bacteria in HGIM and are widely used in the development of probiotic products for infants, children and adults. To develop bifidobacteria-based bioproducts, therefore, it is necessary to develop a large-scale biomass production platform based on a good understanding of the ideal medium and bioprocessing parameters for their growth and viability. In addition, high cell viability should be maintained during downstream processing and storage of probiotic cell powder or the final formulated product. In this work we review the latest information about the biology, therapeutic activities, cultivation and industrial production of bifidobacteria.

[The effect of long-term preservation of microbial cells immobilized in poly(vinyl alcohol) cryogel on their viability and biosynthesis of target metabolites].

PubMed

Efremenko, E N; Tatarinova, N Iu

2007-01-01

The effect of cell storage at -18 degrees C for 18-24 months on reproductive capacity was investigated for various microorganisms (gram-positive and gram-negative bacteria, yeasts, and filamentous fungi) immobilized in poly(vinyl alcohol) cryogel. To examine the viability of immobilized cells after defrosting, the bioluminescent method of intracellular ATP determination was used. A high level of metabolic activity of immobilized cells after various periods of storage was recorded for Streptomyces anulatus, Rhizopus orvzae, and Escherichia coli, which are producers of the antibiotic aurantin, L(+)-lactic acid, and the recombinant enzyme organophosphate hydrolase, respectively. It was shown that the initial concentration of immobilized cells in cryogel granules plays an important role in the survival of Str. anulatus and Pseudomonas putida after 1.5 years of storage. It was found that, after slow defrosting in the storage medium at 50C for 18 h of immobilized cells of the yeast Saccharomvces cerevisiae that had been stored for nine months, the number of reproductive cells increased due to the formation of ascospores.

Microbial lipid extraction from Lipomyces starkeyi using irreversible electroporation.

PubMed

Karim, Ahasanul; Yousuf, Abu; Islam, M Amirul; Naif, Yasir H; Faizal, Che Ku Mohammad; Alam, Md Zahangir; Pirozzi, Domenico

2018-02-21

The aim of the study was to investigate the feasibility of using irreversible electroporation (EP) as a microbial cell disruption technique to extract intracellular lipid within short time and in an eco-friendly manner. An EP circuit was designed and fabricated to obtain 4 kV with frequency of 100 Hz of square waves. The yeast cells of Lipomyces starkeyi (L. starkeyi) were treated by EP for 2-10 min where the distance between electrodes was maintained at 2, 4, and 6 cm. Colony forming units (CFU) were counted to observe the cell viability under the high voltage electric field. The forces of the pulsing electric field caused significant damage to the cell wall of L. starkeyi and the disruption of microbial cells was visualized by field emission scanning electron microscopic (FESEM) image. After breaking the cell wall, lipid was extracted and measured to assess the efficiency of EP over other techniques. The extent of cell inactivation was up to 95% when the electrodes were placed at the distance of 2 cm, which provided high treatment intensity (36.7 kWh m -3 ). At this condition, maximum lipid (63 mg g -1 ) was extracted when the biomass was treated for 10 min. During the comparison, EP could extract 31.88% lipid while the amount was 11.89% for ultrasonic and 16.8% for Fenton's reagent. The results recommend that the EP is a promising technique for lowering the time and solvent usage for lipid extraction from microbial biomass. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018. © 2018 American Institute of Chemical Engineers.

Life in Solid Ice on Earth and Other Planetary Bodies

NASA Astrophysics Data System (ADS)

Price, P. Buford

2004-06-01

Theory and direct observation indicate that micro-organisms exist in liquid veins in ice and permafrost, provided the temperature is above the eutectic for H_2O and soluble impurities present. Microbes can exist and metabolize in glacial ice and permafrost on Earth, Mars, and Europa. One can search directly (with fluorescence microscopy at liquid veins in Vostok ice core samples) or with a biologging instrument (for microbial fluorescence in a borehole in terrestrial or martian permafrost or ice). The viability lifetime against DNA destruction of bacterial spores can be measured with analytical techniques that identify calcium dipicolinate, which is unique to spores.

Potential application of glycerol in the production of plant beneficial microorganisms.

PubMed

Vassilev, Nikolay; Malusa, Eligio; Requena, Antonia Reyes; Martos, Vanessa; López, Ana; Maksimovic, Ivana; Vassileva, Maria

2017-05-01

This review highlights the importance of research for development of biofertilizer and biocontrol products based on the use of glycerol for further process scale-up to industrial microbiology. Glycerol can be used successfully in all stages of production of plant beneficial microorganisms. It serves as an excellent substrate in both submerged and solid-state fermentation processes with free and immobilized microbial cells. Glycerol is also one of the most attractive formulation agents that ensures high cell density and viability including in harsh environmental conditions. Future research is discussed to make this inexpensive material a base for industrial production of plant beneficial microorganisms.

Stability and morphological and molecular-genetic identification of algae in buried soils

NASA Astrophysics Data System (ADS)

Temraleeva, A. D.; Moskalenko, S. V.; El'tsov, M. V.; Vagapov, I. M.; Ovchinnikov, A. Yu.; Gugalinskaya, L. A.; Alifanov, V. M.; Pinskii, D. L.

2017-08-01

Living cultural strains of the green algae `Chlorella' mirabilis and Muriella terrestris have been isolated from buried soils, and their identification has been confirmed by morphological and molecular-genetic analysis. It has been shown that the retention of their viability could be related to their small size and the presence of sporopollenin in cell walls. The effect of methods for the reactivation of dormant microbial forms on the growth of algae in paleosols has been estimated. The total DNA content has been determined in buried and recent background soils, and relationship between DNA and the presence and age of burial has been established.

Super-long Anabiosis of Ancient Microorganisms in Ice and Terrestrial Models for Development of Methods to Search for Life on Mars, Europa and other Planetary Bodies

NASA Technical Reports Server (NTRS)

Abyzov, S. S.; Duxbury, N. S.; Bobin, N. E.; Fukuchi, M.; Hoover, R. B.; Kanda, H.; Mitskevich, I. N.; Mulyukin, A. L.; Naganuma, T.; Poglazova, M. N.;

Short-term differential adaptation to anaerobic stress via genomic mutations by Escherichia coli strains K-12 and B lacking alcohol dehydrogenase

PubMed Central

Kim, Hyun Ju; Jeong, Haeyoung; Hwang, Seungwoo; Lee, Moo-Seung; Lee, Yong-Jik; Lee, Dong-Woo; Lee, Sang Jun

2014-01-01

Microbial adaptations often occur via genomic mutations under adverse environmental conditions. This study used Escherichia coli ΔadhE cells as a model system to investigate adaptation to anaerobic conditions, which we then compared with the adaptive mechanisms of two closely related E. coli strains, K-12 and B. In contrast to K-12 ΔadhE cells, the E. coli B ΔadhE cells exhibited significantly delayed adaptive growth under anaerobic conditions. Adaptation by the K-12 and B strains mainly employed anaerobic lactate fermentation to restore cellular growth. Several mutations were identified in the pta or pflB genes of adapted K-12 cells, but mostly in the pta gene of the B strains. However, the types of mutation in the adapted K-12 and B strains were similar. Cellular viability was affected directly by severe redox imbalance in B ΔadhE cells, which also impaired their ability to adapt to anaerobic conditions. This study demonstrates that closely related microorganisms may undergo different adaptations under the same set of adverse conditions, which might be associated with the specific metabolic characteristics of each strain. This study provides new insights into short-term microbial adaptation to stressful conditions, which may reflect dynamic microbial population changes in nature. PMID:25250024

Screening of industrial wastewaters as feedstock for the microbial production of oils for biodiesel production and high-quality pigments

DOE PAGES

Schneider, Teresa; Graeff-Honninger, Simone; French, William Todd; ...

2012-01-01

The production of biodiesel has notably increased over the past decade. Currently, plant oil is the main feedstock for biodiesel production, but, due to concerns related to the competition with food production, alternative oil feedstocks have to be found. Oleaginous yeasts are known to produce high amounts of lipids, but no integrated process from microbial fermentation to final biodiesel production has reached commercial realization yet due to economic constraints. Therefore, growth and lipid production of red yeast Rhodotorula glutinis was tested on low-cost substrates, namely, wastewaters from potato, fruit juice, and lettuce processing. Additionally, the production of carotenoids as high-valuemore » by-products was examined. All evaluated wastewaters met the general criteria for microbial lipid production. However, no significant increase in lipid content was observed, probably due to lack of available carbon in wastewaters from fruit juice and lettuce processing, and excess of available nitrogen in potato processing wastewater, respectively. During growth on wastewaters from fruit juice and lettuce processing the carotenoid content increased significantly in the first 48 hours. The relations between carbon content, nitrogen content, and carotenoid production need to be further assessed. For economic viability, lipid and carotenoid production needs to be increased significantly. Lastly, the screening of feedstocks should be extended to other wastewaters.« less

Hydrogel-Based Fluorescent Dual pH and Oxygen Sensors Loaded in 96-Well Plates for High-Throughput Cell Metabolism Studies.

PubMed

Wu, Shanshan; Wu, Siying; Yi, Zheyuan; Zeng, Fei; Wu, Weizhen; Qiao, Yuan; Zhao, Xingzhong; Cheng, Xing; Tian, Yanqing

2018-02-13

In this study, we developed fluorescent dual pH and oxygen sensors loaded in multi-well plates for in-situ and high-throughput monitoring of oxygen respiration and extracellular acidification during microbial cell growth for understanding metabolism. Biocompatible PHEMA-co-PAM materials were used as the hydrogel matrix. A polymerizable oxygen probe (OS2) derived from PtTFPP and a polymerizable pH probe (S2) derived from fluorescein were chemically conjugated into the matrix to solve the problem of the probe leaching from the matrix. Gels were allowed to cure directly on the bottom of 96-well plates at room-temperature via redox polymerization. The influence of matrix's composition on the sensing behaviors was investigated to optimize hydrogels with enough robustness for repeatable use with good sensitivity. Responses of the dual sensing hydrogels to dissolved oxygen (DO) and pH were studied. These dual oxygen-pH sensing plates were successfully used for microbial cell-based screening assays, which are based on the measurement of fluorescence intensity changes induced by cellular oxygen consumption and pH changes during microbial growth. This method may provide a real-time monitoring of cellular respiration, acidification, and a rapid kinetic assessment of multiple samples for cell viability as well as high-throughput drug screening. All of these assays can be carried out by a conventional plate reader.

Microbial Attachment Inhibition through Low-Voltage Electrochemical Reactions on Electrically Conducting Membranes.

PubMed

Ronen, Avner; Duan, Wenyan; Wheeldon, Ian; Walker, Sharon; Jassby, David

2015-11-03

Bacterial biofilm formation on membrane surfaces remains a serious challenge in water treatment systems. The impact of low voltages on microbial attachment to electrically conducting ultrafiltration membranes was investigated using a direct observation cross-flow membrane system mounted on a fluorescence microscope. Escherichia coli and microparticle deposition and detachment rates were measured as a function of the applied electrical potential to the membrane surface. Selecting bacteria and particles with low surface charge minimized electrostatic interactions between the bacteria and charged membrane surface. Application of an electrical potential had a significant impact on the detachment of live bacteria in comparison to dead bacteria and particles. Image analysis indicated that when a potential of 1.5 V was applied to the membrane/counter electrode pair, the percent of dead bacteria was 32±2.1 and 67±3.6% when the membrane was used as a cathode or anode, respectively, while at a potential of 1 V, 92±2.4% were alive. The application of low electrical potentials resulted in the production of low (μM) concentrations of hydrogen peroxide (HP) through the electroreduction of oxygen. The electrochemically produced HP reduced microbial cell viability and increased cellular permeability. Exposure to low concentrations of electrochemically produced HP on the membrane surface prevents bacterial attachment, thus ensuring biofilm-free conditions during membrane filtration operations.

Characterization of the microbial diversity in yacon spontaneous fermentation

PubMed Central

Reina, L. D.; Pérez-Díaz, I. M.; Breidt, F.; Azcarate-Peril, M. A.; Medina, E.; Butz, N.V.

2015-01-01

The prebiotic fructooligosaccharides (FOS) content of yacon makes this root an attractive alternative for the supplementation of a variety of food products. The preservation of yacon by fermentation has been proposed as an alternative to increase the probiotic content of the root concomitantly with its shelf life. Thus the fermented yacon could have significant functional content. The objective of this research was to characterize the biochemistry and microbiology of spontaneous yacon fermentation and define the viability of the proposed process. The biochemical analysis of spontaneous heterolactic fermentation of yacon showed a progressive drop in pH with increased lactic and acetic acids, and the production of mannitol during fermentation. The microbial ecology of yacon fermentation was investigated using culture-dependent and culture-independent methods. Bacterial cell counts revealed a dominance of lactic acid bacteria (LAB) over yeasts, which were also present during fermentation. Results showed that the heterofermentative LAB were primarily Leuconostoc species, which dominated the fermentation. The fermentation of yacon by Leuconostoc spp. is thus presented as a viable method to achieve long term preservation of this root. PMID:25777679

Characterization of the microbial diversity in yacon spontaneous fermentation at 20 °C.

PubMed

Reina, L D; Pérez-Díaz, I M; Breidt, F; Azcarate-Peril, M A; Medina, E; Butz, N

2015-06-16

The prebiotic fructooligosaccharide content of yacon makes this root an attractive alternative for the supplementation of a variety of food products. The preservation of yacon by fermentation has been proposed as an alternative to increase the probiotic content of the root concomitantly with its shelf life. Thus the fermented yacon could have significant functional content. The objective of this research was to characterize the biochemistry and microbiology of spontaneous yacon fermentation with 2% NaCl and define the viability of the proposed process. The biochemical analysis of spontaneous heterolactic fermentation of yacon showed a progressive drop in pH with increased lactic and acetic acids, and the production of mannitol during fermentation. The microbial ecology of yacon fermentation was investigated using culture-dependent and culture-independent methods. Bacterial cell counts revealed a dominance of lactic acid bacteria (LAB) over yeasts, which were also present during the first 2 days of the fermentation. Results showed that the heterofermentative LAB were primarily Leuconostoc species, thus it presents a viable method to achieve long term preservation of this root. Copyright © 2015. Published by Elsevier B.V.

Comparative analysis of microbial fuel cell based biosensors developed with a mixed culture and Shewanella loihica PV-4 and underlying biological mechanism.

PubMed

Yi, Yue; Xie, Beizhen; Zhao, Ting; Liu, Hong

2018-06-13

Microbial fuel cell based biosensors (MFC-biosensors) utilize anode biofilms as biological recognition elements to monitor biochemical oxygen demand (BOD) and biotoxicity. However, the relatively poor sensitivity constrains the application of MFC-biosensors. To address this limitation, this study provided a systematic comparison of sensitivity between the MFC-biosensors constructed with two inocula. Higher biomass density and viability were both observed in the anode biofilm of the mixed culture MFC, which resulted in better sensitivity for BOD assessment. Compared with using mixed culture as inoculum, the anode biofilm developed with Shewanella loihica PV-4 presented lower content of extracellular polymeric substances and poorer ability to secrete protein under toxic shocks. Moreover, the looser structure in the S. loihica PV-4 biofilm further facilitated its susceptibilities to toxic agents. Therefore, the MFC-biosensor with a pure culture of S. loihica PV-4 delivered higher sensitivity for biotoxicity monitoring. This study proposed a new perspective to enhance sensor performance. Copyright © 2018 Elsevier Ltd. All rights reserved.

Antimicrobial function of Nd3+-doped anatase titania-coated nickel ferrite composite nanoparticles: a biomaterial system.

PubMed

Rana, S; Rawat, J; Sorensson, M M; Misra, R D K

2006-07-01

The present study describes and makes a relative comparison of the antimicrobial function of undoped and neodymium-doped titania coated-nickel ferrite composite nanoparticles processed by uniquely combining the reverse micelle and chemical hydrolysis approaches. This methodology facilitates the formation of undoped and doped photocatalytic titania shells and a magnetic ferrite core. The ferrite core is needed to help in the removal of particles from the sprayed surface using a small magnetic field. Doping of the titania shell with neodymium significantly enhances the photocatalytic and anti-microbial function of the core-shell composite nanoparticles without influencing the magnetic characteristics of the nickel ferrite core. The increased performance is believed to be related to the inhibition of electron-hole recombination and a decrease in the band gap energy of titania. The retention of magnetic strength ensures controlled movement of the composite nanoparticles by the magnetic field, facilitating their application as removable anti-microbial photocatalyst nanoparticles. The consistent behavior of the composite nanoparticles points to the viability of the synthesis process adopted.

Microbial removals by a novel biofilter water treatment system.

PubMed

Wendt, Christopher; Ives, Rebecca; Hoyt, Anne L; Conrad, Ken E; Longstaff, Stephanie; Kuennen, Roy W; Rose, Joan B

2015-04-01

Two point-of-use drinking water treatment systems designed using a carbon filter and foam material as a possible alternative to traditional biosand systems were evaluated for removal of bacteria, protozoa, and viruses. Two configurations were tested: the foam material was positioned vertically around the carbon filter in the sleeve unit or horizontally in the disk unit. The filtration systems were challenged with Cryptosporidium parvum, Raoultella terrigena, and bacteriophages P22 and MS2 before and after biofilm development to determine average log reduction (ALR) for each organism and the role of the biofilm. There was no significant difference in performance between the two designs, and both designs showed significant levels of removal (at least 4 log10 reduction in viruses, 6 log10 for protozoa, and 8 log10 for bacteria). Removal levels meet or exceeded Environmental Protection Agency (EPA) standards for microbial purifiers. Exploratory test results suggested that mature biofilm formation contributed 1-2 log10 reductions. Future work is recommended to determine field viability. © The American Society of Tropical Medicine and Hygiene.

Microbial degradation of total petroleum hydrocarbons in crude oil: a field-scale study at the low-land rainforest of Ecuador.

PubMed

Maddela, Naga Raju; Scalvenzi, Laura; Venkateswarlu, Kadiyala

2017-10-01

A field-level feasibility study was conducted to determine total petroleum hydrocarbon (TPH)-degrading potential of two bacterial strains, Bacillus thuringiensis B3 and B. cereus B6, and two fungi, Geomyces pannorum HR and Geomyces sp. strain HV, all soil isolates obtained from an oil field located in north-east region of Ecuador. Crude oil-treated soil samples contained in wooden boxes received a mixture of all the four microorganisms and were incubated for 90 days in an open low-land area of Amazon rainforest. The percent removal of TPHs in soil samples that received the mixed microbial inoculum was 87.45, indicating the great potential of the soil isolates in field-scale removal of crude oil. The TPHs-degrading efficiency was verified by determining the toxicity of residues, remained in soil after biodegradation, toward viability of Artemia salina or seed germination and plant growth of cowpea. Our results clearly suggest that the selected soil isolates of bacteria and fungi could be effectively used for large-scale bioremediation of sites contaminated with crude oil.

Immobilization of Microbial Cells for Alcoholic and Malolactic Fermentation of Wine and Cider

NASA Astrophysics Data System (ADS)

Kourkoutas, Yiannis; Manojlović, Verica; Nedović, Viktor A.

Wine- or cider-making is highly associated with biotechnology owing to the traditional nature of must fermentation.. Nowadays, there have been considerable developments in wine- or cider-making techniques affecting all phases of wine or cider production, but more importantly, the fermentation process. It is well-known that the transformation of grape must by microbial activity results in the production of wine, and the fermentation of apples (or sometimes pears) in the production of cider. In this process, a variety of compounds affecting the organoleptic profile of wine or cider are synthesized. It is also common sense that in wine- or cider-making, the main objective is to achieve an adequate quality of the product. The technological progress and the improved quality of the wines or ciders have been associated with the control of technical parameters. Herein, cell immobilization offers numerous advantages, such as enhanced fermentation productivity, ability for cell recycling, application of continuous configurations, enhanced cell stability and viability, and improvement of quality (Margaritis and Merchant 1984; Stewart and Russel 1986; Kourkoutas et al. 2004a).

Protist-Bacteria Associations: Gammaproteobacteria and Alphaproteobacteria Are Prevalent as Digestion-Resistant Bacteria in Ciliated Protozoa

PubMed Central

Gong, Jun; Qing, Yao; Zou, Songbao; Fu, Rao; Su, Lei; Zhang, Xiaoli; Zhang, Qianqian

2016-01-01

Protistan bacterivory, a microbial process involving ingestion and digestion, is ecologically important in the microbial loop in aquatic and terrestrial ecosystems. While bacterial resistance to protistan ingestion has been relatively well understood, little is known about protistan digestion in which some ingested bacteria could not be digested in cells of major protistan grazers in the natural environment. Here we report the phylogenetic identities of digestion-resistant bacteria (DRB) that could survive starvation and form relatively stable associations with 11 marine and one freshwater ciliate species. Using clone library and sequencing of 16S rRNA genes, we found that the protistan predators could host a high diversity of DRB, most of which represented novel bacterial taxa that have not been cultivated. The localization inside host cells, quantity, and viability of these bacteria were checked using fluorescence in situ hybridization. The DRB were affiliated with Actinobacteria, Bacteroidetes, Firmicutes, Parcubacteria (OD1), Planctomycetes, and Proteobacteria, with Gammaproteobacteria and Alphaproteobacteria being the most frequently occurring classes. The dominance of Gamma- and Alphaproteobacteria corresponds well to a previous study of Global Ocean Sampling metagenomic data showing the widespread types of bacterial type VI and IV secretion systems (T6SS and T4SS) in these two taxa, suggesting a putatively significant role of secretion systems in promoting marine protist-bacteria associations. In the DRB assemblages, opportunistic bacteria such as Alteromonadaceae, Pseudoalteromonadaceae, and Vibrionaceae often presented with high proportions, indicating these bacteria could evade protistan grazing thus persist and accumulate in the community, which, however, contrasts with their well-known rarity in nature. This begs the question whether viral lysis is significant in killing these indigestible bacteria in microbial communities. Taken together, our study on the identity of DRB sheds new light on microbial interactions and generates further hypotheses including the potential importance of bacterial protein secretion systems in structuring bacterial community composition and functioning of “microbial black box” in aquatic environments. PMID:27148188

Recently Deglaciated High-Altitude Soils of the Himalaya: Diverse Environments, Heterogenous Bacterial Communities and Long-Range Dust Inputs from the Upper Troposphere

PubMed Central

Stres, Blaz; Sul, Woo Jun; Murovec, Bostjan; Tiedje, James M.

2013-01-01

Background The Himalaya with its altitude and geographical position forms a barrier to atmospheric transport, which produces much aqueous-particle monsoon precipitation and makes it the largest continuous ice-covered area outside polar regions. There is a paucity of data on high-altitude microbial communities, their native environments and responses to environmental-spatial variables relative to seasonal and deglaciation events. Methodology/Principal Findings Soils were sampled along altitude transects from 5000 m to 6000 m to determine environmental, spatial and seasonal factors structuring bacterial communities characterized by 16 S rRNA gene deep sequencing. Dust traps and fresh-snow samples were used to assess dust abundance and viability, community structure and abundance of dust associated microbial communities. Significantly different habitats among the altitude-transect samples corresponded to both phylogenetically distant and closely-related communities at distances as short as 50 m showing high community spatial divergence. High within-group variability that was related to an order of magnitude higher dust deposition obscured seasonal and temporal rearrangements in microbial communities. Although dust particle and associated cell deposition rates were highly correlated, seasonal dust communities of bacteria were distinct and differed significantly from recipient soil communities. Analysis of closest relatives to dust OTUs, HYSPLIT back-calculation of airmass trajectories and small dust particle size (4–12 µm) suggested that the deposited dust and microbes came from distant continental, lacustrine and marine sources, e.g. Sahara, India, Caspian Sea and Tibetan plateau. Cyanobacteria represented less than 0.5% of microbial communities suggesting that the microbial communities benefitted from (co)deposited carbon which was reflected in the psychrotolerant nature of dust-particle associated bacteria. Conclusions/Significance The spatial, environmental and temporal complexity of the high-altitude soils of the Himalaya generates ongoing disturbance and colonization events that subject heterogeneous microniches to stochastic colonization by far away dust associated microbes and result in the observed spatially divergent bacterial communities. PMID:24086740

Organic and inorganic composition and microbiology of produced waters from Pennsylvania shale gas wells

USGS Publications Warehouse

Akob, Denise M.; Cozzarelli, Isabelle M.; Dunlap, Darren S.; Rowan, Elisabeth L.; Lorah, Michelle M.

2015-01-01

Hydraulically fractured shales are becoming an increasingly important source of natural gas production in the United States. This process has been known to create up to 420 gallons of produced water (PW) per day, but the volume varies depending on the formation, and the characteristics of individual hydraulic fracture. PW from hydraulic fracturing of shales are comprised of injected fracturing fluids and natural formation waters in proportions that change over time. Across the state of Pennsylvania, shale gas production is booming; therefore, it is important to assess the variability in PW chemistry and microbiology across this geographical span. We quantified the inorganic and organic chemical composition and microbial communities in PW samples from 13 shale gas wells in north central Pennsylvania. Microbial abundance was generally low (66–9400 cells/mL). Non-volatile dissolved organic carbon (NVDOC) was high (7–31 mg/L) relative to typical shallow groundwater, and the presence of organic acid anions (e.g., acetate, formate, and pyruvate) indicated microbial activity. Volatile organic compounds (VOCs) were detected in four samples (∼1 to 11.7 μg/L): benzene and toluene in the Burket sample, toluene in two Marcellus samples, and tetrachloroethylene (PCE) in one Marcellus sample. VOCs can be either naturally occurring or from industrial activity, making the source of VOCs unclear. Despite the addition of biocides during hydraulic fracturing, H2S-producing, fermenting, and methanogenic bacteria were cultured from PW samples. The presence of culturable bacteria was not associated with salinity or location; although organic compound concentrations and time in production were correlated with microbial activity. Interestingly, we found that unlike the inorganic chemistry, PW organic chemistry and microbial viability were highly variable across the 13 wells sampled, which can have important implications for the reuse and handling of these fluids

New perspective on functional capabilities of microbiome associated with spacecraft assembly facilities

NASA Astrophysics Data System (ADS)

Vaishampayan, Parag

2016-07-01

In compliance with Planetary Protection policy, NASA monitors the total microbial burden of spacecraft and associated environments as a means for minimizing forward contamination. Despite numerous characterizations of microbial populations in spacecraft assembly cleanrooms, understanding the metabolic traits responsible for their persistence and survival remains a significant challenge. The principal objective of this study is to establish functional traits by exploring the entire gene content (metagenome) of the cleanroom microbial community. DNA-based techniques are incapable of distinguishing viable microorganisms from dead microbial cells in samples. Consequently, metagenomic analyses based on total environmental DNA extracts do not render a meaningful understanding of the metabolic and/or functional characteristics of living microorganisms in cleanrooms. A molecular viability marker was applied to samples collected from a cleanroom facility, and subsequent metagenomic sequencing experiments showed considerable differences between the resulting viable-only and total microbiomes. Nevertheless, analyses of sequence abundance suggested that the viable microbiome was influenced by both the human microbiome and the ambient ecosystem external to the facility, which resulted in a complex community profile. Also detected were the first viral signatures ever retrieved from a cleanroom facility: the genomes of human cyclovirus 7078A and Propionibacterium phage P14.4. We also wanted to evaluate if the strict cleaning and decontamination procedures selectively favor survival and growth of hardy microrganisms, such as pathogens. Three geographically distinct cleanrooms were sampled during the assembly of three NASA spacecraft: Dawn, Phoenix, and Mars Science Laboratory. Potential pathogens and their corresponding virulence factors were present in all the samples. Decreased microbial and pathogenic diversity during spacecraft assembly, compared to before and after, indicates that decontamination and preventative measures were effective and well implemented. The findings presented here, as well as the innovative methods that enabled their discovery, promise to have profound implications for the design and interpretation of ongoing and future studies in cleanrooms, indoor environments, and potential future human missions to Mars.

Microbiological indicators for assessing ecosystem soil quality and changes in it at degraded sites treated with compost

NASA Astrophysics Data System (ADS)

Ancona, Valeria; Barra Caracciolo, Anna; Grenni, Paola; Di Lenola, Martina; Calabrese, Angelantonio; Campanale, Claudia; Felice Uricchio, Vito

2014-05-01

Soil quality is defined as the capacity of a soil to function as a vital system, within natural or managed ecosystem boundaries, sustain plant and animal health and productivity, maintain or enhance air and water environment quality and support human health and habitation. Soil organisms are extremely diverse and contribute to a wide range of ecosystem services that are essential to the sustainable functioning of natural and managed ecosystems. In particular, microbial communities provide several ecosystem services, which ensure soil quality and fertility. In fact, they adapt promptly to environmental changes by varying their activity and by increasing the reproduction of populations that have favourable skills. The structure (e.g. cell abundance) and functioning (e.g. viability and activity) of natural microbial communities and changes in them under different environmental conditions can be considered useful indicators of soil quality state. In this work we studied the quality state of three different soils, located in Taranto Province (Southern Italy), affected by land degradation processes, such as organic matter depletion, desertification and contamination (PCB and metals). Moreover, compost, produced from selected organic waste, was added to the soils studied in order to improve their quality state. Soil samples were collected before and after compost addition and both microbial and chemical analyses were performed in order to evaluate the soil quality state at each site at different times. For this purpose, the microbiological indicators evaluated were bacterial abundance (DAPI counts), cell viability (Live/Dead method), dehydrogenase activity (DHA) and soil respiration. At the same time, the main physico-chemical soil characteristics (organic carbon, available phosphorous, total nitrogen, carbonate and water content, texture and pH) were also measured. Moreover, in the contaminated soil samples PCB and inorganic (e.g. Pb, Se, Sn, Zn) contaminants were analysed respectively by GC-MS and ICP-MS. The overall results showed that the bacterial structure and functioning were affected in different ways by the organic carbon availability and quality, and contaminant occurrence (organic or inorganic compounds). The compost treatment contributed to improve soil fertility and to increase cell number and activity after 7 months in the two low organic carbon content soils. At the polluted site a general increase in bacterial activity after compost addition was also observed and this might be related to a decrease in inorganic and organic contamination levels.

Seawater Mg/Ca controls polymorph mineralogy of microbial CaCO3: a potential proxy for calcite-aragonite seas in Precambrian time.

PubMed

Ries, J B; Anderson, M A; Hill, R T

2008-03-01

A previously published hydrothermal brine-river water mixing model driven by ocean crust production suggests that the molar Mg/Ca ratio of seawater (mMg/Ca(sw)) has varied significantly (approximately 1.0-5.2) over Precambrian time, resulting in six intervals of aragonite-favouring seas (mMg/Ca(sw) > 2) and five intervals of calcite-favouring seas (mMg/Ca(sw) < 2) since the Late Archaean. To evaluate the viability of microbial carbonates as mineralogical proxy for Precambrian calcite-aragonite seas, calcifying microbial marine biofilms were cultured in experimental seawaters formulated over the range of Mg/Ca ratios believed to have characterized Precambrian seawater. Biofilms cultured in experimental aragonite seawater (mMg/Ca(sw) = 5.2) precipitated primarily aragonite with lesser amounts of high-Mg calcite (mMg/Ca(calcite) = 0.16), while biofilms cultured in experimental calcite seawater (mMg/Ca(sw) = 1.5) precipitated exclusively lower magnesian calcite (mMg/Ca(calcite) = 0.06). Furthermore, Mg/Ca(calcite )varied proportionally with Mg/Ca(sw). This nearly abiotic mineralogical response of the biofilm CaCO3 to altered Mg/Ca(sw) is consistent with the assertion that biofilm calcification proceeds more through the elevation of , via metabolic removal of CO2 and/or H+, than through the elevation of Ca2+, which would alter the Mg/Ca ratio of the biofilm's calcifying fluid causing its pattern of CaCO3 polymorph precipitation (aragonite vs. calcite; Mg-incorporation in calcite) to deviate from that of abiotic calcification. If previous assertions are correct that the physicochemical properties of Precambrian seawater were such that Mg/Ca(sw) was the primary variable influencing CaCO3 polymorph mineralogy, then the observed response of the biofilms' CaCO3 polymorph mineralogy to variations in Mg/Ca(sw), combined with the ubiquity of such microbial carbonates in Precambrian strata, suggests that the original polymorph mineralogy and Mg/Ca(calcite )of well-preserved microbial carbonates may be an archive of calcite-aragonite seas throughout Precambrian time. These results invite a systematic evaluation of microbial carbonate primary mineralogy to empirically constrain Precambrian seawater Mg/Ca.

Effectiveness of a full-scale horizontal slow sand filter for controlling phytopathogens in recirculating hydroponics: From microbial isolation to full microbiome assessment.

PubMed

Prenafeta-Boldú, Francesc X; Trillas, Isabel; Viñas, Marc; Guivernau, Miriam; Cáceres, Rafaela; Marfà, Oriol

2017-12-01

The microbial disinfestation efficiency of an innovative horizontal-flow slow sand filter (HSSF) for treating nutrient solution spent from an experimental closed-loop nursery was evaluated by means of a combination of culture-dependent and independent molecular techniques. A dense inoculum of the fungal plant pathogen Fusarium oxysporum f.sp. lycopersici was applied in the fertigation system (10 6 cells per mL). Indigenous and introduced populations of eubacteria and fungi were assessed in the nutrient solution, the HSSF influent/effluent, and a sand bed transect by isolation on selective media, as well as by quantitative qPCR and next-generation sequencing (NGS) on target ribosomal genes. The HSSF effectively reduced viable Fusarium propagules and fungal gene content with an efficiency consistently above 99.9% (5 orders of magnitude down). On the other hand, Fusarium cells accumulated in the sand bed, indicating that physical entrapment was the main removal mechanism. The viability of retained Fusarium cells tended to decrease in time, so that treatment efficiency might be enhanced by antagonistic species from the genera Bacillus, Pseudomonas, and Trichoderma, also identified in the sand bed. Indigenous bacterial populations from the HSSF effluent were reduced by 87.2% and 99.9% in terms of colony forming units and gene counts, respectively, when compared to the influent. Furthermore, microbial populations from the HSSF effluent were different from those observed in the sand bed and the influent. In summary, the HSSF microbial disinfestation efficiency is comparable to that reported for other more intensive and costly methodologies, while allowing a significant recovery of water and nutrients. Copyright © 2017 Elsevier B.V. All rights reserved.

CMEIAS color segmentation: an improved computing technology to process color images for quantitative microbial ecology studies at single-cell resolution.

PubMed

Gross, Colin A; Reddy, Chandan K; Dazzo, Frank B

2010-02-01

Quantitative microscopy and digital image analysis are underutilized in microbial ecology largely because of the laborious task to segment foreground object pixels from background, especially in complex color micrographs of environmental samples. In this paper, we describe an improved computing technology developed to alleviate this limitation. The system's uniqueness is its ability to edit digital images accurately when presented with the difficult yet commonplace challenge of removing background pixels whose three-dimensional color space overlaps the range that defines foreground objects. Image segmentation is accomplished by utilizing algorithms that address color and spatial relationships of user-selected foreground object pixels. Performance of the color segmentation algorithm evaluated on 26 complex micrographs at single pixel resolution had an overall pixel classification accuracy of 99+%. Several applications illustrate how this improved computing technology can successfully resolve numerous challenges of complex color segmentation in order to produce images from which quantitative information can be accurately extracted, thereby gain new perspectives on the in situ ecology of microorganisms. Examples include improvements in the quantitative analysis of (1) microbial abundance and phylotype diversity of single cells classified by their discriminating color within heterogeneous communities, (2) cell viability, (3) spatial relationships and intensity of bacterial gene expression involved in cellular communication between individual cells within rhizoplane biofilms, and (4) biofilm ecophysiology based on ribotype-differentiated radioactive substrate utilization. The stand-alone executable file plus user manual and tutorial images for this color segmentation computing application are freely available at http://cme.msu.edu/cmeias/ . This improved computing technology opens new opportunities of imaging applications where discriminating colors really matter most, thereby strengthening quantitative microscopy-based approaches to advance microbial ecology in situ at individual single-cell resolution.

Study of phytochemical, anti-microbial, anti-oxidant, and anti-cancer properties of Allium wallichii.

PubMed

Bhandari, Jaya; Muhammad, BushraTaj; Thapa, Pratiksha; Shrestha, Bhupal Govinda

2017-02-08

There is growing interest in the use of plants for the treatment and prevention of cancer. Medicinal plants are currently being evaluated as source of promising anticancer agents. In this paper, we have investigated the anticancer potential of plant Allium wallichii, a plant native to Nepal and growing at elevations of 2300-4800 m. This is the first study of its kind for the plant mentioned. The dried plant was extracted in aqueous ethanol. Phytochemical screening, anti-microbial assay, anti-oxidant assay, cytotoxicity assay and the flow-cytometric analysis were done for analyzing different phytochemicals present, anti-microbial activity, anti-oxidant activity and anti-cancer properties of Allium wallichii. We observed the presence of steroids, terpenoids, flavonoids, reducing sugars and glycosides in the plant extract and the plant showed moderate anti-microbial and anti-oxidant activity. The IC 50 values of Allium wallichii in different cancer cell lines are 69.69 μg/ml for Prostate cancer (PC3) cell line, 55.29 μg/ml for Breast Cancer (MCF-7) cell line and 46.51 μg/ml for cervical cancer (HeLa) cell line as compared to Doxorubicin (0.85 μg/ml). The cell viability assay using FACS showed that the IC 50 value of Allium wallichii for Burkitt's lymphoma (B-Lymphoma) cell line was 3.817 ± 1.99 mg/ml. Allium wallichii can be an important candidate to be used as an anticancer agent. Separation of pure compounds with bioassay guided extraction, spectrometric analysis and subsequent cytotoxicity assay of the pure bioactive compounds from Allium wallichii is highly recommended as the crude extract itself showed promising cytotoxicity.

Toxicity of essential oil of Satureja khuzistanica: in vitro cytotoxicity and anti-microbial activity.

PubMed

Yousefzadi, Morteza; Riahi-Madvar, Ali; Hadian, Javad; Rezaee, Fatemeh; Rafiee, Roya; Biniaz, Mehdi

2014-01-01

In nature, essential oils play an important role in the protection of the plants by exerting anti-bacterial, -viral, -fungal, -oxidative, -genotoxic, and free radical scavenging properties, as well as in some cases acting as insecticides. Several Satureja species are used in traditional medicine due to recognized therapeutic properties, namely anti-microbial and cytotoxic activities. The purpose of the present work was to determine the biologic activity of the essential oil of S. khuzistanica Jamzad (Lamiaceae) against four human cancer cell lines, as well as its inhibitory effects against a wide array (i.e. n = 11) of pathogenic bacteria and fungi. The essential oil was isolated by hydro-distillation and analyzed by GC-FID and GC-MS. Carvacrol (92.87%) and limonene (1.2%) were found to be the main components of the isolated oil. Anti-microbial activity of the essential oil was assessed using a disc diffusion method; an MTT cytotoxicity assay was employed to test effects of the oil on each cancer cell line. The oil exhibited considerable anti-microbial activity against the majority of the tested bacteria and fungi. The test oil also significantly reduced cell viability of Vero, SW480, MCF7, and JET 3 cells in a dose-dependent manner, with the IC50 values calculated for each cell type being, respectively, 31.2, 62.5, 125, and 125 μg/ml. Based on the findings, it is concluded that the essential oil of S. khuzistanica and its major constituents have a potential for further use in anti-bacterial and anti-cancer applications, pending far more extensive testing of toxicities in normal (i.e. primary) cells.

IODP Expedition 337: Deep Coalbed Biosphere off Shimokita - Microbial processes and hydrocarbon system associated with deeply buried coalbed in the ocean

NASA Astrophysics Data System (ADS)

Inagaki, Fumio; Hinrichs, Kai-Uwe; Kubo, Yusuke; IODP Expedition 337 Scientists

2016-06-01

The Integrated Ocean Drilling Program (IODP) Expedition 337 was the first expedition dedicated to subseafloor microbiology that used riser-drilling technology with the drilling vessel Chikyu. The drilling Site C0020 is located in a forearc basin formed by the subduction of the Pacific Plate off the Shimokita Peninsula, Japan, at a water depth of 1180 m. Primary scientific objectives during Expedition 337 were to study the relationship between the deep microbial biosphere and a series of ˜ 2 km deep subseafloor coalbeds and to explore the limits of life in the deepest horizons ever probed by scientific ocean drilling. To address these scientific objectives, we penetrated a 2.466 km deep sedimentary sequence with a series of lignite layers buried around 2 km below the seafloor. The cored sediments, as well as cuttings and logging data, showed a record of dynamically changing depositional environments in the former forearc basin off the Shimokita Peninsula during the late Oligocene and Miocene, ranging from warm-temperate coastal backswamps to a cool water continental shelf. The occurrence of small microbial populations and their methanogenic activity were confirmed down to the bottom of the hole by microbiological and biogeochemical analyses. The factors controlling the size and viability of ultra-deep microbial communities in those warm sedimentary habitats could be the increase in demand of energy and water expended on the enzymatic repair of biomolecules as a function of the burial depth. Expedition 337 provided a test ground for the use of riser-drilling technology to address geobiological and biogeochemical objectives and was therefore a crucial step toward the next phase of deep scientific ocean drilling.

Reduction of microbial contamination and improvement of germination of sweet basil (Ocimum basilicum L.) seeds via surface dielectric barrier discharge

NASA Astrophysics Data System (ADS)

Ambrico, Paolo F.; Šimek, Milan; Morano, Massimo; De Miccolis Angelini, Rita M.; Minafra, Angelantonio; Trotti, Pasquale; Ambrico, Marianna; Prukner, Václav; Faretra, Francesco

2017-08-01

Naturally contaminated basil seeds were treated by a surface dielectric barrier discharge driven in the humid air by an amplitude modulated AC high voltage to avoid heat shock. In order to avoid direct contact of seeds with microdischarge filaments, the seeds to be treated were placed at sufficient distance from the surface discharge. After treatment, the seeds were analyzed in comparison with control samples for their microbial contamination as well as for the capability of germination and seedling growth. Moreover, chemical modification of seed surface was observed through the elemental energy dispersive x-ray analysis and wettability tests. We found that treatment applied at 20% duty cycle (effective discharge duration up to 20 s) significantly decreases microbial load without reducing the viability of the seeds. On the other side, seedling growth was considerably accelerated after the treatment, and biometric growth parameters of seedlings (total length, weight, leaf extension) considerably increased compared to the controls. Interestingly, scanning electron microscopy images taken for the different duration of treatment revealed that seed radicle micropylar regions underwent significant morphological changes while the coat was substantially undamaged. Inside the seed, the embryo seemed to be well preserved while the endosperm body was detached from the epithelial tegument. A total of 9 different genera of fungi were recovered from the analyzed seeds. Scanning electron microscopy images revealed that conidia were localized especially in the micropylar region, and after plasma treatment, most of them showed substantial damages. Therefore, the overall effect of the treatment of naturally contaminated seeds by reactive oxygen and nitrogen species produced by plasma and the consequent changes in surface chemistry and microbial load can significantly improve seed vigor.

Outgrowth of fibroblast cells from goat skin explants in three different culture media and the establishment of cell lines.

PubMed

Singh, Mahipal; Sharma, Anil K

2011-02-01

Three different commercially available media, known to support human and porcine-specific fibroblast cultures, were tested for their growth potential on goat skin explants. Although outgrowth of fibroblasts was observed in all media tested, irrespective of breed, porcine-specific media exhibited higher rate of growth. Using this media, three fibroblast cell lines (GSF289, GSF737, and GSF2010) from ear skin explants of normal healthy dairy goats of Kiko and Saanen breed were successfully established in culture. Liquid nitrogen stocks of these frozen cells had a viability rate of 96.2% in in vitro cultures. These cells were morphologically indistinguishable from the cell stocks prior to freezing. Analysis of the growth of a fifth passage culture revealed an 'S' shaped growth curve with a population doubling time of 25 h. The cell lines were found negative for microbial, fungal, and mycoplasma contaminations. These goat skin fibroblast lines and the simple method of their isolation and freezing with high rate of viability will provide additional tools to study molecular mechanisms that regulate fibroblast function and for genetic manipulation of small ruminants.

Preparation and characterization of aminoethyl hydroxypropyl starch modified with collagen peptide.

PubMed

Wen, Huigao; Hu, Jin; Ge, Hongyu; Zou, Shengqiong; Xiao, Yao; Li, Ya; Feng, Han; Fan, Lihong

2017-08-01

The preparation of aminoethyl hydroxypropyl starch collagen peptide (AEHPS-COP) was via an enzyme-catalyzed reaction between amino groups in aminoethyl hydroxypropyl starch (AEHPS) and γ-carboxamide groups in collagen peptide (COP) by using microbial transglutaminase (MTGase) as biocatalyst. As an intermediate reactant, AEHPS was synthesized from hydroxypropyl starch (HPS) and 2-chloroethylamine hydrochloride (CEH). The chemical structures of HPS, AEHPS and AEHPS-COP were characterized by Fourier transform infrared spectroscopy (FT-IR) and 13 C nuclear magnetic resonance ( 13 C NMR). The reaction conditions that influenced the degree of substitution (DS) of AEHPS-COP were optimized, which included the reaction temperature, the reaction time, the mass ratio of collagen peptide to aminoethyl hydroxypropyl starch and the pH value. In addition, in vitro antioxidant activities of AEHPS-COP were evaluated through the scavenging activity of hydroxyl and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical. Furthermore, the methylthiazol tetrazolium (MTT) assay was applied to investigate the cell viability of AEHPS-COP. The results indicated that the AEHPS-COP exhibited better cell viability to L929 mouse fibroblast cells. Therefore, the AEHPS-COP showed a promising potential application in cosmetic, biomedical and pharmaceutical fields. Copyright © 2017 Elsevier B.V. All rights reserved.

A Mechanism for the Temporal Potentiation of Genipin to the Cytotoxicity of Cisplatin in Colon Cancer Cells.

PubMed

Wang, Ruihua; MoYung, K C; Zhao, Y J; Poon, Karen

2016-01-01

To investigate the potentiation effect of Genipin to Cisplatin induced cell senescence in HCT-116 colon cancer cells in vitro. Cell viability was estimated by Propidium iodide and Hoechst 3342, reactive oxygen species (ROS) with DHE, mitochondrial membrane potential (MMP) with JC-1 MMP assay Kit and electron current production with microbial fuel cells (MFC). Genipin inhibited the UCP2 mediated anti-oxidative proton leak significantly promoted the Cisplatin induced ROS and subsequent cell death, which was similar to that of UCP2-siRNA. Cells treated with Cisplatin alone or combined with Genipin, ROS negatively, while MMP positively correlated with cell viability. Cisplatin induced ROS was significantly decreased by detouring electrons to MFC, or increased by Genipin combined treatment. Compensatory effects of UCP2 up-regulation with time against Genipin treatment were suggested. Shorter the Genipin treatment before Cisplatin better promoted the Cisplatin induced ROS and subsequent cell death. The interaction of leaked electron with Cisplatin was important during ROS generation. Inhibition of UCP2-mediated proton leak with Genipin potentiated the cytotoxicity of Cisplatin. Owing to the compensatory effects against Genipin, shorter Genipin treatment before Cisplatin was recommended in order to achieve better potentiation effect.

Comparison of dairy desserts produced with a potentially probiotic mixed culture and dispersions obtained from Gracilaria birdiae and Gracilaria domingensis seaweeds used as thickening agents.

PubMed

Tavares Estevam, Adriana Carneiro; de Almeida, Michele Correia; de Oliveira, Tiago Almeida; Florentino, Eliane Rolim; Alonso Buriti, Flávia Carolina; Porto, Ana Lúcia Figueiredo

2017-09-20

Dairy desserts have emerged as interesting options for the incorporation of probiotics, bioactive ingredients and alternative sources of thickeners. This shows an opportunity to investigate the use of Gracilaria seaweeds in the formulation of potentially probiotic dairy desserts. This study aimed to compare the effects of dispersions obtained from Gracilaria domingensis and Gracilaria birdiae used as thickening agents on texture properties of dairy desserts fermented with SAB 440-A, composed of the starter Streptococcus thermophilus and the potential probiotics Bifidobacterium animalis and Lactobacillus acidophilus, and also to study their physicochemical characteristics, microbial viability and sensory acceptability. No significant differences between desserts with G. birdiae or G. domingensis dispersions regarding total solids, ash and fat content, as well as pH, titratable acidity, the viability of the microorganisms of the mixed culture and sensory acceptability were verified (P > 0.05). Nonetheless, the dessert with G. domingensis dispersion showed higher dietary fibre content and significantly increased firmness than the one produced with G. birdiae (P < 0.05). Moreover, B. animalis was able to maintain higher populations, above 7 log cfu g -1 during 21 days of storage of desserts, in the presence of either G. birdiae or G. domingensis dispersions, despite the fact that L. acidophilus has shown low viability in the final products. Therefore, the G. domingensis dispersion is suitable to be used as a thickening agent to produce dairy desserts with enhanced firmness and good sensory acceptability, it being also advisable to use only B. animalis as a probiotic for this product.

Bactericidal effect of photocatalytically-active nanostructured TiO2 surfaces on biofilms of the early oral colonizer, Streptococcus oralis.

PubMed

Westas, Emma; Hayashi, Mariko; Cecchinato, Francesca; Wennerberg, Ann; Andersson, Martin; Jimbo, Ryo; Davies, Julia R

2017-08-01

This study evaluated the photocatalytic bactericidal effect of nanostructured anatase-rich titanium dioxide (TiO 2 ) on microbial biofilms. Commercially pure titanium discs were spin-coated with photocatalytic TiO 2 nanoparticles (P25). Uncoated discs were used as control (CTRL). Half of the CTRL and half of the P25-coated surfaces were coated with purified saliva (SAL) to give four different groups (CTRL, CTRL + SAL, P25 and P25 + SAL). Streptococcus oralis were allowed to form biofilms on the discs for 18 h and non-adherent cells were rinsed off. Bacterial viability was assessed at time 0 with Live/Dead BacLight staining and epifluorescence microscopy. The remaining discs were divided into a non-UV group and UVA-irradiated (+UV) group (irradiation time, 6 or 24 h). Thereafter, viability was assessed as above. Viability at time 0 was high and no dead cells were seen on any of the surfaces, even after 24 h, in the absence of UVA. However, after 24 h of exposure, the proportion of viable cells was reduced by 40% on the P25 discs compared to 0 and 6 h, and this effect was enhanced with a salivary pellicle. Members of mixed species biofilms differ in their susceptibility to the bactericidal effect of the surfaces tested and further investigations are needed to optimize the conditions. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 2321-2328, 2017. © 2017 Wiley Periodicals, Inc.

Enhanced viability of Lactobacillus reuteri for probiotics production in mixed solid-state fermentation in the presence of Bacillus subtilis.

PubMed

Zhang, Yi-Ran; Xiong, Hai-Rong; Guo, Xiao-Hua

2014-01-01

In order to develop a multi-microbe probiotic preparation of Lactobacillus reuteri G8-5 and Bacillus subtilis MA139 in solid-state fermentation, a series of parameters were optimized sequentially in shake flask culture. The effect of supplementation of B. subtilis MA139 as starters on the viability of L. reuteri G8-5 was also explored. The results showed that the optimized process was as follows: water content, 50 %; initial pH of diluted molasses, 6.5; inocula volume, 2 %; flask dry contents, 30∼35 g/250 g without sterilization; and fermentation time, 2 days. The multi-microbial preparations finally provided the maximum concentration of Lactobacillus of about 9.01 ± 0.15 log CFU/g and spores of Bacillus of about 10.30 ± 0.08 log CFU/g. Compared with pure fermentation of L. reuteri G8-5, significantly high viable cells, low value of pH, and reducing sugar in solid substrates were achieved in mixed fermentation in the presence of B. subtilis MA139 (P < 0.05). Meanwhile, the mixed fermentation showed the significantly higher antimicrobial activity against E. coli K88 (P < 0.05). Based on the overall results, the optimized process enhanced the production of multi-microbe probiotics in solid-state fermentation with low cost. Moreover, the viability of L. reuteri G8-5 could be significantly enhanced in the presence of B. subtilis MA139 in solid-state fermentation, which favored the production of probiotics for animal use.

Blue light irradiation triggers the antimicrobial potential of ZnO nanoparticles on drug-resistant Acinetobacter baumannii.

PubMed

Yang, Ming-Yeh; Chang, Kai-Chih; Chen, Liang-Yu; Wang, Po-Ching; Chou, Chih-Chiang; Wu, Zhong-Bin; Hu, Anren

2018-03-01

Photodynamic inactivation (PDI) is a non-invasive and safe therapeutic method for microbial infections. Bacterial antibiotic resistance is caused by antibiotics abuse. Drug-resistant Acinetobacter spp. is a serious problem in hospitals around the world. These pathogens from nosocomial infections have high mortality rates in frailer people, and Acinetobacter spp. is commonly found in immunocompromised patients. Visible light is safer than ultraviolet light (UV) for PDI of nosocomial pathogens with mammalian cells. Zinc oxide nanoparticles (ZnO-NPs) were used in this study as an antimicrobial agent and a photosensitizer. ZnO is recognized as safe and has extensive usage in food additives, medical and cosmetic products. In this study, we used 0.125 mg/ml ZnO-NPs combined with 10.8 J/cm 2 blue light (BL) on Acinetobacter baumannii (A. baumannii) that could significantly reduce microbial survival. However, individual exposure to ZnO-NPs does not affect the viability of A. baumannii. BL irradiation could trigger the antimicrobial ability of ZnO nanoparticles on A. baumannii. The mechanism of photocatalytic ZnO-NPs treatment for sterilization occurs through bacterial membrane disruptions. Otherwise, the photocatalytic ZnO-NPs treatment showed high microbial eradication in nosocomial pathogens, including colistin-resistant and imipenem-resistant A. baumannii and Klebsiella pneumoniae. Based on our results, the photocatalytic ZnO-NPs treatment could support hygiene control and clinical therapies without antibiotics to nosocomial bacterial infections. Copyright © 2018. Published by Elsevier B.V.

Base-Catalyzed Depolymerization of Solid Lignin-Rich Streams Enables Microbial Conversion

DOE PAGES

Rodriguez, Alberto; Salvachúa, Davinia; Katahira, Rui; ...

2017-08-01

Lignin valorization offers significant potential to enhance the economic viability of lignocellulosic biorefineries. However, because of its heterogeneous and recalcitrant nature, conversion of lignin to value-added coproducts remains a considerable technical challenge. Here, we employ base-catalyzed depolymerization (BCD) using a process-relevant solid lignin stream produced via deacetylation, mechanical refining, and enzymatic hydrolysis to enable biological lignin conversion. BCD was conducted with the solid lignin substrate over a range of temperatures at two NaOH concentrations, and the results demonstrate that the lignin can be partially extracted and saponified at temperatures as low as 60 degrees C. At 120 °C and 2%more » NaOH, the high extent of lignin solubility was accompanied by a considerable decrease in the lignin average molecular weight and the release of lignin-derived monomers including hydroxycinnamic acids. BCD liquors were tested for microbial growth using seven aromatic-catabolizing bacteria and two yeasts. Three organisms (Pseudomonas putida KT2440, Rhodotorula mucilaginosa, and Corynebacterium glutamicum) tolerate high BCD liquor concentrations (up to 90% v/v) and rapidly consume the main lignin-derived monomers, resulting in lignin conversion of up to 15%. Furthermore, as a proof of concept, muconic acid production from a representative lignin BCD liquor was demonstrated with an engineered P. putida KT2440 strain. Our results highlight the potential for a mild lignin depolymerization process to enhance the microbial conversion of solid lignin-rich biorefinery streams.« less

Base-Catalyzed Depolymerization of Solid Lignin-Rich Streams Enables Microbial Conversion

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

Rodriguez, Alberto; Salvachúa, Davinia; Katahira, Rui

Lignin valorization offers significant potential to enhance the economic viability of lignocellulosic biorefineries. However, because of its heterogeneous and recalcitrant nature, conversion of lignin to value-added coproducts remains a considerable technical challenge. Here, we employ base-catalyzed depolymerization (BCD) using a process-relevant solid lignin stream produced via deacetylation, mechanical refining, and enzymatic hydrolysis to enable biological lignin conversion. BCD was conducted with the solid lignin substrate over a range of temperatures at two NaOH concentrations, and the results demonstrate that the lignin can be partially extracted and saponified at temperatures as low as 60 degrees C. At 120 °C and 2%more » NaOH, the high extent of lignin solubility was accompanied by a considerable decrease in the lignin average molecular weight and the release of lignin-derived monomers including hydroxycinnamic acids. BCD liquors were tested for microbial growth using seven aromatic-catabolizing bacteria and two yeasts. Three organisms (Pseudomonas putida KT2440, Rhodotorula mucilaginosa, and Corynebacterium glutamicum) tolerate high BCD liquor concentrations (up to 90% v/v) and rapidly consume the main lignin-derived monomers, resulting in lignin conversion of up to 15%. Furthermore, as a proof of concept, muconic acid production from a representative lignin BCD liquor was demonstrated with an engineered P. putida KT2440 strain. Our results highlight the potential for a mild lignin depolymerization process to enhance the microbial conversion of solid lignin-rich biorefinery streams.« less

Starter Culture Selection for Making Chinese Sesame-Flavored Liquor Based on Microbial Metabolic Activity in Mixed-Culture Fermentation

PubMed Central

Wu, Qun; Ling, Jie

2014-01-01

Selection of a starter culture with excellent viability and metabolic activity is important for inoculated fermentation of traditional food. To obtain a suitable starter culture for making Chinese sesame-flavored liquor, the yeast and bacterium community structures were investigated during spontaneous and solid-state fermentations of this type of liquor. Five dominant species in spontaneous fermentation were identified: Saccharomyces cerevisiae, Pichia membranaefaciens, Issatchenkia orientalis, Bacillus licheniformis, and Bacillus amyloliquefaciens. The metabolic activity of each species in mixed and inoculated fermentations of liquor was investigated in 14 different cocultures that used different combinations of these species. The relationships between the microbial species and volatile metabolites were analyzed by partial least-squares (PLS) regression analysis. We found that S. cerevisiae was positively correlated to nonanal, and B. licheniformis was positively associated with 2,3-butanediol, isobutyric acid, guaiacol, and 4-vinyl guaiacol, while I. orientalis was positively correlated to butyric acid, isovaleric acid, hexanoic acid, and 2,3-butanediol. These three species are excellent flavor producers for Chinese liquor. Although P. membranaefaciens and B. amyloliquefaciens were not efficient flavor producers, the addition of them alleviated competition among the other three species and altered their growth rates and flavor production. As a result, the coculture of all five dominant species produced the largest amount of flavor compounds. The result indicates that flavor producers and microbial interaction regulators are important for inoculated fermentation of Chinese sesame-flavored liquor. PMID:24814798

Construction of Viable Soil Defined Media Using Quantitative Metabolomics Analysis of Soil Metabolites

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

Jenkins, Stefan; Swenson, Tami L.; Lau, Rebecca

Exometabolomics enables analysis of metabolite utilization of low molecular weight organic substances by soil bacteria. Thus, environmentally-based defined media are needed to examine ecologically relevant patterns of substrate utilization. Here, we describe an approach for the construction of defined media using untargeted characterization of water soluble soil microbial metabolites from a saprolite soil collected from the Oak Ridge Field Research Center (ORFRC). To broadly characterize metabolites, both liquid chromatography mass spectrometry (LC/MS) and gas chromatography mass spectrometry (GC/MS) were used. With this approach, 96 metabolites were identified, including amino acids, amino acid derivatives, sugars, sugar alcohols, mono- and di-carboxylic acids,more » nucleobases, and nucleosides. From this pool of metabolites, 25 were quantified. Molecular weight cut-off filtration determined the fraction of carbon accounted for by the quantified metabolites and revealed that these soil metabolites have an uneven quantitative distribution (e.g., trehalose accounted for 9.9% of the < 1 kDa fraction). This quantitative information was used to formulate two soil defined media (SDM), one containing 23 metabolites (SDM1) and one containing 46 (SDM2). To evaluate the viability of the SDM, we examined the growth of 30 phylogenetically diverse soil bacterial isolates from the ORFRC field site. The simpler SDM1 supported the growth of 13 isolates while the more complex SDM2 supported 15 isolates. To investigate SDM1 substrate preferences, one isolate, Pseudomonas corrugata strain FW300-N2E2 was selected for a time-series exometabolomics analysis. Interestingly, it was found that this organism preferred lower-abundance substrates such as guanine, glycine, proline and arginine and glucose and did not utilize the more abundant substrates maltose, mannitol, trehalose and uridine. These results demonstrate the viability and utility of using exometabolomics to construct a tractable environmentally relevant media. We anticipate that this approach can be expanded to other environments to enhance isolation and characterization of diverse microbial communities.« less

Construction of Viable Soil Defined Media Using Quantitative Metabolomics Analysis of Soil Metabolites

PubMed Central

Jenkins, Stefan; Swenson, Tami L.; Lau, Rebecca; Rocha, Andrea M.; Aaring, Alex; Hazen, Terry C.; Chakraborty, Romy; Northen, Trent R.

2017-01-01

Exometabolomics enables analysis of metabolite utilization of low molecular weight organic substances by soil bacteria. Environmentally-based defined media are needed to examine ecologically relevant patterns of substrate utilization. Here, we describe an approach for the construction of defined media using untargeted characterization of water soluble soil microbial metabolites from a saprolite soil collected from the Oak Ridge Field Research Center (ORFRC). To broadly characterize metabolites, both liquid chromatography mass spectrometry (LC/MS) and gas chromatography mass spectrometry (GC/MS) were used. With this approach, 96 metabolites were identified, including amino acids, amino acid derivatives, sugars, sugar alcohols, mono- and di-carboxylic acids, nucleobases, and nucleosides. From this pool of metabolites, 25 were quantified. Molecular weight cut-off filtration determined the fraction of carbon accounted for by the quantified metabolites and revealed that these soil metabolites have an uneven quantitative distribution (e.g., trehalose accounted for 9.9% of the <1 kDa fraction). This quantitative information was used to formulate two soil defined media (SDM), one containing 23 metabolites (SDM1) and one containing 46 (SDM2). To evaluate the viability of the SDM, we examined the growth of 30 phylogenetically diverse soil bacterial isolates from the ORFRC field site. The simpler SDM1 supported the growth of 13 isolates while the more complex SDM2 supported 15 isolates. To investigate SDM1 substrate preferences, one isolate, Pseudomonas corrugata strain FW300-N2E2 was selected for a time-series exometabolomics analysis. Interestingly, it was found that this organism preferred lower-abundance substrates such as guanine, glycine, proline and arginine and glucose and did not utilize the more abundant substrates maltose, mannitol, trehalose and uridine. These results demonstrate the viability and utility of using exometabolomics to construct a tractable environmentally relevant media. We anticipate that this approach can be expanded to other environments to enhance isolation and characterization of diverse microbial communities. PMID:29312276

Effects of aqueous uranyl speciation on the kinetics of microbial uranium reduction

DOE PAGES

Belli, Keaton M.; DiChristina, Thomas J.; Van Cappellen, Philippe; ...

2015-02-16

The ability to predict the success of the microbial reduction of soluble U(VI) to highly insoluble U(IV) as an in situ bioremediation strategy is complicated by the wide range of geochemical conditions at contaminated sites and the strong influence of aqueous uranyl speciation on the bioavailability and toxicity of U(VI) to metal-reducing bacteria. In order to determine the effects of aqueous uranyl speciation on uranium bioreduction kinetics, incubations and viability assays with Shewanella putrefaciens strain 200 were conducted over a range of pH and dissolved inorganic carbon (DIC), Ca 2+, and Mg 2+ concentrations. A speciation-dependent kinetic model was developedmore » to reproduce the observed time series of total dissolved uranium concentration over the range of geochemical conditions tested. The kinetic model yielded the highest rate constant for the reduction of uranyl non-carbonate species (i.e., the ‘free’ hydrated uranyl ion, uranyl hydroxides, and other minor uranyl complexes), indicating that they represent the most readily reducible fraction of U(VI) despite being the least abundant uranyl species in solution. In the presence of DIC, Ca 2+, and Mg 2+ is suppressed during the formation of more bioavailable uranyl non-carbonate species and resulted in slower bioreduction rates. At high concentrations of bioavailable U(VI), however, uranium toxicity to S. putrefaciens inhibited bioreduction, and viability assays confirmed that the concentration of non-carbonate uranyl species best predicts the degree of toxicity. The effect of uranium toxicity was accounted for by incorporating the free ion activity model of metal toxicity into the bioreduction rate law. These results demonstrate that, in the absence of competing terminal electron acceptors, uranium bioreduction kinetics can be predicted over a wide range of geochemical conditions based on the bioavailability and toxicity imparted on U(VI) by solution composition. Finally, these findings also imply that the concentration of uranyl non-carbonate species, despite being extremely low, is a determining factor controlling uranium bioreduction at contaminated sites.« less

Construction of Viable Soil Defined Media Using Quantitative Metabolomics Analysis of Soil Metabolites

DOE PAGES

Jenkins, Stefan; Swenson, Tami L.; Lau, Rebecca; ...

2017-12-22

Exometabolomics enables analysis of metabolite utilization of low molecular weight organic substances by soil bacteria. Thus, environmentally-based defined media are needed to examine ecologically relevant patterns of substrate utilization. Here, we describe an approach for the construction of defined media using untargeted characterization of water soluble soil microbial metabolites from a saprolite soil collected from the Oak Ridge Field Research Center (ORFRC). To broadly characterize metabolites, both liquid chromatography mass spectrometry (LC/MS) and gas chromatography mass spectrometry (GC/MS) were used. With this approach, 96 metabolites were identified, including amino acids, amino acid derivatives, sugars, sugar alcohols, mono- and di-carboxylic acids,more » nucleobases, and nucleosides. From this pool of metabolites, 25 were quantified. Molecular weight cut-off filtration determined the fraction of carbon accounted for by the quantified metabolites and revealed that these soil metabolites have an uneven quantitative distribution (e.g., trehalose accounted for 9.9% of the < 1 kDa fraction). This quantitative information was used to formulate two soil defined media (SDM), one containing 23 metabolites (SDM1) and one containing 46 (SDM2). To evaluate the viability of the SDM, we examined the growth of 30 phylogenetically diverse soil bacterial isolates from the ORFRC field site. The simpler SDM1 supported the growth of 13 isolates while the more complex SDM2 supported 15 isolates. To investigate SDM1 substrate preferences, one isolate, Pseudomonas corrugata strain FW300-N2E2 was selected for a time-series exometabolomics analysis. Interestingly, it was found that this organism preferred lower-abundance substrates such as guanine, glycine, proline and arginine and glucose and did not utilize the more abundant substrates maltose, mannitol, trehalose and uridine. These results demonstrate the viability and utility of using exometabolomics to construct a tractable environmentally relevant media. We anticipate that this approach can be expanded to other environments to enhance isolation and characterization of diverse microbial communities.« less

Thermal, High Pressure, and Electric Field Processing Effects on Plant Cell Membrane Integrity and Relevance to Fruit and Vegetable Quality

PubMed Central

Gonzalez, Maria E; Barrett, Diane M

2010-01-01

Advanced food processing methods that accomplish inactivation of microorganisms but minimize adverse thermal exposure are of great interest to the food industry. High pressure (HP) and pulsed electric field (PEF) processing are commercially applied to produce high quality fruit and vegetable products in the United States, Europe, and Japan. Both microbial and plant cell membranes are significantly altered following exposure to heat, HP, or PEF. Our research group sought to quantify the degree of damage to plant cell membranes that occurs as a result of exposure to heat, HP, or PEF, using the same analytical methods. In order to evaluate whether new advanced processing methods are superior to traditional thermal processing methods, it is necessary to compare them. In this review, we describe the existing state of knowledge related to effects of heat, HP, and PEF on both microbial and plant cells. The importance and relevance of compartmentalization in plant cells as it relates to fruit and vegetable quality is described and various methods for quantification of plant cell membrane integrity are discussed. These include electrolyte leakage, cell viability, and proton nuclear magnetic resonance (1H-NMR). PMID:20492210

Recent advances in microbial fermentation for dairy and health

PubMed Central

Arendt, Elke; Hill, Colin; Stanton, Catherine; Ross, R Paul

2017-01-01

Microbial fermentation has been used historically for the preservation of foods, the health benefits of which have since come to light. Early dairy fermentations depended on the spontaneous activity of the indigenous microbiota of the milk. Modern fermentations rely on defined starter cultures with desirable characteristics to ensure consistency and commercial viability. The selection of defined starters depends on specific phenotypes that benefit the product by guaranteeing shelf life and ensuring safety, texture, and flavour. Lactic acid bacteria can produce a number of bioactive metabolites during fermentation, such as bacteriocins, biogenic amines, exopolysaccharides, and proteolytically released peptides, among others. Prebiotics are added to food fermentations to improve the performance of probiotics. It has also been found that prebiotics fermented in the gut can have benefits that go beyond helping probiotic growth. Studies are now looking at how the fermentation of prebiotics such as fructo-oligosaccharides can help in the prevention of diseases such as osteoporosis, obesity, and colorectal cancer. The potential to prevent or even treat disease through the fermentation of food is a medically and commercially attractive goal and is showing increasing promise. However, the stringent regulation of probiotics is beginning to detrimentally affect the field and limit their application. PMID:28649371

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

Schneider, Teresa; Graeff-Honninger, Simone; French, William Todd

The production of biodiesel has notably increased over the past decade. Currently, plant oil is the main feedstock for biodiesel production, but, due to concerns related to the competition with food production, alternative oil feedstocks have to be found. Oleaginous yeasts are known to produce high amounts of lipids, but no integrated process from microbial fermentation to final biodiesel production has reached commercial realization yet due to economic constraints. Therefore, growth and lipid production of red yeast Rhodotorula glutinis was tested on low-cost substrates, namely, wastewaters from potato, fruit juice, and lettuce processing. Additionally, the production of carotenoids as high-valuemore » by-products was examined. All evaluated wastewaters met the general criteria for microbial lipid production. However, no significant increase in lipid content was observed, probably due to lack of available carbon in wastewaters from fruit juice and lettuce processing, and excess of available nitrogen in potato processing wastewater, respectively. During growth on wastewaters from fruit juice and lettuce processing the carotenoid content increased significantly in the first 48 hours. The relations between carbon content, nitrogen content, and carotenoid production need to be further assessed. For economic viability, lipid and carotenoid production needs to be increased significantly. Lastly, the screening of feedstocks should be extended to other wastewaters.« less

Implications of SPION and NBT nanoparticles upon in-vitro and in-situ biodegradation of LDPE film.

PubMed

Kapri, Anil; Zaidi, M G H; Goel, Reeta

2010-06-01

Comparative influence of two nanoparticles viz. superparamagnetic iron oxide nanoparticles (SPION) and nanobarium titanate (NBT) was studied upon the in-vitro and in-situ low-density polyethylene (LDPE) biodegradation efficiency of a potential polymer-degrading microbial consortium. Supplementation of 0.01% concentration (w/v) of the nanoparticles in minimal broth significantly increased the bacterial growth, along with early onset of the exponential phase. Under in-vitro conditions, lambda-max shifts were quicker with nanoparticles and Fourier transform infrared spectroscopy (FTIR) illustrated significant changes in CH/CH2 vibrations, along with introduction of hydroxyl residues in the polymer backbone. Further, simultaneous thermogravimetric-differential thermogravimetry-differential thermal analysis (TG-DTG-DTA) reported multiple-step decomposition of LDPE degraded in the presence of nanoparticles. These findings were supported by scanning electron micrographs (SEM) which revealed greater dissolution of film surface in the presence of nanoparticles. Furthermore, progressive degradation of the film was greatly enhanced when it was incubated under soil conditions for 3 months with the nanoparticles. The study highlights the significance of bacteria-nanoparticle interactions which can dramatically influence key metabolic processes like biodegradation. The authors also propose the exploration of nanoparticles to influence various other microbial processes for commercial viabilities.

Alginic Acid-Aided Dispersion of Carbon Nanotubes, Graphene, and Boron Nitride Nanomaterials for Microbial Toxicity Testing

PubMed Central

Chang, Chong Hyun

2018-01-01

Robust evaluation of potential environmental and health risks of carbonaceous and boron nitride nanomaterials (NMs) is imperative. However, significant agglomeration of pristine carbonaceous and boron nitride NMs due to strong van der Waals forces renders them not suitable for direct toxicity testing in aqueous media. Here, the natural polysaccharide alginic acid (AA) was used as a nontoxic, environmentally relevant dispersant with defined composition to disperse seven types of carbonaceous and boron nitride NMs, including multiwall carbon nanotubes, graphene, boron nitride nanotubes, and hexagonal boron nitride flakes, with various physicochemical characteristics. AA’s biocompatibility was confirmed by examining AA effects on viability and growth of two model microorganisms (the protozoan Tetrahymena thermophila and the bacterium Pseudomonas aeruginosa). Using 400 mg·L−1 AA, comparably stable NM (200 mg·L−1) stock dispersions were obtained by 30-min probe ultrasonication. AA non-covalently interacted with NM surfaces and improved the dispersibility of NMs in water. The dispersion stability varied with NM morphology and size rather than chemistry. The optimized dispersion protocol established here can facilitate preparing homogeneous NM dispersions for reliable exposures during microbial toxicity testing, contributing to improved reproducibility of toxicity results. PMID:29385723

Alginic Acid-Aided Dispersion of Carbon Nanotubes, Graphene, and Boron Nitride Nanomaterials for Microbial Toxicity Testing.

PubMed

Wang, Ying; Mortimer, Monika; Chang, Chong Hyun; Holden, Patricia A

2018-01-30

Robust evaluation of potential environmental and health risks of carbonaceous and boron nitride nanomaterials (NMs) is imperative. However, significant agglomeration of pristine carbonaceous and boron nitride NMs due to strong van der Waals forces renders them not suitable for direct toxicity testing in aqueous media. Here, the natural polysaccharide alginic acid (AA) was used as a nontoxic, environmentally relevant dispersant with defined composition to disperse seven types of carbonaceous and boron nitride NMs, including multiwall carbon nanotubes, graphene, boron nitride nanotubes, and hexagonal boron nitride flakes, with various physicochemical characteristics. AA's biocompatibility was confirmed by examining AA effects on viability and growth of two model microorganisms (the protozoan Tetrahymena thermophila and the bacterium Pseudomonas aeruginosa ). Using 400 mg·L -1 AA, comparably stable NM (200 mg·L -1 ) stock dispersions were obtained by 30-min probe ultrasonication. AA non-covalently interacted with NM surfaces and improved the dispersibility of NMs in water. The dispersion stability varied with NM morphology and size rather than chemistry. The optimized dispersion protocol established here can facilitate preparing homogeneous NM dispersions for reliable exposures during microbial toxicity testing, contributing to improved reproducibility of toxicity results.

Tools for Accurate and Efficient Analysis of Complex Evolutionary Mechanisms in Microbial Genomes. Final Report

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

Nakhleh, Luay

I proposed to develop computationally efficient tools for accurate detection and reconstruction of microbes' complex evolutionary mechanisms, thus enabling rapid and accurate annotation, analysis and understanding of their genomes. To achieve this goal, I proposed to address three aspects. (1) Mathematical modeling. A major challenge facing the accurate detection of HGT is that of distinguishing between these two events on the one hand and other events that have similar "effects." I proposed to develop a novel mathematical approach for distinguishing among these events. Further, I proposed to develop a set of novel optimization criteria for the evolutionary analysis of microbialmore » genomes in the presence of these complex evolutionary events. (2) Algorithm design. In this aspect of the project, I proposed to develop an array of e cient and accurate algorithms for analyzing microbial genomes based on the formulated optimization criteria. Further, I proposed to test the viability of the criteria and the accuracy of the algorithms in an experimental setting using both synthetic as well as biological data. (3) Software development. I proposed the nal outcome to be a suite of software tools which implements the mathematical models as well as the algorithms developed.« less

Thermal, high pressure, and electric field processing effects on plant cell membrane integrity and relevance to fruit and vegetable quality.

PubMed

Gonzalez, Maria E; Barrett, Diane M

2010-09-01

Advanced food processing methods that accomplish inactivation of microorganisms but minimize adverse thermal exposure are of great interest to the food industry. High pressure (HP) and pulsed electric field (PEF) processing are commercially applied to produce high quality fruit and vegetable products in the United States, Europe, and Japan. Both microbial and plant cell membranes are significantly altered following exposure to heat, HP, or PEF. Our research group sought to quantify the degree of damage to plant cell membranes that occurs as a result of exposure to heat, HP, or PEF, using the same analytical methods. In order to evaluate whether new advanced processing methods are superior to traditional thermal processing methods, it is necessary to compare them. In this review, we describe the existing state of knowledge related to effects of heat, HP, and PEF on both microbial and plant cells. The importance and relevance of compartmentalization in plant cells as it relates to fruit and vegetable quality is described and various methods for quantification of plant cell membrane integrity are discussed. These include electrolyte leakage, cell viability, and proton nuclear magnetic resonance (¹H-NMR).

Anti-biofouling function of amorphous nano-Ta2O5 coating for VO2-based intelligent windows.

PubMed

Li, Jinhua; Guo, Geyong; Wang, Jiaxing; Zhou, Huaijuan; Shen, Hao; Yeung, Kelvin W K

2017-04-28

From environmental and health perspectives, the acquisition of a surface anti-biofouling property holds important significance for the usability of VO 2 intelligent windows. Herein, we firstly deposited amorphous Ta 2 O 5 nanoparticles on VO 2 film by the magnetron sputtering method. It was found that the amorphous nano-Ta 2 O 5 coating possessed a favorable anti-biofouling capability against Pseudomonas aeruginosa as an environmental microorganism model, behind which lay the mechanism that the amorphous nano-Ta 2 O 5 could interrupt the microbial membrane electron transport chain and significantly elevate the intracellular reactive oxygen species (ROS) level. A plausible relationship was established between the anti-biofouling activity and physicochemical nature of amorphous Ta 2 O 5 nanoparticles from the perspective of defect chemistry. ROS-induced oxidative damage gave rise to microbial viability loss. In addition, the amorphous nano-Ta 2 O 5 coating can endow VO 2 with favorable cytocompatibility with human skin fibroblasts. This study may provide new insights into understanding the anti-biofouling and antimicrobial actions of amorphous transition metal oxide nanoparticles, which is conducive to expanding their potential applications in environmental fields.

Anti-biofouling function of amorphous nano-Ta2O5 coating for VO2-based intelligent windows

NASA Astrophysics Data System (ADS)

Li, Jinhua; Guo, Geyong; Wang, Jiaxing; Zhou, Huaijuan; Shen, Hao; Yeung, Kelvin W. K.

2017-04-01

From environmental and health perspectives, the acquisition of a surface anti-biofouling property holds important significance for the usability of VO2 intelligent windows. Herein, we firstly deposited amorphous Ta2O5 nanoparticles on VO2 film by the magnetron sputtering method. It was found that the amorphous nano-Ta2O5 coating possessed a favorable anti-biofouling capability against Pseudomonas aeruginosa as an environmental microorganism model, behind which lay the mechanism that the amorphous nano-Ta2O5 could interrupt the microbial membrane electron transport chain and significantly elevate the intracellular reactive oxygen species (ROS) level. A plausible relationship was established between the anti-biofouling activity and physicochemical nature of amorphous Ta2O5 nanoparticles from the perspective of defect chemistry. ROS-induced oxidative damage gave rise to microbial viability loss. In addition, the amorphous nano-Ta2O5 coating can endow VO2 with favorable cytocompatibility with human skin fibroblasts. This study may provide new insights into understanding the anti-biofouling and antimicrobial actions of amorphous transition metal oxide nanoparticles, which is conducive to expanding their potential applications in environmental fields.

In vivo biofilm formation on stainless steel bonded retainers during different oral health-care regimens.

PubMed

Jongsma, Marije A; van der Mei, Henny C; Atema-Smit, Jelly; Busscher, Henk J; Ren, Yijin

2015-03-23

Retention wires permanently bonded to the anterior teeth are used after orthodontic treatment to prevent the teeth from relapsing to pre-treatment positions. A disadvantage of bonded retainers is biofilm accumulation on the wires, which produces a higher incidence of gingival recession, increased pocket depth and bleeding on probing. This study compares in vivo biofilm formation on single-strand and multi-strand retention wires with different oral health-care regimens. Two-centimetre wires were placed in brackets that were bonded to the buccal side of the first molars and second premolars in the upper arches of 22 volunteers. Volunteers used a selected toothpaste with or without the additional use of a mouthrinse containing essential oils. Brushing was performed manually. Regimens were maintained for 1 week, after which the wires were removed and the oral biofilm was collected to quantify the number of organisms and their viability, determine the microbial composition and visualize the bacteria by electron microscopy. A 6-week washout period was employed between regimens. Biofilm formation was reduced on single-strand wires compared with multi-strand wires; bacteria were observed to adhere between the strands. The use of antibacterial toothpastes marginally reduced the amount of biofilm on both wire types, but significantly reduced the viability of the biofilm organisms. Additional use of the mouthrinse did not result in significant changes in biofilm amount or viability. However, major shifts in biofilm composition were induced by combining a stannous fluoride- or triclosan-containing toothpaste with the mouthrinse. These shifts can be tentatively attributed to small changes in bacterial cell surface hydrophobicity after the adsorption of the toothpaste components, which stimulate bacterial adhesion to the hydrophobic oil, as illustrated for a Streptococcus mutans strain.

The effect of sulphur dioxide and oxygen on the viability and culturability of a strain of Acetobacter pasteurianus and a strain of Brettanomyces bruxellensis isolated from wine.

PubMed

du Toit, W J; Pretorius, I S; Lonvaud-Funel, A

2005-01-01

The objective of this study was to investigate the effects of free molecular and bound forms of sulphur dioxide and oxygen on the viability and culturability of a selected strain of Acetobacter pasteurianus and a selected strain of Brettanomyces bruxellensis in wine. Acetic acid bacteria and Brettanomyces/Dekkera yeasts associated with wine spoilage were isolated from bottled commercial red wines. One bacterium, A. pasteurianus strain A8, and one yeast, B. bruxellensis strain B3a, were selected for further study. The resistance to sulphur dioxide and the effect of oxygen addition on these two selected strains were determined by using plating and epifluorescence techniques for monitoring cell viability in wine. Acetobacter pasteurianus A8 was more resistant to sulphur dioxide than B. bruxellensis B3a, with the latter being rapidly affected by a short exposure time to free molecular form of sulphur dioxide. As expected, neither of these microbial strains was affected by the bound form of sulphur dioxide. The addition of oxygen negated the difference observed between plate and epifluorescence counts for A. pasteurianus A8 during storage, while it stimulated growth of B. bruxellensis B3a. Acetobacter pasteurianus A8 can survive under anaerobic conditions in wine in the presence of sulphur dioxide. Brettanomyces bruxellensis B3a is more sensitive to sulphur dioxide than A. pasteurianus A8, but can grow in the presence of oxygen. Care should be taken to exclude oxygen from contact with wine when it is being transferred or moved. Wine spoilage can be avoided by preventing growth of undesirable acetic acid bacteria and Brettanomyces/Dekkera yeasts through the effective use of sulphur dioxide and the management of oxygen throughout the winemaking process.

Effects of a simulated martian UV flux on the cyanobacterium, Chroococcidiopsis sp. 029.

PubMed

Cockell, Charles S; Schuerger, Andrew C; Billi, Daniela; Friedmann, E Imre; Panitz, Corinna

2005-04-01

Dried monolayers of Chroococcidiopsis sp. 029, a desiccation-tolerant, endolithic cyanobacterium, were exposed to a simulated martian-surface UV and visible light flux, which may also approximate to the worst-case scenario for the Archean Earth. After 5 min, there was a 99% loss of cell viability, and there were no survivors after 30 min. However, this survival was approximately 10 times higher than that previously reported for Bacillus subtilis. We show that under 1 mm of rock, Chroococcidiopsis sp. could survive (and potentially grow) under the high martian UV flux if water and nutrient requirements for growth were met. In isolated cells, phycobilisomes and esterases remained intact hours after viability was lost. Esterase activity was reduced by 99% after a 1-h exposure, while 99% loss of autofluorescence required a 4-h exposure. However, cell morphology was not changed, and DNA was still detectable by 4',6-diamidino-2-phenylindole staining after an 8-h exposure (equivalent to approximately 1 day on Mars at the equator). Under 1 mm of simulant martian soil or gneiss, the effect of UV radiation could not be detected on esterase activity or autofluorescence after 4 h. These results show that under the intense martian UV flux the morphological signatures of life can persist even after viability, enzymatic activity, and pigmentation have been destroyed. Finally, the global dispersal of viable, isolated cells of even this desiccation-tolerant, ionizing-radiation-resistant microorganism on Mars is unlikely as they are killed quickly by unattenuated UV radiation when in a desiccated state. These findings have implications for the survival of diverse microbial contaminants dispersed during the course of human exploratory class missions on the surface of Mars.

Evidence that Formation of Protoanemonin from Metabolites of 4-Chlorobiphenyl Degradation Negatively Affects the Survival of 4-Chlorobiphenyl-Cometabolizing Microorganisms

PubMed Central

Blasco, R.; Mallavarapu, M.; Wittich, R.; Timmis, K. N.; Pieper, D. H.

1997-01-01

A rapid decline in cell viability of different PCB-metabolizing organisms was observed in soil microcosms amended with 4-chlorobiphenyl. The toxic effect could not be attributed to 4-chlorobiphenyl but was due to a compound formed from the transformation of 4-chlorobiphenyl by the natural microflora. Potential metabolites of 4-chlorobiphenyl, 4-chlorobenzoate and 4-chlorocatechol, caused similar toxic effects. We tested the hypothesis that the toxic effects are due to the formation of protoanemonin, a plant-derived antibiotic, which is toxic to microorganisms and which has been shown to be formed from 4-chlorocatechol by enzymes of the 3-oxoadipate pathway. Consistent with our hypothesis, addition to soil microcosms of strains able to reroute intermediary 4-chlorocatechol from the 3-oxoadipate pathway and into the meta-cleavage pathway or able to mineralize 4-chlorocatechol by a modified ortho-cleavage pathway resulted in reversal of this toxic effect. Surprisingly, while direct addition of protoanemonin influenced both the viability of fungi and the microbial activity of the soil microcosm, there was little effect on bacterial viability due to its rapid degradation. This rapid degradation accounts for our inability to detect this compound in soils amended with 4-chlorocatechol. However, significant accumulation of protoanemonin was observed by a mixed bacterial community enriched with benzoate or a mixture of benzoate and 4-methylbenzoate, providing the metabolic potential of the soil to form protoanemonin. The effects of soil heterogeneity and microcosm interactions are discussed in relation to the different effects of protoanemonin when applied as a shock load and when it is produced in small amounts from precursors over long periods. PMID:16535507

In vivo biofilm formation on stainless steel bonded retainers during different oral health-care regimens

PubMed Central

Jongsma, Marije A; van der Mei, Henny C; Atema-Smit, Jelly; Busscher, Henk J; Ren, Yijin

2015-01-01

Retention wires permanently bonded to the anterior teeth are used after orthodontic treatment to prevent the teeth from relapsing to pre-treatment positions. A disadvantage of bonded retainers is biofilm accumulation on the wires, which produces a higher incidence of gingival recession, increased pocket depth and bleeding on probing. This study compares in vivo biofilm formation on single-strand and multi-strand retention wires with different oral health-care regimens. Two-centimetre wires were placed in brackets that were bonded to the buccal side of the first molars and second premolars in the upper arches of 22 volunteers. Volunteers used a selected toothpaste with or without the additional use of a mouthrinse containing essential oils. Brushing was performed manually. Regimens were maintained for 1 week, after which the wires were removed and the oral biofilm was collected to quantify the number of organisms and their viability, determine the microbial composition and visualize the bacteria by electron microscopy. A 6-week washout period was employed between regimens. Biofilm formation was reduced on single-strand wires compared with multi-strand wires; bacteria were observed to adhere between the strands. The use of antibacterial toothpastes marginally reduced the amount of biofilm on both wire types, but significantly reduced the viability of the biofilm organisms. Additional use of the mouthrinse did not result in significant changes in biofilm amount or viability. However, major shifts in biofilm composition were induced by combining a stannous fluoride- or triclosan-containing toothpaste with the mouthrinse. These shifts can be tentatively attributed to small changes in bacterial cell surface hydrophobicity after the adsorption of the toothpaste components, which stimulate bacterial adhesion to the hydrophobic oil, as illustrated for a Streptococcus mutans strain. PMID:25572920

Redefining the effect of salt on thermophilic starter cell viability, culturability and metabolic activity in cheese.

PubMed

Hickey, C D; Fallico, V; Wilkinson, M G; Sheehan, J J

2018-02-01

This study investigated the differential effect of salt concentration in the outside and inside layers of brine salted cheeses on viability, culturability and enzyme activity of starter bacteria. The high-salt environment of the outside layer caused a sharp decrease in L. helveticus viability as measured by traditional plate counts. Remarkably, this was associated with lower release of intracellular enzymes (LDH), reduced levels of proteolysis and larger membrane integrity as measured by flow cytometry (FC) following classical Live/Dead staining. FC analysis of light scattering properties highlighted a significant reduction in size and granularity of the microbiota located in the cheese surface, suggestive of cell shrinkage and condensation of internal macromolecules probably due to hyperosmotic stress. The microbiota of the cheese surface were found to experience greater oxidative stress, as measured by FC analysis of the total levels of reactive oxygen species, compared to that of the interior layer. These results lead us to postulate that the physiology and health status of the microbiota were significantly different in the outer and inner layers of the cheese. The hyperosmotic environment of the outer layer resulted in reduced cell lysis, as measurable by assays based upon membrane integrity, but rather triggered cell death via mechanisms involving cell shrinkage and ROS-mediated damage of vital intracellular components. This study challenges the current thinking on how salt controls microbial activity in ripening cheese, especially in cheeses which are brine salted as local variations in biochemical ripening indices can differ significantly from the outside to the inside of a ripening cheese. Copyright © 2017 Elsevier Ltd. All rights reserved.

Natural biopolymer for preservation of microorganisms during sampling and storage.

PubMed

Sorokulova, Iryna; Watt, James; Olsen, Eric; Globa, Ludmila; Moore, Timothy; Barbaree, James; Vodyanoy, Vitaly

2012-01-01

Stability of microbial cultures during sampling and storage is a vital issue in various fields of medicine, biotechnology, food science, and forensics. We have developed a unique bacterial preservation process involving a non-toxic, water-soluble acacia gum polymer that eliminates the need for refrigerated storage of samples. The main goal of this study is to characterize the efficacy of acacia gum polymer for preservation of pathogenic bacteria (Bacillus anthracis and methicillin-resistant Staphylococcus aureus-MRSA) on different materials, used for swabbing and filtration: cotton, wool, polyester, rayon, charcoal cloth, and Whatman paper. Acacia gum polymer used for preservation of two pathogens has been shown to significantly protect bacteria during dehydration and storage in all tested samples at the range of temperatures (5-45°C for MRSA and 40-90°C for B. anthracis). Our results showed higher recovery as well as higher viability during the storage of both bacteria in all materials with acacia gum. Addition of acacia gum polymer to swabbing materials or filters will increase efficacy of sample collection and identification of pathogenic bacteria from locations such as hospitals or the environment. Proposed approach can also be used for long-term storage of culture collections, since acacia gum contributes to viability and stability of bacterial cultures. Copyright © 2011 Elsevier B.V. All rights reserved.

Microencapsulation of Bacterial Cells by Emulsion Technique for Probiotic Application.

PubMed

Mandal, Surajit; Hati, Subrota

2017-01-01

Probiotics are dietary concepts to improve the dynamics of intestinal microbial balance favorably. Careful screening of probiotic strains for their technological suitability can also allow selection of strains with the best manufacturing and food technology characteristics. However, even the most robust probiotic bacteria are currently in the range of food applications to which they can be applied. Additionally, bacteria with exceptional functional heath properties are ruled out due to technological limitations. New process and formulation technologies will enable both expansion of the range of products in to which probiotics can be applied and the use of efficacious stains that currently cannot be manufactured or stored with existing technologies. Viability of probiotics has been both a marketing and technological concern for many industrial produces. Probiotics are difficult to work with, the bacteria often die during processing, and shelf life is unpredictable. Probiotics are extremely susceptible environmental conditions such as oxygen, processing and preservation treatments, acidity, and salt concentration, which collectively affect the overall viability of probiotics. Manufacturers have long been fortifying products with probiotics; they have faced significant processing challenges regarding the stability and survivability of probiotics during processing and preservation treatments, storage as well during their passage through GIT. Application of microencapsulation significantly improves the stability of probiotics during food processing and gastrointestinal transit.

Impact of Moderate Heat, Carvacrol, and Thymol Treatments on the Viability, Injury, and Stress Response of Listeria monocytogenes

PubMed Central

Guevara, L.; Antolinos, V.; Palop, A.; Periago, P. M.

2015-01-01

The microbial safety and stability of minimally processed foods are based on the application of combined preservative factors. Since microorganisms are able to develop adaptive networks to survive under conditions of stress, food safety may be affected, and therefore understanding of stress adaptive mechanisms plays a key role in designing safe food processing conditions. In the present study, the viability and the sublethal injury of Listeria monocytogenes exposed to moderate heat (55°C) and/or essential oil compounds (carvacrol and thymol, 0.3 mM) treatments were studied. Synergistic effects were obtained when combining mild heat (55°C) with one or both essential oil compounds, leading to inactivation kinetics values three to four times lower than when using heat alone. All the treatments applied caused some injury in the population. The injury levels ranged from around 20% of the surviving population under the mildest conditions to more than 99.99% under the most stringent conditions. Protein extracts of cells exposed to these treatments were analysed by two-dimensional gel electrophoresis. The results obtained revealed that stressed cells exhibited differential protein expression to control cells. The proteins upregulated under these stressing conditions were implicated, among other functions, in stress response, metabolism, and protein refolding. PMID:26539510

Survival and germinability of Bacillus subtilis spores exposed to simulated Mars solar radiation: implications for life detection and planetary protection.

PubMed

Tauscher, Courtney; Schuerger, Andrew C; Nicholson, Wayne L

2006-08-01

Bacterial spores have been considered as microbial life that could survive interplanetary transport by natural impact processes or human spaceflight activity. Deposition of terrestrial microbes or their biosignature molecules onto the surface of Mars could negatively impact life detection experiments and planetary protection measures. Simulated Mars solar radiation, particularly the ultraviolet component, has been shown to reduce spore viability, but its effect on spore germination and resulting production of biosignature molecules has not been explored. We examined the survival and germinability of Bacillus subtilis spores exposed to simulated martian conditions that include solar radiation. Spores of B. subtilis that contain luciferase resulting from expression of an sspB-luxAB gene fusion were deposited on aluminum coupons to simulate deposition on spacecraft surfaces and exposed to simulated Mars atmosphere and solar radiation. The equivalent of 42 min of simulated Mars solar radiation exposure reduced spore viability by nearly 3 logs, while germination-induced bioluminescence, a measure of germination metabolism, was reduced by less than 1 log. The data indicate that spores can retain the potential to initiate germination-associated metabolic processes and produce biological signature molecules after being rendered nonviable by exposure to Mars solar radiation.

Viability and adaptation potential of indigenous microorganisms from natural gas field fluids in high pressure incubations with supercritical CO2.

PubMed

Frerichs, Janin; Rakoczy, Jana; Ostertag-Henning, Christian; Krüger, Martin

2014-01-21

Carbon Capture and Storage (CCS) is currently under debate as large-scale solution to globally reduce emissions of the greenhouse gas CO2. Depleted gas or oil reservoirs and saline aquifers are considered as suitable reservoirs providing sufficient storage capacity. We investigated the influence of high CO2 concentrations on the indigenous bacterial population in the saline formation fluids of a natural gas field. Bacterial community changes were closely examined at elevated CO2 concentrations under near in situ pressures and temperatures. Conditions in the high pressure reactor systems simulated reservoir fluids i) close to the CO2 injection point, i.e. saturated with CO2, and ii) at the outer boundaries of the CO2 dissolution gradient. During the incubations with CO2, total cell numbers remained relatively stable, but no microbial sulfate reduction activity was detected. After CO2 release and subsequent transfer of the fluids, an actively sulfate-respiring community was re-established. The predominance of spore-forming Clostridiales provided evidence for the resilience of this taxon against the bactericidal effects of supercritical (sc)CO2. To ensure the long-term safety and injectivity, the viability of fermentative and sulfate-reducing bacteria has to be considered in the selection, design, and operation of CCS sites.

Transcriptome analysis reveals that multidrug efflux genes are upregulated to protect Pseudomonas aeruginosa from pentachlorophenol stress.

PubMed

Muller, Jocelyn Fraga; Stevens, Ann M; Craig, Johanna; Love, Nancy G

2007-07-01

Through chemical contamination of natural environments, microbial communities are exposed to many different types of chemical stressors; however, research on whole-genome responses to this contaminant stress is limited. This study examined the transcriptome response of a common soil bacterium, Pseudomonas aeruginosa, to the common environmental contaminant pentachlorophenol (PCP). Cells were grown in chemostats at a low growth rate to obtain substrate-limited, steady-state, balanced-growth conditions. The PCP stress was administered as a continuous increase in concentration, and samples taken over time were examined for physiological function changes with whole-cell acetate uptake rates (WAURs) and cell viability and for gene expression changes by Affymetrix GeneChip technology and real-time reverse transcriptase PCR. Cell viability, measured by heterotrophic plate counts, showed a moderately steady decrease after exposure to the stressor, but WAURs did not change in response to PCP. In contrast to the physiological data, the microarray data showed significant changes in the expression of several genes. In particular, genes coding for multidrug efflux pumps, including MexAB-OprM, were strongly upregulated. The upregulation of these efflux pumps protected the cells from the potentially toxic effects of PCP, allowing the physiological whole-cell function to remain constant.

Ascaris and Escherichia coli Inactivation in an Ecological Sanitation System in Port-au-Prince, Haiti.

PubMed

Berendes, David; Levy, Karen; Knee, Jackie; Handzel, Thomas; Hill, Vincent R

2015-01-01

The goal of this study was to evaluate the microbial die-off in a latrine waste composting system in Port-au-Prince, Haiti. Temperature data and samples were collected from compost aged 0-12+ months. Samples collected from compost bin centers and corners at two depths were assessed for moisture content, E. coli concentration, and Ascaris spp. viability. Center temperatures in compost bins were all above 58 °C, while corner temperatures were 10 - 20 °C lower. Moisture content was 67 ± 10% in all except the oldest compost. A 4-log reduction in E. coli was observed over the first sixteen weeks of composting at both locations and depths, after which E. coli was undetectable (LOD: 142 MPN g(-1) dry weight). In new compost, 10.4% and 8.3% of Ascaris eggs were viable and fully embryonated, respectively. Percent viability dropped to zero in samples older than six weeks. These findings indicate that the Haitian EcoSan composting process was effective in inactivating E. coli and Ascaris spp. in latrine waste within sixteen weeks. This study is one of the first to document efficacy of an ecological sanitation system under field conditions and provides insight into composting methods and monitoring for other international settings.

In vivo and in vitro evaluation of an Acetobacter xylinum synthesized microbial cellulose membrane intended for guided tissue repair

PubMed Central

Mendes, Péricles Nóbrega; Rahal, Sheila Canevese; Pereira-Junior, Oduvaldo Câmara Marques; Fabris, Viciany Erique; Lenharo, Sara Lais Rahal; de Lima-Neto, João Ferreira; da Cruz Landim-Alvarenga, Fernanda

2009-01-01

Background Barrier materials as cellulose membranes are used for guided tissue repair. However, it is essential that the surrounding tissues accept the device. The present study histologically evaluated tissue reaction to a microbial cellulose membrane after subcutaneous implantation in mice. Furthermore, the interaction between mesenchymal stem cells and the biomaterial was studied in vitro to evaluate its ability to act as cellular scaffold for tissue engineering. Methods Twenty-five Swiss Albino mice were used. A 10 × 10 mm cellulose membrane obtained through biosynthesis using Acetobacter xylinum bacteria was implanted into the lumbar subcutaneous tissue of each mouse. The mice were euthanatized at seven, 15, 30, 60, and 90 days, and the membrane and surrounding tissues were collected and examined by histology. Results A mild inflammatory response without foreign body reaction was observed until 30 days post-surgery around the implanted membrane. Polarized microscopy revealed that the membrane remained intact at all evaluation points. Scanning electron microscopy of the cellulose membrane surface showed absence of pores. The in vitro evaluation of the interaction between cells and biomaterial was performed through viability staining analysis of the cells over the biomaterial, which showed that 95% of the mesenchymal stem cells aggregating to the cellulose membrane were alive and that 5% were necrotic. Scanning electron microscopy showed mesenchymal stem cells with normal morphology and attached to the cellulose membrane surface. Conclusion The microbial cellulose membrane evaluated was found to be nonresorbable, induced a mild inflammatory response and may prove useful as a scaffold for mesenchymal stem cells. PMID:19317903

Long-term exposure of bacterial and protozoan communities to TiO2 nanoparticles in an aerobic-sequencing batch reactor

NASA Astrophysics Data System (ADS)

Supha, Chitpisud; Boonto, Yuphada; Jindakaraked, Manee; Ananpattarachai, Jirapat; Kajitvichyanukul, Puangrat

2015-06-01

Titanium dioxide (TiO2) nanopowders at different concentrations (0-50 mg L-1) were injected into an aerobic-sequencing batch reactor (SBR) to investigate the effects of long-term exposure to nanoparticles on bacterial and protozoan communities. The detection of nanoparticles in the bioflocs was analyzed by scanning electron microscopy, transmission electron microscopy, and energy-dispersive x-ray spectroscopy. The SBR wastewater experiments were conducted under the influence of ultraviolet light with photocatalytic TiO2. The intrusion of TiO2 nanoparticles was found both on the surface and inside of the bioflocs. The change of microbial population in terms of mixed liquor-suspended solids and the sludge volume index was monitored. The TiO2 nanoparticles tentatively exerted an adverse effect on the microbial population, causing the reduction of microorganisms (both bacteria and protozoa) in the SBR. The respiration inhibition rate of the bacteria was increased, and the viability of the microbial population was reduced at the high concentration (50 mg L-1) of TiO2. The decreasing number of protozoa in the presence of TiO2 nanoparticles during 20 days of treatment with 0.5 and 1.0 mg L-1 TiO2 is clearly demonstrated. The measured chemical oxygen demand (COD) in the effluent tends to increase with a long-term operation. The increase of COD in the system suggests a decrease in the efficiency of the wastewater treatment plant. However, the SBR can effectively remove the TiO2 nanoparticles (up to 50 mg L-1) from the effluent.

Microbiological Methodology in Astrobiology

NASA Technical Reports Server (NTRS)

Abyzov, S. S.; Gerasimenko, L. M.; Hoover, R. B.; Mitskevich, I. N.; Mulyukin, A. L.; Poglazova, M. N.; Rozanov, A. Y.

2005-01-01

Searching for life in astromaterials to be delivered from the future missions to extraterrestrial bodies is undoubtedly related to studies of the properties and signatures of living microbial cells and microfossils on Earth. As model terrestrial analogs of Martian polar subsurface layers are often regarded the Antarctic glacier and Earth permafrost habitats where alive microbial cells preserved viability for millennia years due to entering the anabiotic state. For the future findings of viable microorganisms in samples from extraterrestrial objects, it is important to use a combined methodology that includes classical microbiological methods, plating onto nutrient media, direct epifluorescence and electron microscopy examinations, detection of the elemental composition of cells, radiolabeling techniques, PCR and FISH methods. Of great importance is to ensure authenticity of microorganisms (if any in studied samples) and to standardize the protocols used to minimize a risk of external contamination. Although the convincing evidence of extraterrestrial microbial life will may come from the discovery of living cells in astromaterials, biomorphs and microfossils must also be regarded as a target in search of life evidence bearing in mind a scenario that alive microorganisms had not be preserved and underwent mineralization. Under the laboratory conditions, processes that accompanied fossilization of cyanobacteria were reconstructed, and artificially produced cyanobacterial stromatolites resembles by their morphological properties those found in natural Earth habitats. Regarding the vital importance of distinguishing between biogenic and abiogenic signatures and between living and fossil microorganisms in analyzed samples, it is worthwhile to use some previously developed approaches based on electron microscopy examinations and analysis of elemental composition of biomorphs in situ and comparison with the analogous data obtained for laboratory microbial cultures and fossilized microorganisms. This communication will be focused on the analysis of our experience in working with ancient microorganisms and fossils and discussion of some issues that are crucial for development of the program for future finding of extraterrestrial life and its evidence.

Black rhinoceros (Diceros bicornis) and domestic horse (Equus caballus) hindgut microflora demonstrate similar fermentation responses to grape seed extract supplementation in vitro.

PubMed

Huntley, N F; Naumann, H D; Kenny, A L; Kerley, M S

2017-10-01

The domestic horse is used as a nutritional model for rhinoceros maintained under human care. The validity of this model for browsing rhinoceros has been questioned due to high prevalence of iron overload disorder (IOD) in captive black rhinoceros (Diceros bicornis), which is associated with high morbidity and mortality. Iron chelators, such as tannins, are under investigation as dietary supplements to ameliorate or prevent IOD in prone species. Polyphenolic compounds variably affect microbial fermentation, so the first objective of this experiment was to evaluate the effects of grape seed extract (GSE; a concentrated source of condensed tannins; CT) on black rhinoceros hindgut fermentation. Equine nutrition knowledge is used to assess supplements for rhinoceros; therefore, the second objective was to evaluate the domestic horse model for black rhinoceros fermentation and compare fermentation responses to GSE using a continuous single-flow in vitro culture system. Two replicated continuous culture experiments were conducted using horse and black rhinoceros faeces as inoculum sources comparing four diets with increasing GSE inclusion (0.0%, 1.3%, 2.7% and 4.0% of diet dry matter). Diet and GSE polyphenolic compositions were determined, and sodium sulphite effect on neutral detergent fibre extraction of CT-containing forages was tested. Increasing GSE inclusion stimulated microbial growth and fermentation, and proportionally increased diet CT concentration and iron-binding capacity. Horse and black rhinoceros hindgut microflora nutrient digestibility and fermentation responses to GSE did not differ, and results supported equine fermentation as an adequate model for microbial fermentation in the black rhinoceros. Interpretation of these results is limited to hindgut fermentation and further research is needed to compare foregut digestibility and nutrient absorption between these two species. Supplementation of GSE in black rhinoceros diets up to 4% is unlikely to adversely affect hindgut nutrient digestibility or microbial viability and fermentation. Journal of Animal Physiology and Animal Nutrition © 2016 Blackwell Verlag GmbH.

A low-cost procedure for production of fresh autochthonous wine yeast.

PubMed

Maqueda, Matilde; Pérez-Nevado, Francisco; Regodón, José A; Zamora, Emiliano; Alvarez, María L; Rebollo, José E; Ramírez, Manuel

2011-03-01

A low-cost procedure was designed for easy and rapid response-on-demand production of fresh wine yeast for local wine-making. The pilot plant produced fresh yeast culture concentrate with good microbial quality and excellent oenological properties from four selected wine yeasts. The best production yields were obtained using 2% sugar beet molasses and a working culture volume of less than 60% of the fermenter capacity. The yeast yield using 2% sugar grape juice was low and had poor cell viability after freeze storage, although the resulting yeast would be directly available for use in the winery. The performance of these yeasts in commercial wineries was excellent; they dominated must fermentation and improved its kinetics, as well as improving the physicochemical parameters and the organoleptic quality of red and white wines.

Virulence of entomopathogenic bacteria in the bed bug, Cimex lectularius.

PubMed

Pietri, Jose E; Liang, Dangsheng

2018-01-01

Due in part to the development of insecticide resistance, the common bed bug, Cimex lectularius, has overcome human intervention efforts to make a global resurgence. The failure of chemical pesticides has created a need for novel strategies to combat bed bugs. While a number of insect pests are susceptible to the use of entomopathogenic microbes or microbial-derived toxins, biological control methods have not been thoroughly explored in bed bugs. Here, we tested the virulence of three entomopathogenic bacterial species in C. lectularius to determine their potential for bed bug control. We examined bed bug survival after inoculation with live or heat-killed Serratia marcescens, Pseudomonas fluorescens, and Bacillus thuringiensis israelensis at varying temperatures. We also analyzed the viability and growth of the same bacteria in infected bed bugs. All three bacterial species were pathogenic to bed bugs. However, the effects of S. marcescens and P. fluorescens were temperature-dependent while the lethality of B. thuringiensis israelensis was not. In addition, bacterial virulence was partly dependent on the route of infection but was not strongly associated with proliferation. Thus, our results suggest multiple possible mechanisms of microbial pathogenicity in the bed bug and indicate that entomopathogenic bacteria, or products derived from them, may have useful applications for bed bug control. Copyright © 2017 Elsevier Inc. All rights reserved.

Use of spectroscopic and imaging techniques to evaluate pretreated sugarcane bagasse as a substrate for cellulase production under solid-state fermentation.

PubMed

Rodríguez-Zúñiga, Ursula Fabiola; Bertucci Neto, Victor; Couri, Sonia; Crestana, Silvio; Farinas, Cristiane Sanchez

2014-03-01

The enzymatic cocktail of cellulases is one of the most costly inputs affecting the economic viability of the biochemical route for biomass conversion into biofuels and other chemicals. Here, the influence of liquid hot water, dilute acid, alkali, and combined acid/alkali pretreatments on sugarcane bagasse (SCB) used for cellulase production was investigated by means of spectroscopic and imaging techniques. Chemical composition and structural characteristics, such as crystallinity (determined by X-ray diffraction), functional groups (Fourier transform infrared spectroscopy), and microstructure (scanning electron microscopy), were used to correlate SCB pretreatments with enzymatic biosynthesis by a strain of the filamentous fungus Aspergillus niger under solid-state fermentation. The combined acid/alkali pretreatment resulted in a SCB with higher cellulose content (86.7%). However, the high crystallinity (74%) of the resulting biomass was detrimental to microbial uptake and enzyme production. SCB pretreated with liquid hot water yielded the highest filter paper cellulase (FPase), carboxymethyl cellulase (CMCase), and xylanase activities (0.4, 14.9, and 26.1 U g(-1), respectively). The results showed that a suitable pretreatment for SCB to be used as a substrate for cellulase production should avoid severe conditions in order to preserve amorphous cellulose and to enhance the physical properties that assist microbial access.

Modeling of inactivation of surface borne microorganisms occurring on seeds by cold atmospheric plasma (CAP)

NASA Astrophysics Data System (ADS)

Mitra, Anindita; Li, Y.-F.; Shimizu, T.; Klämpfl, Tobias; Zimmermann, J. L.; Morfill, G. E.

2012-10-01

Cold Atmospheric Plasma (CAP) is a fast, low cost, simple, easy to handle technology for biological application. Our group has developed a number of different CAP devices using the microwave technology and the surface micro discharge (SMD) technology. In this study, FlatPlaSter2.0 at different time intervals (0.5 to 5 min) is used for microbial inactivation. There is a continuous demand for deactivation of microorganisms associated with raw foods/seeds without loosing their properties. This research focuses on the kinetics of CAP induced microbial inactivation of naturally growing surface microorganisms on seeds. The data were assessed for log- linear and non-log-linear models for survivor curves as a function of time. The Weibull model showed the best fitting performance of the data. No shoulder and tail was observed. The models are focused in terms of the number of log cycles reduction rather than on classical D-values with statistical measurements. The viability of seeds was not affected for CAP treatment times up to 3 min with our device. The optimum result was observed at 1 min with increased percentage of germination from 60.83% to 89.16% compared to the control. This result suggests the advantage and promising role of CAP in food industry.

Microbial survival in the stratosphere and implications for global dispersal

USGS Publications Warehouse

Smith, David J.; Griffin, Dale W.; McPeters, Richard D.; Ward, Peter D.; Schuerger, Andrew C.

2011-01-01

Spores of Bacillus subtilis were exposed to a series of stratosphere simulations. In total, five distinct treatments measured the effect of reduced pressure, low temperature, high desiccation, and intense ultraviolet (UV) irradiation on stratosphereisolated and ground-isolated B. subtilis strains. Environmental conditions were based on springtime data from a mid-latitude region of the lower stratosphere (20 km). Experimentally, each treatment consisted of the following independent or combined conditions: -70 °C, 56 mb, 10-12%relative humidity and 0.00421, 5.11, and 54.64 W/m2 of UVC (200-280 nm), UVB (280-315 nm), UVA (315-400 nm), respectively. Bacteria were deposited on metal coupon surfaces in monolayers of ~1 x 106 spores and prepared with palagonite (particle size< 20 μm). After 6 h of exposure to the stratosphere environment, 99.9% of B. subtilis spores were killed due to UV irradiation. In contrast, temperature, desiccation, and pressure simulations without UV had no effect on spore viability up through 96 h. There were no differences in survival between the stratosphere-isolated versus ground-isolated B. subtilis strains. Inactivation of most bacteria in our simulation indicates that the stratosphere can be a critical barrier to long-distance microbial dispersal and that survival in the upper atmosphere may be constrained by UV irradiation.

Pyrogallol, an absorbable microbial gallotannins-metabolite and mango polyphenols (Mangifera Indica L.) suppress breast cancer ductal carcinoma in situ proliferation in vitro.

PubMed

Nemec, Matthew J; Kim, Hyemee; Marciante, Alexandria B; Barnes, Ryan C; Talcott, Stephen T; Mertens-Talcott, Susanne U

2016-09-14

Mango is rich in bioactive absorbable polyphenols, but also contains considerable amounts of unabsorbable gallotannins at varying degrees of polymerization. Gallotannins are not absorbable upon consumption and have rarely been considered in the discussion of health benefits of polyphenols. Therefore, the objective of this study was to investigate the anti-proliferative activities of the major microbial metabolite of gallotannins, pyrogallol (PG) and a low molecular weight fraction of mango (Mangifera Indica L.) polyphenols (ML) and involved pathways including the AKT/mTOR signaling axis in an in situ breast cancer cell line, MCF10DCIS.COM. Fluorouracil (5-FU), a widely used genotoxic cancer therapeutic, was used a positive control and in combination with ML and PG to assess potential interactions. Concentrations that were non-cytotoxic in non-cancer cells were identified in non-cancer mammary fibroblasts (MCF-12F) and only non-cytotoxic dietarily relevant concentrations were selected for the investigation in MCF10DCIS.COM cancer cells. In addition to proliferation and viability, mRNA and expression of total and phosphorylated protein were investigated. Results show that both, ML and PG significantly reduced proliferation in MCF10DCIS.COM, but did not significantly reduce viability following a 48 h exposure. ML significantly reduced mRNA expression of mTOR and HIF-1α, while PG significantly reduced mRNA of IGF-1R, AKT, mTOR and HIF-1α. ML and PG reduced total protein expression of IGF-1R, IR, AKT, mTOR, and P70S6K. In addition, PG reduced IRS protein. Both treatments also had an effect on phosphorylated protein levels, with PG significantly reducing IGF-1R, AKT, and P70S6K levels. ML had a similar effect and significantly decreased IR, AKT, and P70S6K phosphorylation levels. Within the low concentration-range, ML and PG did not interact with the cytotoxic activities of 5-FU. Overall, the AKT/mTOR signaling axis appears to be implicated as causal in decreased proliferation induced by diet-relevant concentrations of ML and PG.

Meta-analytic study of organic acids as an alternative performance-enhancing feed additive to antibiotics for broiler chickens.

PubMed

Polycarpo, G V; Andretta, I; Kipper, M; Cruz-Polycarpo, V C; Dadalt, J C; Rodrigues, P H M; Albuquerque, R

2017-10-01

The effect of organic acids as an alternative to antibiotics on the performance of broiler chickens was evaluated by meta-analysis, identifying and quantifying the main factors that influence results. A total of 51,960 broilers from 121 articles published between 1991 and 2016 were used. Interactions of additives [non-supplemented group (control), organic acids, and growth promoter antibiotics] with microbial challenge (with or without inoculation of pathogenic microorganisms) were studied on performance variables. Moreover, the effects of organic acids, used individually or in blends, were evaluated. Relative values of average daily gain (ADG) and average daily feed intake (ADFI) were obtained in relation to control: ΔADG and ΔADFI, respectively. Analysis of variance-covariance revealed lower ADG with organic acids when compared to antibiotics (P < 0.05). There was a significant interaction between the additives and the challenge on feed conversion ratio (FCR) (P < 0.01) and on viability (P < 0.05). Without challenge, organic acids improved broilers' FCR (P < 0.01), presenting results similar to antibiotics (P > 0.05). Under challenge, the organic acids were again effective on FCR (-5.67% in relation to control, P < 0.05), but they did not match antibiotics (-13.40% in relation to control, P < 0.01). Viability was improved only under challenge conditions, and only by antibiotics (+4.39% in relation to control, P < 0.05). ADG (P < 0.05) and FCR (P < 0.01) were increased by blends of organic acids, but not by the organic acids used alone (P > 0.05). ADFI and production factor were not influenced by the treatments (P > 0.05). ΔADFI of organic-acid supplemented group showed a linear influence on ΔADG, which increases 0.64% at every 1% increase in ΔADFI. In conclusion, organic acids can be utilized as performance enhancing, but the results are lower than those found with antibiotics, particularly under microbial challenge. The blends of organic acids provide better results than the utilization of one organic acid alone. © The Author 2017. Published by Oxford University Press on behalf of Poultry Science Association.

Meta-analytic study of organic acids as an alternative performance-enhancing feed additive to antibiotics for broiler chickens

PubMed Central

Polycarpo, G. V.; Kipper, M.; Cruz-Polycarpo, V. C.; Dadalt, J. C.; Rodrigues, P. H. M.; Albuquerque, R.

2017-01-01

Abstract The effect of organic acids as an alternative to antibiotics on the performance of broiler chickens was evaluated by meta-analysis, identifying and quantifying the main factors that influence results. A total of 51,960 broilers from 121 articles published between 1991 and 2016 were used. Interactions of additives [non-supplemented group (control), organic acids, and growth promoter antibiotics] with microbial challenge (with or without inoculation of pathogenic microorganisms) were studied on performance variables. Moreover, the effects of organic acids, used individually or in blends, were evaluated. Relative values of average daily gain (ADG) and average daily feed intake (ADFI) were obtained in relation to control: ΔADG and ΔADFI, respectively. Analysis of variance-covariance revealed lower ADG with organic acids when compared to antibiotics (P < 0.05). There was a significant interaction between the additives and the challenge on feed conversion ratio (FCR) (P < 0.01) and on viability (P < 0.05). Without challenge, organic acids improved broilers’ FCR (P < 0.01), presenting results similar to antibiotics (P > 0.05). Under challenge, the organic acids were again effective on FCR (−5.67% in relation to control, P < 0.05), but they did not match antibiotics (−13.40% in relation to control, P < 0.01). Viability was improved only under challenge conditions, and only by antibiotics (+4.39% in relation to control, P < 0.05). ADG (P < 0.05) and FCR (P < 0.01) were increased by blends of organic acids, but not by the organic acids used alone (P > 0.05). ADFI and production factor were not influenced by the treatments (P > 0.05). ΔADFI of organic-acid supplemented group showed a linear influence on ΔADG, which increases 0.64% at every 1% increase in ΔADFI. In conclusion, organic acids can be utilized as performance enhancing, but the results are lower than those found with antibiotics, particularly under microbial challenge. The blends of organic acids provide better results than the utilization of one organic acid alone. PMID:28938776

Effects of Simulated Human Gastrointestinal Digestion of Two Purple-Fleshed Potato Cultivars on Anthocyanin Composition and Cytotoxicity in Colonic Cancer and Non-Tumorigenic Cells

PubMed Central

Kubow, Stan; Iskandar, Michèle M.; Melgar-Bermudez, Emiliano; Sleno, Lekha; Sabally, Kebba; Azadi, Behnam; How, Emily; Prakash, Satya; Burgos, Gabriela; zum Felde, Thomas

2017-01-01

A dynamic human gastrointestinal (GI) model was used to digest cooked tubers from purple-fleshed Amachi and Leona potato cultivars to study anthocyanin biotransformation in the stomach, small intestine and colonic vessels. Colonic Caco-2 cancer cells and non-tumorigenic colonic CCD-112CoN cells were tested for cytotoxicity and cell viability after 24 h exposure to colonic fecal water (FW) digests (0%, 10%, 25%, 75% and 100% FW in culture media). After 24 h digestion, liquid chromatography-mass spectrometry identified 36 and 15 anthocyanin species throughout the GI vessels for Amachi and Leona, respectively. The total anthocyanin concentration was over thirty-fold higher in Amachi compared to Leona digests but seven-fold higher anthocyanin concentrations were noted for Leona versus Amachi in descending colon digests. Leona FW showed greater potency to induce cytotoxicity and decrease viability of Caco-2 cells than observed with FW from Amachi. Amachi FW at 100% caused cytotoxicity in non-tumorigenic cells while FW from Leona showed no effect. The present findings indicate major variations in the pattern of anthocyanin breakdown and release during digestion of purple-fleshed cultivars. The differing microbial anthocyanin metabolite profiles in colonic vessels between cultivars could play a significant role in the impact of FW toxicity on tumor and non-tumorigenic cells. PMID:28850070

Combining hydrogen evolution and corrosion data - A case study on the economic viability of selected metal cathodes in microbial electrolysis cells

NASA Astrophysics Data System (ADS)

Brown, Robert Keith; Schmidt, Ulrike Christiane; Harnisch, Falk; Schröder, Uwe

2017-07-01

In this study, hydrogen evolution reaction (HER) catalytic and corrosion data is determined for selected metal cathode materials. The HER data was gathered using cyclic voltammetry (CV) in electrolytes with several pH values and varying current densities. Of the tested materials, the stainless steel alloy EN 1.4401/AISI 316 generally had the lowest HER overpotentials at the pH values 0.25, 7 and 9. At the higher pH values of 11 and 14 a custom NiMoFe alloy with a m/m% composition of 60-30-10 showed the lowest overpotentials. After each CV experiment, the electrolyte solution was analyzed to determine the corrosion of the metal cathodes. Results of corrosion measurements showed that the stainless steels EN 1.4401 had the lowest corrosion losses on average across all tested pH values. Combining HER and corrosion data revealed that: In the pH 9 electrolyte solution, EN 1.4401 was not always the best catalyst in terms of its overpotential, but it incurs the least material costs due to its lack of corrosion, this balance thereby making it the "best choice" under the given conditions. The combination of HER and corrosion data provides a more effective framework for discussing economic viability than either data set alone.

Evaluation of Energy Dose and Output Power Optimum of Diode’s Laser of 450 nm and 650 nm in Photoantimicrobial Mechanisms Against Inhibition of C. Albicans Biofilm Cells

NASA Astrophysics Data System (ADS)

Dewi-Astuty, S.; Suhariningsih; Dyah-Astuti, S.; Baktir, A.

2018-03-01

Photoantimicrobial as a pathogenic microbial inhibitory therapy system such as C. albicans in biofilms forms has been studied in vitro. Mechanisms of inhibiting called inactivating used the absorb principles of a dye agents such as chlorophyll against the photon energy of diode laser which any number of ROS product depend on energy doses of a laser, time of irradiation, concentration and time of incubation the dyes agent. The inactivation profile of C. albicans biofilm cells was observed based on cell viability reduction after photoantimicrobial treatment with or without oxygenation by XTT assay test. Results show that the inhibiting significantly with the time incubation of the dye agents and the oxygen degree inside the sample. The inhibition for oxygenation biofilm’s group 10% lower than without oxygenation biofilm’s group at the maximum of reduction of cell viability occurred in the 3hour incubation group. Optimum of inactivation are 89.6% (without oxygenation) and 94.8% (with oxygenation) after irradiation with 450 nm laser (power output 128.73 at energy dose 86.09 J/cm2), While, by 650 nm laser (power output 164.53 mW at energy dose 92.52 J/cm2) irradiation treatment obtained optimum of inactivation are 89.5% (without oxygenation) and 92.3% (with oxygenation).

Promotion of stem cell proliferation by vegetable peptone.

PubMed

Lee, J; Lee, J; Hwang, H; Jung, E; Huh, S; Hyun, J; Park, D

2009-10-01

Technical limitations and evolution of therapeutic applications for cell culture-derived products have accelerated elimination of animal-derived constituents from such products to minimize inadvertent introduction of microbial contaminants, such as fungi, bacteria or viruses. The study described here was conducted to investigate the proliferative effect of vegetable peptone on adult stem cells in the absence of serum, and its possible mechanisms of action. Cell viability and proliferation were determined using the MTT assay and Click-iT EdU flow cytometry, respectively. In addition, changes in expression of cytokine genes were analysed using MILLIPLEX human cytokine enzyme-linked immunosorbent assay kit. Viability of cord blood-derived mesenchymal stem cells (CB-MSC) and adipose tissue-derived stem cells (ADSC) increased significantly when treated with the peptone. In addition, median value of the group treated with peptone shifted to the right when compared to the untreated control group. Furthermore, quantitative analysis of the cytokines revealed that production of vascular endothelial growth factor (VEGF), transforming growth factor-beta1 (TGF-beta1), and interleukin-6 (IL-6) increased significantly in response to treatment with our vegetable peptone in both CB-MSCs and ADSCs. Our findings revealed that the vegetable peptone promotes proliferation of CB-MSCs and ADSCs. In addition, results of this study suggest that induction of stem cell proliferation by vegetable peptone is likely to be related to its induction of VEGF, TGF-beta1, and IL-6 expression.

Cytocompatible antifungal acrylic resin containing silver nanoparticles for dentures

PubMed Central

Acosta-Torres, Laura Susana; Mendieta, Irasema; Nuñez-Anita, Rosa Elvira; Cajero-Juárez, Marcos; Castaño, Víctor M

2012-01-01

Background Inhibition of Candida albicans on denture resins could play a significant role in preventing the development of denture stomatitis. The safety of a new dental material with antifungal properties was analyzed in this work. Methods Poly(methyl methacrylate) [PMMA] discs and PMMA-silver nanoparticle discs were formulated, with the commercial acrylic resin, Nature-CrylTM, used as a control. Silver nanoparticles were synthesized and characterized by ultraviolet-visible spectroscopy, dispersive Raman spectroscopy, and transmission electron microscopy. The antifungal effect was assessed using a luminescent microbial cell viability assay. Biocompatibility tests were carried out using NIH-3T3 mouse embryonic fibroblasts and a Jurkat human lymphocyte cell line. Cells were cultured for 24 or 72 hours in the presence or absence of the polymer formulations and analyzed using three different tests, ie, cellular viability by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and cell proliferation by enzyme-linked immunosorbent assay BrdU, and genomic DNA damage (Comet assay). Finally, the samples were evaluated mechanically, and the polymer-bearing silver nanoparticles were analyzed microscopically to evaluate dispersion of the nanoparticles. Results The results show that PMMA-silver nanoparticle discs significantly reduce adherence of C. albicans and do not affect metabolism or proliferation. They also appear not to cause genotoxic damage to cells. Conclusion The present work has developed a new biocompatible antifungal PMMA denture base material. PMID:22969297

In vitro assessment of stainless steel orthodontic brackets coated with titanium oxide mixed Ag for anti-adherent and antibacterial properties against Streptococcus mutans and Porphyromonas gingivalis.

PubMed

Fatani, Eman Jameel; Almutairi, Hamed H; Alharbi, Ali O; Alnakhli, Yasser Obaidallah; Divakar, Darshan Devang; Muzaheed; Alkheraif, Abdulaziz Abdullah; Khan, Aftab Ahmed

2017-11-01

Orthodontic brackets made from stainless steel were introduced in dentistry, though they have less ability in reducing enamel demineralization and are not successful in preventing microbial as well as biofilm growth. In this study, we evaluated the significant role of different brackets in reducing enamel demineralization indirectly. Results from different tests indicate the significant reduction in adhesion, biofilm formation and slow growth of tested bacterial species on brackets coated with Ag + TiO2 and found to be statistically significant lower than control. There was no loss in cell viability in all brackets indicating that the cells are biocompatible with different bracket materials. Scanning electron microscopy showed less bacteria attached with the surface coated with Ag + TiO2 indicated that bacteria were losing adherent nature on coated surface. In conclusion, TiO2+Ag coating on stainless steel brackets possessed anti-adherent properties and also have demonstrable antibacterial properties therefore helps in preventing dental caries and plaque accumulation indirectly. The cell compatibility of TiO2+Ag coated brackets is superior to the uncoated samples therefore can be used in orthodontics as it not only provide suitable antimicrobial activity and resistance to biofilm formation but also sustained the cell viability of human gingival fibroblast (HGF) cell lines. Copyright © 2017 Elsevier Ltd. All rights reserved.

An innovative approach to bioremediation of mercury contaminated soils from industrial mining operations.

PubMed

McCarthy, Damien; Edwards, Grant C; Gustin, Mae S; Care, Andrew; Miller, Matthieu B; Sunna, Anwar

2017-10-01

Soils contaminated with mercury (Hg) have proved expensive and logistically difficult to remediate. Research continues into finding suitable environmentally-friendly and efficient ways of achieving this end. Bioremediation is an option, which employs the strategies microorganisms have evolved to deal with Hg. One microbial strategy involves uptake and intracellular volatilisation of mercuric ions, which passively diffuse from the cell and back into the atmosphere. In this work, Pseudomonas veronii cells grown to stationary phase were immobilised in a xanthan gum-based biopolymer via encapsulation. The P. veronii-biopolymer mix was then coated onto natural zeolite granules. Zeolite immobilised cells remained viable for at least 16 weeks stored under ambient room temperature. Furthermore, the immobilised cells were shown to retain both viability and Hg volatilisation functionality after transportation from Australia to the USA, where they were applied to Hg contaminated soil. Maximum flux rates exceeded 10 μg Hg m 2  h -1 from mine tailings (≈7 mg kg -1  Hg with 50% v/v water). This was 4 orders of magnitude above background flux levels. It is envisioned that emitted gaseous elemental mercury (GEM) can be readily captured, and transformed back into metallic Hg, which can then be stored appropriately or recycled. This breaks the Hg cycle, as GEM is no longer translocated back to the atmospheric compartment. The immobilising excipients used in this research overcome many logistical issues with delivery of suitable microbial loads to locations of mercury contamination and presents a facile and inexpensive method of augmenting contaminated sites with selected microbial consortia for bioremediation. Copyright © 2017 Elsevier Ltd. All rights reserved.

Wheat bran extract alters colonic fermentation and microbial composition, but does not affect faecal water toxicity: a randomised controlled trial in healthy subjects.

PubMed

Windey, Karen; De Preter, Vicky; Huys, Geert; Broekaert, Willem F; Delcour, Jan A; Louat, Thierry; Herman, Jean; Verbeke, Kristin

2015-01-28

Wheat bran extract (WBE), containing arabinoxylan-oligosaccharides that are potential prebiotic substrates, has been shown to modify bacterial colonic fermentation in human subjects and to beneficially affect the development of colorectal cancer (CRC) in rats. However, it is unclear whether these changes in fermentation are able to reduce the risk of developing CRC in humans. The aim of the present study was to evaluate the effects of WBE on the markers of CRC risk in healthy volunteers, and to correlate these effects with colonic fermentation. A total of twenty healthy subjects were enrolled in a double-blind, cross-over, randomised, controlled trial in which the subjects ingested WBE (10 g/d) or placebo (maltodextrin, 10 g/d) for 3 weeks, separated by a 3-week washout period. At the end of each study period, colonic handling of NH3 was evaluated using the biomarker lactose[15N, 15N']ureide, colonic fermentation was characterised through a metabolomics approach, and the predominant microbial composition was analysed using denaturing gradient gel electrophoresis. As markers of CRC risk, faecal water genotoxicity was determined using the comet assay and faecal water cytotoxicity using a colorimetric cell viability assay. Intake of WBE induced a shift from urinary to faecal 15N excretion, indicating a stimulation of colonic bacterial activity and/or growth. Microbial analysis revealed a selective stimulation of Bifidobacterium adolescentis. In addition, WBE altered the colonic fermentation pattern and significantly reduced colonic protein fermentation compared with the run-in period. However, faecal water cytotoxicity and genotoxicity were not affected. Although intake of WBE clearly affected colonic fermentation and changed the composition of the microbiota, these changes were not associated with the changes in the markers of CRC risk.

The Significance of the Enteric Microbiome on the Development of Childhood Disease: A Review of Prebiotic and Probiotic Therapies in Disorders of Childhood.

PubMed

Slattery, John; MacFabe, Derrick F; Frye, Richard E

2016-01-01

Recent studies have highlighted the fact that the enteric microbiome, the trillions of microbes that inhabit the human digestive tract, has a significant effect on health and disease. Methods for manipulating the enteric microbiome, particularly through probiotics and microbial ecosystem transplantation, have undergone some study in clinical trials. We review some of the evidence for microbiome alteration in relation to childhood disease and discuss the clinical trials that have examined the manipulation of the microbiome in an effort to prevent or treat childhood disease with a primary focus on probiotics, prebiotics, and/or synbiotics (ie, probiotics + prebiotics). Studies show that alterations in the microbiome may be a consequence of events occurring during infancy and/or childhood such as prematurity, C-sections, and nosocomial infections. In addition, certain childhood diseases have been associated with microbiome alterations, namely necrotizing enterocolitis, infantile colic, asthma, atopic disease, gastrointestinal disease, diabetes, malnutrition, mood/anxiety disorders, and autism spectrum disorders. Treatment studies suggest that probiotics are potentially protective against the development of some of these diseases. Timing and duration of treatment, the optimal probiotic strain(s), and factors that may alter the composition and function of the microbiome are still in need of further research. Other treatments such as prebiotics, fecal microbial transplantation, and antibiotics have limited evidence. Future translational work, in vitro models, long-term and follow-up studies, and guidelines for the composition and viability of probiotic and microbial therapies need to be developed. Overall, there is promising evidence that manipulating the microbiome with probiotics early in life can help prevent or reduce the severity of some childhood diseases, but further research is needed to elucidate biological mechanisms and determine optimal treatments.

An investigation into the stability and sterility of citric acid solutions used for cough reflex testing.

PubMed

Falconer, James R; Wu, Zimei; Lau, Hugo; Suen, Joanna; Wang, Lucy; Pottinger, Sarah; Lee, Elaine; Alazawi, Nawar; Kallesen, Molly; Gargiulo, Derryn A; Swift, Simon; Svirskis, Darren

2014-10-01

Citric acid is used in cough reflex testing in clinical and research settings to assess reflexive cough in patients at risk of swallowing disorders. To address a lack of knowledge in this area, this study investigated the stability and sterility of citric acid solutions. Triplicate solutions of citric acid (0.8 M) in isotonic saline were stored at 4 ± 2 °C for up to 28 days and analysed by high-performance liquid chromatography. Microbiological sterility of freshly prepared samples and bulk samples previously used for 2 weeks within the hospital was determined using a pour plate technique. Microbial survival in citric acid was determined by inoculating Staphylococcus aureus, Escherichia coli, or Candida albicans into citric acid solution and monitoring the number of colony-forming units/mL over 40 min. Citric acid solutions remained stable at 4 °C for 28 days (98.4 ± 1.8 % remained). The freshly prepared and clinical samples tested were sterile. However, viability studies revealed that citric acid solution allows for the survival of C. albicans but not for S. aureus or E. coli. The microbial survival study showed that citric acid kills S. aureus and E. coli but has no marked effect on C. albicans after 40 min. Citric acid samples at 0.8 M remained stable over the 4-week testing period, with viable microbial cells absent from samples tested. However, C. albicans has the ability to survive in citric acid solution if inadvertently introduced in practice. For this reason, in clinical and research practice it is suggested to use single-use aliquots prepared aseptically which can be stored for up to 28 days at 4 °C.

The effect of ozonization on furniture dust: microbial content and immunotoxicity in vitro.

PubMed

Huttunen, Kati; Kauhanen, Eeva; Meklin, Teija; Vepsäläinen, Asko; Hirvonen, Maija-Riitta; Hyvärinen, Anne; Nevalainen, Aino

2010-05-01

Moisture and mold problems in buildings contaminate also the furniture and other movable property. If cleaning of the contaminated furniture is neglected, it may continue to cause problems to the occupants even after the moisture-damage repairs. The aim of this study was to determine the effectiveness of high-efficiency ozone treatment in cleaning of the furniture from moisture-damaged buildings. In addition, the effectiveness of two cleaning methods was compared. Samples were vacuumed from the padded areas before and after the treatment. The microbial flora and concentrations in the dust sample were determined by quantitative cultivation and QPCR-methods. The immunotoxic potential of the dust samples was analyzed by measuring effects on cell viability and production of inflammatory mediators in vitro. Concentrations of viable microbes decreased significantly in most of the samples after cleaning. Cleaning with combined steam wash and ozonisation was more effective method than ozonising alone, but the difference was not statistically significant. Detection of fungal species with PCR showed a slight but nonsignificant decrease in concentrations after the cleaning. The immunotoxic potential of the collected dust decreased significantly in most of the samples. However, in a small subgroup of samples, increased concentrations of microbes and immunotoxicological activity were detected. This study shows that a transportable cleaning unit with high-efficiency ozonising is in most cases effective in decreasing the concentrations of viable microbes and immunotoxicological activity of the furniture dust. However, the method does not destroy or remove all fungal material present in the dust, as detected with QPCR analysis, and in some cases the cleaning procedure may increase the microbial concentrations and immunotoxicity of the dust. Copyright 2010 Elsevier B.V. All rights reserved.

The limits of extremophilic life expanded under extraterrestrial environment-simulated experiments

NASA Astrophysics Data System (ADS)

Lage, C.; Dalmaso, G.; Teixeira, L.; Bendia, A.; Rosado, A.

2012-09-01

Astrobiology is a brand new area of science that seeks to understand the origin and dynamics of life in the universe. Several hypotheses to explain life in the cosmic context have been developed throughout human history, but only now technology has allowed many of them to be tested. Laboratory experiments have been able to show how chemical elements essential to life, carbon, nitrogen, oxygen and hydrogen combine in biologically important compounds. Interestingly, these compounds are found universally. As these compounds were combined to the point of originating cells and complex organisms is still a challenge to be unveiled by science. However, our 4.5 billion years-old solar system was born within a 10-billion years-old universe. Thus, simple cells like microorganisms may have had time to form in planets older than ours or other suitable molecular places in the universe. One hypothesis to explain the origin of life on Earth is called panspermia, which predicts that microbial life could have been formed in the universe billions of years ago, traveling between planets, and inseminating units of life that could have become more complex in habitable planets like ours. A project designed to test the viability of extremophile microorganisms exposed to simulated extraterrestrial environments is ongoing at the Carlos Chagas Filho Institute of Biophysics to test whether microbial life could withstand those inhospitable environments. Ultra-resistant (known or novel ones) microorganisms collected from terrestrial extreme environments, extremophiles, have been exposed to intense radiation sources simulating solar radiation (at synchrotron accelerators), capable of emitting in a few hours radiation equivalent of million years accumulated doses. The results obtained in these experiments reveal the interesting possibility of the existence of microbial life beyond Earth.

Mini-Review: Probing the limits of extremophilic life in extraterrestrial environment-simulated experiments

NASA Astrophysics Data System (ADS)

Lage, Claudia A. S.; Dalmaso, Gabriel Z. L.; Teixeira, Lia C. R. S.; Bendia, Amanda G.; Paulino-Lima, Ivan G.; Galante, Douglas; Janot-Pacheco, Eduardo; Abrevaya, Ximena C.; Azúa-Bustos, Armando; Pelizzari, Vivian H.; Rosado, Alexandre S.

2012-10-01

Astrobiology is a relatively recent scientific field that seeks to understand the origin and dynamics of life in the Universe. Several hypotheses have been proposed to explain life in the cosmic context throughout human history, but only now, technology has allowed many of them to be tested. Laboratory experiments have been able to show how chemical elements essential to life, such as carbon, nitrogen, oxygen and hydrogen combine in biologically important compounds. Interestingly, these compounds are ubiquitous. How these compounds were combined to the point of originating cells and complex organisms is still to be unveiled by science. However, our 4.5 billion years old Solar system appeared in a 10 billion years old Universe. Thus, simple cells such as micro-organisms may have had time to form in planets older than ours or in other suitable places in the Universe. One hypothesis related to the appearance of life on Earth is called panspermia, which predicts that microbial life could have been formed in the Universe billions of years ago, travelling between planets, and inseminating units of life that could have become more complex in habitable planets such as Earth. A project designed to test the viability of extremophile micro-organisms exposed to simulated extraterrestrial environments is in progress at the Carlos Chagas Filho Institute of Biophysics (UFRJ, Brazil) to test whether microbial life could withstand inhospitable environments. Radiation-resistant (known or novel ones) micro-organisms collected from extreme terrestrial environments have been exposed (at synchrotron accelerators) to intense radiation sources simulating Solar radiation, capable of emitting radiation in a few hours equivalent to many years of accumulated doses. The results obtained in these experiments reveal an interesting possibility of the existence of microbial life beyond Earth.

QAC modified PVDF membranes: Antibiofouling performance, mechanisms, and effects on microbial communities in an MBR treating municipal wastewater.

PubMed

Chen, Mei; Zhang, Xingran; Wang, Zhiwei; Wang, Liang; Wu, Zhichao

2017-09-01

Biofouling remains as a critical issue limiting the widespread applications of membrane bioreactors (MBRs). The use of antibiofouling membranes is an emerging method to tackle this issue. In this study, a polyvinylidene fluoride (PVDF) membrane was modified using a quaternary ammonium compound (QAC) to create an antibiofouling membrane. The membrane was used in an MBR and the performance, mechanisms, and effects on microbial communities of this membrane were compared to a control operated in parallel. Results showed that the membrane exhibited a significantly reduced transmembrane pressure increase rate of 0.29 kPa/d compared with 0.91 kPa/d of the control. Analysis using a confocal laser scanning microscope (CLSM) revealed almost complete lack of living microbes on the antibiofouling membrane in contrast to the control. However, specific oxygen uptake rate and dehydrogenase activity analyses demonstrated no adverse impacts on microbial viability of the bulk activated sludge. Bacterial population analysis using the Illumina Miseq platform added further evidence that the use of antibiofouling membrane did not exert negative influences on richness, diversity and structure of the bacterial community. Effluent quality of the test MBR also exhibited minimal difference from that of the control reactor. The amount of polysaccharides and proteins in the biofouling layer was also significantly reduced. Quartz crystal microbalance with dissipation monitoring suggested that the antibiofouling membrane only allowed organic matter with strong adhesion properties to attach onto the membrane surfaces. These findings highlight the potential of the antibiofouling membrane to be used in MBRs for wastewater treatment and reclamation. Copyright © 2017 Elsevier Ltd. All rights reserved.

Vapor Hydrogen Peroxide Sterilization Certification

NASA Astrophysics Data System (ADS)

Chen, Fei; Chung, Shirley; Barengoltz, Jack

For interplanetary missions landing on a planet of potential biological interest, United States NASA planetary protection currently requires that the flight system must be assembled, tested and ultimately launched with the intent of minimizing the bioload taken to and deposited on the planet. Currently the only NASA approved microbial reduction method is dry heat sterilization process. However, with utilization of such elements as highly sophisticated electronics and sensors in modern spacecraft, this process presents significant materials challenges and is thus an undesirable bioburden reduction method to design engineers. The objective of this work is to introduce vapor hydrogen peroxide (VHP) as an alternative to dry heat microbial reduction to meet planetary protection requirements. The VHP sterilization technology is widely used by the medical industry, but high doses of VHP may degrade the performance of flight hardware, or compromise material compatibility. The goal of our study is determine the minimum VHP process conditions for PP acceptable microbial reduction levels. A series of experiments were conducted using Geobacillus stearothermophilus to determine VHP process parameters that provided significant reductions in spore viability while allowing survival of sufficient spores for statistically significant enumeration. In addition to the obvious process parameters -hydrogen peroxide concentration, number of pulses, and exposure duration -the investigation also considered the possible effect of environmental pa-rameters. Temperature, relative humidity, and material substrate effects on lethality were also studied. Based on the results, a most conservative D value was recommended. This recom-mended D value was also validated using VHP "hardy" strains that were isolated from clean-rooms and environmental populations collected from spacecraft relevant areas. The efficiency of VHP at ambient condition as well as VHP material compatibility will also be presented.

Rapid Microbial Sample Preparation from Blood Using a Novel Concentration Device

PubMed Central

Boardman, Anna K.; Campbell, Jennifer; Wirz, Holger; Sharon, Andre; Sauer-Budge, Alexis F.

2015-01-01

Appropriate care for bacteremic patients is dictated by the amount of time needed for an accurate diagnosis. However, the concentration of microbes in the blood is extremely low in these patients (1–100 CFU/mL), traditionally requiring growth (blood culture) or amplification (e.g., PCR) for detection. Current culture-based methods can take a minimum of two days, while faster methods like PCR require a sample free of inhibitors (i.e., blood components). Though commercial kits exist for the removal of blood from these samples, they typically capture only DNA, thereby necessitating the use of blood culture for antimicrobial testing. Here, we report a novel, scaled-up sample preparation protocol carried out in a new microbial concentration device. The process can efficiently lyse 10 mL of bacteremic blood while maintaining the microorganisms’ viability, giving a 30‑μL final output volume. A suite of six microorganisms (Staphylococcus aureus, Streptococcus pneumoniae, Escherichia coli, Haemophilus influenzae, Pseudomonas aeruginosa, and Candida albicans) at a range of clinically relevant concentrations was tested. All of the microorganisms had recoveries greater than 55% at the highest tested concentration of 100 CFU/mL, with three of them having over 70% recovery. At the lowest tested concentration of 3 CFU/mL, two microorganisms had recoveries of ca. 40–50% while the other four gave recoveries greater than 70%. Using a Taqman assay for methicillin-sensitive S. aureus (MSSA)to prove the feasibility of downstream analysis, we show that our microbial pellets are clean enough for PCR amplification. PCR testing of 56 spiked-positive and negative samples gave a specificity of 0.97 and a sensitivity of 0.96, showing that our sample preparation protocol holds great promise for the rapid diagnosis of bacteremia directly from a primary sample. PMID:25675242

A Comprehensive Assessment of Biologicals Contained Within Commercial Airliner Cabin Air

NASA Technical Reports Server (NTRS)

LaDuc, Myron T.; Osman, Shariff; Dekas, Anne; Stuecker, Tara; Newcombe, Dave; Piceno, Yvette; Fuhrman, J.; Andersen, Gary; Venkateswaran, Kasthuri; Bearman, Greg

2006-01-01

Both culture-based and culture-independent, biomarker-targeted microbial enumeration and identification technologies were employed to estimate total microbial and viral burden and diversity within the cabin air of commercial airliners. Samples from each of twenty flights spanning three commercial carriers were collected via air-impingement. When the total viable microbial population was estimated by assaying relative concentrations of the universal energy carrier ATP, values ranged from below detection limits (BDL) to 4.1 x 106 cells/cubic m of air. The total viable microbial population was extremely low in both of Airline A (approximately 10% samples) and C (approximately 18% samples) compared to the samples collected aboard flights on Airline A and B (approximately 70% samples). When samples were collected as a function of time over the course of flights, a gradual accumulation of microbes was observed from the time of passenger boarding through mid-flight, followed by a sharp decline in microbial abundance and viability from the initiation of descent through landing. It is concluded in this study that only 10% of the viable microbes of the cabin air were cultivable and suggested a need to employ state-of-the art molecular assay that measures both cultivable and viable-but-non-cultivable microbes. Among the cultivable bacteria, colonies of Acinetobacter sp. were by far the most profuse in Phase I, and Gram-positive bacteria of the genera Staphylococcus and Bacillus were the most abundant during Phase II. The isolation of the human pathogens Acinetobacter johnsonii, A. calcoaceticus, Janibacter melonis, Microbacterium trichotecenolyticum, Massilia timonae, Staphylococcus saprophyticus, Corynebacterium lipophiloflavum is concerning, as these bacteria can cause meningitis, septicemia, and a handful of sometimes fatal diseases and infections. Molecular microbial community analyses exhibited presence of the alpha-, beta-, gamma-, and delta- proteobacteria, as well as Gram-positive bacteria, Fusobacteria, Cyanobacteria, Deinococci, Bacterioidetes, Spirochetes, and Planctomyces in varying abundance. Neisseria meningitidis rDNA sequences were retrieved in great abundance from Airline A followed by Streptococcus oralis/mitis sequences. Pseudomonas synxantha sequences dominated Airline B clone libraries, followed by those of N. meningitidis and S. oralis/mitis. In Phase II, Airline C, sequences representative of more than 113 species, enveloping 12 classes of bacteria, were retrieved. Proteobacterial sequences were retrieved in greatest frequency (58% of all clone sequences), followed in short order by those stemming from Gram-positives bacteria (31% of all clone sequences). As for overall phylogenetic breadth, Gram-positive and alpha-proteobacteria seem to have a higher affinity for international flights, whereas beta-and gamma-proteobacteria are far more common about domestic cabin air parcels in Airline C samples. Ultimately, the majority of microbial species circulating throughout the cabin airs of commercial airliners are commensal, infrequently pathogenic normal flora of the human nasopharynx and respiratory system. Many of these microbes likely originate from the oral and nasal cavities, and lungs of passengers and flight crew and are disseminated unknowingly via routine conversation, coughing, sneezing, and stochastic passing of fomites. The data documented in this study will be useful to generate a baseline microbial population database and can be utilized to develop biosensor instrumentation for monitoring microbial quality of cabin or urban air.

Probiotics and clinical effects: is the number what counts?

PubMed

Bertazzoni, Elisa; Donelli, Gianfranco; Midtvedt, Tore; Nicoli, Jacques; Sanz, Yolanda

2013-08-01

Probiotics are defined as 'live microorganisms that when administered in adequate amounts confer health benefits on the host', underlining the need of microbial viability and the requirement of a suitable dose to obtain a health benefit. The dose and the administration regimen are critical issues for probiotics either ingested as foods claiming health benefits or used as drugs in clinics. In fact, regulatory authorities demand to guarantee consumers that a probiotic is effective in the recommended conditions of use and responds to its specific claims. Thus, a proper identification of probiotic strain(s), a definition of the amount of microorganisms surviving by the end of the product shelf-life, and a demonstration of their beneficial effects by appropriate human trials are required. The current knowledge on the effective dose of different probiotic strains used for several disorders is here reviewed.

Nanoscopic morphological changes in yeast cell surfaces caused by oxidative stress: an atomic force microscopic study.

PubMed

Canetta, Elisabetta; Walker, Graeme M; Adya, Ashok K

2009-06-01

Nanoscopic changes in the cell surface morphology of the yeasts Saccharomyces cerevisiae (strain NCYC 1681) and Schizosaccharomyces pombe (strain DVPB 1354), due to their exposure to varying concentrations of hydrogen peroxide (oxidative stress), were investigated using an atomic force microscope (AFM). Increasing hydrogen peroxide concentration led to a decrease in cell viabilities and mean cell volumes, and an increase in the surface roughness of the yeasts. In addition, AFM studies revealed that oxidative stress caused cell compression in both S. cerevisiae and Schiz. pombe cells and an increase in the number of aged yeasts. These results confirmed the importance and usefulness of AFM in investigating the morphology of stressed microbial cells at the nanoscale. The results also provided novel information on the relative oxidative stress tolerance of S. cerevisiae and Schiz. pombe.

Effect of photoactivated disinfection with a light-emitting diode on bacterial species and biofilms associated with periodontitis and peri-implantitis.

PubMed

Eick, Sigrun; Markauskaite, Giedre; Nietzsche, Sandor; Laugisch, Oliver; Salvi, Giovanni E; Sculean, Anton

2013-05-01

To determine the effect of photoactivated disinfection (PAD) using toluidine blue and a light-emitting diode (LED) in the red spectrum (wave length at 625-635 nm) on species associated with periodontitis and peri-implantitis and bacteria within a periodontopathic biofilm. Sixteen single microbial species including 2 Porphyromonas gingivalis and 2 Aggregatibacter actinomycetemcomitans and a multispecies mixture consisting of 12 species suspended in saline without and with 25% human serum were exposed to PAD. Moreover, single-species biofilms consisting of 2 P. gingivalis and 2 A. actinomycetemcomitans strains and a multi-species biofilm on 24-well-plates, grown on titanium discs and in artificial periodontal pockets were exposed to PAD with and without pretreatment with 0.25% hydrogen peroxide. Changes in the viability were determined by counting the colony forming units (cfu). PAD reduced the cfu counts in saline by 1.42 log₁₀ after LED application for 30s and by 1.99 log₁₀ after LED application for 60s compared with negative controls (each p<0.001). Serum did not inhibit the efficacy of PAD. PAD reduced statistically significantly (p<0.05) the cfu counts of the P. gingivalis biofilms. The viability of the A. actinomycetemcomitans biofilms and the multi-species biofilms was statistically significantly decreased when PAD was applied after a pretreatment with 0.25% hydrogen peroxide. The biofilm formed in artificial pockets was more sensitive to PAD with and without pretreatment with hydrogen peroxide compared with those formed on titanium discs. PAD using a LED was effective against periodontopathic bacterial species and reduced viability in biofilms but was not able to completely destroy complex biofilms. The use of PAD following pretreatment with hydrogen peroxide resulted in an additional increase in the antimicrobial activity which may represent a new alternative to treat periodontal and peri-implant infections thus warranting further testing in clinical studies. Copyright © 2012 Elsevier B.V. All rights reserved.

Antimicrobial Efficacy of a Silver Impregnated Hydrophilic PU Foam.

PubMed

Percival, Steven L

2018-06-01

A novel hydrophilic polyurethane (PU) foam dressing which is impregnated with silver chloride, Optifoam® Gentle (OG) Ag+ (Medline Industries Inc., Chicago, Illinois), was evaluated in this study. The aims of this study were to determine the rate of elution of silver from the foam dressing over a period of 168 hours into simulated wound fluid and an evaluation of antimicrobial efficacy using zone of inhibition (ZOI), direct kill, and time-kill viability. Thirty-two microorganisms associated with wounds including Pseudomonas aeruginosa, Methicillin sensitive Staphylococcus aureus (MSSA), Acinetobacter baumannii, Candida albicans, and antibiotic-resistant strains (Methicillin-resistant S. aureus [MRSA] and Vancomycin-resistant Enterococci [VRE]) were evaluated. Silver release from the wound dressing showed an exponential curve with a stable sustained release of 25ppm achieved after 24 hours, which was maintained for the full duration of the study. OG Ag+ caused inhibition zones ranging from 4-16mm after a 24-hour contact time. In the direct kill assay, OG Ag+ reduced the microbial numbers below the limit of detection and reduced viability by a log of four within 24 hours. For the time-kill viability studies, the results support the use of this hydrophilic polyurethane foam as a wound dressing for use in wounds at risk of infection or infected by achieving a four log kill within six hours and a six log kill in 16 hours. In conclusion, OG Ag+ was shown to be an effective wound dressing in the killing of a range of important opportunistic pathogens of relevance to wound healing and infections. Achieving a six log kill against S. aureus and E.coli, within 16 hours in the time kill assay, (ASTM E2315-03) demonstrates that OG Ag+ should be an important addition to the armoury available for the management of acute and chronic wounds at risk of infection or clinically infected.

A Comparison of Microbial Water Quality and Diversity for Ballast and Tropical Harbor Waters

PubMed Central

Goh, Shin Giek; Liang, Liang; Kim, Yiseul; Rose, Joan B.; Yew-Hoong, Karina Gin

2015-01-01

Indicator organisms and antibiotic resistance were used as a proxy to measure microbial water quality of ballast tanks of ships, and surface waters in a tropical harbor. The survival of marine bacteria in ballast tanks appeared to diminish over longer water retention time, with a reduction of cell viability observed after a week based on heterotrophic plate counts. Pyrosequencing of 16S rRNA genes showed distinct differences in microbial composition of ballast and harbor waters. The harbor waters had a higher abundance of operational taxonomic units (OTUs) assigned to Cyanobacteria (Synechococcus spp.) and α-proteobacteria (SAR11 members), while marine hydrocarbon degraders such as γ-proteobacteria (Ocenspirillaes spp., Thiotrchales spp.) and Bacteroidetes (Flavobacteriales spp.) dominated the ballast water samples. Screening of indicator organisms found Escherichia coli (E. coli), Enterococcus and Pseudomonas aeruginosa (P. aeruginosa) in two or more of the ballast and harbor water samples tested. Vibrio spp. and Salmonella spp. were detected exclusively in harbor water samples. Using quantitative PCR (qPCR), we screened for 13 antibiotic resistant gene (ARG) targets and found higher abundances of sul1 (4.13–3.44 x 102 copies/mL), dfrA (0.77–1.80 x10 copies/mL) and cfr (2.00–5.21 copies/mL) genes compared to the other ARG targets selected for this survey. These genes encode for resistance to sulfonamides, trimethoprim and chloramphenicol-florfenicol antibiotics, which are also known to persist in sediments of aquaculture farms and coastal environments. Among the ARGs screened, we found significant correlations (P<0.05) between ereA, ermG, cfr and tetO genes to one or more of the indicator organisms detected in this study, which may suggest that these members contribute to the environmental resistome. This study provides a baseline water quality survey, quantitatively assessing indicators of antibiotic resistance, potentially pathogenic organisms and a broad-brush description of difference in microbial composition and diversity between open oceans and tropical coastal environments through the use of next generation sequencing technology. PMID:26575481

A Comparison of Microbial Water Quality and Diversity for Ballast and Tropical Harbor Waters.

PubMed

Ng, Charmaine; Le, Thai-Hoang; Goh, Shin Giek; Liang, Liang; Kim, Yiseul; Rose, Joan B; Yew-Hoong, Karina Gin

2015-01-01

Indicator organisms and antibiotic resistance were used as a proxy to measure microbial water quality of ballast tanks of ships, and surface waters in a tropical harbor. The survival of marine bacteria in ballast tanks appeared to diminish over longer water retention time, with a reduction of cell viability observed after a week based on heterotrophic plate counts. Pyrosequencing of 16S rRNA genes showed distinct differences in microbial composition of ballast and harbor waters. The harbor waters had a higher abundance of operational taxonomic units (OTUs) assigned to Cyanobacteria (Synechococcus spp.) and α-proteobacteria (SAR11 members), while marine hydrocarbon degraders such as γ-proteobacteria (Ocenspirillaes spp., Thiotrchales spp.) and Bacteroidetes (Flavobacteriales spp.) dominated the ballast water samples. Screening of indicator organisms found Escherichia coli (E. coli), Enterococcus and Pseudomonas aeruginosa (P. aeruginosa) in two or more of the ballast and harbor water samples tested. Vibrio spp. and Salmonella spp. were detected exclusively in harbor water samples. Using quantitative PCR (qPCR), we screened for 13 antibiotic resistant gene (ARG) targets and found higher abundances of sul1 (4.13-3.44 x 102 copies/mL), dfrA (0.77-1.80 x10 copies/mL) and cfr (2.00-5.21 copies/mL) genes compared to the other ARG targets selected for this survey. These genes encode for resistance to sulfonamides, trimethoprim and chloramphenicol-florfenicol antibiotics, which are also known to persist in sediments of aquaculture farms and coastal environments. Among the ARGs screened, we found significant correlations (P<0.05) between ereA, ermG, cfr and tetO genes to one or more of the indicator organisms detected in this study, which may suggest that these members contribute to the environmental resistome. This study provides a baseline water quality survey, quantitatively assessing indicators of antibiotic resistance, potentially pathogenic organisms and a broad-brush description of difference in microbial composition and diversity between open oceans and tropical coastal environments through the use of next generation sequencing technology.

Survival of microbial isolates from clouds toward simulated atmospheric stress factors

NASA Astrophysics Data System (ADS)

Joly, Muriel; Amato, Pierre; Sancelme, Martine; Vinatier, Virginie; Abrantes, Magali; Deguillaume, Laurent; Delort, Anne-Marie

2015-09-01

In the atmosphere, airborne microbial cells are exposed to conditions that are thought to affect their survival. Here, we investigated the survival of 5 microorganisms among the most represented in the cultivable community of clouds (4 bacteria affiliated to Pseudomonas, Sphingomonas and Arthrobacter and 1 yeast of Dioszegia) after exposition to different atmospheric factors generally considered stressful for cells: artificial solar light (10 h), oxidant (hydrogen peroxide: 0-1 mM for 90 min), osmotic shocks (0.1-2.5 M NaCl) and freeze-thaw cycles (6 cycles of 5 °C/-40 °C). Each condition was applied separately to cell suspensions, and survival rates were examined by culture. Survival was highly strain and stress dependent, with no relationship with pigmentation or ice nucleation activity. In all strains, solar light had no or mitigated influence, and exposition to H2O2 at the concentration measured in cloud water only slightly impacted viability (>70% of the cells survived). The strain Sphingomonas sp. was particularly impacted by osmotic shocks while repeated freeze-thaw was particularly damaging for Arthrobacter and Pseudomonas species. Overall, our results tend to indicate that in the atmosphere, the most stringent selection factors on living organisms are probably freeze-thaw and condensation/evaporation (osmotic shocks) cycles, whereas the impacts of oxidants and of solar light are limited.

Friend or Foe-Light Availability Determines the Relationship between Mycorrhizal Fungi, Rhizobia and Lima Bean (Phaseolus lunatus L.).

PubMed

Ballhorn, Daniel J; Schädler, Martin; Elias, Jacob D; Millar, Jess A; Kautz, Stefanie

2016-01-01

Plant associations with root microbes represent some of the most important symbioses on earth. While often critically promoting plant fitness, nitrogen-fixing rhizobia and arbuscular mycorrhizal fungi (AMF) also demand significant carbohydrate allocation in exchange for key nutrients. Though plants may often compensate for carbon loss, constraints may arise under light limitation when plants cannot extensively increase photosynthesis. Under such conditions, costs for maintaining symbioses may outweigh benefits, turning mutualist microbes into parasites, resulting in reduced plant growth and reproduction. In natural systems plants commonly grow with different symbionts simultaneously which again may interact with each other. This might add complexity to the responses of such multipartite relationships. We experimented with lima bean (Phaseolus lunatus), which efficiently forms associations with both types of root symbionts. We applied full light and low-light to each of four treatments of microbial inoculation. After an incubation period of 14 weeks, we quantified vegetative aboveground and belowground biomass and number and viability of seeds to determine effects of combined inoculant and light treatment on plant fitness. Under light-limited conditions, vegetative and reproductive traits were inhibited in AMF and rhizobia inoculated lima bean plants relative to controls (un-colonized plants). Strikingly, reductions in seed production were most critical in combined treatments with rhizobia x AMF. Our findings suggest microbial root symbionts create additive costs resulting in decreased plant fitness under light-limited conditions.

Thermo-and pH-sensitive hydrogel membranes composed of poly(N-isopropylacrylamide)-hyaluronan for biomedical applications: Influence of hyaluronan incorporation on the membrane properties.

PubMed

Kamoun, Elbadawy A; Fahmy, Alaa; Taha, Tarek H; El-Fakharany, Esmail M; Makram, Mohamed; Soliman, Hesham M A; Shehata, Hassan

2018-01-01

Interpenetrating hydrogel membranes consisting of pH-sensitive hyaluronan (HA) and thermo-sensitive poly(N-isopropylacrylamide) (PNIPAAM) were synthesized using redox polymerization, followed by N,N-methylenebisacrylamide (BIS) and epichlorohydrin (EPI) were added as chemical crosslinkers. The interaction between membrane compositions has been characterized by FTIR spectroscopy and discussed intensively. The result indicates that HA incorporation in membranes increase the gel fraction, swelling uptake, and the flexibility/elasticity of crosslinked membranes, however it reduced oppositely the mechanical elongation of membranes. PNIPAAm-HA hydrogels responded to both temperature and pH changes and the stimuli-responsiveness was reversible. However, in vitro bioevaluation results revealed that the released ampicillin during the burst release time was sharply influenced and increased with increasing HA contents in membranes; afterwards it became sustainable. Whereas, high HA contents in hydrogels unexpectedly impacted negatively on the cells viability, owing to the viscosity of cell culture media changed. A big resistance was observed against microbial growth of Staphylococcus aureus, Salmonella typhi, and Candida albicans in case of pure PNIPAAm hydrogel membranes without HA or ampicillin. However, HA incorporation or the loaded ampicillin in membranes showed unexpected easily microbial growth. The fast release performance with dual pH-thermo-sensitive hydrogels were suggested as promising materials for quick drug carrier in the biomedical field. Copyright © 2017 Elsevier B.V. All rights reserved.

Cross-tolerance between osmotic and freeze-thaw stress in microbial assemblages from temperate lakes.

PubMed

Wilson, Sandra L; Frazer, Corey; Cumming, Brian F; Nuin, Paulo A S; Walker, Virginia K

2012-11-01

Osmotic stress can accompany increases in solute concentrations because of freezing or high-salt environments. Consequently, microorganisms from environments with a high-osmotic potential may exhibit cross-tolerance to freeze stress. To test this hypothesis, enrichments derived from the sediment and water of temperate lakes with a range of salt concentrations were subjected to multiple freeze-thaw cycles. Surviving isolates were identified and metagenomes were sampled prior to and following selection. Enrichments from alkali lakes were typically the most freeze-thaw resistant with only 100-fold losses in cell viability, and those from freshwater lakes were most susceptible, with cell numbers reduced at least 100,000-fold. Metagenomic analysis suggested that selection reduced assemblage diversity more in freshwater samples than in those from saline lakes. Survivors included known psychro-, halo- and alkali-tolerant bacteria. Characterization of freeze-thaw-resistant isolates from brine and alkali lakes showed that few isolates had ice-associating activities such as antifreeze or ice nucleation properties. However, all brine- and alkali-derived isolates had high intracellular levels of osmolytes and/or appeared more likely to form biofilms. Conversely, these phenotypes were infrequent amongst the freshwater-derived isolates. These observations are consistent with microbial cross-tolerance between osmotic and freeze-thaw stresses. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Ultraviolet radiation-induced limitation to epilithic microbial growth in arid deserts--dosimetric experiments in the hyperarid core of the Atacama Desert.

PubMed

Cockell, Charles S; McKay, Christopher P; Warren-Rhodes, Kim; Horneck, Gerda

2008-02-27

Experiments were conducted during November 2003 in the dry core of the Atacama Desert, Yungay, Chile to test the hypothesis that UV radiation, in environments where liquid water is not available, and thus enzymatic repair of UV-induced damage is inhibited, can prevent epilithic colonization. Novel dosimeters made from the cryptoendolithic, desiccation and radiation-resistant cyanobacterium Chroococcidiopsis sp. isolated from the dry Negev desert, Israel, showed that monolayers of this organism were killed within one day. The diurnal profile of microbial loss of viability was investigated with dosimeters of Bacillus subtilis, which similarly showed cell death within one day. Soil grains obtained from south of Yungay where liquid water is more abundant and transported to the hyperarid core showed killing of indigenous vegetative organisms within one day. Gypsum and mineral grain coverings of 1mm were sufficient to prevent measurable UV-induced damage of Chroococcidiopsis and B. subtilis after 8d exposure. These results show that under extreme desiccation and an ambient UV flux the surface of rocks can potentially be rendered sterile, but that millimetre thick mineral coverings can protect organisms from UV-induced killing, consistent with the observed patterns of lithophytic colonization in the Atacama Desert. These data further show that UV radiation can be an important limiting factor in surface biological rock weathering in arid regions.

Louis pasteur, the father of immunology?

PubMed

Smith, Kendall A

2012-01-01

Louis Pasteur is traditionally considered as the progenitor of modern immunology because of his studies in the late nineteenth century that popularized the germ theory of disease, and that introduced the hope that all infectious diseases could be prevented by prophylactic vaccination, as well as also treated by therapeutic vaccination, if applied soon enough after infection. However, Pasteur was working at the dawn of the appreciation of the microbial world, at a time when the notion of such a thing as an immune system did not exist, certainly not as we know it today, more than 130 years later. Accordingly, why was Pasteur such a genius as to discern how the immune system functions to protect us against invasion by the microbial world when no one had even made the distinction between fungi, bacteria, or viruses, and no one had formulated any theories of immunity. A careful reading of Pasteur's presentations to the Academy of Sciences reveals that Pasteur was entirely mistaken as to how immunity occurs, in that he reasoned, as a good microbiologist would, that appropriately attenuated microbes would deplete the host of vital trace nutrients absolutely required for their viability and growth, and not an active response on the part of the host. Even so, he focused attention on immunity, preparing the ground for others who followed. This review chronicles Pasteur's remarkable metamorphosis from organic chemist to microbiologist to immunologist, and from basic science to medicine.

Effect of Aqueous Extract of the Seaweed Gracilaria domingensis on the Physicochemical, Microbiological, and Textural Features of Fermented Milks.

PubMed

Tavares Estevam, Adriana Carneiro; Alonso Buriti, Flávia Carolina; de Oliveira, Tiago Almeida; Pereira, Elainy Virginia Dos Santos; Florentino, Eliane Rolim; Porto, Ana Lúcia Figueiredo

2016-04-01

The effects of the Gracilaria domingensis seaweed aqueous extract in comparison with gelatin on the physicochemical, microbial, and textural characteristics of fermented milks processed with the mixed culture SAB 440 A, composed of Streptococcus thermophilus, Lactobacillus acidophilus, and Bifidobacterium animalis ssp. lactis, were investigated. The addition of G. domingensis aqueous extract did not affect pH, titratable acidity, and microbial viability of fermented milks when compared with the control (with no texture modifier) and the products with added gelatin. Fermented milk with added the seaweed aqueous extract showed firmness, consistency, cohesiveness, and viscosity index at least 10% higher than those observed for the control product (P < 0.05). At 4 h of fermentation, the fermented milks with only G. domingensis extract showed a texture comparable to that observed for products containing only gelatin. At 5 h of fermentation, firmness and consistency increased significantly (P < 0.05) in products with only seaweed extract added, a behavior not observed in products with the full amount of gelatin, probably due to the differences between the interactions of these ingredients with casein during the development of the gel network throughout the acidification of milk. The G. domingensis aqueous extract appears as a promising gelatin alternative to be used as texture modifier in fermented milks and related dairy products. © 2016 Institute of Food Technologists®

Engineering microbes for tolerance to next-generation biofuels

PubMed Central

2011-01-01

A major challenge when using microorganisms to produce bulk chemicals such as biofuels is that the production targets are often toxic to cells. Many biofuels are known to reduce cell viability through damage to the cell membrane and interference with essential physiological processes. Therefore, cells must trade off biofuel production and survival, reducing potential yields. Recently, there have been several efforts towards engineering strains for biofuel tolerance. Promising methods include engineering biofuel export systems, heat shock proteins, membrane modifications, more general stress responses, and approaches that integrate multiple tolerance strategies. In addition, in situ recovery methods and media supplements can help to ease the burden of end-product toxicity and may be used in combination with genetic approaches. Recent advances in systems and synthetic biology provide a framework for tolerance engineering. This review highlights recent targeted approaches towards improving microbial tolerance to next-generation biofuels with a particular emphasis on strategies that will improve production. PMID:21936941

Probiotics, prebiotics, and microencapsulation: A review.

PubMed

Sarao, Loveleen Kaur; Arora, M

2017-01-22

The development of a suitable technology for the production of probiotics is a key research for industrial production, which should take into account the viability and the stability of the organisms involved. Microbial criteria, stress tolerance during processing, and storage of the product constitute the basis for the production of probiotics. Generally, the bacteria belonging to the genera Lactobacillus and Bifidobacterium have been used as probiotics. Based on their positive qualities, probiotic bacteria are widely used in the production of food. Interest in the incorporation of the probiotic bacteria into other products apart from dairy products has been increasing and represents a great challenge. The recognition of dose delivery systems for probiotic bacteria has also resulted in research efforts aimed at developing probiotic food outside the dairy sector. Producing probiotic juices has been considered more in the recent years, due to an increased concern in personal health of consumers. This review focuses on probiotics, prebiotics, and the microencapsulation of living cells.

Oxidative stress in bacteria (Pseudomonas putida) exposed to nanostructures of silicon carbide.

PubMed

Borkowski, Andrzej; Szala, Mateusz; Kowalczyk, Paweł; Cłapa, Tomasz; Narożna, Dorota; Selwet, Marek

2015-09-01

Silicon carbide (SiC) nanostructures produced by combustion synthesis can cause oxidative stress in the bacterium Pseudomonas putida. The results of this study showed that SiC nanostructures damaged the cell membrane, which can lead to oxidative stress in living cells and to the loss of cell viability. As a reference, micrometric SiC was also used, which did not exhibit toxicity toward cells. Oxidative stress was studied by analyzing the activity of peroxidases, and the expression of the glucose-6-phosphate dehydrogenase gene (zwf1) using real-time PCR and northern blot techniques. Damage to nucleic acid was studied by isolating and hydrolyzing plasmids with the formamidopyrimidine [fapy]-DNA glycosylase (also known as 8-oxoguanine DNA glycosylase) (Fpg), which is able to detect damaged DNA. The level of viable microbial cells was investigated by propidium iodide and acridine orange staining. Copyright © 2015 Elsevier Ltd. All rights reserved.

Encapsulation of Aloe Vera extract into natural Tragacanth Gum as a novel green wound healing product.

PubMed

Ghayempour, Soraya; Montazer, Majid; Mahmoudi Rad, Mahnaz

2016-12-01

Application of natural materials in wound healing is an interest topic due to effective treatment with no side effects. In this paper, Aloe Vera extract was encapsulated into Tragacanth Gum through a sonochemical microemulsion process to prepare a wound healing product. FESEM/EDX and FT-IR proved the successfully formation of the nanocapsules with spherical shape by cross-linking aluminum ions with Tragacanth Gum. The therapeutic characteristics of the prepared wound healing product were investigated using antimicrobial, cytotoxicity and wound healing assays. Relative high antimicrobial activities with the microbial reduction of 84, 91 and 80% against E. coli, S. aureus and C. albicans, a cell viability of 98% against human fibroblast cells and a good wound healing activity with considerable migration rate of fibroblast cells are the important advantages of the new formed wound healing product. Copyright © 2016 Elsevier B.V. All rights reserved.

Molecular and chemical dialogues in bacteria-protozoa interactions.

PubMed

Song, Chunxu; Mazzola, Mark; Cheng, Xu; Oetjen, Janina; Alexandrov, Theodore; Dorrestein, Pieter; Watrous, Jeramie; van der Voort, Menno; Raaijmakers, Jos M

2015-08-06

Protozoan predation of bacteria can significantly affect soil microbial community composition and ecosystem functioning. Bacteria possess diverse defense strategies to resist or evade protozoan predation. For soil-dwelling Pseudomonas species, several secondary metabolites were proposed to provide protection against different protozoan genera. By combining whole-genome transcriptome analyses with (live) imaging mass spectrometry (IMS), we observed multiple changes in the molecular and chemical dialogues between Pseudomonas fluorescens and the protist Naegleria americana. Lipopeptide (LP) biosynthesis was induced in Pseudomonas upon protozoan grazing and LP accumulation transitioned from homogeneous distributions across bacterial colonies to site-specific accumulation at the bacteria-protist interface. Also putrescine biosynthesis was upregulated in P. fluorescens upon predation. We demonstrated that putrescine induces protozoan trophozoite encystment and adversely affects cyst viability. This multifaceted study provides new insights in common and strain-specific responses in bacteria-protozoa interactions, including responses that contribute to bacterial survival in highly competitive soil and rhizosphere environments.

Antimicrobial activity of biopolymer-antibiotic thin films fabricated by advanced pulsed laser methods

NASA Astrophysics Data System (ADS)

Cristescu, R.; Popescu, C.; Dorcioman, G.; Miroiu, F. M.; Socol, G.; Mihailescu, I. N.; Gittard, S. D.; Miller, P. R.; Narayan, R. J.; Enculescu, M.; Chrisey, D. B.

2013-08-01

We report on thin film deposition by matrix assisted pulsed laser evaporation (MAPLE) of two polymer-drug composite thin film systems. A pulsed KrF* excimer laser source (λ = 248 nm, τ = 25 ns, ν = 10 Hz) was used to deposit composite thin films of poly(D,L-lactide) (PDLLA) containing several gentamicin concentrations. FTIR spectroscopy was used to demonstrate that MAPLE-transferred materials exhibited chemical structures similar to those of drop cast materials. Scanning electron microscopy data indicated that MAPLE may be used to fabricate thin films of good morphological quality. The activity of PDLLA-gentamicin composite thin films against Staphylococcus aureus bacteria was demonstrated using drop testing. The influence of drug concentration on microbial viability was also assessed. Our studies indicate that polymer-drug composite thin films prepared by MAPLE may be used to impart antimicrobial activity to implants, medical devices, and other contact surfaces.

A historical overview of bacteriophage therapy as an alternative to antibiotics for the treatment of bacterial pathogens

PubMed Central

Wittebole, Xavier; De Roock, Sophie; Opal, Steven M

2014-01-01

The seemingly inexorable spread of antibiotic resistance genes among microbial pathogens now threatens the long-term viability of our current antimicrobial therapy to treat severe bacterial infections such as sepsis. Antibiotic resistance is reaching a crisis situation in some bacterial pathogens where few therapeutic alternatives remain and pan-resistant strains are becoming more prevalent. Non-antibiotic therapies to treat bacterial infections are now under serious consideration and one possible option is the therapeutic use of specific phage particles that target bacterial pathogens. Bacteriophage therapy has essentially been re-discovered by modern medicine after widespread use of phage therapy in the pre-antibiotic era lost favor, at least in Western countries, after the introduction of antibiotics. We review the current therapeutic rationale and clinical experience with phage therapy as a treatment for invasive bacterial infection as novel alternative to antimicrobial chemotherapy. PMID:23973944

Microstencils to generate defined, multi-species patterns of bacteria

DOE PAGES

Timm, Collin M.; Hansen, Ryan R.; Doktycz, Mitchel J.; ...

2015-11-12

Microbial communities are complex heterogeneous systems that are influenced by physical and chemical interactions with their environment, host, and community members. Techniques that facilitate the quantitative evaluation of how microscale organization influences the morphogenesis of multispecies communities could provide valuable insights into the dynamic behavior and organization of natural communities, the design of synthetic environments for multispecies culture, and the engineering of artificial consortia. In this work, we demonstrate a method for patterning microbes into simple arrangements that allow the quantitative measurement of growth dynamics as a function of their proximity to one another. The method combines parylene-based liftoff techniquesmore » with microfluidic delivery to simultaneously pattern multiple bacterial species with high viability using low-cost, customizable methods. Furthermore, quantitative measurements of bacterial growth for two competing isolates demonstrate that spatial coordination can play a critical role in multispecies growth and structure.« less

Infectious microbial diseases and host defense responses in Sydney rock oysters

PubMed Central

Raftos, David A.; Kuchel, Rhiannon; Aladaileh, Saleem; Butt, Daniel

2014-01-01

Aquaculture has long been seen as a sustainable solution to some of the world's growing food shortages. However, experience over the past 50 years indicates that infectious diseases caused by viruses, bacteria, and eukaryotes limit the productivity of aquaculture. In extreme cases, these types of infectious agents threaten the viability of entire aquaculture industries. This article describes the threats from infectious diseases in aquaculture and then focuses on one example (QX disease in Sydney rock oysters) as a case study. QX appears to be typical of many emerging diseases in aquaculture, particularly because environmental factors seem to play a crucial role in disease outbreaks. Evidence is presented that modulation of a generic subcellular stress response pathway in oysters is responsible for both resistance and susceptibility to infectious microbes. Understanding and being able to manipulate this pathway may be the key to sustainable aquaculture. PMID:24795701

Efficient estimation of the maximum metabolic productivity of batch systems

DOE PAGES

St. John, Peter C.; Crowley, Michael F.; Bomble, Yannick J.

2017-01-31

Production of chemicals from engineered organisms in a batch culture involves an inherent trade-off between productivity, yield, and titer. Existing strategies for strain design typically focus on designing mutations that achieve the highest yield possible while maintaining growth viability. While these methods are computationally tractable, an optimum productivity could be achieved by a dynamic strategy in which the intracellular division of resources is permitted to change with time. New methods for the design and implementation of dynamic microbial processes, both computational and experimental, have therefore been explored to maximize productivity. However, solving for the optimal metabolic behavior under the assumptionmore » that all fluxes in the cell are free to vary is a challenging numerical task. Here, previous studies have therefore typically focused on simpler strategies that are more feasible to implement in practice, such as the time-dependent control of a single flux or control variable.« less

Cell death versus cell survival instructed by supramolecular cohesion of nanostructures

NASA Astrophysics Data System (ADS)

Newcomb, Christina J.; Sur, Shantanu; Ortony, Julia H.; Lee, One-Sun; Matson, John B.; Boekhoven, Job; Yu, Jeong Min; Schatz, George C.; Stupp, Samuel I.

2014-02-01

Many naturally occurring peptides containing cationic and hydrophobic domains have evolved to interact with mammalian cell membranes and have been incorporated into materials for non-viral gene delivery, cancer therapy or treatment of microbial infections. Their electrostatic attraction to the negatively charged cell surface and hydrophobic interactions with the membrane lipids enable intracellular delivery or cell lysis. Although the effects of hydrophobicity and cationic charge of soluble molecules on the cell membrane are well known, the interactions between materials with these molecular features and cells remain poorly understood. Here we report that varying the cohesive forces within nanofibres of supramolecular materials with nearly identical cationic and hydrophobic structure instruct cell death or cell survival. Weak intermolecular bonds promote cell death through disruption of lipid membranes, while materials reinforced by hydrogen bonds support cell viability. These findings provide new strategies to design biomaterials that interact with the cell membrane.

Effects of Low-Temperature Plasma-Sterilization on Mars Analog Soil Samples Mixed with Deinococcus radiodurans.

PubMed

Schirmack, Janosch; Fiebrandt, Marcel; Stapelmann, Katharina; Schulze-Makuch, Dirk

2016-05-26

We used Ar plasma-sterilization at a temperature below 80 °C to examine its effects on the viability of microorganisms when intermixed with tested soil. Due to a relatively low temperature, this method is not thought to affect the properties of a soil, particularly its organic component, to a significant degree. The method has previously been shown to work well on spacecraft parts. The selected microorganism for this test was Deinococcus radiodurans R1, which is known for its remarkable resistance to radiation effects. Our results showed a reduction in microbial counts after applying a low temperature plasma, but not to a degree suitable for a sterilization of the soil. Even an increase of the treatment duration from 1.5 to 45 min did not achieve satisfying results, but only resulted in in a mean cell reduction rate of 75% compared to the untreated control samples.

Temperature sensitivity of organic-matter decay in tidal marshes

USGS Publications Warehouse

Kirwan, Matthew L.; Guntenspergen, Glenn R.; Langley, J.A.

2014-01-01

Approximately half of marine carbon sequestration takes place in coastal wetlands, including tidal marshes, where organic matter contributes to soil elevation and ecosystem persistence in the face of sea-level rise. The long-term viability of marshes and their carbon pools depends, in part, on how the balance between productivity and decay responds to climate change. Here, we report the sensitivity of labile soil organic-matter decay in tidal marshes to seasonal and latitudinal variations in temperature measured over a 3-year period. We find a moderate increase in decay rate at warmer temperatures (3-6% per °C, Q10 = 1.3-1.5). Despite the profound differences between microbial metabolism in wetlands and uplands, our results indicate a strong conservation of temperature sensitivity. Moreover, simple comparisons with organic-matter production suggest that elevated atmospheric CO2 and warmer temperatures will accelerate carbon accumulation in marsh soils, and potentially enhance their ability to survive sea-level rise.

Microstencils to generate defined, multi-species patterns of bacteria

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

Timm, Collin M.; Hansen, Ryan R.; Doktycz, Mitchel J.

Microbial communities are complex heterogeneous systems that are influenced by physical and chemical interactions with their environment, host, and community members. Techniques that facilitate the quantitative evaluation of how microscale organization influences the morphogenesis of multispecies communities could provide valuable insights into the dynamic behavior and organization of natural communities, the design of synthetic environments for multispecies culture, and the engineering of artificial consortia. In this work, we demonstrate a method for patterning microbes into simple arrangements that allow the quantitative measurement of growth dynamics as a function of their proximity to one another. The method combines parylene-based liftoff techniquesmore » with microfluidic delivery to simultaneously pattern multiple bacterial species with high viability using low-cost, customizable methods. Furthermore, quantitative measurements of bacterial growth for two competing isolates demonstrate that spatial coordination can play a critical role in multispecies growth and structure.« less

Survival or Revival: Long-Term Preservation Induces a Reversible Viable but Non-Culturable State in Methane-Oxidizing Bacteria

PubMed Central

Hoefman, Sven; Van Hoorde, Koenraad; Boon, Nico; Vandamme, Peter; De Vos, Paul; Heylen, Kim

2012-01-01

Knowledge on long-term preservation of micro-organisms is limited and research in the field is scarce despite its importance for microbial biodiversity and biotechnological innovation. Preservation of fastidious organisms such as methane-oxidizing bacteria (MOB) has proven difficult. Most MOB do not survive lyophilization and only some can be cryopreserved successfully for short periods. A large-scale study was designed for a diverse set of MOB applying fifteen cryopreservation or lyophilization conditions. After three, six and twelve months of preservation, the viability (via live-dead flow cytometry) and culturability (via most-probable number analysis and plating) of the cells were assessed. All strains could be cryopreserved without a significant loss in culturability using 1% trehalose in 10-fold diluted TSB (TT) as preservation medium and 5% DMSO as cryoprotectant. Several other cryopreservation and lyophilization conditions, all of which involved the use of TT medium, also allowed successful preservation but showed a considerable loss in culturability. We demonstrate here that most of these non-culturables survived preservation according to viability assessment indicating that preservation induces a viable but non-culturable (VBNC) state in a significant fraction of cells. Since this state is reversible, these findings have major implications shifting the emphasis from survival to revival of cells in a preservation protocol. We showed that MOB cells could be significantly resuscitated from the VBNC state using the TT preservation medium. PMID:22539945

Trace detection of specific viable bacteria using tetracysteine-tagged bacteriophages.

PubMed

Wu, Lina; Luan, Tian; Yang, Xiaoting; Wang, Shuo; Zheng, Yan; Huang, Tianxun; Zhu, Shaobin; Yan, Xiaomei

2014-01-07

Advanced methods are urgently needed to determine the identity and viability of trace amounts of pathogenic bacteria in a short time. Existing approaches either fall short in the accurate assessment of microbial viability or lack specificity in bacterial identification. Bacteriophages (or phages for short) are viruses that exclusively infect bacterial host cells with high specificity. As phages infect and replicate only in living bacterial hosts, here we exploit the strategy of using tetracysteine (TC)-tagged phage in combination with biarsenical dye to the discriminative detection of viable target bacteria from dead target cells and other viable but nontarget bacterial cells. Using recombinant M13KE-TC phage and Escherichia coli ER2738 as a model system, distinct differentiation between individual viable target cells from dead target cells was demonstrated by flow cytometry and fluorescence microscopy. As few as 1% viable E. coli ER2738 can be accurately quantified in a mix with dead E. coli ER2738 by flow cytometry. With fluorescence microscopic measurement, specific detection of as rare as 1 cfu/mL original viable target bacteria was achieved in the presence of a large excess of dead target cells and other viable but nontarget bacterial cells in 40 mL artificially contaminated drinking water sample in less than 3 h. This TC-phage-FlAsH approach is sensitive, specific, rapid, and simple, and thus shows great potential in water safety monitoring, health surveillance, and clinical diagnosis of which trace detection and identification of viable bacterial pathogens is highly demanded.

Polycyclic Aromatic Hydrocarbon Affects Acetic Acid Production during Anaerobic Fermentation of Waste Activated Sludge by Altering Activity and Viability of Acetogen.

PubMed

Luo, Jingyang; Chen, Yinguang; Feng, Leiyu

2016-07-05

Till now, almost all the studies on anaerobic fermentation of waste activated sludge (WAS) for bioproducts generation focused on the influences of operating conditions, pretreatment methods and sludge characteristics, and few considered those of widespread persistent organic pollutants (POPs) in sludge, for example, polycyclic aromatic hydrocarbons (PAHs). Herein, phenanthrene, which was a typical PAH and widespread in WAS, was selected as a model compound to investigate its effect on WAS anaerobic fermentation for short-chain fatty acids (SCFAs) accumulation. Experimental results showed that the concentration of SCFAs derived from WAS was increased in the presence of phenanthrene during anaerobic fermentation. The yield of acetic acid which was the predominant SCFA in the fermentation reactor with the concentration of 100 mg/kg dry sludge was 1.8 fold of that in the control. Mechanism exploration revealed that the present phenanthrene mainly affected the acidification process of anaerobic fermentation and caused the shift of the microbial community to benefit the accumulation of acetic acid. Further investigation showed that both the activities of key enzymes (phosphotransacetylase and acetate kinase) involved in acetic acid production and the quantities of their corresponding encoding genes were enhanced in the presence of phenanthrene. Viability tests by determining the adenosine 5'-triphosphate content and membrane potential confirmed that the acetogens were more viable in anaerobic fermentation systems with phenanthrene, which resulted in the increased production of acetic acid.

Application of green fluorescent protein to measure antimicrobial efficacy and the kinetics of cell death against Escherichia coli.

PubMed

Greenhalgh, Richard; Greenhalgh, Malcolm; Alshareef, Fadwa; Robson, Geoffrey D

2017-10-01

Industrial antimicrobials have been extensively used to control unwanted microbial growth by incorporation into a variety of products such as plastics and paints, reducing biodeterioration and biofouling and extending the lifespan of the product. Industrial antimicrobials generally have broad sites of action affecting core cellular functions such as central metabolism, enzyme function, cell wall or DNA synthesis and can either be biocidal or biostatic. In addition, susceptibility can be affected by the metabolic state of the microbe, with metabolically inactive cells generally more resistant than metabolically active cells. Previously it was demonstrated that cytosolically expressed green fluorescent protein could be used as a real-time viability indicator in the yeast Aureobasidium pullulans based on the pH dependent fluorescence of GFP and the collapse of the proton gradient across the cell membrane during cell death. In this study we report on the development and validation of an equivalent GFP fluorescence viability assay in Escherichia coli and used this assay to study the effect of five antimicrobials commonly used in plastics; 4,5-dichloro-2-octyl-isothiazol-3-one (DCOIT), sodium pyrithione, 1,2-benzisothiazol-3-one (BIT), 2-octyl-isothiazol-3-one (OIT) and n-butyl-1,2-benzisothiazol-3-one (BBIT). The results demonstrate broad differences amongst the antimicrobials in both relative efficacy, rate of effect and for some antimicrobials, marked differences in sensitivity toward growing and non-growing cells. Copyright © 2017 Elsevier B.V. All rights reserved.

Mineral Influence on Microbial Survival During Carbon Sequestration

NASA Astrophysics Data System (ADS)

Santillan, E. U.; Shanahan, T. M.; Wolfe, W. W.; Bennett, P.

2012-12-01

CO2 sequestered in a deep saline aquifer will perturb subsurface biogeochemistry by acidifying the groundwater and accelerating mineral diagenesis. Subsurface microbial communities heavily influence geochemistry through their metabolic processes, such as with dissimilatory iron reducing bacteria (DIRB). However, CO2 also acts as a sterilant and will perturb these communities. We investigated the role of mineralogy and its effect on the survival of microbes at high PCO2 conditions using the model DIRB Shewanella oneidensis MR-1. Batch cultures of Shewanella were grown to stationary phase and exposed to high PCO2 using modified Parr reactors. Cell viability was then determined by plating cultures after exposure. Results indicate that at low PCO2 (2 bar), growth and iron reduction are decreased and cell death occurs within 1 hour when exposed to CO2 pressures of 10 bar or greater. Further, fatty acid analysis indicates microbial lipid degradation with C18 fatty acids being the slowest lipids to degrade. When cultures were grown in the presence of rocks or minerals representative of the deep subsurface such as carbonates and silicates and exposed to 25 bar CO2, survival lasted beyond 2 hours. The most effective protecting substratum was quartz sandstone, with cultures surviving beyond 8 hours of CO2 exposure. Scanning electron microscope images reveal biofilm formation on the mineral surfaces with copious amounts of extracellular polymeric substances (EPS) present. EPS from these biofilms acts as a reactive barrier to the CO2, slowing the penetration of CO2 into cells and resulting in increased survival. When biofilm cultures were grown with Al and As to simulate the release of toxic metals from minerals such as feldspars and clays, survival time decreased, indicating mineralogy may also enhance microbial death. Biofilms were then grown on iron-coated quartz sand to determine conversely what influence biofilms may have on mineral dissolution during CO2 perturbation. Growth media was allowed to flow through a sand-packed column at a constant flow rate with pulses of liquid CO2 injected directly into the column. Preliminary data of dissolved iron measured from the effluent indicates that biofilm columns show a slight increase in dissolved iron concentrations before and after CO2 exposure in comparison to abiotic columns. These findings imply the important relationship between microbes and minerals during CO2 sequestration. The ability minerals have to contribute to the selection of microbes has important consequences to the survival of different microbial populations in the subsurface and the consequent biogeochemical changes that may happen.

Influence of methylene blue-mediated photodynamic therapy on the resistance to detachment of streptococcus mutans biofilms from titanium substrata

NASA Astrophysics Data System (ADS)

Sharab, Lina Y.

In dental settings, as well as in other natural systems, plaque-forming microorganisms develop biofilms in which the microbes become protected via their own phenotypic changes and their polymeric exudates from disinfection by washes and antibiotics. Photodynamic Therapy (PDT) is variably effective against these microorganisms, depending on such factors as whether the bacteria are Gram positive or Gram negative, plaque age and thickness, and internal biofilm oxygen concentration. This investigation applied a novel combination of PDT and water-jet impingement techniques to Streptococcus mutans (ATCC strain 27351)-formed biofilms on commercially pure titanium (cpTi) starting with three different phases (ages) of the bacteria, to examine whether the detachment shear stress --as a signature for the work required for removal of the biofilms- would be affected by prior PDT treatment independently from microbial viability. Biofilms were grown with sucrose addition to Brain Heart Infusion media, producing visible thick films and nearly invisible thin films (within the same piece) having the same numbers of culturable microorganisms, the thicker films having greater susceptibility to detachment by water--jet impingement. Colony-forming-unit (CFU) counts routinely correlated well with results from a spectrophotometric Alamar Blue (AB) assay. Use of Methylene Blue (MB) as a photosensitizer (PS) for PDT of biofilms did not interfere with the AB assay, but did mask AB reduction spectral changes when employed with planktonic organisms. It was discovered in this work that PD-treated microbial biofilms, independently from starting or PS-influenced microorganism viability, were significantly (p<0.05) and differentially more easily delaminated and ultimately removed from their substrata biomaterials by the hydrodynamic forces of water-jet impingement. Control biofilms of varying thickness, not receiving PDT treatment, required between 144 and 228 dynes/cm2 of shear stress to delaminate from titanium while PDT-treated companion biofilms were removed at 90 to 140 dynes/cm2, depending on water flow rate. In comparison, it required only between 57 and 68 dynes/cm2 shear stress to separate microbial layers from within the exopolymer matrix of control biofilms, and between 39 and 51 dynes/cm2 to delaminate PDT-treated matrix sections of varying thickness biofilms, again depending on water flow rate. Multiple Attenuated Internal Reflection InfraRed spectra of identical biofilms, grown on germanium prisms having surface properties similar to those of cpTi, confirmed these differences in film-removal susceptibility for shear stresses as low as 10 dynes/cm2, and illustrated the PDT-induced preferential removal of predominantly the polysaccharide biofilm components. Scanning Electron Microscopy of Control and PDT-treated biofilms before and after water-jet impingement also confirmed these findings. These results are consistent with proposals that PDT induces oxidative embrittlement and fragmentation of plaque/biofilm matrix biopolymers, allowing easier release by hydrodynamic (rinsing) forces.

Effects of High Hydrostatic Pressure on Bacterial Growth on Human Ossicles Explanted from Cholesteatoma Patients

PubMed Central

Ostwald, Jürgen; Lindner, Tobias; Zautner, Andreas Erich; Arndt, Kathleen; Pau, Hans Wilhelm; Podbielski, Andreas

2012-01-01

Background High hydrostatic pressure (HHP) treatment can eliminate cholesteatoma cells from explanted human ossicles prior to re-insertion. We analyzed the effects of HHP treatment on the microbial flora on ossicles and on the planktonic and biofilm states of selected isolates. Methodology Twenty-six ossicles were explanted from cholesteatoma patients. Five ossicles were directly analyzed for microbial growth without further treatment. Fifteen ossicles were cut into two pieces. One piece was exposed to HHP of 350 MPa for 10 minutes. Both the treated and untreated (control) pieces were then assessed semi-quantitatively. Three ossicles were cut into two pieces and exposed to identical pressure conditions with or without the addition of one of two different combinations of antibiotics to the medium. Differential effects of 10-minute in vitro exposure of planktonic and biofilm bacteria to pressures of 100 MPa, 250 MPa, 400 MPa and 540 MPa in isotonic and hypotonic media were analyzed using two patient isolates of Staphylococcus epidermidis and Neisseria subflava. Bacterial cell inactivation and biofilm destruction were assessed by colony counting and electron microscopy. Principal Findings A variety of microorganisms were isolated from the ossicles. Irrespective of the medium, HHP treatment at 350 MPa for 10 minutes led to satisfying but incomplete inactivation especially of Gram-negative bacteria. The addition of antibiotics increased the efficacy of elimination. A comparison of HHP treatment of planktonic and biofilm cells showed that the effects of HPP were reduced by about one decadic logarithmic unit when HPP was applied to biofilms. High hydrostatic pressure conditions that are suitable to inactivate cholesteatoma cells fail to completely sterilize ossicles even if antibiotics are added. As a result of the reduced microbial load and the viability loss of surviving bacteria, however, there is a lower risk of re-infection after re-insertion. PMID:22291908

Biodegradability of carbon nanotube/polymer nanocomposites under aerobic mixed culture conditions.

PubMed

Phan, Duc C; Goodwin, David G; Frank, Benjamin P; Bouwer, Edward J; Fairbrother, D Howard

2018-10-15

The properties and commercial viability of biodegradable polymers can be significantly enhanced by the incorporation of carbon nanotubes (CNTs). The environmental impact and persistence of these carbon nanotube/polymer nanocomposites (CNT/PNCs) after disposal will be strongly influenced by their microbial interactions, including their biodegradation rates. At the end of consumer use, CNT/PNCs will encounter diverse communities of microorganisms in landfills, surface waters, and wastewater treatment plants. To explore CNT/PNC biodegradation under realistic environmental conditions, the effect of multi-wall CNT (MWCNT) incorporation on the biodegradation of polyhydroxyalkanoates (PHA) was investigated using a mixed culture of microorganisms from wastewater. Relative to unfilled PHA (0% w/w), the MWCNT loading (0.5-10% w/w) had no statistically significant effect on the rate of PHA matrix biodegradation. Independent of the MWCNT loading, the extent of CNT/PNC mass remaining closely corresponded to the initial mass of CNTs in the matrix suggesting a lack of CNT release. CNT/PNC biodegradation was complete in approximately 20 days and resulted in the formation of a compressed CNT mat that retained the shape of the initial CNT/PNC. This study suggests that although CNTs have been shown to be cytotoxic towards a range of different microorganisms, this does not necessarily impact the biodegradation of the surrounding polymer matrix in mixed culture, particularly in situations where the polymer type and/or microbial population favor rapid polymer biodegradation. Copyright © 2018 Elsevier B.V. All rights reserved.

Evaluation of the effects of different liquid inoculant formulations on the survival and plant-growth-promoting efficiency of Rhodopseudomonas palustris strain PS3.

PubMed

Lee, Sook-Kuan; Lur, Huu-Sheng; Lo, Kai-Jiun; Cheng, Kuan-Chen; Chuang, Chun-Chao; Tang, Shiueh-Jung; Yang, Zhi-Wei; Liu, Chi-Te

2016-09-01

Biofertilizers can help improve soil quality, promote crop growth, and sustain soil health. The photosynthetic bacterium Rhodopseudomonas palustris strain PS3 (hereafter, PS3), which was isolated from Taiwanese paddy soil, can not only exert beneficial effects on plant growth but also enhance the efficiency of nutrient uptake from applied fertilizer. To produce this elite microbial isolate for practical use, product development and formulation are needed to permit the maintenance of the high quality of the inoculant during storage. The aim of this study was to select a suitable formulation that improves the survival and maintains the beneficial effects of the PS3 inoculant. Six additives (alginate, polyethylene glycol [PEG], polyvinylpyrrolidone-40 [PVP], glycerol, glucose, and horticultural oil) were used in liquid-based formulations, and their capacities for maintaining PS3 cell viability during storage in low, medium, and high temperature ranges were evaluated. Horticultural oil (0.5 %) was chosen as a potential additive because it could maintain a relatively high population and conferred greater microbial vitality under various storage conditions. Furthermore, the growth-promoting effects exerted on Chinese cabbage by the formulated inoculants were significantly greater than those of the unformulated treatments. The fresh and dry weights of the shoots were significantly increased, by 10-27 and 22-40 %, respectively. Horticultural oil is considered a safe, low-cost, and easy-to-process material, and this formulation would facilitate the practical use of strain PS3 in agriculture.

Cell growth and protein expression of Shewanella oneidensis in biofilms and hydrogel-entrapped cultures.

PubMed

Zhang, Yingdan; Ng, Chun Kiat; Cohen, Yehuda; Cao, Bin

2014-05-01

The performance of biofilm-based bioprocesses is difficult to predict and control because of the intrinsic heterogeneous and dynamic properties of microbial biofilms. Biofilm mimics, such as microbial cells entrapped in polymeric scaffolds that are permeable for nutrients, have been proposed to replace real biofilms to achieve long-term robust performance in engineering applications. However, the physiological differences between cells that are physically entrapped in a synthetic polymeric matrix and biofilm cells that are encased in a self-produced polymeric matrix remain unknown. In this study, using Shewanella oneidensis as a model organism and alginate hydrogel as a model synthetic matrix, we compared the cell growth and protein expression in entrapped cultures and biofilms. The hydrogel-entrapped cultures were found to exhibit a growth rate comparable with biofilms. There was no substantial difference in cell viability, surface charge, as well as hydrophobicity between the cells grown in alginate hydrogel and those grown in biofilms. However, the gel-entrapped cultures were found to be physiologically different from biofilms. The gel-entrapped cultures had a higher demand for metabolic energy. The siderophore-mediated iron uptake was repressed in the gel-entrapped cells. The presence of the hydrogel matrix decreased the expression of proteins involved in biofilm formation, while inducing the production of extracellular DNA (eDNA) in the gel-entrapped cultures. These results advance the fundamental understanding of the physiology of hydrogel-entrapped cells, which can lead to more efficient biofilm mimic-based applications.

Biochemical Traits, Survival and Biological Properties of the Probiotic Lactobacillus plantarum Grown in the Presence of Prebiotic Inulin and Pectin as Energy Source

PubMed Central

Nazzaro, Filomena; Fratianni, Florinda; Orlando, Pierangelo; Coppola, Raffaele

2012-01-01

The viability of the probiotic strain Lactobacillus plantarum subsp. plantarum, after its passage through simulated gastric and pancreatic juices, was evaluated as function of its pre-growth in a medium containing the known prebiotics pectin or inulin, and was compared to glucose used as control. The presence of pectin or inulin did not markedly affect the growth (10.07 log10 colony forming units/mL and 10.28 log10 colony forming units/mL for pectin and inulin respectively versus 10.42 log10 colony forming units/mL obtained for glucose). Pectin and inulin, in contrast to glucose, induced cell stress resistance against gastrointestinal juices (Δ log101.5 and 2.4 colony forming units/mL respectively, versus Δ log10 4.0 for glucose). The data were corroborated by the analysis of the protein pattern following stress treatments which, in the case of microbial cells grown with glucose, revealed a more marked protein degradation after the double passage through simulated gastric and intestinal juices. Inulin stimulated the production of the relevant healthy bio-molecule butyrate, which amount was 30% higher respect of growth in the presence of glucose. Inulin and pectin improved cell DPPH scavenging activity, and an impressive hydrophobicity (35.28% and 34.81%, respectively) was observed with respect to the microbial growth in presence of glucose (3.39%). PMID:24281559

Louis Pasteur, the Father of Immunology?

PubMed Central

Smith, Kendall A.

2012-01-01

Louis Pasteur is traditionally considered as the progenitor of modern immunology because of his studies in the late nineteenth century that popularized the germ theory of disease, and that introduced the hope that all infectious diseases could be prevented by prophylactic vaccination, as well as also treated by therapeutic vaccination, if applied soon enough after infection. However, Pasteur was working at the dawn of the appreciation of the microbial world, at a time when the notion of such a thing as an immune system did not exist, certainly not as we know it today, more than 130 years later. Accordingly, why was Pasteur such a genius as to discern how the immune system functions to protect us against invasion by the microbial world when no one had even made the distinction between fungi, bacteria, or viruses, and no one had formulated any theories of immunity. A careful reading of Pasteur’s presentations to the Academy of Sciences reveals that Pasteur was entirely mistaken as to how immunity occurs, in that he reasoned, as a good microbiologist would, that appropriately attenuated microbes would deplete the host of vital trace nutrients absolutely required for their viability and growth, and not an active response on the part of the host. Even so, he focused attention on immunity, preparing the ground for others who followed. This review chronicles Pasteur’s remarkable metamorphosis from organic chemist to microbiologist to immunologist, and from basic science to medicine. PMID:22566949

The macrophage soluble receptor AIM/Api6/CD5L displays a broad pathogen recognition spectrum and is involved in early response to microbial aggression

PubMed Central

Martinez, Vanesa G.; Escoda-Ferran, Cristina; Tadeu Simões, Inês; Arai, Satoko; Orta Mascaró, Marc; Carreras, Esther; Martínez-Florensa, Mario; Yelamos, José; Miyazaki, Toru; Lozano, Francisco

2014-01-01

Apoptosis inhibitor of macrophages (AIMs), a homologue of human Spα, is a mouse soluble member of the scavenger receptor cysteine-rich superfamily (SRCR-SF). This family integrates a group of proteins expressed by innate and adaptive immune cells for which no unifying function has yet been described. Pleiotropic functions have been ascribed to AIM, from viability support in lymphocytes during thymic selection to lipid metabolism and anti-inflammatory effects in autoimmune pathologies. In the present report, the pathogen binding properties of AIM have been explored. By using a recombinant form of AIM (rAIM) expressed in mammalian cells, it is shown that this protein is able to bind and aggregate Gram-positive and Gram-negative bacteria, as well as pathogenic and saprophytic fungal species. Importantly, endogenous AIM from mouse serum also binds to microorganisms and secretion of AIM was rapidly induced in mouse spleen macrophages following exposure to conserved microbial cell wall components. Cytokine release induced by well-known bacterial and fungal Toll-like receptor (TLR) ligands on mouse splenocytes was also inhibited in the presence of rAIM. Furthermore, mouse models of pathogen-associated molecular patterns (PAMPs)-induced septic shock of bacterial and fungal origin showed that serum AIM levels changed in a time-dependent manner. Altogether, these data suggest that AIM plays a general homeostatic role by supporting innate humoral defense during pathogen aggression. PMID:24583716

Effects of Background Fluid on the Efficiency of Inactivating Yeast with Non-Thermal Atmospheric Pressure Plasma

PubMed Central

Ryu, Young-Hyo; Kim, Yong-Hee; Lee, Jin-Young; Shim, Gun-Bo; Uhm, Han-Sup; Park, Gyungsoon; Choi, Eun Ha

2013-01-01

Non-thermal plasma at atmospheric pressure has been actively applied to sterilization. However, its efficiency for inactivating microorganisms often varies depending on microbial species and environments surrounding the microorganisms. We investigated the influence of environmental factors (surrounding media) on the efficiency of microbial inactivation by plasma using an eukaryotic model microbe, Saccharomyces cerevisiae, to elucidate the mechanisms for differential efficiency of sterilization by plasma. Yeast cells treated with plasma in water showed the most severe damage in viability and cell morphology as well as damage to membrane lipids, and genomic DNA. Cells in saline were less damaged compared to those in water, and those in YPD (Yeast extract, Peptone, Dextrose) were least impaired. HOG1 mitogen activated protein kinase was activated in cells exposed to plasma in water and saline. Inactivation of yeast cells in water and saline was due to the acidification of the solutions by plasma, but higher survival of yeast cells treated in saline may have resulted from the additional effect related to salt strength. Levels of hydroxyl radical (OH.) produced by plasma were the highest in water and the lowest in YPD. This may have resulted in differential inactivation of yeast cells in water, saline, and YPD by plasma. Taken together, our data suggest that the surrounding media (environment) can crucially affect the outcomes of yeast cell plasma treatment because plasma modulates vital properties of media, and the toxic nature of plasma can also be altered by the surrounding media. PMID:23799081

Biological properties and therapeutic activities of honey in wound healing: A narrative review and meta-analysis.

PubMed

Oryan, Ahmad; Alemzadeh, Esmat; Moshiri, Ali

2016-05-01

For thousands of years, honey has been used for medicinal applications. The beneficial effects of honey, particularly its anti-microbial activity represent it as a useful option for management of various wounds. Honey contains major amounts of carbohydrates, lipids, amino acids, proteins, vitamin and minerals that have important roles in wound healing with minimum trauma during redressing. Because bees have different nutritional behavior and collect the nourishments from different and various plants, the produced honeys have different compositions. Thus different types of honey have different medicinal value leading to different effects on wound healing. This review clarifies the mechanisms and therapeutic properties of honey on wound healing. The mechanisms of action of honey in wound healing are majorly due to its hydrogen peroxide, high osmolality, acidity, non-peroxide factors, nitric oxide and phenols. Laboratory studies and clinical trials have shown that honey promotes autolytic debridement, stimulates growth of wound tissues and stimulates anti-inflammatory activities thus accelerates the wound healing processes. Compared with topical agents such as hydrofiber silver or silver sulfadiazine, honey is more effective in elimination of microbial contamination, reduction of wound area, promotion of re-epithelialization. In addition, honey improves the outcome of the wound healing by reducing the incidence and excessive scar formation. Therefore, application of honey can be an effective and economical approach in managing large and complicated wounds. Copyright © 2015 Tissue Viability Society. Published by Elsevier Ltd. All rights reserved.

Depuration of striped venus clam (Chamelea gallina L.): effects on microorganisms, sand content, and mortality.

PubMed

Maffei, M; Vernocchi, P; Lanciotti, R; Guerzoni, M E; Belletti, N; Gardini, F

2009-01-01

This study was focused on the evaluation of the microbiological indices, defined by European legislation, before and after the depuration of clams (Chamelea gallina) landed in category B seawater. The survival of depurated clams and the meat yield were also evaluated. The results obtained from October 2002 to September 2003 evidenced a mean microbial reduction during depuration of 62% for Escherichia coli and 54% for fecal coliforms (FC). All the samples had FC counts below the limit after 24 h depuration with the exception of the August samples. E. coli was found in concentration slightly higher than the legal limit only in the samples of December and January. In August, the E. coli concentration did not decrease during the depuration, while in the other samples significant reduction of E. coli concentration was observed. Salmonella spp. and V. parahaemolyticus were never detected in the clams harvested between March and September. Vibrio alginolyticus was found in the clams harvested in May and September both before and after the depuration process. The viability of clam was not negatively affected by depuration, in fact, an increase of viability was observed with the exception of the samples of April. The meat yield was not influenced by the depurative treatment in C. gallina; the mean value found before depuration, 10.47% (with 1.95 SD), did not significantly vary after the treatment (10.58%, SD 2.07). In conclusion, the depuration conditions can improve the quality of C. gallina; however, its effects on microbiological quality depended on environmental conditions.

Microbial nitrogen cycling response to forest-based bioenergy production.

PubMed

Minick, Kevan J; Strahm, Brian D; Fox, Thomas R; Sucre, Eric B; Leggett, Zakiya H

2015-12-01

Concern over rising atmospheric CO2 and other greenhouse gases due to fossil fuel combustion has intensified research into carbon-neutral energy production. Approximately 15.8 million ha of pine plantations exist across the southeastern United States, representing a vast land area advantageous for bioenergy production without significant landuse change or diversion of agricultural resources from food production. Furthermore, intercropping of pine with bioenergy grasses could provide annually harvestable, lignocellulosic biomass feedstocks along with production of traditional wood products. Viability of such a system hinges in part on soil nitrogen (N) availability and effects of N competition between pines and grasses on ecosystem productivity. We investigated effects of intercropping loblolly pine (Pinus taeda) with switchgrass (Panicum virgatum) on microbial N cycling processes in the Lower Coastal Plain of North Carolina, USA. Soil samples were collected from bedded rows of pine and interbed space of two treatments, composed of either volunteer native woody and herbaceous vegetation (pine-native) or pure switchgrass (pine-switchgrass) in interbeds. An in vitro 15N pool-dilution technique was employed to quantify gross N transformations at two soil depths (0-5 and 5-15 cm) on four dates in 2012-2013. At the 0-5 cm depth in beds of the pine-switchgrass treatment, gross N mineralization was two to three times higher in November and February compared to the pine-native treatment, resulting in increased NH4(+) availability. Gross and net nitrification were also significantly higher in February in the same pine beds. In interbeds of the pine-switchgrass treatment, gross N mineralization was lower from April to November, but higher in February, potentially reflecting positive effects of switchgrass root-derived C inputs during dormancy on microbial activity. These findings indicate soil N cycling and availability has increased in pine beds of the pine-switchgrass treatment compared to those of the pine-native treatment, potentially alleviating any negative effects of N competition between pine and switchgrass. We expect that reduced soil C in the pine-switchgrass treatment, effects of pine and switchgrass rooting on soil C availability, and plant N demand are major factors influencing soil N transformations. Future research should examine rooting architecture in-intercropped systems and the effects on soil microbial communities and function.

Vapor Hydrogen Peroxide as Alternative to Dry Heat Microbial Reduction

NASA Technical Reports Server (NTRS)

Cash, Howard A.; Kern, Roger G.; Chung, Shirley Y.; Koukol, Robert C.; Barengoltz, Jack B.

2006-01-01

The Jet Propulsion Laboratory, in conjunction with the NASA Planetary Protection Officer, has selected vapor phase hydrogen peroxide (VHP) sterilization process for continued development as a NASA approved sterilization technique for spacecraft subsystems and systems. The goal is to include this technique, with appropriate specification, in NPG8020.12C as a low temperature complementary technique to the dry heat sterilization process. A series of experiments were conducted in vacuum to determine VHP process parameters that provided significant reductions in spore viability while allowing survival of sufficient spores for statistically significant enumeration. With this knowledge of D values, sensible margins can be applied in a planetary protection specification. The outcome of this study provided an optimization of test sterilizer process conditions: VHP concentration, process duration, a process temperature range for which the worst case D value may be imposed, a process humidity range for which the worst case D value may be imposed, and robustness to selected spacecraft material substrates.

Air plasma effect on dental disinfection

NASA Astrophysics Data System (ADS)

Duarte, S.; Kuo, S. P.; Murata, R. M.; Chen, C. Y.; Saxena, D.; Huang, K. J.; Popovic, S.

2011-07-01

A nonthermal low temperature air plasma jet is characterized and applied to study the plasma effects on oral pathogens and biofilms. Experiments were performed on samples of six defined microorganisms' cultures, including those of gram-positive bacteria and fungi, and on a cultivating biofilm sample of Streptococcus mutans UA159. The results show that the plasma jet creates a zone of microbial growth inhibition in each treated sample; the zone increases with the plasma treatment time and expands beyond the entire region directly exposed to the plasma jet. With 30s plasma treatment twice daily during 5 days of biofilm cultivation, its formation was inhibited. The viability of S. mutans cells in the treated biofilms dropped to below the measurable level and the killed bacterial cells concentrated to local regions as manifested by the fluorescence microscopy via the environmental scanning electron microscope. The emission spectroscopy of the jet indicates that its plasma effluent carries an abundance of reactive atomic oxygen, providing catalyst for the observed plasma effect.

Assessment of Plant-Probiotic Performance of Novel Endophytic Bacillus sp. in Talc-Based Formulation.

PubMed

Basheer, Jasim; Ravi, Aswani; Mathew, Jyothis; Krishnankutty, Radhakrishnan Edayileveettil

2018-01-25

Endophytic bacteria are considered to have a plethora of plant growth promoting and anti-phytopathogenic traits to live within the plants. Hence, they have immense promises for plant probiotic development. In the current study, plant probiotic endophytic Bacillus sp. CaB5 which has been previously isolated from Capsicum annuum was investigated for its performance in talc-based formulation. For this, CaB5 was made into formulation with sterile talc, calcium carbonate, and carboxymethyl cellulose. The viability analysis of the formulation by standard plate count and fluorescence methods has confirmed the stable microbial count up to 45 days. Plant probiotic performance of the prepared formulation was analyzed on cowpea (Vigna unguiculata) and lady's finger (Abelmoschus esculentus). The results showed the formulation treatment to have enhancement effect on seed germination as well as plant growth in both selected plants. The results highlight the potential of CaB5-based formulation for field application to enhance growth of economically important plants.

Antimicrobial polycaprolactone/polyethylene glycol embedded lysozyme coatings of Ti implants for osteoblast functional properties in tissue engineering

NASA Astrophysics Data System (ADS)

Visan, A.; Cristescu, R.; Stefan, N.; Miroiu, M.; Nita, C.; Socol, M.; Florica, C.; Rasoga, O.; Zgura, I.; Sima, L. E.; Chiritoiu, M.; Chifiriuc, M. C.; Holban, A. M.; Mihailescu, I. N.; Socol, G.

2017-09-01

In this study, coatings based on lysozyme embedded into a matrix of polyethylene glycol (PEG) and polycaprolactone (PCL) were fabricated by two different methods (Matrix Assisted Pulsed Laser Evaporation - MAPLE and Dip Coating) for obtaining antimicrobial coatings envisaged for long term medical applications. Coatings with different PEG:PCL compositions (3:1; 1:1; 1:3) were synthesized in order to evaluate the antimicrobial activity of lysozyme embedded into the polymeric matrix. The main surface features, such as roughness and wettability, with impact on the microbial adhesion as well as on the eukaryote cell function were measured. The obtained composite coatings exhibited a significant antibacterial activity against Escherichia coli, Bacillus subtilis, Enterococcus faecalis and Staphylococcus aureus strains. As well, specific blended coatings showed appropriate viability, good spreading and normal cell morphology of SaOs2 human osteoblasts and mesenchymal stem cells (MSCs). These investigations highlight the suitability of biodegradable composites as implant coatings for decreasing the risk of bacterial contamination associated with prosthetic procedures.

Photodynamic inactivation of multiresistant bacteria (KPC) using zinc(II)phthalocyanines.

PubMed

Miretti, Mariana; Clementi, Romina; Tempesti, Tomas C; Baumgartner, María T

2017-09-15

The worldwide increase in antibiotic resistance has led to search of alternatives anti-microbial therapies such as photodynamic inactivation. The aim of this paper was to evaluate the photodynamic activity in vitro of a neutral and two cationic Zn phthalocyanines. Their photokilling activity was tested on Escherichia coli ATCC 25922 and Klebsiella pneumoniae Carbapenemase (KPC)-producing. After treating bacteria with phthalocyanines, the cultures were irradiated with white light. As a result, the bacteria were inactivated in presence of cationic phthalocyanines. The photoinactivation was dependent of the irradiation time and phthalocyanine concentration. The most effective photosensitizer on KPC-producing was Zinc(II)tetramethyltetrapyridino[2,3-b:2',3'-g:2″,3″-l:2‴,3‴-q]porphyrazinium methylsulfate (ZnTM2,3PyPz). After irradiation using the water soluble ZnTM2,3PyPz (3μM) the viability of KPC (30min of irradiation) and E. coli (10min of irradiation) decreased ≈99.995%. Copyright © 2017 Elsevier Ltd. All rights reserved.

Air plasma effect on dental disinfection

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

Duarte, S.; Murata, R. M.; Saxena, D.

2011-07-15

A nonthermal low temperature air plasma jet is characterized and applied to study the plasma effects on oral pathogens and biofilms. Experiments were performed on samples of six defined microorganisms' cultures, including those of gram-positive bacteria and fungi, and on a cultivating biofilm sample of Streptococcus mutans UA159. The results show that the plasma jet creates a zone of microbial growth inhibition in each treated sample; the zone increases with the plasma treatment time and expands beyond the entire region directly exposed to the plasma jet. With 30s plasma treatment twice daily during 5 days of biofilm cultivation, its formationmore » was inhibited. The viability of S. mutans cells in the treated biofilms dropped to below the measurable level and the killed bacterial cells concentrated to local regions as manifested by the fluorescence microscopy via the environmental scanning electron microscope. The emission spectroscopy of the jet indicates that its plasma effluent carries an abundance of reactive atomic oxygen, providing catalyst for the observed plasma effect.« less

Caprylic and Polygalacturonic Acid Combinations for Eradication of Microbial Organisms Embedded in Biofilm

PubMed Central

Rosenblatt, Joel; Reitzel, Ruth A.; Vargas-Cruz, Nylev; Chaftari, Anne-Marie; Hachem, Ray; Raad, Issam

2017-01-01

There is a need for non-antibiotic, antimicrobial compositions with low toxicity capable of broad-spectrum eradication of pathogenic biofilms in food preparation and healthcare settings. In this study we demonstrated complete biofilm eradication within 60 min with synergistic combinations of caprylic and polygalacturonic (PG) acids in an in vitro biofilm eradication model against representative hospital and foodborne infectious pathogen biofilms (methicillin-resistant Staphylococcus aureus, multidrug-resistant Pseudomonas aeruginosa, Candida albicans, Escherichia coli, and Salmonella enteritidis). Antimicrobial synergy against biofilms was demonstrated by quantifying viable organisms remaining in biofilms exposed to caprylic acid alone, PG acid alone, or combinations of the two. The combinations also synergistically inhibited growth of planktonic organisms. Toxicity of the combination was assessed in vitro on L929 fibroblasts incubated with extracts of caprylic and PG acid combinations using the Alamar Blue metabolic activity assay and the Trypan Blue exclusion cell viability assay. The extracts did not produce cytotoxic responses relative to untreated control fibroblasts. PMID:29093703

Harvesting energy from the marine sediment--water interface.

PubMed

Reimers, C E; Tender, L M; Fertig, S; Wang, W

2001-01-01

Pairs of platinum mesh or graphite fiber-based electrodes, one embedded in marine sediment (anode), the other in proximal seawater (cathode), have been used to harvest low-level power from natural, microbe established, voltage gradients at marine sediment-seawater interfaces in laboratory aquaria. The sustained power harvested thus far has been on the order of 0.01 W/m2 of electrode geometric area but is dependent on electrode design, sediment composition, and temperature. It is proposed that the sediment/anode-seawater/cathode configuration constitutes a microbial fuel cell in which power results from the net oxidation of sediment organic matter by dissolved seawater oxygen. Considering typical sediment organic carbon contents, typical fluxes of additional reduced carbon by sedimentation to sea floors < 1,000 m deep, and the proven viability of dissolved seawater oxygen as an oxidant for power generation by seawater batteries, it is calculated that optimized power supplies based on the phenomenon demonstrated here could power oceanographic instruments deployed for routine long-term monitoring operations in the coastal ocean.

Non-thermal atmospheric-pressure plasma possible application in wound healing.

PubMed

Haertel, Beate; von Woedtke, Thomas; Weltmann, Klaus-Dieter; Lindequist, Ulrike

2014-11-01

Non-thermal atmospheric-pressure plasma, also named cold plasma, is defined as a partly ionized gas. Therefore, it cannot be equated with plasma from blood; it is not biological in nature. Non-thermal atmospheric-pressure plasma is a new innovative approach in medicine not only for the treatment of wounds, but with a wide-range of other applications, as e.g. topical treatment of other skin diseases with microbial involvement or treatment of cancer diseases. This review emphasizes plasma effects on wound healing. Non-thermal atmospheric-pressure plasma can support wound healing by its antiseptic effects, by stimulation of proliferation and migration of wound relating skin cells, by activation or inhibition of integrin receptors on the cell surface or by its pro-angiogenic effect. We summarize the effects of plasma on eukaryotic cells, especially on keratinocytes in terms of viability, proliferation, DNA, adhesion molecules and angiogenesis together with the role of reactive oxygen species and other components of plasma. The outcome of first clinical trials regarding wound healing is pointed out.

Development of an integrated electrochemical system for in vitro yeast viability testing.

PubMed

Adami, Andrea; Ress, Cristina; Collini, Cristian; Pedrotti, Severino; Lorenzelli, Leandro

2013-02-15

This work describes the development and testing of a microfabricated sensor for rapid cell growth monitoring, especially focused on yeast quality assessment for wine applications. The device consists of a NMOS ISFET sensor with Si(3)N(4) gate, able to indirectly monitor extracellular metabolism through pH variation of the medium, and a solid-state reference electrode implemented with PVC membranes doped with lipophilic salts (tetrabutylammonium-tetrabutylborate (TBA-TBB) and Potassium tetrakis(4-chlorphenyl)borate (KTClpB)). The use of a solid state reference electrode enables the implementation of a large number of cell assays in parallel, without the need of external conventional reference electrodes. Microbial growth testing has been performed both in standard culture conditions and on chip at different concentrations of ethanol in order to carry out a commonly used screening of wine yeast strains. Cell growth tests can be performed in few hours, providing a fast, sensitive and low cost analysis with respect to the conventional procedures. Copyright © 2012 Elsevier B.V. All rights reserved.

Role of Geitlerinema sp. DE2011 and Scenedesmus sp. DE2009 as Bioindicators and Immobilizers of Chromium in a Contaminated Natural Environment.

PubMed

Millach, Laia; Solé, Antoni; Esteve, Isabel

2015-01-01

The aim of this work was to study the potential of the two phototrophic microorganisms, both isolated from Ebro Delta microbial mats, to be used as bioindicators and immobilizers of chromium. The results obtained indicated that (i) the Minimum Metal Concentration (MMC) significantly affecting Chlorophyll a intensity in Geitlerinema sp. DE2011 and Scenedesmus sp. DE2009 was 0.25 µM and 0.75 µM, respectively, these values being lower than those established by current legislation, and (ii) Scenedesmus sp. DE2009 was able to immobilize chromium externally in extracellular polymeric substances (EPS) and intracellularly in polyphosphate (PP) inclusions. Additionally, this microorganism maintained high viability, including at 500 µM. Based on these results, we postulate that Geitlerinema sp. DE2011 and Scenedesmus sp. DE2009 are good chromium-indicators of cytotoxicity and, further, that Scenedesmus sp. DE2009 plays an important role in immobilizing this metal in a contaminated natural environment.

Role of Geitlerinema sp. DE2011 and Scenedesmus sp. DE2009 as Bioindicators and Immobilizers of Chromium in a Contaminated Natural Environment

PubMed Central

2015-01-01

The aim of this work was to study the potential of the two phototrophic microorganisms, both isolated from Ebro Delta microbial mats, to be used as bioindicators and immobilizers of chromium. The results obtained indicated that (i) the Minimum Metal Concentration (MMC) significantly affecting Chlorophyll a intensity in Geitlerinema sp. DE2011 and Scenedesmus sp. DE2009 was 0.25 µM and 0.75 µM, respectively, these values being lower than those established by current legislation, and (ii) Scenedesmus sp. DE2009 was able to immobilize chromium externally in extracellular polymeric substances (EPS) and intracellularly in polyphosphate (PP) inclusions. Additionally, this microorganism maintained high viability, including at 500 µM. Based on these results, we postulate that Geitlerinema sp. DE2011 and Scenedesmus sp. DE2009 are good chromium-indicators of cytotoxicity and, further, that Scenedesmus sp. DE2009 plays an important role in immobilizing this metal in a contaminated natural environment. PMID:26167488

Non-Thermal Atmospheric-Pressure Plasma Possible Application in Wound Healing

PubMed Central

Haertel, Beate; von Woedtke, Thomas; Weltmann, Klaus-Dieter; Lindequist, Ulrike

2014-01-01

Non-thermal atmospheric-pressure plasma, also named cold plasma, is defined as a partly ionized gas. Therefore, it cannot be equated with plasma from blood; it is not biological in nature. Non-thermal atmospheric-pressure plasma is a new innovative approach in medicine not only for the treatment of wounds, but with a wide-range of other applications, as e.g. topical treatment of other skin diseases with microbial involvement or treatment of cancer diseases. This review emphasizes plasma effects on wound healing. Non-thermal atmospheric-pressure plasma can support wound healing by its antiseptic effects, by stimulation of proliferation and migration of wound relating skin cells, by activation or inhibition of integrin receptors on the cell surface or by its pro-angiogenic effect. We summarize the effects of plasma on eukaryotic cells, especially on keratinocytes in terms of viability, proliferation, DNA, adhesion molecules and angiogenesis together with the role of reactive oxygen species and other components of plasma. The outcome of first clinical trials regarding wound healing is pointed out. PMID:25489414

Long-term autonomous resistivity monitoring of oil-contaminated sediments from the Deepwater Horizon spill

NASA Astrophysics Data System (ADS)

Heenan, J. W.; Slater, L. D.; Ntarlagiannis, D.; Atekwana, E. A.; Ross, C.; Nolan, J. T.; Atekwana, E. A.; Werkema, D. D.; Fathepure, B.

2012-12-01

We conducted a long-term electrical resistivity survey at Grand Terre 1 (GT1) Island off the coast of Louisiana, a site contaminated with crude oil associated with the April 2010 BP Deepwater Horizon oil spill. Electrical resistivity has proven sensitivity to biogeochemical processes associated with the biodegradation of hydrocarbons in the subsurface. However, most of these studies have been in freshwater environments and for aged spills. The BP Deepwater Horizon oil spill therefore provided an unprecedented opportunity to capture the early time biogeophysical signals resulting from the physical, chemical and microbial transformation of crude oil in highly saline environments. We used a multi-channel resistivity system powered by solar panels to obtain continuous measurements twice a day on both a surface array and two shallow borehole arrays. This system operated for approximately 1.5 years and provided a unique long-term dataset of resistivity changes. Temperature and specific conductance values for the shallow groundwater were continuously logged. . Resistivity changes likely associated with biodegradation processes were then isolated from these environmental factors by modeling. In addition, groundwater was sampled for geochemical analyses from wells installed at the study site and soil samples were collected for microbial analyses at several locations, including both contaminated and uncontaminated locations. Microcosms were set up to determine the biodegradation potential of indigenous populations, and microbial diversity analysis was used to determine microbial community composition. Surface and borehole resistivity arrays revealed an initial resistive anomaly co-located with the known contamination. Pixel time series analysis of an inverted time sequence of resistivity sections highlighted differing responses between contaminated and uncontaminated locations. The contaminated locations exhibit persistent resistivity decreases over time, whereas areas outside of the contaminated location exhibit relatively uniform resistivity or show clear evidence of seasonal effect. Temperature-corrected resistivity changes show no direct correlation with pore fluid specific conductance changes, suggesting that specific conductance changes (e.g. due to tides) have little influence on imaged resistivity structure. Microbial data suggest that resistivity changes within the contaminated location resulted from biodegradation, showing the presence of native populations capable of degrading aromatic hydrocarbons at salinities ranging from 6 to 15 % NaCl within the contaminated location. Aqueous geochemical measurements performed on samples from the site further indicate that at depth intervals coincident with the resistivity anomaly, marked increases in the concentration of dissolved inorganic carbon (DIC) were observed suggesting biodegradation of petroleum hydrocarbon although other DIC generating processes such as organic matter degradation coupled to sulfate and iron reduction were also prominent. This experiment demonstrates the potential viability of long-term autonomous electrical monitoring as a means of decreasing the frequency of more costly and invasive chemical analysis of natural attenuation.

Recovery of soil unicellular eukaryotes: an efficiency and activity analysis on the single cell level.

PubMed

Lentendu, Guillaume; Hübschmann, Thomas; Müller, Susann; Dunker, Susanne; Buscot, François; Wilhelm, Christian

2013-12-01

Eukaryotic unicellular organisms are an important part of the soil microbial community, but they are often neglected in soil functional microbial diversity analysis, principally due to the absence of specific investigation methods in the special soil environment. In this study we used a method based on high-density centrifugation to specifically isolate intact algal and yeast cells, with the aim to analyze them with flow cytometry and sort them for further molecular analysis such as deep sequencing. Recovery efficiency was tested at low abundance levels that fit those in natural environments (10(4) to 10(6) cells per g soil). Five algae and five yeast morphospecies isolated from soil were used for the testing. Recovery efficiency was between 1.5 to 43.16% and 2 to 30.2%, respectively, and was dependent on soil type for three of the algae. Control treatments without soil showed that the majority of cells were lost due to the method itself (58% and 55.8% respectively). However, the cell extraction technique did not much compromise cell vitality because a fluorescein di-acetate assay indicated high viability percentages (73.3% and 97.2% of cells, respectively). The low abundant algae and yeast morphospecies recovered from soil were cytometrically analyzed and sorted. Following, their DNA was isolated and amplified using specific primers. The developed workflow enables isolation and enrichment of intact autotrophic and heterotrophic soil unicellular eukaryotes from natural environments for subsequent application of deep sequencing technologies. Copyright © 2013 Elsevier B.V. All rights reserved.

Alleviating monoterpene toxicity using a two-phase extractive fermentation for the bioproduction of jet fuel mixtures in Saccharomyces cerevisiae.

PubMed

Brennan, Timothy C R; Turner, Christopher D; Krömer, Jens O; Nielsen, Lars K

2012-10-01

Monoterpenes are a diverse class of compounds with applications as flavors and fragrances, pharmaceuticals and more recently, jet fuels. Engineering biosynthetic pathways for monoterpene production in microbial hosts has received increasing attention. However, monoterpenes are highly toxic to many microorganisms including Saccharomyces cerevisiae, a widely used industrial biocatalyst. In this work, the minimum inhibitory concentration (MIC) for S. cerevisiae was determined for five monoterpenes: β-pinene, limonene, myrcene, γ-terpinene, and terpinolene (1.52, 0.44, 2.12, 0.70, 0.53 mM, respectively). Given the low MIC for all compounds tested, a liquid two-phase solvent extraction system to alleviate toxicity during fermentation was evaluated. Ten solvents were tested for biocompatibility, monoterpene distribution, phase separation, and price. The solvents dioctyl phthalate, dibutyl phthalate, isopropyl myristate, and farnesene showed greater than 100-fold increase in the MIC compared to the monoterpenes in a solvent-free system. In particular, the MIC for limonene in dibutyl phthalate showed a 702-fold (308 mM, 42.1 g L(-1) of limonene) improvement while cell viability was maintained above 90%, demonstrating that extractive fermentation is a suitable tool for the reduction of monoterpene toxicity. Finally, we estimated that a limonane to farnesane ratio of 1:9 has physicochemical properties similar to traditional Jet-A aviation fuel. Since farnesene is currently produced in S. cerevisiae, its use as a co-product and extractant for microbial terpene-based jet fuel production in a two-phase system offers an attractive bioprocessing option. Copyright © 2012 Wiley Periodicals, Inc.

Impact of the morphology and reactivity of nanoscale zero-valent iron (NZVI) on dechlorinating bacteria.

PubMed

Rónavári, Andrea; Balázs, Margit; Tolmacsov, Péter; Molnár, Csaba; Kiss, István; Kukovecz, Ákos; Kónya, Zoltán

2016-05-15

Nanoscale zero-valent iron (NZVI) is increasingly used for reducing chlorinated organic contaminants in soil or groundwater. However, little is known about what impact the particles will have on the biochemical processes and the indigenous microbial communities. Nanoiron reactivity is affected by the structure and morphology of nanoparticles that complicates the applicability in bioremediation. In this study, the effect of precursors (ferrous sulfate and ferric chloride) and reducing agents (sodium dithionite and sodium borohydride) on the morphology and the reactivity of NZVIs was investigated. We also studied the impact of differently synthesized NZVIs on microbial community, which take part in reductive dechlorination. We demonstrated that both the applied iron precursor and the reducing agent had influence on the structure of the nanoparticles. Spherical nanoparticles with higher Fe(0) content (>90%) was observed by using sodium borohydride as reducing agent, while application of sodium dithionite as reducing agent resulted nanostructures with lower Fe(0) content (between 68,7 and 85,5%). To determine the influence of differently synthesized NZVIs on cell viability anaerobic enriched microcosm were used. NVZI was used in 0.1 g/L concentration in all batch experiments. Relative amount of Dehalococcoides, sulfate reducers (SRBs) and methanogens were measured by quantitative PCR. We found that the relative amount of Dehalococcoides slowly decreased in all experiments independently from the precursor and reducing agent, whereas the total amount of microbes increased. The only clear distinction was in relative amount of sulfate reducers which were higher in the presence of NZVIs synthesized from sodium dithionite. Copyright © 2016 Elsevier Ltd. All rights reserved.

Role of the competitive microbial flora in the radiation-induced enhancement of ochratoxin production by Aspergillus alutaceus var. alutaceus NRRL 3174

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

Chelack, W.S.; Borsa, J.; Marquardt, R.R.

1991-09-01

The radiation sensitivity and the toxigenic potential of conidiospores of the fungus Aspergillus alutaceus var alutaceus were determined after irradiation with {sup 60}Co gamma rays and high-energy electrons. Over the pH range of 3.6 to 8.8, the doses required for a 1 log{sup 10} reduction in viability based on the exponential portion of the survival curve ranged from 0.21 to 0.22 kGy, with extrapolation numbers (extrapolation of the exponential portion of the survival curve to zero dose) of 1.01 to 1.33, for electron irradiation, and from 0.24 to 0.27 kGy, with extrapolation numbers of 2.26 to 5.13, for gamma irradiation.more » Nonsterile barley that was inoculated with conidia of the fungus and then irradiated with either electrons or gamma rays and incubated for prolonged periods at 28C and at a moisture content of 25% produced less ochratoxin levels compared with unirradiated controls. In these experiments, inoculation with 10{sup 2} spores per g produced greater radiation-induced enhancement than inoculation with 10{sup 5} spores per g. There was no radiation-induced enhancement when the barley was surface sterilized by chemical means prior to irradiation. These results are consistent with the hypothesis that a reduction in the competing microbial flora by irradiation is responsible for the enhanced mycotoxin production observed when nonsterile barley is inoculated with the toxigenic fungus A. alutaceus var. alutaceus after irradiation.« less

Giardia duodenalis-induced alterations of commensal bacteria kill Caenorhabditis elegans: a new model to study microbial-microbial interactions in the gut

PubMed Central

Gerbaba, Teklu K.; Gupta, Pratyush; Rioux, Kevin; Hansen, Dave

2015-01-01

Giardia duodenalis is the most common cause of parasitic diarrhea worldwide and a well-established risk factor for postinfectious irritable bowel syndrome. We hypothesized that Giardia-induced disruptions in host-microbiota interactions may play a role in the pathogenesis of giardiasis and in postgiardiasis disease. Functional changes induced by Giardia in commensal bacteria and the resulting effects on Caenorhabditis elegans were determined. Although Giardia or bacteria alone did not affect worm viability, combining commensal Escherichia coli bacteria with Giardia became lethal to C. elegans. Giardia also induced killing of C. elegans with attenuated Citrobacter rodentium espF and map mutant strains, human microbiota from a healthy donor, and microbiota from inflamed colonic sites of ulcerative colitis patient. In contrast, combinations of Giardia with microbiota from noninflamed sites of the same patient allowed for worm survival. The synergistic lethal effects of Giardia and E. coli required the presence of live bacteria and were associated with the facilitation of bacterial colonization in the C. elegans intestine. Exposure to C. elegans and/or Giardia altered the expression of 172 genes in E. coli. The genes affected by Giardia included hydrogen sulfide biosynthesis (HSB) genes, and deletion of a positive regulator of HSB genes, cysB, was sufficient to kill C. elegans even in the absence of Giardia. Our findings indicate that Giardia induces functional changes in commensal bacteria, possibly making them opportunistic pathogens, and alters host-microbe homeostatic interactions. This report describes the use of a novel in vivo model to assess the toxicity of human microbiota. PMID:25573177

Microbiological quality and bacterial diversity of the tropical oyster Crassostrea rhizophorae in a monitored farming system and from natural stocks.

PubMed

Silva Neta, M T; Maciel, B M; Lopes, A T S; Marques, E L S; Rezende, R P; Boehs, G

2015-12-02

Microbiological evaluation is one of the most important parameters for analyzing the viability of an oyster farming system, which addresses public health and ecological concerns. Here, the microbiological quality of the oyster Crassostrea rhizophorae cultivated in a monitored environment and from natural beds in Bahia, northeastern Brazil, was determined. Bacterial diversity in oysters was measured by polymerase chain reaction-denaturing gradient gel electrophoresis. Sequence analysis revealed that most bacterial species showed similarity with uncultured or unidentified bacteria from environmental samples, and were clustered into the phylum Proteobacteria. Diverse bacteria from cultivated (monitored) oyster samples were grouped in the same cluster with a high similarity index (above 79%). Microbiological analyses revealed that these oysters did not contain pathogens. These results reflect the natural balance of the microbial communities essential to the maintenance of health and in inhibiting pathogen colonization in the oyster. On the other hand, bacterial diversity of samples from native stocks in extractive areas displayed a similarity index varying between 55 and 77%, and all samples were clustered separately from each other and from the cluster of samples derived from the cultivation area. Microbiological analyses showed that oysters from the extractive area were not fit for human consumption. This reflected a different composition of the microbial community in this area, probably resulting from anthropic impact. Our study also demonstrated that low temperatures and high rainfall limits the bacterial concentration in tropical oysters. This is the first study analyzing the total bacterial community profiles of the oyster C. rhizophorae.

Semiquantitative Performance and Mechanism Evaluation of Carbon Nanomaterials as Cathode Coatings for Microbial Fouling Reduction.

PubMed

Zhang, Qiaoying; Nghiem, Joanne; Silverberg, Gregory J; Vecitis, Chad D

2015-07-01

In this study, we examine bacterial attachment and survival on a titanium (Ti) cathode coated with various carbon nanomaterials (CNM): pristine carbon nanotubes (CNT), oxidized carbon nanotubes (O-CNT), oxidized-annealed carbon nanotubes (OA-CNT), carbon black (CB), and reduced graphene oxide (rGO). The carbon nanomaterials were dispersed in an isopropyl alcohol-Nafion solution and were then used to dip-coat a Ti substrate. Pseudomonas fluorescens was selected as the representative bacterium for environmental biofouling. Experiments in the absence of an electric potential indicate that increased nanoscale surface roughness and decreased hydrophobicity of the CNM coating decreased bacterial adhesion. The loss of bacterial viability on the noncharged CNM coatings ranged from 22% for CB to 67% for OA-CNT and was dependent on the CNM dimensions and surface chemistry. For electrochemical experiments, the total density and percentage of inactivation of the adherent bacteria were analyzed semiquantitatively as functions of electrode potential, current density, and hydrogen peroxide generation. Electrode potential and hydrogen peroxide generation were the dominant factors with regard to short-term (3-h) bacterial attachment and inactivation, respectively. Extended-time electrochemical experiments (12 h) indicated that in all cases, the density of total deposited bacteria increased almost linearly with time and that the rate of bacterial adhesion was decreased 8- to 10-fold when an electric potential was applied. In summary, this study provides a fundamental rationale for the selection of CNM as cathode coatings and electric potential to reduce microbial fouling. Copyright © 2015, American Society for Microbiology. All Rights Reserved.

Did Mineral Surface Chemistry and Toxicity Contribute to Evolution of Microbial Extracellular Polymeric Substances?

PubMed Central

Campbell, Jay M.; Zhang, Nianli; Hickey, William J.

2012-01-01

Abstract Modern ecological niches are teeming with an astonishing diversity of microbial life in biofilms closely associated with mineral surfaces, which highlights the remarkable success of microorganisms in conquering the challenges and capitalizing on the benefits presented by the mineral–water interface. Biofilm formation capability likely evolved on early Earth because biofilms provide crucial cell survival functions. The potential toxicity of mineral surfaces toward cells and the complexities of the mineral–water–cell interface in determining the toxicity mechanisms, however, have not been fully appreciated. Here, we report a previously unrecognized role for extracellular polymeric substances (EPS), which form biofilms in shielding cells against the toxicity of mineral surfaces. Using colony plating and LIVE/DEAD staining methods in oxide suspensions versus oxide-free controls, we found greater viability of wild-type, EPS-producing strains of Pseudomonas aeruginosa PAO1 compared to their isogenic knockout mutant with defective biofilm-producing capacity. Oxide toxicity was specific to its surface charge and particle size. High resolution transmission electron microscopy (HRTEM) images and assays for highly reactive oxygen species (hROS) on mineral surfaces suggested that EPS shield via both physical and chemical mechanisms. Intriguingly, qualitative as well as quantitative measures of EPS production showed that toxic minerals induced EPS production in bacteria. By determining the specific toxicity mechanisms, we provide insight into the potential impact of mineral surfaces in promoting increased complexity of cell surfaces, including EPS and biofilm formation, on early Earth. Key Words: Mineral toxicity—Bacteria—EPS evolution—Biofilms—Cytotoxicity—Silica—Anatase—Alumina. Astrobiology 12, 785–798. PMID:22934560

Antimicrobial efficacy of a novel povidone iodine contact lens disinfection system.

PubMed

Yamasaki, Katsuhide; Saito, Fumio; Ota, Ritsue; Kilvington, Simon

2018-06-01

Contact lens (CL) wear is a risk factor for the acquisition of microbial keratitis. Accordingly, compliance to manufacturers' recommended hygiene and disinfection procedures are vital to safe (CL) use. In this study we evaluated a novel povidone-iodine (PI) (CL) disinfection system (cleadew, Ophtecs Corporation, Japan) against a range of bacterial, fungal and Acanthamoeba. Antimicrobial assays were conducted according to ISO 14729 using the recommended strains of bacteria and fungi, with and without the presence of organic soil. Regrowth of bacteria and fungi in the disinfection system was also examined. The activity on biofilms formed from Stenotrophomonas maltophilia and Achromobacter sp. was evaluated. Efficacy against A. castellanii trophozoites and cysts was also investigated. The PI system gave >4 log 10 kill of all bacteria and fungi following the manufacturer's recommended disinfection and cleaning time of 4h, with or without the presence of organic soil. No regrowth of organisms was found after 14days in the neutralized solution. In the biofilm studies the system resulted in at least a 7 log 10 reduction in viability of bacteria. For Acanthamoeba, >3 log 10 kill of trophozoites and 1.1-2.8 log 10 kill for the cyst stage was obtained. The PI system effective against a variety of pathogenic microorganisms under a range of test conditions. Strict compliance to recommended CL hygiene procedures is essential for safe CL wear. The use of care systems such as PI, with broad spectrum antimicrobial activity, may aid in the prevention of potentially sight threatening microbial keratitis. Copyright © 2017. Published by Elsevier Ltd.

Survival of akinetes (resting-state cells of cyanobacteria) in low earth orbit and simulated extraterrestrial conditions.

PubMed

Olsson-Francis, Karen; de la Torre, Rosa; Towner, Martin C; Cockell, Charles S

2009-12-01

Cyanobacteria are photosynthetic organisms that have been considered for space applications, such as oxygen production in bioregenerative life support systems, and can be used as a model organism for understanding microbial survival in space. Akinetes are resting-state cells of cyanobacteria that are produced by certain genera of heterocystous cyanobacteria to survive extreme environmental conditions. Although they are similar in nature to endospores, there have been no investigations into the survival of akinetes in extraterrestrial environments. The aim of this work was to examine the survival of akinetes from Anabaena cylindrica in simulated extraterrestrial conditions and in Low Earth Orbit (LEO). Akinetes were dried onto limestone rocks and sent into LEO for 10 days on the ESA Biopan VI. In ground-based experiments, the rocks were exposed to periods of desiccation, vacuum (0.7×10(-3) kPa), temperature extremes (-80 to 80°C), Mars conditions (-27°C, 0.8 kPa, CO(2)) and UV radiation (325-400 nm). A proportion of the akinete population was able to survive a period of 10 days in LEO and 28 days in Mars simulated conditions, when the rocks were not subjected to UV radiation. Furthermore, the akinetes were able to survive 28 days of exposure to desiccation and low temperature with high viability remaining. Yet long periods of vacuum and high temperature were lethal to the akinetes. This work shows that akinetes are extreme-tolerating states of cyanobacteria that have a practical use in space applications and yield new insight into the survival of microbial resting-state cells in space conditions.

Usage of humic materials for formulation of stable microbial inoculants

NASA Astrophysics Data System (ADS)

Kydralieva, K. A.; Khudaibergenova, B. M.; Elchin, A. A.; Gorbunova, N. V.; Muratov, V. S.; Jorobekova, Sh. J.

2009-04-01

Some microbes have been domesticated for environment service, for example in a variety of novel applications, including efforts to reduce environmental problems. For instance, antagonistic organisms can be used as biological control agents to reduce the use of chemical pesticides, or efficient degraders can be applied as bioprophylactics to minimise the spread of chemical pollutants. Microorganisms can also be used for the biological clean-up of polluted soil or as plant growth-promoting bacteria that stimulate nutrient uptake. Many microbial applications require large-scale cultivation of the organisms. The biomass production must then be followed by formulation steps to ensure long-term stability and convenient use. However, there remains a need to further develop knowledge on how to optimise fermentation of "non-conventional microorganisms" for environmental applications involving the intact living cells. The goal of presented study is to develop fermentation and formulation techniques for termolabile rhizobacteria isolates - Pseudomonas spp. with major biotechnical potential. Development of efficient and cost-effective media and process parameters giving high cell yields are important priorities. This also involves establishing fermentation parameters yielding cells well adapted to subsequent formulation procedures. Collectively, these strategies will deliver a high proportion of viable cells with good long-term survival. Our main efforts were focused on development of more efficient drying techniques for microorganisms, particularly spray drying and fluidised bed-drying. The advantages of dry formulations are that storage and delivery costs are much lower than for liquid formulations and that long-term survival can be very high if initial packaging is carefully optimised. In order to improve and optimise formulations various kinds of humics-based excipients have been added that have beneficial effects on the viability of the organisms and the storage stability of the product. It is known that humic substances can increase of live organism resistance to stress loads, in particular to chemical stress, low and high temperature. Spray- and fluidized-bed drying and addition of humate-based drying protectants were evaluated for the development of dry formulations of biocontrol and plant growth promoting rhizobacteria. The drying protectants - humic acids and sodium humate gave the highest initial survival rates and the most stable formulations, without significant losses of viability after storage for 1 month at 30oC. As a result, the specific plant growth promoting effect is retained. Thus, humic materials have an unfulfilled potential for biotechnology industries based on such applications. Acknowledgement. This research was supported by the grant of ISTC KR-993.2.

Use of ATP to characterize biomass viability in freely suspended and immobilized cell bioreactors

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

Gikas, P.; Livingston, A.G.

1993-12-01

This work describes investigations into the viability of cells growing on 3,4-dichloroaniline (34DCA). Two bio-reactors are employed for microbial growth, a continuous stirred tank (CST) bioreactor with a 2-L working volume, and a three-phase air lift (TPAL) bioreactor with a 3-L working volume. Experiments have been performed at several dilution rates between 0.027 and 0.115 h[sup [minus]1] in the CST bioreactor and between 0.111 and 0.500h[sup [minus]1] in the TPAL bioreactor. The specific ATP concentration was calculated at each dilution rate in the suspended biomass in both bioreactors as well as in the immobilized biomass in the TPAL bioreactor. Themore » cultures were inspected under an electron microscope to monitor compositional changes. Results from the CST bioreactor showed that the biomass-specific ATP concentration increases from 0.44 to 1.86 mg ATP g[sup [minus]1] dry weight (dw) as dilution rate increases from 0.027 to 0.115 h[sup [minus]1]. At this upper dilution rate the cells were washed out. The specific ATP concentration reached a limiting average value of 1.73 mg ATP g[sup [minus]1] dw, which is assumed to be the quantity of ATP in 100% viable biomass, In the TPAL bioreactor, the ATP level increased with dilution rat in both the immobilized and suspended biomass. The specific ATP concentration in the immobilized biomass increased from approximately 0.051 mg ATP g[sup [minus]1] dw at dilution rates between 0.111 and 0.200 h[sup [minus]1] to approximately 0.119 mg ATP g[sup [minus]1] dw at dilution rates between 0.300 and 0.500 h[sup [minus]1].« less

Detection and Viability of Lactococcus lactis throughout Cheese Ripening

PubMed Central

Cocolin, Luca

2014-01-01

Recent evidences highlighted the presence of Lactococcus lactis during late cheese ripening. For this reason, the role of this microorganism, well known as dairy starter, should be reconsidered throughout cheese manufacturing and ripening. Thus, the main objective of this study was to develop a RT-qPCR protocol for the detection, quantification and determination of the viability of L. lactis in ripened cheese samples by direct analysis of microbial nucleic acids. Standard curves were constructed for the specific quantification of L. lactis in cheese matrices and good results in terms of selectivity, correlation coefficient and efficiency were obtained. Thirty-three ripened cheeses were analyzed and, on the basis of RNA analysis, twelve samples showed 106 to 108 CFU of L. lactis per gram of product, thirteen from 103 to 105 CFU/g, and in eight cheeses, L. lactis was not detected. Traditional plating on M17 medium led to loads ranging from 105 to 109 CFU/g, including the cheese samples where no L. lactis was found by RT-qPCR. From these cheeses, none of the colonies isolated on M17 medium was identified as L. lactis species. These data could be interpreted as a lack of selectivity of M17 medium where colony growth is not always related to lactococcal species. At the same time, the absence or low abundance of L. lactis isolates on M17 medium from cheese where L. lactis was detected by RT-qPCR support the hypothesis that L. lactis starter populations are mainly present in viable but not culturable state during ripening and, for this reason, culture-dependent methods have to be supplemented with direct analysis of cheese. PMID:25503474

Investigation of Structural, Compositional and Anti-Microbial Properties of Copper Thin Film Using Direct Current Magnetron Sputtering for Surgical Instruments

NASA Astrophysics Data System (ADS)

Kalaiselvam, S.; Sandhya, J.; Krishnan, K. V. Hari; Kedharnath, A.; Arulkumar, G.; Roseline, A. Ameelia

Surgical instruments and other bioimplant devices, owing to their importance in the biomedical industry require high biocompatibility to be used in the human body. Nevertheless, issues of compatibility, bacterial infections are quite common in such devices. Hence development of surface coatings on various substrates for implant applications is a promising technique to combat the issues arising in these implant materials. The present investigation aims at coating copper on stainless steel substrate using DC Magnetron sputtering which is used to achieve film of required thickness (0.5-8μm). The deposition pressure, substrate temperature, power supply, distance between the specimen and target are optimized and maintained constant, while the sputtering time (30-110min) is varied. The sputtered copper thin film’s morphology, composition are characterized by SEM and EDAX. X-ray diffraction analysis shows copper oriented on (111) and (002) and copper oxide on (111) planes. The contact angle of copper thin film is 92∘ while AISI 316L shows 73∘. The antimicrobial studies carried in Staphylococcus aureus, Escherichia Coli, Klebsiella pneumonia and Candida albicans show that the maximum reduction was seen upto 35, 26, 54, 39CFU/mL, respectively after 24h. The cell viability is studied by MTT assay test on Vero cell line for 24h, 48h and 72h and average cell viability is 43.85%. The copper release from the thin film to the culture medium is 6691μg/L (maximum) is estimated from AAS studies. The copper coated substrate does not show much reaction with living Vero cells whereas the bacteria and fungi are found to be destroyed.

Survival of Spoilage and Pathogenic Microorganisms on Cardboard and Plastic Packaging Materials

PubMed Central

Siroli, Lorenzo; Patrignani, Francesca; Serrazanetti, Diana I.; Chiavari, Cristiana; Benevelli, Marzia; Grazia, Luigi; Lanciotti, Rosalba

2017-01-01

The aim of this work was to study the interaction of corrugated and plastic materials with pathogenic and spoiling microorganisms frequently associated to fresh produce. The effect of the two packaging materials on the survival during the storage of microorganisms belonging to the species Escherichia coli, Listeria monocytogenes, Salmonella enteritidis, Saccharomyces cerevisiae, Lactobacillus plantarum, Pseudomonas fluorescens, and Aspergillus flavus was studied through traditional plate counting and scanning electron microscopy (SEM). The results obtained showed that cardboard materials, if correctly stored, reduced the potential of packaging to cross-contaminate food due to a faster viability loss by spoilage and pathogenic microorganisms compared to the plastic ones. In fact, the cell loads of the pathogenic species considered decreased over time independently on the inoculation level and packaging material used. However, the superficial viability losses were significantly faster in cardboard compared to plastic materials. The same behavior was observed for the spoilage microorganisms considered. The SEM microphotographs indicate that the reduction of superficial contamination on cardboard surfaces was due to the entrapping of the microbial cells within the fibers and the pores of this material. In addition, SEM data showed that the entrapped cells were subjected to more or less rapid lyses, depending on the species, due to the absence of water and nutrients, with the exception of molds. The latter spoilers were able to proliferate inside the cardboard fibers only when the absorption of water was not prevented during the storage. In conclusion, the findings of this work showed the reduction of cross-contamination potential of corrugated compared to plastic packaging materials used in fruit and vegetable sector. However, the findings outlined the importance of hygiene and low humidity during cardboard storage to prevent the mold growth on packaging. PMID:29312271

Aspergillus fumigatus viability drives allergic responses to inhaled conidia.

PubMed

Nayak, Ajay P; Croston, Tara L; Lemons, Angela R; Goldsmith, W T; Marshall, Nikki B; Kashon, Michael L; Germolec, Dori R; Beezhold, Donald H; Green, Brett J

2018-04-13

Aspergillus fumigatus induced allergic airway disease has been shown to involve conidial germination in vivo but the immunological mechanisms remain uncharacterized. A subchronic murine exposure model was used to examine the immunological mediators that are regulated in response to either culturable or non-culturable A. fumigatus conidia. Female B6C3F1/N mice were repeatedly dosed via inhalation with 1 x 105 viable or heat inactivated conidia (HIC), twice a week for 13 weeks (26 exposures). Control mice inhaled HEPA-filtered air. The influence of A. fumigatus conidial germination on the pulmonary immunopathological outcomes was evaluated by flow cytometry analysis of cellular infiltration in the airways, assessment of lung mRNA expression, and quantitative proteomics and histopathology of whole lung tissue. Repeated inhalation of viable conidia, but not HIC, resulted in allergic inflammation marked by vascular remodeling, extensive eosinophilia, and accumulation of alternatively activated macrophages (AAMs) in the murine airways. More specifically, mice that inhaled viable conidia resulted in a mixed TH1 and TH2 (IL-13) cytokine response. Recruitment of eosinophils corresponded with increased Ccl11 transcripts. Furthermore, genes associated with M2 or alternatively activated macrophage polarization (e.g. Arg1, Chil3 and Retnla) were significantly upregulated in viable A. fumigatus exposed mice. In mice inhaling HIC, CD4+ T cells expressing IFN-γ (TH1) dominated the lymphocytic infiltration. Quantitative proteomics of the lung revealed metabolic reprogramming accompanied by mitochondrial dysfunction and endoplasmic reticulum stress stimulated by oxidative stress from repetitive microbial insult. Our studies demonstrate that A. fumigatus conidial viability in vivo is critical to the immunopathological presentation of chronic fungal allergic disease. Copyright © 2018. Published by Elsevier Inc.

Comparison of two methodologies for the enrichment of mononuclear cells from thawed cord blood products: The automated Sepax system versus the manual Ficoll method.

PubMed

Kaur, Indreshpal; Zulovich, Jane M; Gonzalez, Marissa; McGee, Kara M; Ponweera, Nirmali; Thandi, Daljit; Alvarez, Enrique F; Annandale, Kathy; Flagge, Frank; Rezvani, Katayoun; Shpall, Elizabeth

2017-03-01

Umbilical cord blood (CB) is being used as a source of hematopoietic stem cells (HSCs) and immune cells to treat many disorders. Because these cells are present in low numbers in CB, investigators have developed strategies to expand HSCs and other immune cells such as natural killer (NK) cells. The initial step in this process is to enrich mononuclear cells (MNCs) while depleting unwanted cells. The manual method of MNC enrichment is routinely used by many centers; however, it is an open system, time-consuming and operator dependent. For clinical manufacturing, it is important to have a closed system to avoid microbial contamination. In this study, we optimized an automated, closed system (Sepax) for enriching MNCs from cryopreserved CB units. Using Sepax, we observed higher recovery of total nucleated cells (TNC), CD34 + cells, NK cells and monocytes when compared to manual enrichment, despite similar TNC and CD34 + viability with the two methods. Even though the depletion of red blood cells, granulocytes and platelets was superior using the manual method, significantly higher CFU-GM were obtained in MNCs enriched using Sepax compared to the manual method. This is likely related to the fact that the automated Sepax significantly shortened the processing time (Sepax: 74 - 175 minutes versus manual method: 180 - 290 minutes). The use of DNAse and MgCl 2 during the Sepax thaw and wash procedure prevents clumping of cells and loss of viability, resulting in improved post-thaw cell recovery. We optimized enrichment of MNCs from cryopreserved CB products in a closed system using the Sepax which is a walk away and automated processing system. Copyright © 2017 International Society for Cellular Therapy. Published by Elsevier Inc. All rights reserved.

Mixed culture models for predicting intestinal microbial interactions between Escherichia coli and Lactobacillus in the presence of probiotic Bacillus subtilis.

PubMed

Yang, J J; Niu, C C; Guo, X H

2015-01-01

Bacillus has been proposed as a probiotic due to its in vivo effectiveness in the gastrointestinal tract through antimicrobial activities. The present study investigates the effects of Lactobacillus alone or in the presence of Bacillus subtilis MA139 on the inhibition of pathogenic Escherichia coli K88. Mixed cultures were used to predict the possible interactions among these bacteria within the intestinal tract of animals. B. subtilis MA139 was first assayed for its inhibition against E. coli K88 both under shaking and static culture conditions. A co-culture assay was employed under static conditions to test the inhibitory effects of Lactobacillus reuteri on E. coli K88, with or without addition of B. subtilis MA139. The results showed that B. subtilis MA139 had marked inhibition against E. coli K88 under shaking conditions and weak inhibition under static conditions. Lactobacillus alone as well as in combination with B. subtilis MA139 spores exerted strong inhibition against E. coli K88 under static conditions. However, the inhibition by Lactobacillus in combination with B. subilis spores was much higher than that by Lactobacillus alone (P<0.01). B. subtilis MA139 significantly decreased the pH and oxidation-reduction potential values of the co-culture broth compared to that of Lactobacillus alone (P<0.05). The viability of Lactobacillus increased when co-cultured with B. subtilis MA139 because of significantly higher Lactobacillus counts and lower pH values in the broth (P<0.05). The role of Bacillus in the mixed culture models suggests that Bacillus may produce beneficial effects by increasing the viability of lactobacilli and subsequently inhibiting the growth of pathogenic E. coli. Therefore, the combination of Bacillus and Lactobacillus species as a probiotic is recommended.

Persistence of biomarker ATP and ATP-generating capability in bacterial cells and spores contaminating spacecraft materials under earth conditions and in a simulated martian environment.

PubMed

Fajardo-Cavazos, Patricia; Schuerger, Andrew C; Nicholson, Wayne L

2008-08-01

Most planetary protection research has concentrated on characterizing viable bioloads on spacecraft surfaces, developing techniques for bioload reduction prior to launch, and studying the effects of simulated martian environments on microbial survival. Little research has examined the persistence of biogenic signature molecules on spacecraft materials under simulated martian surface conditions. This study examined how endogenous adenosine-5'-triphosphate (ATP) would persist on aluminum coupons under simulated martian conditions of 7.1 mbar, full-spectrum simulated martian radiation calibrated to 4 W m(-2) of UV-C (200 to 280 nm), -10 degrees C, and a Mars gas mix of CO(2) (95.54%), N(2) (2.7%), Ar (1.6%), O(2) (0.13%), and H(2)O (0.03%). Cell or spore viabilities of Acinetobacter radioresistens, Bacillus pumilus, and B. subtilis were measured in minutes to hours, while high levels of endogenous ATP were recovered after exposures of up to 21 days. The dominant factor responsible for temporal reductions in viability and loss of ATP was the simulated Mars surface radiation; low pressure, low temperature, and the Mars gas composition exhibited only slight effects. The normal burst of endogenous ATP detected during spore germination in B. pumilus and B. subtilis was reduced by 1 or 2 orders of magnitude following, respectively, 8- or 30-min exposures to simulated martian conditions. The results support the conclusion that endogenous ATP will persist for time periods that are likely to extend beyond the nominal lengths of most surface missions on Mars, and planetary protection protocols prior to launch may require additional rigor to further reduce the presence and abundance of biosignature molecules on spacecraft surfaces.

Computer keyboard covers impregnated with a novel antimicrobial polymer significantly reduce microbial contamination.

PubMed

D'Antonio, Natalie N; Rihs, John D; Stout, Janet E; Yu, Victor L

2013-04-01

Contaminated computer keyboards have been acknowledged as a potential source for bacterial transmission between health care providers and patients. Biosafe HM 4100 is an antimicrobial polymer that can be incorporated into the polyurethane material used to make keyboard covers. This study aimed to determine whether plastic keyboard covers containing HM 4100 effectively minimize the survival of bacterial species commonly present on health care environmental surfaces. Polyurethane material that contained 0.5% HM 4100, 1% HM 4100, and 1% HM 4100 with spray coating of 1% HM 4100 were tested. In 2 separate experiments, the surfaces of test materials were inoculated with suspensions of methicillin-resistant Staphylococcus aureus (MRSA), vancomycin-resistant Enterococcus faecalis (VREF), Escherichia coli, or Pseudomonas aeruginosa. Viability was assessed on the materials at 0, 10, 30, 60, 120, and 240 minutes after inoculation. Maximum reductions in viability were observed for all 4 organisms at the longest tested time period on each test material. Mean reductions on the 0.5% HM 4100 material at 240 minutes were 99.99% for E coli, 97.8% for MRSA, 95.0% for VREF, and 92.1% for P aeruginosa. Mean reductions on the 1% HM 4100 at 120 minutes were 99.9% for VREF, 99.9% for MRSA, 99.9% for P aeruginosa, and 99.5% for E coli. Mean reductions on the 1% HM 4100 plus spray coating at 30 minutes were 99.9% for E coli, 99.8% for VREF, 98.8% for P aeruginosa, and 97.2% for MRSA. Incorporation of the HM 4100 antimicrobial polymer into polyurethane keyboard material may reduce the hand carriage of bacteria between health care providers and patients. Copyright © 2013 Association for Professionals in Infection Control and Epidemiology, Inc. Published by Mosby, Inc. All rights reserved.

Confocal laser scanning microscopy analysis of S. epidermidis biofilms exposed to farnesol, vancomycin and rifampicin.

PubMed

Cerca, Nuno; Gomes, Fernanda; Pereira, Sofia; Teixeira, Pilar; Oliveira, Rosário

2012-05-16

Staphylococcus epidermidis is the major bacterial species found in biofilm-related infections on indwelling medical devices. Microbial biofilms are communities of bacteria adhered to a surface and surrounded by an extracellular polymeric matrix. Biofilms have been associated with increased antibiotic tolerance to the immune system. This increased resistance to conventional antibiotic therapy has lead to the search for new antimicrobial therapeutical agents. Farnesol, a quorum-sensing molecule in Candida albicans, has been described as impairing growth of several different microorganisms and we have previously shown its potential as an adjuvant in antimicrobial therapy against S. epidermidis. However, its mechanism of action in S. epidermidis is not fully known. In this work we better elucidate the role of farnesol against S: epidermidis biofilms using confocal laser scanning microscopy (CLSM). 24 h biofilms were exposed to farnesol, vancomycin or rifampicin and were analysed by CLSM, after stained with a Live/Dead stain, a known indicator of cell viability, related with cell membrane integrity. Biofilms were also disrupted by sonication and viable and cultivable cells were quantified by colony forming units (CFU) plating. Farnesol showed a similar effect as vancomycin, both causing little reduction of cell viability but at the same time inducing significant changes in the biofilm structure. On the other hand, rifampicin showed a distinct action in S. epidermidis biofilms, by killing a significant proportion of biofilm bacteria. While farnesol is not very efficient at killing biofilm bacteria, it damages cell membrane, as determined by the live/dead staining, in a similar way as vancomycin. Furthermore, farnesol might induce biofilm detachment, as determined by the reduced biofilm biomass, which can partially explain the previous findings regarding its role as a possible chemotherapy adjuvant.

Licoricidin inhibits the growth of SW480 human colorectal adenocarcinoma cells in vitro and in vivo by inducing cycle arrest, apoptosis and autophagy

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

Ji, Shuai

Licorice (Glycyrrhiza uralensis Fisch.) possesses significant anti-cancer activities, but the active ingredients and underlying mechanisms have not been revealed. By screening the cytotoxic activities of 122 licorice compounds against SW480 human colorectal adenocarcinoma cells, we found that licoricidin (LCD) inhibited SW480 cell viability with an IC{sub 50} value of 7.2 μM. Further studies indicated that LCD significantly induced G1/S cell cycle arrest and apoptosis in SW480 cells, accompanied by inhibition of cyclins/CDK1 expression and activation of caspase-dependent pro-apoptotic signaling. Meanwhile, LCD promoted autophagy in SW480 cells, and activated AMPK signaling and inhibited Akt/mTOR pathway. Overexpression of a dominant-negative AMPKα2 abolishedmore » LCD-induced inhibition of Akt/mTOR, autophagic and pro-apoptotic signaling pathways, and significantly reversed loss of cell viability, suggesting activation of AMPK is essential for the anti-cancer activity of LCD. In vivo anti-tumor experiments indicated that LCD (20 mg/kg, i.p.) significantly inhibited the growth of SW480 xenografts in nude mice with an inhibitory rate of 43.5%. In addition, we obtained the glycosylated product LCDG by microbial transformation, and found that glycosylation slightly enhanced the in vivo anti-cancer activities of LCD. This study indicates that LCD could inhibit SW480 cells by inducing cycle arrest, apoptosis and autophagy, and is a potential chemopreventive or chemotherapeutic agent against colorectal cancer. - Highlights: • Molecular mechanisms for cytotoxic activity of licoricidin (LCD) were investigated. • LCD promoted autophagy of SW480 cells through AMPK and Akt/mTOR signaling pathways. • Both LCD and its glucoside showed in vivo anti-colorectal cancer activities.« less

Morphological observation and analysis using automated image cytometry for the comparison of trypan blue and fluorescence-based viability detection method.

PubMed

Chan, Leo Li-Ying; Kuksin, Dmitry; Laverty, Daniel J; Saldi, Stephanie; Qiu, Jean

2015-05-01

The ability to accurately determine cell viability is essential to performing a well-controlled biological experiment. Typical experiments range from standard cell culturing to advanced cell-based assays that may require cell viability measurement for downstream experiments. The traditional cell viability measurement method has been the trypan blue (TB) exclusion assay. However, since the introduction of fluorescence-based dyes for cell viability measurement using flow or image-based cytometry systems, there have been numerous publications comparing the two detection methods. Although previous studies have shown discrepancies between TB exclusion and fluorescence-based viability measurements, image-based morphological analysis was not performed in order to examine the viability discrepancies. In this work, we compared TB exclusion and fluorescence-based viability detection methods using image cytometry to observe morphological changes due to the effect of TB on dead cells. Imaging results showed that as the viability of a naturally-dying Jurkat cell sample decreased below 70 %, many TB-stained cells began to exhibit non-uniform morphological characteristics. Dead cells with these characteristics may be difficult to count under light microscopy, thus generating an artificially higher viability measurement compared to fluorescence-based method. These morphological observations can potentially explain the differences in viability measurement between the two methods.

Efficient estimation of the maximum metabolic productivity of batch systems.

PubMed

St John, Peter C; Crowley, Michael F; Bomble, Yannick J

2017-01-01

Production of chemicals from engineered organisms in a batch culture involves an inherent trade-off between productivity, yield, and titer. Existing strategies for strain design typically focus on designing mutations that achieve the highest yield possible while maintaining growth viability. While these methods are computationally tractable, an optimum productivity could be achieved by a dynamic strategy in which the intracellular division of resources is permitted to change with time. New methods for the design and implementation of dynamic microbial processes, both computational and experimental, have therefore been explored to maximize productivity. However, solving for the optimal metabolic behavior under the assumption that all fluxes in the cell are free to vary is a challenging numerical task. Previous studies have therefore typically focused on simpler strategies that are more feasible to implement in practice, such as the time-dependent control of a single flux or control variable. This work presents an efficient method for the calculation of a maximum theoretical productivity of a batch culture system using a dynamic optimization framework. The proposed method follows traditional assumptions of dynamic flux balance analysis: first, that internal metabolite fluxes are governed by a pseudo-steady state, and secondly that external metabolite fluxes are dynamically bounded. The optimization is achieved via collocation on finite elements, and accounts explicitly for an arbitrary number of flux changes. The method can be further extended to calculate the complete Pareto surface of productivity as a function of yield. We apply this method to succinate production in two engineered microbial hosts, Escherichia coli and Actinobacillus succinogenes , and demonstrate that maximum productivities can be more than doubled under dynamic control regimes. The maximum theoretical yield is a measure that is well established in the metabolic engineering literature and whose use helps guide strain and pathway selection. We present a robust, efficient method to calculate the maximum theoretical productivity: a metric that will similarly help guide and evaluate the development of dynamic microbial bioconversions. Our results demonstrate that nearly optimal yields and productivities can be achieved with only two discrete flux stages, indicating that near-theoretical productivities might be achievable in practice.

The effects of space relevant environmental factors on halophilic Archaea

NASA Astrophysics Data System (ADS)

Leuko, Stefan; Moeller, Ralf; Rettberg, Petra

Within the last 50 years, space technology has provided tools for transporting terrestrial (microbial) life beyond Earth's protective shield in order to study its responses to selected conditions of space. Microorganisms are ubiquitous and can be found in almost every environment on Earth. They thrive and survive in a broad spectrum of environments and are true masters in adapting to rapidly changing external conditions. Although microorganisms cannot actively grow under the harsh conditions of outer space or other known planets, some microorganisms might be able to survive for a time in space or other planets as dormant, inactive spores or in similar desiccation-resistant resting states, e.g., enclosed in halite crystals or biofilms. Halite crystals are the realm of halophilic Archaea as they have adapted to life at extreme salt concentrations. They can stay entrapped in such crystals for millions of years without losing viability and therefore the family Halobacteriaceae belongs to the group of microorganisms which may survive space travel or may even be found on other planets. Several members of this family have been utilized in space relevant experiments where they were exposed to detrimental environmental conditions such as UV-C radiation, vacuum, temperature cycles (+60(°) C and -25(°) C) and heavy iron bombardment (150 MeV He, 500 MeV Ar and 500 MeV Fe ions). The viability was evaluated by colony forming unit (cfu) counts as well as with the LIFE/DEAD kit. Results revealed that UV-C radiation (up to 1.000 J/m (2) ) has a considerable effect on the viability, whereas the other tested parameters inflict little damage onto the organisms. Repair of UV-C inflicted damage is efficient and several DNA damage repair genes are up-regulated following exposure. Halophilic archaea display a strong resistance against heavy iron bombardment, with dosages of up to 2.000 Gy 500 MeV Fe ions needed to establish a visible effect on the vitality. Genomic integrity after exposure was investigated by several different methods e.g. RAPD - PCR, a technique that elucidates damages within the genome by different amplification patterns. Overall experimental results indicate that halophilic Archaea are able to withstand the exposure to space related environmental factors for a considerable time. This work in combined with others will lead to a detailed understanding of the response of extraterrestrial conditions to halophilic Archaea for astrobiological considerations.

Algae viability over time in a ballast water sample

NASA Astrophysics Data System (ADS)

Gollasch, Stephan; David, Matej

2018-03-01

The biology of vessels' ballast water needs to be analysed for several reasons, one of these being performance tests of ballast water management systems. This analysis includes a viability assessment of phytoplankton. To overcome logistical problems to get algae sample processing gear on board of a vessel to document algae viability, samples may be transported to land-based laboratories. Concerns were raised how the storage conditions of the sample may impact algae viability over time and what the most appropriate storage conditions were. Here we answer these questions with a long-term algae viability study with daily sample analysis using Pulse-Amplitude Modulated (PAM) fluorometry. The sample was analysed over 79 days. We tested different storage conditions: fridge and room temperature with and without light. It seems that during the first two weeks of the experiment the viability remains almost unchanged with a slight downwards trend. In the continuing period, before the sample was split, a slightly stronger downwards viability trend was observed, which occurred at a similar rate towards the end of the experiment. After the sample was split, the strongest viability reduction was measured for the sample stored without light at room temperature. We concluded that the storage conditions, especially regarding temperature and light exposure, have a stronger impact on algae viability compared to the storage duration and that inappropriate storage conditions reduce algal viability. A sample storage time of up to two weeks in a dark and cool environment has little influence on the organism viability. This indicates that a two week time duration between sample taking on board a vessel and the viability measurement in a land-based laboratory may not be very critical.

Effectiveness of chitosan against wine-related microorganisms.

PubMed

Bağder Elmaci, Simel; Gülgör, Gökşen; Tokatli, Mehmet; Erten, Hüseyin; İşci, Asli; Özçelik, Filiz

2015-03-01

The antimicrobial action of chitosan against wine related microorganisms, including Lactobacillus plantarum, Saccharomyces cerevisiae, Oeonococcus oeni, Lactobacillus hilgardii, Brettanomyces bruxellensis, Hanseniaspora uvarum and Zygosaccharomyces bailii was examined in laboratory media. In order to assess the potential applicability of chitosan as a microbial control agent for wine, the effect of chitosan, applied individually and/or in combination with sulphur dioxide (SO2), on the growth of microorganisms involved in various stages of winemaking and on the fermentative performance of S. cerevisiae was investigated. Of the seven wine-related microorganisms studied, S. cerevisiae exhibited the strongest resistance to antimicrobial action of chitosan in laboratory media with a minimum inhibitory concentration (MIC) greater than 2 g/L. L. hilgardii, O. oeni and B. bruxellensis were the most susceptible to chitosan since they were completely inactivated by chitosan at 0.2 g/L. The MIC of chitosan for L. plantarum, H. uvarum and Z. bailii was 2, 0.4 and 0.4 g/L, respectively. In wine experiments, it was found that chitosan had a retarding effect on alcoholic fermentation without significantly altering the viability and the fermentative performance of S. cerevisiae. With regard to non-Saccharomyces yeasts (H. uvarum and Z. bailii) involved in winemaking, the early deaths of these yeasts in mixed cultures with S. cerevisiae were not probably due to the antimicrobial action of chitosan but rather due to ethanol produced by the yeasts. The complex interactions between chitosan and wine ingredients as well as microbial interactions during wine fermentation considerably affect the efficacy of chitosan. It was concluded that chitosan was worthy of further investigation as an alternative or complementary preservative to SO2 in wine industry.

Flux balance modeling to predict bacterial survival during pulsed-activity events

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

Jose, Nicholas A.; Lau, Rebecca; Swenson, Tami L.

Desert biological soil crusts (BSCs) are cyanobacteria-dominated surface soil microbial communities common to plant interspaces in arid environments. The capability to significantly dampen their metabolism allows them to exist for extended periods in a desiccated dormant state that is highly robust to environmental stresses. However, within minutes of wetting, metabolic functions reboot, maximizing activity during infrequent permissive periods. Microcoleus vaginatus, a primary producer within the crust ecosystem and an early colonizer, initiates crust formation by binding particles in the upper layer of soil via exopolysaccharides, making microbial dominated biological soil crusts highly dependent on the viability of this organism. Previousmore » studies have suggested that biopolymers play a central role in the survival of this organism by powering resuscitation, rapidly forming compatible solutes, and fueling metabolic activity in dark, hydrated conditions. To elucidate the mechanism of this phenomenon and provide a basis for future modeling of BSCs, we developed a manually curated, genome-scale metabolic model of Microcoleus vaginatus (iNJ1153). To validate this model, gas chromatography–mass spectroscopy (GC–MS) and liquid chromatography–mass spectroscopy (LC–MS) were used to characterize the rate of biopolymer accumulation and depletion in in hydrated Microcoleus vaginatus under light and dark conditions. Constraint-based flux balance analysis showed agreement between model predictions and experimental reaction fluxes. A significant amount of consumed carbon and light energy is invested into storage molecules glycogen and polyphosphate, while β-polyhydroxybutyrate may function as a secondary resource. Pseudo-steady-state modeling suggests that glycogen, the primary carbon source with the fastest depletion rate, will be exhausted if M. vaginatus experiences dark wetting events 4 times longer than light wetting events.« less

Biological Stability of Drinking Water: Controlling Factors, Methods, and Challenges.

PubMed

Prest, Emmanuelle I; Hammes, Frederik; van Loosdrecht, Mark C M; Vrouwenvelder, Johannes S

2016-01-01

Biological stability of drinking water refers to the concept of providing consumers with drinking water of same microbial quality at the tap as produced at the water treatment facility. However, uncontrolled growth of bacteria can occur during distribution in water mains and premise plumbing, and can lead to hygienic (e.g., development of opportunistic pathogens), aesthetic (e.g., deterioration of taste, odor, color) or operational (e.g., fouling or biocorrosion of pipes) problems. Drinking water contains diverse microorganisms competing for limited available nutrients for growth. Bacterial growth and interactions are regulated by factors, such as (i) type and concentration of available organic and inorganic nutrients, (ii) type and concentration of residual disinfectant, (iii) presence of predators, such as protozoa and invertebrates, (iv) environmental conditions, such as water temperature, and (v) spatial location of microorganisms (bulk water, sediment, or biofilm). Water treatment and distribution conditions in water mains and premise plumbing affect each of these factors and shape bacterial community characteristics (abundance, composition, viability) in distribution systems. Improved understanding of bacterial interactions in distribution systems and of environmental conditions impact is needed for better control of bacterial communities during drinking water production and distribution. This article reviews (i) existing knowledge on biological stability controlling factors and (ii) how these factors are affected by drinking water production and distribution conditions. In addition, (iii) the concept of biological stability is discussed in light of experience with well-established and new analytical methods, enabling high throughput analysis and in-depth characterization of bacterial communities in drinking water. We discussed, how knowledge gained from novel techniques will improve design and monitoring of water treatment and distribution systems in order to maintain good drinking water microbial quality up to consumer's tap. A new definition and methodological approach for biological stability is proposed.

Biological Stability of Drinking Water: Controlling Factors, Methods, and Challenges

PubMed Central

Prest, Emmanuelle I.; Hammes, Frederik; van Loosdrecht, Mark C. M.; Vrouwenvelder, Johannes S.

2016-01-01

Biological stability of drinking water refers to the concept of providing consumers with drinking water of same microbial quality at the tap as produced at the water treatment facility. However, uncontrolled growth of bacteria can occur during distribution in water mains and premise plumbing, and can lead to hygienic (e.g., development of opportunistic pathogens), aesthetic (e.g., deterioration of taste, odor, color) or operational (e.g., fouling or biocorrosion of pipes) problems. Drinking water contains diverse microorganisms competing for limited available nutrients for growth. Bacterial growth and interactions are regulated by factors, such as (i) type and concentration of available organic and inorganic nutrients, (ii) type and concentration of residual disinfectant, (iii) presence of predators, such as protozoa and invertebrates, (iv) environmental conditions, such as water temperature, and (v) spatial location of microorganisms (bulk water, sediment, or biofilm). Water treatment and distribution conditions in water mains and premise plumbing affect each of these factors and shape bacterial community characteristics (abundance, composition, viability) in distribution systems. Improved understanding of bacterial interactions in distribution systems and of environmental conditions impact is needed for better control of bacterial communities during drinking water production and distribution. This article reviews (i) existing knowledge on biological stability controlling factors and (ii) how these factors are affected by drinking water production and distribution conditions. In addition, (iii) the concept of biological stability is discussed in light of experience with well-established and new analytical methods, enabling high throughput analysis and in-depth characterization of bacterial communities in drinking water. We discussed, how knowledge gained from novel techniques will improve design and monitoring of water treatment and distribution systems in order to maintain good drinking water microbial quality up to consumer’s tap. A new definition and methodological approach for biological stability is proposed. PMID:26870010

A critical assessment of the "sterile womb" and "in utero colonization" hypotheses: implications for research on the pioneer infant microbiome.

PubMed

Perez-Muñoz, Maria Elisa; Arrieta, Marie-Claire; Ramer-Tait, Amanda E; Walter, Jens

2017-04-28

After more than a century of active research, the notion that the human fetal environment is sterile and that the neonate's microbiome is acquired during and after birth was an accepted dogma. However, recent studies using molecular techniques suggest bacterial communities in the placenta, amniotic fluid, and meconium from healthy pregnancies. These findings have led many scientists to challenge the "sterile womb paradigm" and propose that microbiome acquisition instead begins in utero, an idea that would fundamentally change our understanding of gut microbiota acquisition and its role in human development. In this review, we provide a critical assessment of the evidence supporting these two opposing hypotheses, specifically as it relates to (i) anatomical, immunological, and physiological characteristics of the placenta and fetus; (ii) the research methods currently used to study microbial populations in the intrauterine environment; (iii) the fecal microbiome during the first days of life; and (iv) the generation of axenic animals and humans. Based on this analysis, we argue that the evidence in support of the "in utero colonization hypothesis" is extremely weak as it is founded almost entirely on studies that (i) used molecular approaches with an insufficient detection limit to study "low-biomass" microbial populations, (ii) lacked appropriate controls for contamination, and (iii) failed to provide evidence of bacterial viability. Most importantly, the ability to reliably derive axenic animals via cesarean sections strongly supports sterility of the fetal environment in mammals. We conclude that current scientific evidence does not support the existence of microbiomes within the healthy fetal milieu, which has implications for the development of clinical practices that prevent microbiome perturbations after birth and the establishment of future research priorities.

A new strategy for umbilical cord blood collection developed at the first Colombian public cord blood bank increases total nucleated cell content.

PubMed

Vanegas, Diana; Triviño, Lady; Galindo, Cristian; Franco, Leidy; Salguero, Gustavo; Camacho, Bernardo; Perdomo-Arciniegas, Ana-María

2017-09-01

The total nucleated cell dosage of umbilical cord blood (UCB) is an important factor in determining successful allogeneic hematopoietic stem cell transplantation after a minimum human leukocyte antigen donor-recipient match. The northern South American population is in need of a new-generation cord blood bank that cryopreserves only units with high total nucleated cell content, thereby increasing the likelihood of use. Colombia set up a public cord blood bank in 2014; and, as a result of its research for improving high total nucleated cell content, a new strategy for UCB collection was developed. Data from 2933 collected and 759 cryopreserved cord blood units between 2014 and 2015 were analyzed. The correlation of donor and collection variables with cellularity was evaluated. Moreover, blood volume, cell content, CD34+ count, clonogenic capacity, and microbial contamination were assessed comparing the new method, which combines in utero and ex utero techniques, with the conventional strategies. Multivariate analysis confirmed a correlation between neonatal birth weight and cell content. The new collection method increased total nucleated cell content in approximately 26% and did not alter pre-cryopreservation and post-thaw cell recovery, viability, or clonogenic ability. Furthermore, it showed a remarkably low microbial contamination rate (1.2%). The strategy for UCB collection developed at the first Colombian public cord blood bank increases total nucleated cell content and does not affect unit quality. The existence of this bank is a remarkable breakthrough for Latin-American patients in need of this kind of transplantation. © 2017 The Authors Transfusion published by Wiley Periodicals, Inc. on behalf of AABB.

Acetogenic and Sulfate-Reducing Bacteria Inhabiting the Rhizoplane and Deep Cortex Cells of the Sea Grass Halodule wrightii†

PubMed Central

Küsel, Kirsten; Pinkart, Holly C.; Drake, Harold L.; Devereux, Richard

1999-01-01

Recent declines in sea grass distribution underscore the importance of understanding microbial community structure-function relationships in sea grass rhizospheres that might affect the viability of these plants. Phospholipid fatty acid analyses showed that sulfate-reducing bacteria and clostridia were enriched in sediments colonized by the sea grasses Halodule wrightii and Thalassia testudinum compared to an adjacent unvegetated sediment. Most-probable-number analyses found that in contrast to butyrate-producing clostridia, acetogens and acetate-utilizing sulfate reducers were enriched by an order of magnitude in rhizosphere sediments. Although sea grass roots are oxygenated in the daytime, colorimetric root incubation studies demonstrated that acetogenic O-demethylation and sulfidogenic iron precipitation activities were tightly associated with washed, sediment-free H. wrightii roots. This suggests that the associated anaerobes are able to tolerate exposure to oxygen. To localize and quantify the anaerobic microbial colonization, root thin sections were hybridized with newly developed 33P-labeled probes that targeted (i) low-G+C-content gram-positive bacteria, (ii) cluster I species of clostridia, (iii) species of Acetobacterium, and (iv) species of Desulfovibrio. Microautoradiography revealed intercellular colonization of the roots by Acetobacterium and Desulfovibrio species. Acetogenic bacteria occurred mostly in the rhizoplane and outermost cortex cell layers, and high numbers of sulfate reducers were detected on all epidermal cells and inward, colonizing some 60% of the deepest cortex cells. Approximately 30% of epidermal cells were colonized by bacteria that hybridized with an archaeal probe, strongly suggesting the presence of methanogens. Obligate anaerobes within the roots might contribute to the vitality of sea grasses and other aquatic plants and to the biogeochemistry of the surrounding sediment. PMID:10543830

Nitrate-Dependent Iron Oxidation: A Potential Mars Metabolism

PubMed Central

Price, Alex; Pearson, Victoria K.; Schwenzer, Susanne P.; Miot, Jennyfer; Olsson-Francis, Karen

2018-01-01

This work considers the hypothetical viability of microbial nitrate-dependent Fe2+ oxidation (NDFO) for supporting simple life in the context of the early Mars environment. This draws on knowledge built up over several decades of remote and in situ observation, as well as recent discoveries that have shaped current understanding of early Mars. Our current understanding is that certain early martian environments fulfill several of the key requirements for microbes with NDFO metabolism. First, abundant Fe2+ has been identified on Mars and provides evidence of an accessible electron donor; evidence of anoxia suggests that abiotic Fe2+ oxidation by molecular oxygen would not have interfered and competed with microbial iron metabolism in these environments. Second, nitrate, which can be used by some iron oxidizing microorganisms as an electron acceptor, has also been confirmed in modern aeolian and ancient sediment deposits on Mars. In addition to redox substrates, reservoirs of both organic and inorganic carbon are available for biosynthesis, and geochemical evidence suggests that lacustrine systems during the hydrologically active Noachian period (4.1–3.7 Ga) match the circumneutral pH requirements of nitrate-dependent iron-oxidizing microorganisms. As well as potentially acting as a primary producer in early martian lakes and fluvial systems, the light-independent nature of NDFO suggests that such microbes could have persisted in sub-surface aquifers long after the desiccation of the surface, provided that adequate carbon and nitrates sources were prevalent. Traces of NDFO microorganisms may be preserved in the rock record by biomineralization and cellular encrustation in zones of high Fe2+ concentrations. These processes could produce morphological biosignatures, preserve distinctive Fe-isotope variation patterns, and enhance preservation of biological organic compounds. Such biosignatures could be detectable by future missions to Mars with appropriate instrumentation. PMID:29616015

Modeling Bacteria Surface Acid-Base Properties: The Overprint Of Biology

NASA Astrophysics Data System (ADS)

Amores, D. R.; Smith, S.; Warren, L. A.

2009-05-01

Bacteria are ubiquitous in the environment and are important repositories for metals as well as nucleation templates for a myriad of secondary minerals due to an abundance of reactive surface binding sites. Model elucidation of whole cell surface reactivity simplifies bacteria as viable but static, i.e., no metabolic activity, to enable fits of microbial data sets from models derived from mineral surfaces. Here we investigate the surface proton charging behavior of live and dead whole cell cyanobacteria (Synechococcus sp.) harvested from a single parent culture by acid-base titration using a Fully Optimized ContinUouS (FOCUS) pKa spectrum method. Viability of live cells was verified by successful recultivation post experimentation, whereas dead cells were consistently non-recultivable. Surface site identities derived from binding constants determined for both the live and dead cells are consistent with molecular analogs for organic functional groups known to occur on microbial surfaces: carboxylic (pKa = 2.87-3.11), phosphoryl (pKa = 6.01-6.92) and amine/hydroxyl groups (pKa = 9.56-9.99). However, variability in total ligand concentration among the live cells is greater than those between the live and dead. The total ligand concentrations (LT, mol- mg-1 dry solid) derived from the live cell titrations (n=12) clustered into two sub-populations: high (LT = 24.4) and low (LT = 5.8), compared to the single concentration for the dead cell titrations (LT = 18.8; n=5). We infer from these results that metabolic activity can substantively impact surface reactivity of morphologically identical cells. These results and their modeling implications for bacteria surface reactivities will be discussed.

Vapor hydrogen peroxide as alternative to dry heat microbial reduction

NASA Astrophysics Data System (ADS)

Chung, S.; Kern, R.; Koukol, R.; Barengoltz, J.; Cash, H.

2008-09-01

The Jet Propulsion Laboratory (JPL), in conjunction with the NASA Planetary Protection Officer, has selected vapor phase hydrogen peroxide (VHP) sterilization process for continued development as a NASA approved sterilization technique for spacecraft subsystems and systems. The goal was to include this technique, with an appropriate specification, in NASA Procedural Requirements 8020.12 as a low-temperature complementary technique to the dry heat sterilization process. The VHP process is widely used by the medical industry to sterilize surgical instruments and biomedical devices, but high doses of VHP may degrade the performance of flight hardware, or compromise material compatibility. The goal for this study was to determine the minimum VHP process conditions for planetary protection acceptable microbial reduction levels. Experiments were conducted by the STERIS Corporation, under contract to JPL, to evaluate the effectiveness of vapor hydrogen peroxide for the inactivation of the standard spore challenge, Geobacillus stearothermophilus. VHP process parameters were determined that provide significant reductions in spore viability while allowing survival of sufficient spores for statistically significant enumeration. In addition to the obvious process parameters of interest: hydrogen peroxide concentration, number of injection cycles, and exposure duration, the investigation also considered the possible effect on lethality of environmental parameters: temperature, absolute humidity, and material substrate. This study delineated a range of test sterilizer process conditions: VHP concentration, process duration, a process temperature range for which the worst case D-value may be imposed, a process humidity range for which the worst case D-value may be imposed, and the dependence on selected spacecraft material substrates. The derivation of D-values from the lethality data permitted conservative planetary protection recommendations.

Nitrate-Dependent Iron Oxidation: A Potential Mars Metabolism.

PubMed

Price, Alex; Pearson, Victoria K; Schwenzer, Susanne P; Miot, Jennyfer; Olsson-Francis, Karen

2018-01-01

This work considers the hypothetical viability of microbial nitrate-dependent Fe 2+ oxidation (NDFO) for supporting simple life in the context of the early Mars environment. This draws on knowledge built up over several decades of remote and in situ observation, as well as recent discoveries that have shaped current understanding of early Mars. Our current understanding is that certain early martian environments fulfill several of the key requirements for microbes with NDFO metabolism. First, abundant Fe 2+ has been identified on Mars and provides evidence of an accessible electron donor; evidence of anoxia suggests that abiotic Fe 2+ oxidation by molecular oxygen would not have interfered and competed with microbial iron metabolism in these environments. Second, nitrate, which can be used by some iron oxidizing microorganisms as an electron acceptor, has also been confirmed in modern aeolian and ancient sediment deposits on Mars. In addition to redox substrates, reservoirs of both organic and inorganic carbon are available for biosynthesis, and geochemical evidence suggests that lacustrine systems during the hydrologically active Noachian period (4.1-3.7 Ga) match the circumneutral pH requirements of nitrate-dependent iron-oxidizing microorganisms. As well as potentially acting as a primary producer in early martian lakes and fluvial systems, the light-independent nature of NDFO suggests that such microbes could have persisted in sub-surface aquifers long after the desiccation of the surface, provided that adequate carbon and nitrates sources were prevalent. Traces of NDFO microorganisms may be preserved in the rock record by biomineralization and cellular encrustation in zones of high Fe 2+ concentrations. These processes could produce morphological biosignatures, preserve distinctive Fe-isotope variation patterns, and enhance preservation of biological organic compounds. Such biosignatures could be detectable by future missions to Mars with appropriate instrumentation.

Flux balance modeling to predict bacterial survival during pulsed-activity events

DOE PAGES

Jose, Nicholas A.; Lau, Rebecca; Swenson, Tami L.; ...

2018-04-16

Desert biological soil crusts (BSCs) are cyanobacteria-dominated surface soil microbial communities common to plant interspaces in arid environments. The capability to significantly dampen their metabolism allows them to exist for extended periods in a desiccated dormant state that is highly robust to environmental stresses. However, within minutes of wetting, metabolic functions reboot, maximizing activity during infrequent permissive periods. Microcoleus vaginatus, a primary producer within the crust ecosystem and an early colonizer, initiates crust formation by binding particles in the upper layer of soil via exopolysaccharides, making microbial dominated biological soil crusts highly dependent on the viability of this organism. Previousmore » studies have suggested that biopolymers play a central role in the survival of this organism by powering resuscitation, rapidly forming compatible solutes, and fueling metabolic activity in dark, hydrated conditions. To elucidate the mechanism of this phenomenon and provide a basis for future modeling of BSCs, we developed a manually curated, genome-scale metabolic model of Microcoleus vaginatus (iNJ1153). To validate this model, gas chromatography–mass spectroscopy (GC–MS) and liquid chromatography–mass spectroscopy (LC–MS) were used to characterize the rate of biopolymer accumulation and depletion in in hydrated Microcoleus vaginatus under light and dark conditions. Constraint-based flux balance analysis showed agreement between model predictions and experimental reaction fluxes. A significant amount of consumed carbon and light energy is invested into storage molecules glycogen and polyphosphate, while β-polyhydroxybutyrate may function as a secondary resource. Pseudo-steady-state modeling suggests that glycogen, the primary carbon source with the fastest depletion rate, will be exhausted if M. vaginatus experiences dark wetting events 4 times longer than light wetting events.« less

Flux balance modeling to predict bacterial survival during pulsed-activity events

NASA Astrophysics Data System (ADS)

Jose, Nicholas A.; Lau, Rebecca; Swenson, Tami L.; Klitgord, Niels; Garcia-Pichel, Ferran; Bowen, Benjamin P.; Baran, Richard; Northen, Trent R.

2018-04-01

Desert biological soil crusts (BSCs) are cyanobacteria-dominated surface soil microbial communities common to plant interspaces in arid environments. The capability to significantly dampen their metabolism allows them to exist for extended periods in a desiccated dormant state that is highly robust to environmental stresses. However, within minutes of wetting, metabolic functions reboot, maximizing activity during infrequent permissive periods. Microcoleus vaginatus, a primary producer within the crust ecosystem and an early colonizer, initiates crust formation by binding particles in the upper layer of soil via exopolysaccharides, making microbial dominated biological soil crusts highly dependent on the viability of this organism. Previous studies have suggested that biopolymers play a central role in the survival of this organism by powering resuscitation, rapidly forming compatible solutes, and fueling metabolic activity in dark, hydrated conditions. To elucidate the mechanism of this phenomenon and provide a basis for future modeling of BSCs, we developed a manually curated, genome-scale metabolic model of Microcoleus vaginatus (iNJ1153). To validate this model, gas chromatography-mass spectroscopy (GC-MS) and liquid chromatography-mass spectroscopy (LC-MS) were used to characterize the rate of biopolymer accumulation and depletion in in hydrated Microcoleus vaginatus under light and dark conditions. Constraint-based flux balance analysis showed agreement between model predictions and experimental reaction fluxes. A significant amount of consumed carbon and light energy is invested into storage molecules glycogen and polyphosphate, while β-polyhydroxybutyrate may function as a secondary resource. Pseudo-steady-state modeling suggests that glycogen, the primary carbon source with the fastest depletion rate, will be exhausted if M. vaginatus experiences dark wetting events 4 times longer than light wetting events.

H2O2 modulates the energetic metabolism of the cloud microbiome

NASA Astrophysics Data System (ADS)

Wirgot, Nolwenn; Vinatier, Virginie; Deguillaume, Laurent; Sancelme, Martine; Delort, Anne-Marie

2017-12-01

Chemical reactions in clouds lead to oxidation processes driven by radicals (mainly HO⚫, NO3⚫, or HO2⚫) or strong oxidants such as H2O2, O3, nitrate, and nitrite. Among those species, hydrogen peroxide plays a central role in the cloud chemistry by driving its oxidant capacity. In cloud droplets, H2O2 is transformed by microorganisms which are metabolically active. Biological activity can therefore impact the cloud oxidant capacity. The present article aims at highlighting the interactions between H2O2 and microorganisms within the cloud system. First, experiments were performed with selected strains studied as a reference isolated from clouds in microcosms designed to mimic the cloud chemical composition, including the presence of light and iron. Biotic and abiotic degradation rates of H2O2 were measured and results showed that biodegradation was the most efficient process together with the photo-Fenton process. H2O2 strongly impacted the microbial energetic state as shown by adenosine triphosphate (ATP) measurements in the presence and absence of H2O2. This ATP depletion was not due to the loss of cell viability. Secondly, correlation studies were performed based on real cloud measurements from 37 cloud samples collected at the PUY station (1465 m a.s.l., France). The results support a strong correlation between ATP and H2O2 concentrations and confirm that H2O2 modulates the energetic metabolism of the cloud microbiome. The modulation of microbial metabolism by H2O2 concentration could thus impact cloud chemistry, in particular the biotransformation rates of carbon compounds, and consequently can perturb the way the cloud system is modifying the global atmospheric chemistry.

Preparation of a standardised faecal slurry for ex-vivo microbiota studies which reduces inter-individual donor bias.

PubMed

O'Donnell, Michelle M; Rea, Mary C; O'Sullivan, Órla; Flynn, Cal; Jones, Beth; McQuaid, Albert; Shanahan, Fergus; Ross, R Paul

2016-10-01

In-vitro gut fermentation systems provide suitable models for studying gut microbiota composition and functionality. However, such methods depend on the availability of donors and the assumption of reproducibility between microbial communities before experimental treatments commence. The aim of this study was to develop a frozen standardised inoculum (FSI) which minimizes inter-individual variation and to determine its stability over time using culture-dependent and culture-independent techniques. A method for the preparation difference of a FSI is described which involves pooling the faecal samples, centrifugation and pelleting of the cell biomass and finally homogenising the cell pellets with phosphate buffer and glycerol. Using this approach, no significant difference in total anaerobe cell viability was observed between the fresh standardised inoculum (before freezing) and the 12days post freezing FSI. Moreover, Quantitative PCR revealed no significant alterations in the estimated bacterial numbers in the FSI preparations for any of the phyla. MiSeq sequencing revealed minute differences in the relative abundance at phylum, family and genus levels between the FSI preparations. Differences in the microbiota denoted as significant were limited between preparations in the majority of cases to changes in percentage relative abundance of ±0.5%. The independently prepared FSIs revealed a high degree of reproducibility in terms of microbial composition between the three preparations. This study provides a method to produce a standardised human faecal inoculum suitable for freezing. Based on culture-dependent and independent analysis, the method ensures a degree of reproducibility between preparations by lessening the effect of inter-individual variation among the donors, thereby making the system more suitable for the accurate interpretation of the effects of experimental treatments. Copyright © 2016 Elsevier B.V. All rights reserved.

Rapidly Evolving Toll-3/4 Genes Encode Male-Specific Toll-Like Receptors in Drosophila.

PubMed

Levin, Tera C; Malik, Harmit S

2017-09-01

Animal Toll-like receptors (TLRs) have evolved through a pattern of duplication and divergence. Whereas mammalian TLRs directly recognize microbial ligands, Drosophila Tolls bind endogenous ligands downstream of both developmental and immune signaling cascades. Here, we find that most Toll genes in Drosophila evolve slowly with little gene turnover (gains/losses), consistent with their important roles in development and indirect roles in microbial recognition. In contrast, we find that the Toll-3/4 genes have experienced an unusually rapid rate of gene gains and losses, resulting in lineage-specific Toll-3/4s and vastly different gene repertoires among Drosophila species, from zero copies (e.g., D. mojavensis) to nineteen copies (e.g., D. willistoni). In D. willistoni, we find strong evidence for positive selection in Toll-3/4 genes, localized specifically to an extracellular region predicted to overlap with the binding site of Spätzle, the only known ligand of insect Tolls. However, because Spätzle genes are not experiencing similar selective pressures, we hypothesize that Toll-3/4s may be rapidly evolving because they bind to a different ligand, akin to TLRs outside of insects. We further find that most Drosophila Toll-3/4 genes are either weakly expressed or expressed exclusively in males, specifically in the germline. Unlike other Toll genes in D. melanogaster, Toll-3, and Toll-4 have apparently escaped from essential developmental roles, as knockdowns have no substantial effects on viability or male fertility. Based on these findings, we propose that the Toll-3/4 genes represent an exceptionally rapidly evolving lineage of Drosophila Toll genes, which play an unusual, as-yet-undiscovered role in the male germline. © The Author 2017. Published by Oxford University Press on behalf of the Society for Molecular Biology and Evolution.

Rapidly Evolving Toll-3/4 Genes Encode Male-Specific Toll-Like Receptors in Drosophila

PubMed Central

Levin, Tera C.; Malik, Harmit S.

2017-01-01

Abstract Animal Toll-like receptors (TLRs) have evolved through a pattern of duplication and divergence. Whereas mammalian TLRs directly recognize microbial ligands, Drosophila Tolls bind endogenous ligands downstream of both developmental and immune signaling cascades. Here, we find that most Toll genes in Drosophila evolve slowly with little gene turnover (gains/losses), consistent with their important roles in development and indirect roles in microbial recognition. In contrast, we find that the Toll-3/4 genes have experienced an unusually rapid rate of gene gains and losses, resulting in lineage-specific Toll-3/4s and vastly different gene repertoires among Drosophila species, from zero copies (e.g., D. mojavensis) to nineteen copies (e.g., D. willistoni). In D. willistoni, we find strong evidence for positive selection in Toll-3/4 genes, localized specifically to an extracellular region predicted to overlap with the binding site of Spätzle, the only known ligand of insect Tolls. However, because Spätzle genes are not experiencing similar selective pressures, we hypothesize that Toll-3/4s may be rapidly evolving because they bind to a different ligand, akin to TLRs outside of insects. We further find that most Drosophila Toll-3/4 genes are either weakly expressed or expressed exclusively in males, specifically in the germline. Unlike other Toll genes in D. melanogaster, Toll-3, and Toll-4 have apparently escaped from essential developmental roles, as knockdowns have no substantial effects on viability or male fertility. Based on these findings, we propose that the Toll-3/4 genes represent an exceptionally rapidly evolving lineage of Drosophila Toll genes, which play an unusual, as-yet-undiscovered role in the male germline. PMID:28541576

Monitoring viability of seeds in gene banks: developing software tools to increase efficiency

USDA-ARS?s Scientific Manuscript database

Monitoring the decline of seed viability is essential for effective long term seed storage in ex situ collections. Recent FAO Genebank Standards recommend monitoring intervals at one-third the time predicted for viability to fall to 85% of initial viability. This poster outlines the development of ...

Production of probiotic bovine salami using Lactobacillus plantarum 299v as adjunct.

PubMed

Blaiotta, Giuseppe; Murru, Nicoletta; Di Cerbo, Alessandro; Romano, Raffaele; Aponte, Maria

2018-04-01

Five probiotic lactobacilli were tested, alone or in combination with two commercial starters, to select the most suitable strain for a probiotic bovine salami production. Lactobacillus plantarum 299v was used with both starters, to make salami according to a traditional recipe. Salami obtained by using just the starters and by spontaneous fermentation, served as control. Microbial dynamics, as well as the main physico-chemical parameters, were monitored throughout ripening. The survival of probiotic 299v was confirmed by strains' tracking by means of RAPD-PCR coupled to a culture-independent approach PCR-DGGE-based. The results showed a remarkable viability of the probiotic strain even after 60 days of storage. Experimental salami exhibited the same level of sensory acceptance of control salami, were hygienically safe, and characterised by pH, weight loss and microbiological loads within the ranges conventionally advocated for optimal fermented sausages. Outcomes indicate the workable possibility of using second-quality beef cuts for probiotic salami production. © 2017 Society of Chemical Industry. © 2017 Society of Chemical Industry.

Insight into the effect of organic and inorganic draw solutes on the flux stability and sludge characteristics in the osmotic membrane bioreactor.

PubMed

Siddique, Muhammad Saboor; Khan, Sher Jamal; Shahzad, Muhammad Aamir; Nawaz, Muhammad Saqib; Hankins, Nicholas P

2018-02-01

In this study, chloride based (CaCl 2 and MgCl 2 ) and acetate based (NaOAc and MgOAc) salts in comparison with NaCl were investigated as draw solutions (DS) to evaluate their viability in the osmotic membrane bioreactor (OMBR). Membrane distillation was coupled with an OMBR setup to develop a hybrid OMBR-MD system, for the production of clean water and DS recovery. Results demonstrate that organic DS were able to mitigate the salinity buildup in the bioreactor as compared to inorganic salts. Prolonged filtration runs were observed with MgCl 2 and MgOAc in contrast with other draw solutes at the same molar concentration. Significant membrane fouling was observed with NaOAc while rapid flux decline due to increased salinity build-up was witnessed with NaCl and CaCl 2 . Improved characteristics of mixed liquor in terms of sludge filterability, particle size, and biomass growth along with the degradation of soluble microbial products (SMP) were found with organic DS. Copyright © 2017 Elsevier Ltd. All rights reserved.

Acetone-butanol-ethanol fermentation of corn stover: current production methods, economic viability and commercial use.

PubMed

Baral, Nawa R; Slutzky, Lauren; Shah, Ajay; Ezeji, Thaddeus C; Cornish, Katrina; Christy, Ann

2016-03-01

Biobutanol is a next-generation liquid biofuel with properties akin to those of gasoline. There is a widespread effort to commercialize biobutanol production from agricultural residues, such as corn stover, which do not compete with human and animal foods. This pursuit is backed by extensive government mandates to expand alternative energy sources. This review provides an overview of research on biobutanol production using corn stover feedstock. Structural composition, pretreatment, sugar yield (following pretreatment and hydrolysis) and generation of lignocellulose-derived microbial inhibitory compounds (LDMICs) from corn stover are discussed. The review also discusses different Clostridium species and strains employed for biobutanol production from corn stover-derived sugars with respect to solvent yields, tolerance to LDMICs and in situ solvent recovery (integrated fermentation). Further, the economics of cellulosic biobutanol production are highlighted and compared to corn starch-derived ethanol and gasoline. As discussed herein, the economic competitiveness of biobutanol production from corn stover largely depends on feedstock processing and fermentation process design. © FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

O-Acetylation of Plant Cell Wall Polysaccharides

PubMed Central

Gille, Sascha; Pauly, Markus

2011-01-01

Plant cell walls are composed of structurally diverse polymers, many of which are O-acetylated. How plants O-acetylate wall polymers and what its function is remained elusive until recently, when two protein families were identified in the model plant Arabidopsis that are involved in the O-acetylation of wall polysaccharides – the reduced wall acetylation (RWA) and the trichome birefringence-like (TBL) proteins. This review discusses the role of these two protein families in polysaccharide O-acetylation and outlines the differences and similarities of polymer acetylation mechanisms in plants, fungi, bacteria, and mammals. Members of the TBL protein family had been shown to impact pathogen resistance, freezing tolerance, and cellulose biosynthesis. The connection of TBLs to polysaccharide O-acetylation thus gives crucial leads into the biological function of wall polymer O-acetylation. From a biotechnological point understanding the O-acetylation mechanism is important as acetyl-substituents inhibit the enzymatic degradation of wall polymers and released acetate can be a potent inhibitor in microbial fermentations, thus impacting the economic viability of, e.g., lignocellulosic based biofuel production. PMID:22639638

Development of a microbial process for the recovery of petroleum oil from depleted reservoirs at 91-96°C.

PubMed

Arora, Preeti; Ranade, Dilip R; Dhakephalkar, Prashant K

2014-08-01

A consortium of bacteria growing at 91°C and above (optimally at 96°C) was developed for the recovery of crude oil from declining/depleted oil reservoirs having temperature of more than 91°C. PCR-DGGE-Sequencing analysis of 16S rRNA gene fragments of NJS-4 consortium revealed the presence of four strains identified as members of the genus Clostridium. The metabolites produced by NJS-4 consortium included volatile fatty acids, organic acids, surfactants, exopolysaccarides and CO2, which reduced viscosity, emulsified crude oil and increased the pressure that facilitated displacement of emulsified oil towards the surface. NJS-4 enhanced oil recovery by 26.7% and 10.1% in sand pack trials and core flood studies respectively in optimized nutrient medium comprised of sucrose and sodium acetate as carbon/energy source and urea as nitrogen source (pH 7-9, 96°C, and 4% salinity). Nutrient medium for MEOR was constituted using commercial grade cheap nutrients to improve the economic viability of MEOR process. Copyright © 2014 Elsevier Ltd. All rights reserved.

Microorganisms as tracers in groundwater injection and recovery experiments: A review

USGS Publications Warehouse

Harvey, R.W.

1997-01-01

Modern day injection and recovery techniques designed to examine the transport behavior of microorganisms in groundwater have evolved from experiments conducted in the late 1800s, in which bacteria that form red or yellow pigments were used to trace flow paths through karst and fractured- rock aquifers. A number of subsequent groundwater hydrology studies employed bacteriophage that can be injected into aquifers at very high concentrations (e g., 1013 phage ml-1) and monitored through many log units of dilution to follow groundwater flow paths for great distances, particularly in karst terrain. Starting in the 1930s, microbial indicators of fecal contamination (particularly coliform bacteria and their coliphages) were employed as tracers to determine potential migration of pathogens in groundwater. Several injection and recovery experiments performed in the 1990s employed indigenous groundwater microorganisms (both cultured and uncultured) that are better able to survive under in situ conditions. Better methods for labeling native bacteria (e.g by stable isotope labeling or inserting genetic markers; such as the ability to cause ice nucleation) are being developed that will not compromise the organisms' viability during the experimental time course.

Four Considerations Regarding the Viability of Small Rural Schools in New Zealand.

ERIC Educational Resources Information Center

Stevens, Ken

1993-01-01

Reviews a New Zealand report titled, "Report of the Economic and Educational Viability of Small Schools Review." Discusses the four aspects of small schools considered in the report: (1) educational viability; (2) economic viability; (3) relationships of small schools with their communities; and (4) conflict between availability and…

Viability analysis in biological evaluations: Concepts of population viability analysis, biological population, and ecological scale

Treesearch

Gregory D. Hayward; John R. Squires

1994-01-01

Environmental protection strategies often rely on environmental impact assessments. As part of the assessment process biologists are routinely asked to evaluate the effects of management actions on plants and animals. This evaluation often requires that biologists make judgments about the viability of affected populations. However, population viability...

Survey of the Definition of Fetal Viability and the Availability, Indications, and Decision Making Processes for Post-Viability Termination of Pregnancy for Fetal Abnormalities and Health Conditions in Canada.

PubMed

Hull, Danna; Davies, Gregory; Armour, Christine M

2016-06-01

The purpose of this study was to explore the definition of fetal viability and the availability, indications, and decision making processes for post-viability termination of pregnancy for fetal abnormalities and health conditions in Canada. An online survey of members of the Canadian Association of Genetic Counsellors, the Canadian College of Medical Geneticists, and the Canadian Society for Maternal-Fetal Medicine who provide direct counselling to, or management of, prenatal patients in Canada (total sample size 815). Results of this study showed that the majority of respondents indicated that their centre will offer post-viability termination of pregnancy (98/123; 80 %). Sixty-seven percent (68/101) of respondents reported the definition of fetal viability to be 24 weeks' gestation. Most respondents reported that a collaborative decision making process was used to determine if post-viability termination of pregnancy would be offered (136/170; 80 %). For conditions presumed to be lethal/likely lethal, the majority of respondents would "sometimes" or "always" offer post-viability termination of pregnancy, whereas for conditions presumed to have a mild effect, the majority of respondents would "rarely" or "never" offer post-viability termination of pregnancy. Ninety percent (77/86) of respondents reported that perinatal hospice is offered as an alternative to termination of pregnancy. In conclusion, this study suggests that although post-viability termination is available in many provinces in Canada, variation in the definition of fetal viability and indications appear to exist. While these variations may lead to unequal access to post-viability termination of pregnancy across Canada, they might also represent the complexity of the decision making process and the importance of examining individual factors to ensure that the most appropriate decision is made in each case.

Retinoids, retinoid analogs, and lactoferrin interact and differentially affect cell viability of 2 bovine mammary cell types in vitro.

PubMed

Wang, Y; Baumrucker, C R

2010-07-01

Two bovine mammary cell types (BME-UV1 and MeBo cells) were used to evaluate the effect of natural retinoids, retinoid analogs, and bovine lactoferrin (bLf) on cell viability in vitro. Experiments with Alamar Blue showed a linear relationship between fluorescence and cell viability index. The BME-UV1 cells exhibited twice the metabolic activity but required half the doubling time of the MeBo cells. The BME-UV1 cells were very sensitive to all-trans retinoic acid (atRA) inhibition of cell viability (P<0.05) and exhibited a dose-dependent inhibition with 9-cisRA (9cRA; P<0.05). The MeBo cells exhibited some inhibition with these natural ligands (P<0.05), but they were not as sensitive. The addition of bLf had similar inhibitory effects (P<0.05) on cell viability of the 2 mammary cell types. Applications of RA receptor (RAR) agonist indicated that the stimulation of the RAR in both mammary cell types was highly effective in inhibition of cell viability (P<0.05), whereas the application of an RAR antagonist stimulated MeBo cell viability (P<0.05) and inhibited BME-UV1 cell viability (P<0.05). Finally, the use of the RAR antagonist in conjunction with bLf indicated a rescue of the bLf effect in the MeBo cells, suggesting that bLf is acting through the RAR receptor. Conversely, bLf reverted inhibition of cell viability by 9cRA in the BME-UV1 cell type (P<0.05). We conclude that RAR interaction in bovine mammary cell types regulates cell viability in vitro; we hypothesize that the natural ligands mediate regulation of bovine mammary cell viability in vivo and that bLf can either enhance or reverse the retinoid-induced inhibition of cell viability, depending on the type of bovine mammary cell studied.

The effect of ultrasound-related stimuli on cell viability in microfluidic channels

PubMed Central

2013-01-01

Background In ultrasonic micro-devices, contrast agent micro-bubbles are known to initiate cavitation and streaming local to cells, potentially compromising cell viability. Here we investigate the effects of US alone by omitting contrast agent and monitoring cell viability under moderate-to-extreme ultrasound-related stimuli. Results Suspended H9c2 cardiac myoblasts were exposed to ultrasonic fields within a glass micro-capillary and their viability monitored under different US-related stimuli. An optimal injection flow rate of 2.6 mL/h was identified in which, high viability was maintained (~95%) and no mechanical stress towards cells was evident. This flow rate also allowed sufficient exposure of cells to US in order to induce bioeffects (~5 sec), whilst providing economical sample collection and processing times. Although the transducer temperature increased from ambient 23°C to 54°C at the maximum experimental voltage (29 Vpp), computational fluid dynamic simulations and controls (absence of US) revealed that the cell medium temperature did not exceed 34°C in the pressure nodal plane. Cells exposed to US amplitudes ranging from 0–29 Vpp, at a fixed frequency sweep period (tsw = 0.05 sec), revealed that viability was minimally affected up to ~15 Vpp. There was a ~17% reduction in viability at 21 Vpp, corresponding to the onset of Rayleigh-like streaming and a ~60% reduction at 29 Vpp, corresponding to increased streaming velocity or the potential onset of cavitation. At a fixed amplitude (29 Vpp) but with varying frequency sweep period (tsw = 0.02-0.50 sec), cell viability remained relatively constant at tsw ≥ 0.08 sec, whilst viability reduced at tsw < 0.08 sec and minimum viability recorded at tsw = 0.05 sec. Conclusion The absence of CA has enabled us to investigate the effect of US alone on cell viability. Moderate-to-extreme US-related stimuli of cells have allowed us to discriminate between stimuli that maintain high viability and stimuli that significantly reduce cell viability. Results from this study may be of potential interest to researchers in the field of US-induced intracellular drug delivery and ultrasonic manipulation of biological cells. PMID:23809777

The effect of ultrasound-related stimuli on cell viability in microfluidic channels.

PubMed

Ankrett, Dyan N; Carugo, Dario; Lei, Junjun; Glynne-Jones, Peter; Townsend, Paul A; Zhang, Xunli; Hill, Martyn

2013-06-28

In ultrasonic micro-devices, contrast agent micro-bubbles are known to initiate cavitation and streaming local to cells, potentially compromising cell viability. Here we investigate the effects of US alone by omitting contrast agent and monitoring cell viability under moderate-to-extreme ultrasound-related stimuli. Suspended H9c2 cardiac myoblasts were exposed to ultrasonic fields within a glass micro-capillary and their viability monitored under different US-related stimuli. An optimal injection flow rate of 2.6 mL/h was identified in which, high viability was maintained (~95%) and no mechanical stress towards cells was evident. This flow rate also allowed sufficient exposure of cells to US in order to induce bioeffects (~5 sec), whilst providing economical sample collection and processing times. Although the transducer temperature increased from ambient 23°C to 54°C at the maximum experimental voltage (29 Vpp), computational fluid dynamic simulations and controls (absence of US) revealed that the cell medium temperature did not exceed 34°C in the pressure nodal plane. Cells exposed to US amplitudes ranging from 0-29 Vpp, at a fixed frequency sweep period (tsw = 0.05 sec), revealed that viability was minimally affected up to ~15 Vpp. There was a ~17% reduction in viability at 21 Vpp, corresponding to the onset of Rayleigh-like streaming and a ~60% reduction at 29 Vpp, corresponding to increased streaming velocity or the potential onset of cavitation. At a fixed amplitude (29 Vpp) but with varying frequency sweep period (tsw = 0.02-0.50 sec), cell viability remained relatively constant at tsw ≥ 0.08 sec, whilst viability reduced at tsw < 0.08 sec and minimum viability recorded at tsw = 0.05 sec. The absence of CA has enabled us to investigate the effect of US alone on cell viability. Moderate-to-extreme US-related stimuli of cells have allowed us to discriminate between stimuli that maintain high viability and stimuli that significantly reduce cell viability. Results from this study may be of potential interest to researchers in the field of US-induced intracellular drug delivery and ultrasonic manipulation of biological cells.

Colony Failure Linked to Low Sperm Viability in Honey Bee (Apis mellifera) Queens and an Exploration of Potential Causative Factors.

PubMed

Pettis, Jeffery S; Rice, Nathan; Joselow, Katie; vanEngelsdorp, Dennis; Chaimanee, Veeranan

2016-01-01

Queen health is closely linked to colony performance in honey bees as a single queen is normally responsible for all egg laying and brood production within the colony. In the U. S. in recent years, queens have been failing at a high rate; with 50% or greater of queens replaced in colonies within 6 months when historically a queen might live one to two years. This high rate of queen failure coincides with the high mortality rates of colonies in the US, some years with >50% of colonies dying. In the current study, surveys of sperm viability in US queens were made to determine if sperm viability plays a role in queen or colony failure. Wide variation was observed in sperm viability from four sets of queens removed from colonies that beekeepers rated as in good health (n = 12; average viability = 92%), were replacing as part of normal management (n = 28; 57%), or where rated as failing (n = 18 and 19; 54% and 55%). Two additional paired set of queens showed a statistically significant difference in viability between colonies rated by the beekeeper as failing or in good health from the same apiaries. Queens removed from colonies rated in good health averaged high viability (ca. 85%) while those rated as failing or in poor health had significantly lower viability (ca. 50%). Thus low sperm viability was indicative of, or linked to, colony performance. To explore the source of low sperm viability, six commercial queen breeders were surveyed and wide variation in viability (range 60-90%) was documented between breeders. This variability could originate from the drones the queens mate with or temperature extremes that queens are exposed to during shipment. The role of shipping temperature as a possible explanation for low sperm viability was explored. We documented that during shipment queens are exposed to temperature spikes (<8 and > 40°C) and these spikes can kill 50% or more of the sperm stored in queen spermathecae in live queens. Clearly low sperm viability is linked to colony performance and laboratory and field data provide evidence that temperature extremes are a potential causative factor.

30 CFR 203.85 - What is in an economic viability and relief justification report?

Code of Federal Regulations, 2012 CFR

2012-07-01

... 30 Mineral Resources 2 2012-07-01 2012-07-01 false What is in an economic viability and relief... Sulfur General § 203.85 What is in an economic viability and relief justification report? This report... economic viability and relief justification report must contain the following items for an oil and gas...

30 CFR 203.68 - What pre-application costs will MMS consider in determining economic viability?

Code of Federal Regulations, 2010 CFR

2010-07-01

... determining economic viability? 203.68 Section 203.68 Mineral Resources MINERALS MANAGEMENT SERVICE... § 203.68 What pre-application costs will MMS consider in determining economic viability? (a) We will not consider ineligible costs as set forth in § 203.89(h) in determining economic viability for purposes of...

30 CFR 203.68 - What pre-application costs will BSEE consider in determining economic viability?

Code of Federal Regulations, 2012 CFR

2012-07-01

... in determining economic viability? 203.68 Section 203.68 Mineral Resources BUREAU OF SAFETY AND... determining economic viability? (a) We will not consider ineligible costs as set forth in § 203.89(h) in determining economic viability for purposes of royalty relief. (b) We will consider sunk costs according to...

30 CFR 203.85 - What is in an economic viability and relief justification report?

Code of Federal Regulations, 2011 CFR

2011-07-01

... 30 Mineral Resources 2 2011-07-01 2011-07-01 false What is in an economic viability and relief... Oil, Gas, and Sulfur General Required Reports § 203.85 What is in an economic viability and relief... your own model and results. The economic viability and relief justification report must contain the...

30 CFR 203.68 - What pre-application costs will BSEE consider in determining economic viability?

Code of Federal Regulations, 2013 CFR

2013-07-01

... in determining economic viability? 203.68 Section 203.68 Mineral Resources BUREAU OF SAFETY AND... determining economic viability? (a) We will not consider ineligible costs as set forth in § 203.89(h) in determining economic viability for purposes of royalty relief. (b) We will consider sunk costs according to...

30 CFR 203.68 - What pre-application costs will MMS consider in determining economic viability?

Code of Federal Regulations, 2011 CFR

2011-07-01

... determining economic viability? 203.68 Section 203.68 Mineral Resources BUREAU OF OCEAN ENERGY MANAGEMENT... economic viability? (a) We will not consider ineligible costs as set forth in § 203.89(h) in determining economic viability for purposes of royalty relief. (b) We will consider sunk costs according to the...

30 CFR 203.85 - What is in an economic viability and relief justification report?

Code of Federal Regulations, 2013 CFR

2013-07-01

... 30 Mineral Resources 2 2013-07-01 2013-07-01 false What is in an economic viability and relief... Sulfur General § 203.85 What is in an economic viability and relief justification report? This report... economic viability and relief justification report must contain the following items for an oil and gas...

30 CFR 203.68 - What pre-application costs will BSEE consider in determining economic viability?

Code of Federal Regulations, 2014 CFR

2014-07-01

... in determining economic viability? 203.68 Section 203.68 Mineral Resources BUREAU OF SAFETY AND... determining economic viability? (a) We will not consider ineligible costs as set forth in § 203.89(h) in determining economic viability for purposes of royalty relief. (b) We will consider sunk costs according to...

30 CFR 203.85 - What is in an economic viability and relief justification report?

Code of Federal Regulations, 2014 CFR

2014-07-01

... 30 Mineral Resources 2 2014-07-01 2014-07-01 false What is in an economic viability and relief... Sulfur General § 203.85 What is in an economic viability and relief justification report? This report... economic viability and relief justification report must contain the following items for an oil and gas...

30 CFR 203.85 - What is in an economic viability and relief justification report?

Code of Federal Regulations, 2010 CFR

2010-07-01

... 30 Mineral Resources 2 2010-07-01 2010-07-01 false What is in an economic viability and relief... Required Reports § 203.85 What is in an economic viability and relief justification report? This report... economic viability and relief justification report must contain the following items for an oil and gas...

Business Reengineering - Government Viability

DTIC Science & Technology

2000-04-01

BUSINESS REENGINEERING - GOVERNMENT VIABILITY BY LIEUTENANT COLONEL GRAY K. COYNER United States Air Force Reserve DISTRIBUTION STATEMENT A: Approved...PROJECT Business Reengineering - Government Viability by Lt Col Gray K. Coyner USAFR Col Harry E. LeBoeuf Jr. Project Advisor The views expressed in...release. Distribution is unlimited. ii ABSTRACT AUTHOR: Gray K. Coyner TITLE: Business Reengineering - Government Viability FORMAT: Strategy Research

45 CFR 1302.23 - Suspension or termination of grantee which shows legal status but not financial viability.

Code of Federal Regulations, 2010 CFR

2010-10-01

... legal status but not financial viability. 1302.23 Section 1302.23 Public Welfare Regulations Relating to....23 Suspension or termination of grantee which shows legal status but not financial viability. (a) If the date of change of financial viability precedes or will precede the end of the grantee's program...

45 CFR 1302.22 - Suspension or termination of grantee which shows financial viability but not legal status.

Code of Federal Regulations, 2010 CFR

2010-10-01

... financial viability but not legal status. 1302.22 Section 1302.22 Public Welfare Regulations Relating to....22 Suspension or termination of grantee which shows financial viability but not legal status. If a... may show financial viability, the grant shall be suspended or terminated or refunding shall be denied...

Storage and Viability Assessment of Date Palm Pollen.

PubMed

Maryam; Jaskani, Muhammad J; Naqvi, Summar A

2017-01-01

Pollen storage and viability are very important for pollination, breeding, biodiversity, biotechnology, conservation, and other biological and non-biological studies of the date palm. Optimizing procedures and duration of storage are important for effective and long-term date palm pollen storage and viability. Here we describe pollen storage methods, such as room temperature (25-30 °C), refrigeration (4 °C), storage at 4 °C in desiccators, deep freezer (-20 °C), and cryopreservation (-196 °C). Based on pollen viability by staining and in vitro germination methods, cryopreservation is the best method for long-term storage without any significant effect on pollen viability (75-84%); however, the percentage of pollen viability depends on the storage period.

The combined influence of sub-optimal temperature and salinity on the in vitro viability of Perkinsus marinus, a protistan parasite of the eastern oyster Crassostrea virginica

USGS Publications Warehouse

La Peyre, M.K.; Casas, S.M.; Gayle, W.; La Peyre, Jerome F.

2010-01-01

Perkinsus marinus is a major cause of mortality in eastern oysters along the Gulf of Mexico and Atlantic coasts. It is also well documented that temperature and salinity are the primary environmental factors affecting P. marinus viability and proliferation. However, little is known about the effects of combined sub-optimal temperatures and salinities on P. marinus viability. This in vitro study examined those effects by acclimating P. marinus at three salinities (7, 15, 25. ppt) to 10 ??C to represent the lowest temperatures generally reached in the Gulf of Mexico, and to 2 ??C to represent the lowest temperatures reached along the mid-Atlantic coasts and by measuring changes in cell viability and density on days 1, 30, 60 and 90 following acclimation. Cell viability and density were also measured in 7. ppt cultures acclimated to each temperature and then transferred to 3.5. ppt. The largest decreases in cell viability occurred only with combined low temperature and salinity, indicating that there is clearly a synergistic effect. The largest decreases in cell viability occurred only with both low temperature and salinity after 30. days (3.5. ppt, 2 ??C: 0% viability), 60. days (3.5. ppt, 10 ??C: 0% viability) and 90. days (7. ppt, 2 ??C: 0.6 ?? 0.7%; 7. ppt, 10 ??C: 0.2 ?? 0.2%). ?? 2010 .

A real-time, non-invasive, micro-optrode technique for detecting seed viability by using oxygen influx.

PubMed

Xin, Xia; Wan, Yinglang; Wang, Wenjun; Yin, Guangkun; McLamore, Eric S; Lu, Xinxiong

2013-10-28

Quantifying seed viability is required for seed bank maintenance. The classical methods for detecting seed viability are time consuming and frequently cause seed damage and unwanted germination. We have established a novel micro-optrode technique (MOT) to measure seed viability in a quick and non-invasive manner by measuring the oxygen influxes of intact seeds, approximately 10 seconds to screen one seed. Here, we used soybean, wheat, and oilseed rape as models to test our method. After 3-hour imbibition, oxygen influxes were recorded in real-time with the total measurement taking less than 5 minutes. The results indicated a significantly positive correlation between oxygen influxes and viability in all 3 seed types. We also established a linear equation between oxygen influxes and seed viability for each seed type. For measurements, seeds were kept in the early imbibition stage without germination. Thus, MOT is a reliable, quick, and low-cost seed viability detecting technique.

From "Gut Feeling" to Objectivity: Machine Preservation of the Liver as a Tool to Assess Organ Viability.

PubMed

Watson, Christopher J E; Jochmans, Ina

2018-01-01

The purpose of this review was to summarise how machine perfusion could contribute to viability assessment of donor livers. In both hypothermic and normothermic machine perfusion, perfusate transaminase measurement has allowed pretransplant assessment of hepatocellular damage. Hypothermic perfusion permits transplantation of marginal grafts but as yet has not permitted formal viability assessment. Livers undergoing normothermic perfusion have been investigated using parameters similar to those used to evaluate the liver in vivo. Lactate clearance, glucose evolution and pH regulation during normothermic perfusion seem promising measures of viability. In addition, bile chemistry might inform on cholangiocyte viability and the likelihood of post-transplant cholangiopathy. While the use of machine perfusion technology has the potential to reduce and even remove uncertainty regarding liver graft viability, analysis of large datasets, such as those derived from large multicenter trials of machine perfusion, are needed to provide sufficient information to enable viability parameters to be defined and validated .

Colony Failure Linked to Low Sperm Viability in Honey Bee (Apis mellifera) Queens and an Exploration of Potential Causative Factors

PubMed Central

Pettis, Jeffery S.; Rice, Nathan; Joselow, Katie; vanEngelsdorp, Dennis; Chaimanee, Veeranan

2016-01-01

Queen health is closely linked to colony performance in honey bees as a single queen is normally responsible for all egg laying and brood production within the colony. In the U. S. in recent years, queens have been failing at a high rate; with 50% or greater of queens replaced in colonies within 6 months when historically a queen might live one to two years. This high rate of queen failure coincides with the high mortality rates of colonies in the US, some years with >50% of colonies dying. In the current study, surveys of sperm viability in US queens were made to determine if sperm viability plays a role in queen or colony failure. Wide variation was observed in sperm viability from four sets of queens removed from colonies that beekeepers rated as in good health (n = 12; average viability = 92%), were replacing as part of normal management (n = 28; 57%), or where rated as failing (n = 18 and 19; 54% and 55%). Two additional paired set of queens showed a statistically significant difference in viability between colonies rated by the beekeeper as failing or in good health from the same apiaries. Queens removed from colonies rated in good health averaged high viability (ca. 85%) while those rated as failing or in poor health had significantly lower viability (ca. 50%). Thus low sperm viability was indicative of, or linked to, colony performance. To explore the source of low sperm viability, six commercial queen breeders were surveyed and wide variation in viability (range 60–90%) was documented between breeders. This variability could originate from the drones the queens mate with or temperature extremes that queens are exposed to during shipment. The role of shipping temperature as a possible explanation for low sperm viability was explored. We documented that during shipment queens are exposed to temperature spikes (<8 and > 40°C) and these spikes can kill 50% or more of the sperm stored in queen spermathecae in live queens. Clearly low sperm viability is linked to colony performance and laboratory and field data provide evidence that temperature extremes are a potential causative factor. PMID:26863438

Development of a viability standard curve for microencapsulated probiotic bacteria using confocal microscopy and image analysis software.

PubMed

Moore, Sarah; Kailasapathy, Kasipathy; Phillips, Michael; Jones, Mark R

2015-07-01

Microencapsulation is proposed to protect probiotic strains from food processing procedures and to maintain probiotic viability. Little research has described the in situ viability of microencapsulated probiotics. This study successfully developed a real-time viability standard curve for microencapsulated bacteria using confocal microscopy, fluorescent dyes and image analysis software. Copyright © 2015 Elsevier B.V. All rights reserved.

Molecular Assays for Determining Mycobacterium leprae Viability in Tissues of Experimentally Infected Mice

PubMed Central

Davis, Grace L.; Ray, Nashone A.; Lahiri, Ramanuj; Gillis, Thomas P.; Krahenbuhl, James L.; Williams, Diana L.; Adams, Linda B.

2013-01-01

Background The inability of Mycobacterium leprae to grow on axenic media has necessitated specialized techniques in order to determine viability of this organism. The purpose of this study was to develop a simple and sensitive molecular assay for determining M. leprae viability directly from infected tissues. Methodology/Principle Findings Two M. leprae-specific quantitative reverse transcription PCR (qRT-PCR) assays based on the expression levels of esxA, encoding the ESAT-6 protein, and hsp18, encoding the heat shock 18 kDa protein, were developed and tested using infected footpad (FP) tissues of both immunocompetent and immunocompromised (athymic nu/nu) mice. In addition, the ability of these assays to detect the effects of anti-leprosy drug treatment on M. leprae viability was determined using rifampin and rifapentine, each at 10 mg/kg for 1, 5, or 20 daily doses, in the athymic nu/nu FP model. Molecular enumeration (RLEP PCR) and viability determinations (qRT-PCR) were performed via Taqman methodology on DNA and RNA, respectively, purified from ethanol-fixed FP tissue and compared with conventional enumeration (microscopic counting of acid fast bacilli) and viability assays (radiorespirometry, viability staining) which utilized bacilli freshly harvested from the contralateral FP. Both molecular and conventional assays demonstrated growth and high viability of M. leprae in nu/nu FPs over a 4 month infection period. In contrast, viability was markedly decreased by 8 weeks in immunocompetent mice. Rifapentine significantly reduced bacterial viability after 5 treatments, whereas rifampin required up to 20 treatments for the same efficacy. Neither drug was effective after a single treatment. In addition, host gene expression was monitored with the same RNA preparations. Conclusions hsp18 and esxA qRT-PCR are sensitive molecular indicators, reliably detecting viability of M. leprae in tissues without the need for bacterial isolation or immediate processing, making these assays applicable for in vivo drug screening and promising for clinical and field applications. PMID:24179562

Molecular assays for determining Mycobacterium leprae viability in tissues of experimentally infected mice.

PubMed

Davis, Grace L; Ray, Nashone A; Lahiri, Ramanuj; Gillis, Thomas P; Krahenbuhl, James L; Williams, Diana L; Adams, Linda B

2013-01-01

The inability of Mycobacterium leprae to grow on axenic media has necessitated specialized techniques in order to determine viability of this organism. The purpose of this study was to develop a simple and sensitive molecular assay for determining M. leprae viability directly from infected tissues. Two M. leprae-specific quantitative reverse transcription PCR (qRT-PCR) assays based on the expression levels of esxA, encoding the ESAT-6 protein, and hsp18, encoding the heat shock 18 kDa protein, were developed and tested using infected footpad (FP) tissues of both immunocompetent and immunocompromised (athymic nu/nu) mice. In addition, the ability of these assays to detect the effects of anti-leprosy drug treatment on M. leprae viability was determined using rifampin and rifapentine, each at 10 mg/kg for 1, 5, or 20 daily doses, in the athymic nu/nu FP model. Molecular enumeration (RLEP PCR) and viability determinations (qRT-PCR) were performed via Taqman methodology on DNA and RNA, respectively, purified from ethanol-fixed FP tissue and compared with conventional enumeration (microscopic counting of acid fast bacilli) and viability assays (radiorespirometry, viability staining) which utilized bacilli freshly harvested from the contralateral FP. Both molecular and conventional assays demonstrated growth and high viability of M. leprae in nu/nu FPs over a 4 month infection period. In contrast, viability was markedly decreased by 8 weeks in immunocompetent mice. Rifapentine significantly reduced bacterial viability after 5 treatments, whereas rifampin required up to 20 treatments for the same efficacy. Neither drug was effective after a single treatment. In addition, host gene expression was monitored with the same RNA preparations. hsp18 and esxA qRT-PCR are sensitive molecular indicators, reliably detecting viability of M. leprae in tissues without the need for bacterial isolation or immediate processing, making these assays applicable for in vivo drug screening and promising for clinical and field applications.

Fluorescence Microscopy Methods for Determining the Viability of Bacteria in Association with Mammalian Cells

PubMed Central

Johnson, M. Brittany; Criss, Alison K.

2013-01-01

Central to the field of bacterial pathogenesis is the ability to define if and how microbes survive after exposure to eukaryotic cells. Current protocols to address these questions include colony count assays, gentamicin protection assays, and electron microscopy. Colony count and gentamicin protection assays only assess the viability of the entire bacterial population and are unable to determine individual bacterial viability. Electron microscopy can be used to determine the viability of individual bacteria and provide information regarding their localization in host cells. However, bacteria often display a range of electron densities, making assessment of viability difficult. This article outlines protocols for the use of fluorescent dyes that reveal the viability of individual bacteria inside and associated with host cells. These assays were developed originally to assess survival of Neisseria gonorrhoeae in primary human neutrophils, but should be applicable to any bacterium-host cell interaction. These protocols combine membrane-permeable fluorescent dyes (SYTO9 and 4',6-diamidino-2-phenylindole [DAPI]), which stain all bacteria, with membrane-impermeable fluorescent dyes (propidium iodide and SYTOX Green), which are only accessible to nonviable bacteria. Prior to eukaryotic cell permeabilization, an antibody or fluorescent reagent is added to identify extracellular bacteria. Thus these assays discriminate the viability of bacteria adherent to and inside eukaryotic cells. A protocol is also provided for using the viability dyes in combination with fluorescent antibodies to eukaryotic cell markers, in order to determine the subcellular localization of individual bacteria. The bacterial viability dyes discussed in this article are a sensitive complement and/or alternative to traditional microbiology techniques to evaluate the viability of individual bacteria and provide information regarding where bacteria survive in host cells. PMID:24056524

Comparisons of human amniotic mesenchymal stem cell viability in FDA-approved collagen-based scaffolds: Implications for engineered diaphragmatic replacement.

PubMed

Shieh, Hester F; Graham, Christopher D; Brazzo, Joseph A; Zurakowski, David; Fauza, Dario O

2017-06-01

We sought to examine amniotic fluid mesenchymal stem cell (afMSC) viability within two FDA-approved collagen-based scaffolds, as a prerequisite to clinical translation of afMSC-based engineered diaphragmatic repair. Human afMSCs were seeded in a human-derived collagen hydrogel and in a bovine-derived collagen sheet at 3 matching densities. Cell viability was analyzed at 1, 3, and 5days using an ATP-based 3D bioluminescence assay. Statistical comparisons were by ANOVA (P<0.05). There was a highly significant 3-way interaction between scaffold type, seeding density, and time in 3D culture as determinants of cell viability, clearly favoring the human hydrogel (P<0.001). In both scaffolds, cell viability was highest at the highest seeding density of 150,000 cells/mL. Time in 3D culture impacted cell viability at the optimal seeding density in the human hydrogel, with the highest levels on days 1 (P<0.001) and 5 (P=0.05) with no significant effect in the bovine sheet (P=0.39-0.96). Among clinically-approved cell delivery vehicles, mesenchymal stem cell viability is significantly enhanced in a collagen hydrogel when compared with a collagen sheet. Cell viability can be further optimized by seeding density and time in 3D culture. These data further support the regulatory viability of clinical trials of engineered diaphragmatic repair. N/A (animal and laboratory study). Copyright © 2017 Elsevier Inc. All rights reserved.

Noninvasive Real-Time Assessment of Cell Viability in a Three-Dimensional Tissue.

PubMed

Mahfouzi, Seyed Hossein; Amoabediny, Ghassem; Doryab, Ali; Safiabadi-Tali, Seyed Hamid; Ghanei, Mostafa

2018-04-01

Maintaining cell viability within 3D tissue engineering scaffolds is an essential step toward a functional tissue or organ. Assessment of cell viability in 3D scaffolds is necessary to control and optimize tissue culture process. Monitoring systems based on respiration activity of cells (e.g., oxygen consumption) have been used in various cell cultures. In this research, an online monitoring system based on respiration activity was developed to monitor cell viability within acellular lung scaffolds. First, acellular lung scaffolds were recellularized with human umbilical cord vein endothelial cells, and then, cell viability was monitored during a 5-day period. The real-time monitoring system generated a cell growth profile representing invaluable information on cell viability and proliferative states during the culture period. The cell growth profile obtained by the monitoring system was consistent with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide analysis and glucose consumption measurement. This system provided a means for noninvasive, real-time, and repetitive investigation of cell viability. Also, we showed the applicability of this monitoring system by introducing shaking as an operating parameter in a long-term culture.

Assessing Subsurface Bioaugmentation of a Mixed Culture Capable of Chlorinated Solvent Cometabolism via Molecular Methods

NASA Astrophysics Data System (ADS)

Dolan, M. E.; Lim, H. K.; Semprini, L.; Giovanonni, S.; Vergin, K.; McCarty, P. L.; Hopkins, G. D.

2001-12-01

The goal of this project is the successful bioaugmentation of a mixed culture capable of aerobic cometabolism of chlorinated solvent mixtures into an aquifer test zone at Moffett Federal Airfield, CA (Moffett). The test zone consists of two parallel well legs both fed butane and oxygen. One leg will be bioaugmented and the other will serve as an indigenous control. Injection and extraction wells and six (3 per leg) intermediately placed groundwater monitoring points will be frequently monitored for chlorinated solvents, butane, dissolved oxygen, and pH. Groundwater will also be periodically analyzed for microbial content using terminal restriction fragment length polymorphism (T-RFLP) and fluorescence in-situ hybridization (FISH) analyses. In each well leg, two fully-penetrating wells containing solid media will be periodically analyzed for microbial colonization (T-RFLP). The mixed bioaugmentation culture originated from environmental samples taken from Hanford, WA. The culture was enriched on butane and tested for viability in Moffett groundwater and aquifer solids. A clone library was created from the 16S rDNA in the mixed culture and 86 clones were sorted based on RFLP patterns. Complete sequencing of the 16S rDNA gene from the three most prevalent clones revealed 45 clones similar to Acidovorax or Hydrogenophaga, gram negative proteobacterium, and 12 clones similar to Rhodococcus, a gram positive filamentous organism. Fluorescently-labeled rRNA probes were designed for FISH analyses and appropriate restriction enzymes were chosen for T-RFLP analyses based upon the sequence information. Microcosm tests were conducted prior to the initiation of the field study to evaluate butane, 1,1-dichloroethylene (1,1-DCE), and 1,1,1-trichloroethane (TCA) degradation kinetics and microbial community composition. Bioaugmented microcosms began butane utilization sooner than non-bioaugmented ones in the presence and absence of 1,1-DCE, and were able to degrade more 1,1-DCE (up to 500 Yg/L) faster than non-bioaugmented microcosms. T-RFLP analyses of triplicate bottles produced very consistent results. An organism(s) with a T-RFLP signature of 183 bp was found to dominate in bioaugmented microcosms and was consistently absent from non-bioaugmented microcosms. T-RFLP and FISH analyses of groundwater and solid media during the bioaugmentation field demonstration are expected to reveal the extent of transport and subsurface colonization of the bioaugmentation culture.

Far from superficial: microbial diversity associated with the skin and mucus of fish

USGS Publications Warehouse

Cipriano, Rocco C.; Dove, Alistair; Cipriano, R.C.; Bruckner, A.W.; Shchelkunov, I.S.

2011-01-01

During horizontal or water-borne infection involving an obligate pathogen (e.g. – Aeromonas salmonicida, cause of furunculosis), the pathogen interacted with and influenced the microbial diversity of the dermal mucus of fish. Prior to infection, the prevalent bacterial flora cultured from juvenile Atlantic salmon (Salmo salar) included Pseudomonas fluorescens, Comomonas terrigenia, Acinetobacter sp., Moraxella sp., Pseudomonas dimunita, Alcaligenes denitrificans, Pseudomonas pseudoalcaligenes, and Pseudomonas alcaligenes, Serratia liquefaciens, Aeromonas hydrophila, other motile Aeromonas spp., and Corynebacterium aquaticum. After A. salmonicida was initially detected in this population as an external mucus infection, Acinetobacter sp., Moraxella sp., C. terrigenia, P. fluorescens, and P. dimunita, Staphylococcus sp., and A. hydrophila, were also present in appreciable numbers. Within several weeks, however, the A. salmonicida infection amplified and composed 78% of the total flora in the mucus. Only P. dimunita (4%). P. fluorescens (2%), and C. terrigenia (1%) were cultured at that time and more than a third of these fish showed evidence of a systemic A. salmonicida infection within their kidneys. Eight weeks after oral oxytetracycline treatments, A. salmonicida was no longer isolated from the mucus or kidneys of any fish and glucose inert or other oxidative microbes (e.g., P. fluorescens, C. terrigenia, Acinetobacter sp., Moraxella sp.) were beginning to repopulate the external surface of the salmon in increasing frequency. Still present and composing fairly large percentages of the total flora were A. hydrophila, as well as Enterobacter sp., and P. putrefaciens. A normal microbial diversity was re-established as the fish recovered. In another investigation, reduced biological diversity was noted in the dermal mucus among smallmouth bass that were sampled from the Jackson River (Covington, VA). In these fish, A. hydrophila and P. putrefaciens were the two predominant microorganisms composing 49.5% and 31.2% of the total bacterial flora, despite the absence of systemic infection or any other clinical signs of disease. In another instance, P. fluorescens was the sole bacterium associated with the surface of Atlantic salmon eggs regardless of their viability at the eyed stage of development. Collectively, these results indicate that the kinetics and distributions of the surface bacterial flora on aquatic organisms is affected by numerous factors including pathogen invasion, environmental conditions, and fish culture practices.

Exogenous lactobacilli mitigate microbial changes associated with grain fermentation (corn, oats, and wheat) by equine fecal microflora ex vivo.

PubMed

Harlow, Brittany E; Lawrence, Laurie M; Harris, Patricia A; Aiken, Glen E; Flythe, Michael D

2017-01-01

Cereal grains are often included in equine diets. When starch intake exceeds foregut digestion starch will reach the hindgut, impacting microbial ecology. Probiotics (e.g., lactobacilli) are reported to mitigate GI dysbioses in other species. This study was conducted to determine the effect of exogenous lactobacilli on pH and the growth of amylolytic and lactate-utilizing bacteria. Feces were collected from 3 mature geldings fed grass hay with access to pasture. Fecal microbes were harvested by differential centrifugation, washed, and re-suspended in anaerobic media containing ground corn, wheat, or oats at 1.6% (w/v) starch and one of five treatments: Control (substrate only), L. acidophilus, L. buchneri, L. reuteri, or an equal mixture of all three (107 cells/mL, final concentration). After 24 h of incubation (37°C, 160 rpm), samples were collected for pH and enumerations of total amylolytics, Group D Gram-positive cocci (GPC; Enterococci, Streptococci), lactobacilli, and lactate-utilizing bacteria. Enumeration data were log transformed prior to ANOVA (SAS, v. 9.3). Lactobacilli inhibited pH decline in corn and wheat fermentations (P < 0.0001). Specifically, addition of either L. reuteri or L. acidophilus was most effective at mitigating pH decline with both corn and wheat fermentation, in which the greatest acidification occurred (P < 0.05). Exogenous lactobacilli decreased amylolytics, while increasing lactate-utilizers in corn and wheat fermentations (P < 0.0001). In oat fermentations, L. acidophilus and L. reuteri inhibited pH decline and increased lactate-utilizers while decreasing amylolytics (P < 0.0001). For all substrates, L. reuteri additions (regardless of viability) had the lowest number of GPC and the highest number of lactobacilli and lactate-utilizers (P < 0.05). There were no additive effects when lactobacilli were mixed. Exogenous lactobacilli decreased the initial (first 8 h) rate of starch catalysis when wheat was the substrate, but did not decrease total (24 h) starch utilization in any case. These results indicate that exogenous lactobacilli can impact the microbial community and pH of cereal grain fermentations by equine fecal microflora ex vivo. Additionally, dead (autoclaved) exogenous lactobacilli had similar effects as live lactobacilli on fermentation. This latter result indicates that the mechanism by which lactobacilli impact other amylolytic bacteria is not simple resource competition.

Exogenous lactobacilli mitigate microbial changes associated with grain fermentation (corn, oats, and wheat) by equine fecal microflora ex vivo

PubMed Central

Harlow, Brittany E.; Lawrence, Laurie M.; Harris, Patricia A.; Aiken, Glen E.

2017-01-01

Cereal grains are often included in equine diets. When starch intake exceeds foregut digestion starch will reach the hindgut, impacting microbial ecology. Probiotics (e.g., lactobacilli) are reported to mitigate GI dysbioses in other species. This study was conducted to determine the effect of exogenous lactobacilli on pH and the growth of amylolytic and lactate-utilizing bacteria. Feces were collected from 3 mature geldings fed grass hay with access to pasture. Fecal microbes were harvested by differential centrifugation, washed, and re-suspended in anaerobic media containing ground corn, wheat, or oats at 1.6% (w/v) starch and one of five treatments: Control (substrate only), L. acidophilus, L. buchneri, L. reuteri, or an equal mixture of all three (107 cells/mL, final concentration). After 24 h of incubation (37°C, 160 rpm), samples were collected for pH and enumerations of total amylolytics, Group D Gram-positive cocci (GPC; Enterococci, Streptococci), lactobacilli, and lactate-utilizing bacteria. Enumeration data were log transformed prior to ANOVA (SAS, v. 9.3). Lactobacilli inhibited pH decline in corn and wheat fermentations (P < 0.0001). Specifically, addition of either L. reuteri or L. acidophilus was most effective at mitigating pH decline with both corn and wheat fermentation, in which the greatest acidification occurred (P < 0.05). Exogenous lactobacilli decreased amylolytics, while increasing lactate-utilizers in corn and wheat fermentations (P < 0.0001). In oat fermentations, L. acidophilus and L. reuteri inhibited pH decline and increased lactate-utilizers while decreasing amylolytics (P < 0.0001). For all substrates, L. reuteri additions (regardless of viability) had the lowest number of GPC and the highest number of lactobacilli and lactate-utilizers (P < 0.05). There were no additive effects when lactobacilli were mixed. Exogenous lactobacilli decreased the initial (first 8 h) rate of starch catalysis when wheat was the substrate, but did not decrease total (24 h) starch utilization in any case. These results indicate that exogenous lactobacilli can impact the microbial community and pH of cereal grain fermentations by equine fecal microflora ex vivo. Additionally, dead (autoclaved) exogenous lactobacilli had similar effects as live lactobacilli on fermentation. This latter result indicates that the mechanism by which lactobacilli impact other amylolytic bacteria is not simple resource competition. PMID:28358885

Evaluation of cytotoxicity and antimicrobial activity of Acticoat Burn Dressing for management of microbial contamination in cultured skin substitutes grafted to athymic mice.

PubMed

Supp, Andrew P; Neely, Alice N; Supp, Dorothy M; Warden, Glenn D; Boyce, Steven T

2005-01-01

Cultured skin substitutes (CSS) have become a useful adjunctive treatment for closure of burn wounds, but CSS are avascular and remain susceptible to microbial destruction longer than split-thickness skin grafts. Irrigation of CSS grafted to burn wounds with a topical antimicrobial solution (TAS) has been shown to promote engraftment of CSS, but TAS usage has potential limitations. Acticoat Burn Dressing (Acticoat; Westaim Biomedical, Exeter, NH) is a silver-coated barrier dressing reported to exhibit antimicrobial activity and to reduce infection in partial-thickness and full-thickness wounds. This study evaluated the cytotoxicity of Acticoat with CSS and the efficacy of Acticoat for the management of microbial contamination in CSS grafted to full-thickness wounds in athymic mice. The cytotoxicity of Acticoat was assessed in preliminary studies after 1 week of exposure to CSS during in vitro maturation or healing on wounds in athymic mice. Histologies were analyzed and cellular viability in the CSS was determined by MTT conversion on days 0, 1, and 7 of Acticoat exposure. At 1, 2, 3, and 4 weeks after grafting, wounds were traced, and areas of healing CSS were calculated by image analysis. At 4 weeks, wound biopsies were evaluated and scored for engraftment of human cells. In a subsequent study, wounds were inoculated with strain SBI-N of Pseudomonas aeruginosa at 1 x 10(5) cfu/wound before the application of CSS or inoculated onto the surface of Acticoat. At 4 weeks, swab cultures were collected from the surface of CSS and scored for the presence of SBI-N. Statistical significance was accepted at the 95% confidence level (P <.05). The data show that exposure in vitro of CSS to Acticoat was cytotoxic within 1 day, but 1 week of exposure in vivo did not injure CSS or inhibit wound healing. Contaminated wounds treated with Acticoat healed similarly to control treatments, with comparable rates of engraftment, and detection of SBI-N on the surface of only one graft. No SBI-N was detected on CSS after inoculation onto the surface of Acticoat. These results suggest that Acticoat may be suitable as a protective dressing to reduce environmental contamination of CSS, if used in conjunction with additional antimicrobials to control organisms present in the wound.

Differential effects of herbicides atrazine and fenoxaprop-ethyl, and insecticides diazinon and malathion, on viability and maturation of porcine oocytes in vitro.

PubMed

Casas, Eduardo; Bonilla, Edmundo; Ducolomb, Yvonne; Betancourt, Miguel

2010-02-01

Exposure to pesticides may be a major cause of reproductive dysfunction in humans and animals. Atrazine and fenoxaprop-ethyl, widely used herbicides, and malathion and diazinon, organophosphate insecticides, are considered only slightly toxic to vertebrates; however, there is evidence of greater effects on reproductive function. The aim of this study was to evaluate the effect of these pesticides on oocyte viability and in vitro maturation. Gametes were matured in increasing concentrations of the pesticides and then stained with MTT to evaluate viability and bisbenzimide to assess the maturation stage, in the same oocyte. Atrazine had no effect on viability but maturation was significantly reduced, while fenoxaprop-ethyl affected both parameters. The insecticides affected viability and maturation but to a different degree. The four pesticides showed a more pronounced effect on maturation than on viability, due to a blockage at germinal vesicle stage.

Glucose-6-Phosphate Dehydrogenase (G6PD)-Deficient Epithelial Cells Are Less Tolerant to Infection by Staphylococcus aureus

PubMed Central

Ho, Hung-Yao; Chen, Lei-Chin; Chen, Chien-Cheng; Shu, Jwu-Ching

2013-01-01

Glucose-6-phosphate dehydrogenase (G6PD) is a key enzyme in the pentose phosphate pathway and provides reducing energy to all cells by maintaining redox balance. The most common clinical manifestations in patients with G6PD deficiency are neonatal jaundice and acute hemolytic anemia. The effects of microbial infection in patients with G6PD deficiency primarily relate to the hemolytic anemia caused by Plasmodium or viral infections and the subsequent medication that is required. We are interested in studying the impact of bacterial infection in G6PD-deficient cells. G6PD knock down A549 lung carcinoma cells, together with the common pathogen Staphylococcus aureus, were employed in our cell infection model. Here, we demonstrate that a lower cell viability was observed among G6PD-deficient cells when compared to scramble controls upon bacterial infection using the MTT assay. A significant increase in the intracellular ROS was detected among S. aureus-infected G6PD-deficient cells by observing dichlorofluorescein (DCF) intensity within cells under a fluorescence microscope and quantifying this signal using flow cytometry. The impairment of ROS removal is predicted to enhance apoptotic activity in G6PD-deficient cells, and this enhanced apoptosis was observed by annexin V/PI staining under a confocal fluorescence microscope and quantified by flow cytometry. A higher expression level of the intrinsic apoptotic initiator caspase-9, as well as the downstream effector caspase-3, was detected by Western blotting analysis of G6PD-deficient cells following bacterial infection. In conclusion, we propose that bacterial infection, perhaps the secreted S. aureus α-hemolysin in this case, promotes the accumulation of intracellular ROS in G6PD-deficient cells. This would trigger a stronger apoptotic activity through the intrinsic pathway thereby reducing cell viability when compared to wild type cells. PMID:24223971

Fungal-Induced Cell Cycle Impairment, Chromosome Instability and Apoptosis via Differential Activation of NF-κB

PubMed Central

Ben-Abdallah, Mariem; Sturny-Leclère, Aude; Avé, Patrick; Louise, Anne; Moyrand, Frédérique; Weih, Falk; Janbon, Guilhem; Mémet, Sylvie

2012-01-01

Microbial pathogens have developed efficient strategies to compromise host immune responses. Cryptococcus neoformans is a facultative intracellular pathogen, recognised as the most common cause of systemic fungal infections leading to severe meningoencephalitis, mainly in immunocompromised patients. This yeast is characterized by a polysaccharide capsule, which inhibits its phagocytosis. Whereas phagocytosis escape and macrophage intracellular survival have been intensively studied, extracellular survival of this yeast and restraint of host innate immune response are still poorly understood. In this study, we have investigated whether C. neoformans affected macrophage cell viability and whether NF-κB (nuclear factor-κB), a key regulator of cell growth, apoptosis and inflammation, was involved. Using wild-type (WT) as well as mutant strains of C. neoformans for the pathogen side, and WT and mutant cell lines with altered NF-κB activity or signalling as well as primary macrophages for the host side, we show that C. neoformans manipulated NF-κB-mediated signalling in a unique way to regulate macrophage cell fate and viability. On the one hand, serotype A strains reduced macrophage proliferation in a capsule-independent fashion. This growth decrease, which required a critical dosage of NF-κB activity, was caused by cell cycle disruption and aneuploidy, relying on fungal-induced modification of expression of several cell cycle checkpoint regulators in S and G2/M phases. On the other hand, C. neoformans infection induced macrophage apoptosis in a capsule-dependent manner with a differential requirement of the classical and alternative NF-κB signalling pathways, the latter one being essential. Together, these findings shed new light on fungal strategies to subvert host response through uncoupling of NF-κB activity in pathogen-controlled apoptosis and impairment of cell cycle progression. They also provide the first demonstration of induction of aneuploidy by a fungal pathogen, which may have wider implications for human health as aneuploidy is proposed to promote tumourigenesis. PMID:22396644

Fungal-induced cell cycle impairment, chromosome instability and apoptosis via differential activation of NF-κB.

PubMed

Ben-Abdallah, Mariem; Sturny-Leclère, Aude; Avé, Patrick; Louise, Anne; Moyrand, Frédérique; Weih, Falk; Janbon, Guilhem; Mémet, Sylvie

2012-01-01

Microbial pathogens have developed efficient strategies to compromise host immune responses. Cryptococcus neoformans is a facultative intracellular pathogen, recognised as the most common cause of systemic fungal infections leading to severe meningoencephalitis, mainly in immunocompromised patients. This yeast is characterized by a polysaccharide capsule, which inhibits its phagocytosis. Whereas phagocytosis escape and macrophage intracellular survival have been intensively studied, extracellular survival of this yeast and restraint of host innate immune response are still poorly understood. In this study, we have investigated whether C. neoformans affected macrophage cell viability and whether NF-κB (nuclear factor-κB), a key regulator of cell growth, apoptosis and inflammation, was involved. Using wild-type (WT) as well as mutant strains of C. neoformans for the pathogen side, and WT and mutant cell lines with altered NF-κB activity or signalling as well as primary macrophages for the host side, we show that C. neoformans manipulated NF-κB-mediated signalling in a unique way to regulate macrophage cell fate and viability. On the one hand, serotype A strains reduced macrophage proliferation in a capsule-independent fashion. This growth decrease, which required a critical dosage of NF-κB activity, was caused by cell cycle disruption and aneuploidy, relying on fungal-induced modification of expression of several cell cycle checkpoint regulators in S and G2/M phases. On the other hand, C. neoformans infection induced macrophage apoptosis in a capsule-dependent manner with a differential requirement of the classical and alternative NF-κB signalling pathways, the latter one being essential. Together, these findings shed new light on fungal strategies to subvert host response through uncoupling of NF-κB activity in pathogen-controlled apoptosis and impairment of cell cycle progression. They also provide the first demonstration of induction of aneuploidy by a fungal pathogen, which may have wider implications for human health as aneuploidy is proposed to promote tumourigenesis.

Proteomic Analysis of Silk Viability in Maize Inbred Lines and Their Corresponding Hybrids

PubMed Central

Wang, Yafei; Zhao, Xiaofeng; Zhang, Fangfang; Tang, Jihua; Fu, Zhiyuan

2015-01-01

A long period of silk viability is critical for a good seed setting rate in maize (Zea mays L.), especially for inbred lines and hybrids with a long interval between anthesis and silking. To explore the molecular mechanism of silk viability and its heterosis, three inbred lines with different silk viability characteristics (Xun928, Lx9801, and Zong3) and their two hybrids (Xun928×Zong3 and Lx9801×Zong3) were analyzed at different developmental stages by a proteomic method. The differentially accumulated proteins were identified by mass spectrometry and classified into metabolism, protein biosynthesis and folding, signal transduction and hormone homeostasis, stress and defense responses, and cellular processes. Proteins involved in nutrient (methionine) and energy (ATP) supply, which support the pollen tube growth in the silk, were important for silk viability and its heterosis. The additive and dominant effects at a single locus, as well as complex epistatic interactions at two or more loci in metabolic pathways, were the primary contributors for mid-parent heterosis of silk viability. Additionally, the proteins involved in the metabolism of anthocyanins, which indirectly negatively regulate local hormone accumulation, were also important for the mid-parent heterosis of silk viability. These results also might imply the developmental dependence of heterosis, because many of the differentially accumulated proteins made distinct contributions to the heterosis of silk viability at specific developmental stages. PMID:26630375

Proteomic Analysis of Silk Viability in Maize Inbred Lines and Their Corresponding Hybrids.

PubMed

Ma, Zhihui; Qin, Yongtian; Wang, Yafei; Zhao, Xiaofeng; Zhang, Fangfang; Tang, Jihua; Fu, Zhiyuan

2015-01-01

A long period of silk viability is critical for a good seed setting rate in maize (Zea mays L.), especially for inbred lines and hybrids with a long interval between anthesis and silking. To explore the molecular mechanism of silk viability and its heterosis, three inbred lines with different silk viability characteristics (Xun928, Lx9801, and Zong3) and their two hybrids (Xun928×Zong3 and Lx9801×Zong3) were analyzed at different developmental stages by a proteomic method. The differentially accumulated proteins were identified by mass spectrometry and classified into metabolism, protein biosynthesis and folding, signal transduction and hormone homeostasis, stress and defense responses, and cellular processes. Proteins involved in nutrient (methionine) and energy (ATP) supply, which support the pollen tube growth in the silk, were important for silk viability and its heterosis. The additive and dominant effects at a single locus, as well as complex epistatic interactions at two or more loci in metabolic pathways, were the primary contributors for mid-parent heterosis of silk viability. Additionally, the proteins involved in the metabolism of anthocyanins, which indirectly negatively regulate local hormone accumulation, were also important for the mid-parent heterosis of silk viability. These results also might imply the developmental dependence of heterosis, because many of the differentially accumulated proteins made distinct contributions to the heterosis of silk viability at specific developmental stages.

[Viability and germination of Hechtia perotensis (Bromeliaceae) seed].

PubMed

Elizalde, Violeta; García, José Rodolfo; Peña-Valdivia, Cecilia Beatriz; Ybarra, Ma Carmen; Leyva, Otto Raúl; Trejo, Carlos

2017-03-01

Endemic populations of Hechtia perotensis have been described in Puebla and Veracruz, Mexico. Good quality seed collections can be used in conservation, research and ecological restoration. To evaluate seed quality of wild and endemic species, some compounds are used as effective promoters of germination, such as potassium nitrate (KNO3) and gibberellic acid (AG3), because they increase seed germination capacity and reduce latency. The triphenyl tetrazolium chloride (tetrazolium) test correlates seed viability because it is based on the activity of dehydrogenases in live tissues that catalyze mitochondrial respiration. The objective of this study was to obtain information on size and weight of capsules and seeds and seed germination and viability of H. perotensis, collected in Veracruz in the year 2012 and 2015. The hypotheses were 1) that seed germination and viability are independent of the year of collection, 2) that there is a tetrazolium concentration that can identify seed viability better than others, and 3) that pretreatment with KNO3 or AG3 improves seed germination. Seed germination was assessed using a completely randomized design with three treatments (control and the germination promoters 0.2 % KNO3 and 500 mg/L AG3), four treatments for the viability test (control, 0.2, 0.5 and 1.0 % of tetrazolium) and six replicates for each treatment. A total of one hundred seeds for germination experiments, and 25 seeds for the viability test were used. The results between and within years were analyzed with ANOVA and multiple comparison with the Tukey test. The proportion of non-germinated seeds was quantified along with the number of normal and abnormal seedlings, seeds with viable embryo, seeds without embryo, and seeds with low or no viability. On average, for the 2012 collected sample, 36 % had viable embryos, 7 % had low viability, 24 % were not viable and 33 % had no embryo. This result was significantly different from the 2015 sample, for which 87 % of seed showed viable embryos, 10 % had low viability, 0 % was not viable and 3 % had no embryo. Seed germination was also significantly different between years (22 and 92 %) Pregerminative treatments did not improve germination. Seed germination and viability of H. perotensis significantly varied between years of seed collection.

Viability and Virulence of Entomopathogenic Nematodes Exposed to Ultraviolet Radiation.

PubMed

Shapiro-Ilan, David I; Hazir, Selcuk; Lete, Luis

2015-09-01

Entomopathogenic nematodes (EPNs) can be highly effective biocontrol agents, but their efficacy can be reduced due to exposure to environmental stress such as from ultraviolet (UV) radiation. Our objectives were to 1) compare UV tolerance among a broad array of EPN species, and 2) investigate the relationship between reduced nematode viability (after exposure to UV) and virulence. Nematodes exposed to a UV radiation (254 nm) for 10 or 20 min were assessed separately for viability (survival) and virulence to Galleria mellonella. We compared 9 different EPN species and 15 strains: Heterorhabditis bacteriophora (Baine, fl11, Oswego, and Vs strains), H. floridensis (332), H. georgiana (Kesha), H. indica (HOM1), H. megidis (UK211), Steinernema carpocapsae (All, Cxrd, DD136, and Sal strains), S. feltiae (SN), S. rarum (17C&E), and S. riobrave (355). In viability assessments, steinernematids, particularly strains of S. carpocapsae, generally exhibited superior UV tolerance compared with the heterorhabditids. However, some heterorhabditids tended to be more tolerant than others, e.g., H. megidis and H. bacteriophora (Baine) were most susceptible and H. bacteriophora (Vs) was the only heterorhabditid that did not exhibit a significant effect after 10 min of exposure. All heterorhabditids experienced reduced viability after 20 min exposure though several S. carpocapsae strains did not. In total, after 10 or 20 min exposure, the viability of seven nematode strains did not differ from their non-UV exposed controls. In virulence assays, steinernematids (particularly S. carpocapsae strains) also tended to exhibit higher UV tolerance. However, in contrast to the viability measurements, all nematodes experienced a reduction in virulence relative to their controls. Correlation analysis revealed that viability among nematode strains is not necessarily related to virulence. In conclusion, our results indicate that the impact of UV varies substantially among EPNs, and viability alone is not a sufficient measure for potential impact on biocontrol efficacy as other characters such as virulence may be severely affected even when viability remains high.

The CCR4-NOT Complex Is Implicated in the Viability of Aneuploid Yeasts

PubMed Central

Tange, Yoshie; Kurabayashi, Atsushi; Goto, Bunshiro; Hoe, Kwang-Lae; Kim, Dong-Uk; Park, Han-Oh; Hayles, Jacqueline; Chikashige, Yuji; Tsutumi, Chihiro; Hiraoka, Yasushi; Yamao, Fumiaki; Nurse, Paul; Niwa, Osami

2012-01-01

To identify the genes required to sustain aneuploid viability, we screened a deletion library of non-essential genes in the fission yeast Schizosaccharomyces pombe, in which most types of aneuploidy are eventually lethal to the cell. Aneuploids remain viable for a period of time and can form colonies by reducing the extent of the aneuploidy. We hypothesized that a reduction in colony formation efficiency could be used to screen for gene deletions that compromise aneuploid viability. Deletion mutants were used to measure the effects on the viability of spores derived from triploid meiosis and from a chromosome instability mutant. We found that the CCR4-NOT complex, an evolutionarily conserved general regulator of mRNA turnover, and other related factors, including poly(A)-specific nuclease for mRNA decay, are involved in aneuploid viability. Defective mutations in CCR4-NOT complex components in the distantly related yeast Saccharomyces cerevisiae also affected the viability of spores produced from triploid cells, suggesting that this complex has a conserved role in aneuploids. In addition, our findings suggest that the genes required for homologous recombination repair are important for aneuploid viability. PMID:22737087

Chondrotoxicity of Liposomal Bupivacaine in Articular Chondrocytes: Preliminary Findings.

PubMed

Shaw, K Aaron; Johnson, Peter C; Zumbrun, Steve; Chuang, Augustine H; Cameron, Craig D

2017-03-01

The chondrotoxicity of local anesthetics has been previously recognized. Recent introduction of a liposomal formulation of bupivacaine has been found to significantly improve postoperative pain control but its effect on chondrocyte viability has yet to be investigated with this new formulation. We sought to assess the in vitro chondrotoxicity of liposomal bupivacaine. Chondrocytes were isolated from articular cartilage from fresh stifle joints and grown in culture medium. Cultured chondrocyte-derived cells (CDCs) were treated with 0.9% normal saline solution, 0.5%, 0.25%, and 0.13% bupivacaine and ropivacaine, 1.3% liposomal bupivacaine for 1 hour. Following treatment, cells were washed and incubated in media for 23 hours. The CDCs were then harvested and viability was assessed by flow cytometry using SYTOX green dead cell stain. Treated CDCs demonstrated a dose-response effect for chondrocyte viability when treated with bupivacaine, ropivacaine, and liposomal bupivacaine. Liposomal bupivacaine demonstrated the highest chondrocyte viability following treatment. Ropivacaine demonstrated higher chondrocyte viability than bupivacaine. Following 1 hour of treatment, liposomal bupivacaine demonstrated the highest chondrocyte viability. Chondrocyte viability was inversely proportional to anesthetic concentration. Reprint & Copyright © 2017 Association of Military Surgeons of the U.S.

36 CFR 219.20 - Ecological sustainability.

Code of Federal Regulations, 2010 CFR

2010-07-01

... PLANNING National Forest System Land and Resource Management Planning Ecological, Social, and Economic..., as appropriate, assessments of the risks to species viability and the identification of ecological conditions needed to maintain species viability over time based on the following: (A) The viability of each...

36 CFR 219.20 - Ecological sustainability.

Code of Federal Regulations, 2011 CFR

2011-07-01

... PLANNING National Forest System Land and Resource Management Planning Ecological, Social, and Economic..., as appropriate, assessments of the risks to species viability and the identification of ecological conditions needed to maintain species viability over time based on the following: (A) The viability of each...

Efficient Intracellular Delivery of Molecules with High Cell Viability Using Nanosecond-Pulsed Laser-Activated Carbon Nanoparticles

PubMed Central

2015-01-01

Conventional physical and chemical methods that efficiently deliver molecules into cells are often associated with low cell viability. In this study, we evaluated the cellular effects of carbon nanoparticles believed to emit photoacoustic waves due to nanosecond-pulse laser activation to test the hypothesis that this method could achieve efficient intracellular delivery while maintaining high cell viability. Suspensions of DU145 human prostate carcinoma cells, carbon black (CB) nanoparticles, and calcein were exposed to 5–9 ns long laser pulses of near-infrared (1064 nm wavelength) light and then analyzed by flow cytometry for intracellular uptake of calcein and cell viability by propidium iodide staining. We found that intracellular uptake increased and in some cases saturated at high levels with only small losses in cell viability as a result of increasing laser fluence, laser exposure time, and as a unifying parameter, the total laser energy. Changing interpulse spacing between 0.1 and 10 s intervals showed no significant change in bioeffects, suggesting that the effects of each pulse were independent when spaced by at least 0.1 s intervals. Pretreatment of CB nanoparticles to intense laser exposure followed by mixing with cells also had no significant effect on uptake or viability. Similar uptake and viability were seen when CB nanoparticles were substituted with India ink, when DU145 cells were substituted with H9c2 rat cardiomyoblast cells, and when calcein was substituted with FITC-dextran. The best laser exposure conditions tested led to 88% of cells with intracellular uptake and close to 100% viability, indicating that nanosecond-pulse laser-activated carbon nanoparticles can achieve efficient intracellular delivery while maintaining high cell viability. PMID:24547946

Heavy metal-immobilizing organoclay facilitates polycyclic aromatic hydrocarbon biodegradation in mixed-contaminated soil.

PubMed

Biswas, Bhabananda; Sarkar, Binoy; Mandal, Asit; Naidu, Ravi

2015-11-15

Soils contaminated with a mixture of heavy metals and polycyclic aromatic hydrocarbons (PAHs) pose toxic metal stress to native PAH-degrading microorganisms. Adsorbents such as clay and modified clay minerals can bind the metal and reduce its toxicity to microorganisms. However, in a mixed-contaminated soil, an adsorption process more specific to the metals without affecting the bioavailability of PAHs is desired for effective degradation. Furthermore, the adsorbent should enhance the viability of PAH-degrading microorganisms. A metal-immobilizing organoclay (Arquad(®) 2HT-75-bentonite treated with palmitic acid) (MIOC) able to reduce metal (cadmium (Cd)) toxicity and enhance PAH (phenanthrene) biodegradation was developed and characterized in this study. The MIOC differed considerably from the parent clay in terms of its ability to reduce metal toxicity (MIOC>unmodified bentonite>Arquad-bentonite). The MIOC variably increased the microbial count (10-43%) as well as activities (respiration 3-44%; enzymatic activities up to 68%), and simultaneously maintained phenanthrene in bioavailable form in a Cd-phenanthrene mixed-contaminated soil over a 21-day incubation period. This study may lead to a new MIOC-assisted bioremediation technique for PAHs in mixed-contaminated soils. Copyright © 2015 Elsevier B.V. All rights reserved.

How to sustainably feed a microbe: Strategies for biological production of carbon-based commodities with renewable electricity

DOE PAGES

Butler, Caitlyn S.; Lovley, Derek R.

2016-11-28

As interest and application of renewable energy grows, strategies are needed to align the asynchronous supply and demand. Microbial metabolisms are a potentially sustainable mechanism for transforming renewable electrical energy into biocommodities that are easily stored and transported. Acetogens and methanogens can reduce carbon dioxide to organic products including methane, acetic acid, and ethanol. The library of biocommodities is expanded when engineered metabolisms of acetogens are included. Typically, electrochemical systems are employed to integrate renewable energy sources with biological systems for production of carbon-based commodities. Within these systems, there are three prevailing mechanisms for delivering electrons to microorganisms for themore » conversion of carbon dioxide to reduce organic compounds: (1) electrons can be delivered to microorganisms via H 2 produced separately in a electrolyzer, (2) H 2 produced at a cathode can convey electrons to microorganisms supported on the cathode surface, and (3) a cathode can directly feed electrons to microorganisms. Each of these strategies has advantages and disadvantages that must be considered in designing full-scale processes. Furthermore, this review considers the evolving understanding of each of these approaches and the state of design for advancing these strategies toward viability.« less

Enhancement of Survival and Electricity Production in an Engineered Bacterium by Light-Driven Proton Pumping▿ †

PubMed Central

Johnson, Ethan T.; Baron, Daniel B.; Naranjo, Belén; Bond, Daniel R.; Schmidt-Dannert, Claudia; Gralnick, Jeffrey A.

2010-01-01

Microorganisms can use complex photosystems or light-dependent proton pumps to generate membrane potential and/or reduce electron carriers to support growth. The discovery that proteorhodopsin is a light-dependent proton pump that can be expressed readily in recombinant bacteria enables development of new strategies to probe microbial physiology and to engineer microbes with new light-driven properties. Here, we describe functional expression of proteorhodopsin and light-induced changes in membrane potential in the bacterium Shewanella oneidensis strain MR-1. We report that there were significant increases in electrical current generation during illumination of electrochemical chambers containing S. oneidensis expressing proteorhodopsin. We present evidence that an engineered strain is able to consume lactate at an increased rate when it is illuminated, which is consistent with the hypothesis that proteorhodopsin activity enhances lactate uptake by increasing the proton motive force. Our results demonstrate that there is coupling of a light-driven process to electricity generation in a nonphotosynthetic engineered bacterium. Expression of proteorhodopsin also preserved the viability of the bacterium under nutrient-limited conditions, providing evidence that fulfillment of basic energy needs of organisms may explain the widespread distribution of proteorhodopsin in marine environments. PMID:20453141

Enhancement of survival and electricity production in an engineered bacterium by light-driven proton pumping.

PubMed

Johnson, Ethan T; Baron, Daniel B; Naranjo, Belén; Bond, Daniel R; Schmidt-Dannert, Claudia; Gralnick, Jeffrey A

2010-07-01

Microorganisms can use complex photosystems or light-dependent proton pumps to generate membrane potential and/or reduce electron carriers to support growth. The discovery that proteorhodopsin is a light-dependent proton pump that can be expressed readily in recombinant bacteria enables development of new strategies to probe microbial physiology and to engineer microbes with new light-driven properties. Here, we describe functional expression of proteorhodopsin and light-induced changes in membrane potential in the bacterium Shewanella oneidensis strain MR-1. We report that there were significant increases in electrical current generation during illumination of electrochemical chambers containing S. oneidensis expressing proteorhodopsin. We present evidence that an engineered strain is able to consume lactate at an increased rate when it is illuminated, which is consistent with the hypothesis that proteorhodopsin activity enhances lactate uptake by increasing the proton motive force. Our results demonstrate that there is coupling of a light-driven process to electricity generation in a nonphotosynthetic engineered bacterium. Expression of proteorhodopsin also preserved the viability of the bacterium under nutrient-limited conditions, providing evidence that fulfillment of basic energy needs of organisms may explain the widespread distribution of proteorhodopsin in marine environments.

Apolar Bioactive Fraction of Melipona scutellaris Geopropolis on Streptococcus mutans Biofilm.

PubMed

da Cunha, Marcos Guilherme; Franchin, Marcelo; Galvão, Lívia Câmara de Carvalho; Bueno-Silva, Bruno; Ikegaki, Masaharu; de Alencar, Severino Matias; Rosalen, Pedro Luiz

2013-01-01

The aim of this study was to evaluate the influence of the bioactive nonpolar fraction of geopropolis on Streptococcus mutans biofilm. The ethanolic extract of Melipona scutellaris geopropolis was subjected to a liquid-liquid partition, thus obtaining the bioactive hexane fraction (HF) possessing antimicrobial activity. The effects of HF on S. mutans UA159 biofilms generated on saliva-coated hydroxyapatite discs were analyzed by inhibition of formation, killing assay, and glycolytic pH-drop assays. Furthermore, biofilms treated with vehicle control and HF were analyzed by scanning electron microscopy (SEM). HF at 250  μ g/mL and 400  μ g/mL caused 38% and 53% reduction in the biomass of biofilm, respectively, when compared to vehicle control (P < 0.05) subsequently observed at SEM images, and this reduction was noticed in the amounts of extracellular alkali-soluble glucans, intracellular iodophilic polysaccharides, and proteins. In addition, the S. mutans viability (killing assay) and acid production by glycolytic pH drop were not affected (P > 0.05). In conclusion, the bioactive HF of geopropolis was promising to control the S. mutans biofilm formation, without affecting the microbial population but interfering with its structure by reducing the biochemical content of biofilm matrix.

Natural marine bacteria as model organisms for the hazard-assessment of consumer products containing silver nanoparticles.

PubMed

Echavarri-Bravo, Virginia; Paterson, Lynn; Aspray, Thomas J; Porter, Joanne S; Winson, Michael K; Hartl, Mark G J

2017-09-01

Scarce information is available regarding the fate and toxicology of engineered silver nanoparticles (AgNPs) in the marine environment, especially when compared to other environmental compartments. Hence, the antibacterial activity of the NM-300 AgNPs (OECD programme) and a household product containing colloidal AgNPs (Mesosilver) was investigated using marine bacteria, pure cultures and natural mixed populations (microcosm approach). Bacterial susceptibility to AgNPs was species-specific, with Gram negative bacteria being more resistant than the Gram positive species (NM-300 concentration used ranged between 0.062 and 1.5 mg L -1 ), and the Mesosilver product was more toxic than the NM-300. Bacterial viability and the physiological status (O 2 uptake measured by respirometry) of the microbial community in the microcosm was negatively affected at an initial concentration of 1 mg L -1 NM-300. The high chloride concentrations in the media/seawater led to the formation of silver-chloro complexes thus enhancing AgNP toxicity. We recommend the use of natural marine bacteria as models when assessing the environmental relevant antibacterial properties of products containing nanosilver. Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.

Spatially confined photoinactivation of bacteria: towards novel tools for detailed mechanistic studies

NASA Astrophysics Data System (ADS)

Thomsen, Hanna; James, Jeemol; Farewell, Anne; Ericson, Marica B.

2018-02-01

Antimicrobial resistance is a serious global threat fueling an accelerated field of research aimed at developing novel antimicrobial therapies. A particular challenge is the treatment of microbial biofilms formed upon bacterial growth and often associated with chronic infections. Biofilms comprise bacteria that have adhered to a surface and formed 3D microcolonies, and demonstrate significantly increased antimicrobial resistance compared to the planktonic counterpart. A challenge in developing novel strategies for fighting these chronic infections is a lack of mechanistic understanding of what primarily contributes to enhanced drug resistance. Tools for noninvasive study of live biofilms are necessary to begin to understand these mechanisms on both a single cell and 3D level. Herein, a method by which multiphoton microscopy is implemented to study a biofilm model of Staphylococcus epidermidis to noninvasively visualize and measure penetration of compounds in 3D biofilm structure and two photon excitation was exploited for spatially confined photoinactivation and microscopy optimized for evaluation of microbiological viability at a microscopic level. Future studies are aimed at future development of the proposed techniques for detailed studies of, e.g., quorum sensing and mechanisms contributing to antimicrobial resistance.

Techno-economic analysis of horseradish peroxidase production using a transient expression system in Nicotiana benthamiana.

PubMed

Walwyn, David Richard; Huddy, Suzanne M; Rybicki, Edward P

2015-01-01

Despite the advantages of plant-based transient expression systems relative to microbial or mammalian cell systems, the commercial production of recombinant proteins using plants has not yet been achieved to any significant extent. One of the challenges has been the lack of published data on the costs of manufacture for products other than biopharmaceuticals. In this study, we report on the techno-economic analysis of the production of a standard commercial enzyme, namely, horseradish peroxidase (HRP), using a transient expression system in Nicotiana benthamiana. Based on the proven plant yield of 240 mg HRP/kg biomass, a biomass productivity of 15-kg biomass/m(2)/year and a process yield of 54 % (mg HRP product/mg HRP in biomass), it is apparent that HRP can be manufactured economically via transient expression in plants in a large-scale facility (>5 kg HRP/year). At this level, the process is competitive versus the existing technology (extraction of the enzyme from horseradish), and the product is of comparable or improved activity, containing only the preferred isoenzyme C. Production scale, protein yield and biomass productivity are found to be the most important determinants of overall viability.

Lipid-Based Immuno-Magnetic Separation of Archaea from a Mixed Community

NASA Astrophysics Data System (ADS)

Frickle, C. M.; Bailey, J.; Lloyd, K. G.; Shumaker, A.; Flood, B.

2014-12-01

Despite advancing techniques in microbiology, an estimated 98% of all microbial species on Earth have yet to be isolated in pure culture. Natural samples, once transferred to the lab, are commonly overgrown by "weed" species whose metabolic advantages enable them to monopolize available resources. Developing new methods for the isolation of thus-far uncultivable microorganisms would allow us to better understand their ecology, physiology and genetic potential. Physically separating target organisms from a mixed community is one approach that may allow enrichment and growth of the desired strain. Here we report on a novel method that uses known physiological variations between taxa, in this case membrane lipids, to segregate the desired organisms while keeping them alive and viable for reproduction. Magnetic antibodies bound to the molecule squalene, which is found in the cell membranes of certain archaea, but not bacteria, enable separation of archaea from bacteria in mixed samples. Viability of cells was tested by growing the separated fractions in batch culture. Efficacy and optimization of the antibody separation technique are being evaluated using qPCR and cell counts. Future work will apply this new separation technique to natural samples.

Experimental evaluation of new chitin-chitosan graft for duraplasty.

PubMed

Pogorielov, M; Kravtsova, A; Reilly, G C; Deineka, V; Tetteh, G; Kalinkevich, O; Pogorielova, O; Moskalenko, R; Tkach, G

2017-02-01

Natural materials such as collagen and alginate have promising applications as dural graft substitutes. These materials are able to restore the dural defect and create optimal conditions for the development of connective tissue at the site of injury. A promising material for biomedical applications is chitosan-a linear polysaccharide obtained by the deacetylation of chitin. It has been found to be nontoxic, biodegradable, biofunctional and biocompatible in addition to having antimicrobial characteristics. In this study we designed new chitin-chitosan substitutes for dura mater closure and evaluated their effectiveness and safety. Chitosan films were produced from 3 % of chitosan (molar mass-200, 500 or 700 kDa, deacetylation rate 80-90%) with addition of 20% of chitin. Antimicrobial effictively and cell viability were analysed for the different molar masses of chitosan. The film containing chitosan of molar mass 200 kDa, had the best antimicrobial and biological activity and was successfully used for experimental duraplasty in an in vivo model. In conclusion the chitin-chitosan membrane designed here met the requirements for a dura matter graft exhibiting the ability to support cell growth, inhibit microbial growth and biodegradade at an appropriate rate. Therefore this is a promising material for clinical duroplasty.

Sublethal detergent concentrations increase metabolization of recalcitrant polyphosphonates by the cyanobacterium Spirulina platensis.

PubMed

Forlani, Giuseppe; Bertazzini, Michele; Giberti, Samuele; Wieczorek, Dorota; Kafarski, Paweł; Lipok, Jacek

2013-05-01

As a consequence of increasing industrial applications, thousand tons of polyphosphonates are introduced every year into the environment. The inherent stability of the C-P bond results in a prolonged half-life. Moreover, low uptake rates limit further their microbial metabolization. To assess whether low detergent concentrations were able to increase polyphosphonate utilization by the cyanobacterium Spirulina platensis, tolerance limits to the exposure to various detergents were determined by measuring the growth rate in the presence of graded levels below the critical micellar concentration. Then, the amount of hexamethylenediamine-N,N,N',N'-tetrakis(methylphosphonic acid) that is metabolized in the absence or in the presence of sublethal detergent concentrations was quantified by (31)P NMR analysis on either P-starved or P-fed cyanobacterial cultures. The strain tolerated the presence of detergents in the order: nonionic > anionic > cationic. When added to the culture medium at the highest concentrations showing no detrimental effects upon cell viability, detergents either improved or decreased polyphosphonate utilization, the anionic sodium dodecyl sulfate being the most beneficial. Metabolization was not lower in P-fed cells--a result that strengthens the possibility of using, in the future, this strain for bioremediation purposes.

Use of immunomagnetic separation for the detection of Desulfovibrio vulgaris from environmental samples

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

Chakraborty, R.; Hazen, T.C.; Joyner, D.C.

2011-04-15

Immunomagnetic separation (IMS) has proved highly efficient for recovering microorganisms from heterogeneous samples. Current investigation targeted the separation of viable cells of the sulfate-reducing bacterium, Desulfovibrio vulgaris. Streptavidin-coupled paramagnetic beads and biotin labeled antibodies raised against surface antigens of this microorganism were used to capture D. vulgaris cells in both bioreactor grown laboratory samples and from extremely low-biomass environmental soil and subsurface drilling samples. Initial studies on detection, recovery efficiency and viability for IMS were performed with laboratory grown D. vulgaris cells using various cell densities. Efficiency of cell isolation and recovery (i.e., release of the microbial cells from themore » beads following separation) was followed by microscopic imaging and acridine orange direct counts (AODC). Excellent recovery efficiency encouraged the use of IMS to capture Desulfovibrio spp. cells from low-biomass environmental samples. The environmental samples were obtained from a radionuclide-contaminated site in Germany and the chromium (VI)-contaminated Hanford site, an ongoing bioremediation project of the U.S. Department of Energy. Field deployable IMS technology may greatly facilitate environmental sampling and bioremediation process monitoring and enable transcriptomics and proteomics/metabolomics-based studies directly on cells collected from the field.« less

Following the mechanisms of bacteriostatic versus bactericidal action using Raman spectroscopy.

PubMed

Bernatová, Silvie; Samek, Ota; Pilát, Zdeněk; Serý, Mojmír; Ježek, Jan; Jákl, Petr; Siler, Martin; Krzyžánek, Vladislav; Zemánek, Pavel; Holá, Veronika; Dvořáčková, Milada; Růžička, Filip

2013-10-24

Antibiotics cure infections by influencing bacterial growth or viability. Antibiotics can be divided to two groups on the basis of their effect on microbial cells through two main mechanisms, which are either bactericidal or bacteriostatic. Bactericidal antibiotics kill the bacteria and bacteriostatic antibiotics suppress the growth of bacteria (keep them in the stationary phase of growth). One of many factors to predict a favorable clinical outcome of the potential action of antimicrobial chemicals may be provided using in vitro bactericidal/bacteriostatic data (e.g., minimum inhibitory concentrations-MICs). Consequently, MICs are used in clinical situations mainly to confirm resistance, and to determine the in vitro activities of new antimicrobials. We report on the combination of data obtained from MICs with information on microorganisms' "fingerprint" (e.g., DNA/RNA, and proteins) provided by Raman spectroscopy. Thus, we could follow mechanisms of the bacteriostatic versus bactericidal action simply by detecting the Raman bands corresponding to DNA. The Raman spectra of Staphylococcus epidermidis treated with clindamycin (a bacteriostatic agent) indeed show little effect on DNA which is in contrast with the action of ciprofloxacin (a bactericidal agent), where the Raman spectra show a decrease in strength of the signal assigned to DNA, suggesting DNA fragmentation.

Can a novel silver nano coating reduce infections and maintain cell viability in vitro?

PubMed

Qureshi, Ammar T; Landry, Jace P; Dasa, Vinod; Janes, Marlene; Hayes, Daniel J

2014-03-01

Herein we report a facile layer-by-layer method for creating an antimicrobial coating composed of silver nanoparticles on medical grade titanium test discs. Nanoscale silver nanoparticle layers are attached to the titanium orthopedic implant material via aminopropyltriethoxy silane crosslinker that reacts with neighboring silane moieties to create an interconnected network. A monolayer of silane, followed by a monolayer of silver nanoparticles would form one self-assembled layer and this process can be repeated serially, resulting in increased silver nanoparticles deposition. The release rate of silver ion increases predictably with increasing numbers of layers and at appropriate thicknesses these coatings demonstrate 3-4 log reduction of viable Escherichia coli and Staphylococcus aureus bacteria. Increasing the thickness of the coatings resulted in reduced bacterial colonization as determined by fluorescent staining and image analysis. Interestingly, the cytotoxicity of murine 3T3 cells as quantified by fluorescent staining and flow cytometry, was minimal and did not vary significantly with the coating thickness. Additionally, these coatings are mechanically stable and resist delamination by orthogonal stress test. This simple layer-by-layer coating technique may provide a cost-effective and biocompatible method for reducing microbial colonization of implantable orthopedic devices.

Metabolic Recruitment and Directed Evolution of Nucleoside Triphosphate Uptake in Escherichia coli.

PubMed

Pezo, Valérie; Hassan, Camille; Louis, Dominique; Sargueil, Bruno; Herdewijn, Piet; Marlière, Philippe

2018-05-18

We report the design and elaboration of a selection protocol for importing a canonical substrate of DNA polymerase, thymidine triphosphate (dTTP) in Escherichia coli. Bacterial strains whose growth depend on dTTP uptake, through the action of an algal plastid transporter expressed from a synthetic gene inserted in the chromosome, were constructed and shown to withstand the simultaneous loss of thymidylate synthase and thymidine kinase. Such thyA tdk dual deletant strains provide an experimental model of tight nutritional containment for preventing dissemination of microbial GMOs. Our strains transported the four canonical dNTPs, in the following order of preference: dCTP > dATP ≥ dGTP > dTTP. Prolonged cultivation under limitation of exogenous dTTP led to the enhancement of dNTP transport by adaptive evolution. We investigated the uptake of dCTP analogues with altered sugar or nucleobase moieties, which were found to cause a loss of cell viability and an increase of mutant frequency, respectively. E. coli strains equipped with nucleoside triphosphate transporters should be instrumental for evolving organisms whose DNA genome is morphed chemically by fully substituting its canonical nucleotide components.

How to sustainably feed a microbe: Strategies for biological production of carbon-based commodities with renewable electricity

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

Butler, Caitlyn S.; Lovley, Derek R.

As interest and application of renewable energy grows, strategies are needed to align the asynchronous supply and demand. Microbial metabolisms are a potentially sustainable mechanism for transforming renewable electrical energy into biocommodities that are easily stored and transported. Acetogens and methanogens can reduce carbon dioxide to organic products including methane, acetic acid, and ethanol. The library of biocommodities is expanded when engineered metabolisms of acetogens are included. Typically, electrochemical systems are employed to integrate renewable energy sources with biological systems for production of carbon-based commodities. Within these systems, there are three prevailing mechanisms for delivering electrons to microorganisms for themore » conversion of carbon dioxide to reduce organic compounds: (1) electrons can be delivered to microorganisms via H 2 produced separately in a electrolyzer, (2) H 2 produced at a cathode can convey electrons to microorganisms supported on the cathode surface, and (3) a cathode can directly feed electrons to microorganisms. Each of these strategies has advantages and disadvantages that must be considered in designing full-scale processes. Furthermore, this review considers the evolving understanding of each of these approaches and the state of design for advancing these strategies toward viability.« less

How to Sustainably Feed a Microbe: Strategies for Biological Production of Carbon-Based Commodities with Renewable Electricity.

PubMed

Butler, Caitlyn S; Lovley, Derek R

2016-01-01

As interest and application of renewable energy grows, strategies are needed to align the asynchronous supply and demand. Microbial metabolisms are a potentially sustainable mechanism for transforming renewable electrical energy into biocommodities that are easily stored and transported. Acetogens and methanogens can reduce carbon dioxide to organic products including methane, acetic acid, and ethanol. The library of biocommodities is expanded when engineered metabolisms of acetogens are included. Typically, electrochemical systems are employed to integrate renewable energy sources with biological systems for production of carbon-based commodities. Within these systems, there are three prevailing mechanisms for delivering electrons to microorganisms for the conversion of carbon dioxide to reduce organic compounds: (1) electrons can be delivered to microorganisms via H 2 produced separately in a electrolyzer, (2) H 2 produced at a cathode can convey electrons to microorganisms supported on the cathode surface, and (3) a cathode can directly feed electrons to microorganisms. Each of these strategies has advantages and disadvantages that must be considered in designing full-scale processes. This review considers the evolving understanding of each of these approaches and the state of design for advancing these strategies toward viability.

Effect of storage time on the viability of cryopreserved bovine spermatozoa

USDA-ARS?s Scientific Manuscript database

Long term cryopreserved semen viability can impact the National Animal Germplasm Program’s (NAGP) sampling strategy and ability to reconstitute livestock populations. Therefore, the purpose of this project was to determine if prolonged storage of cryopreserved sperm impacts cell viability. Cryoprese...

Potential of coconut water and soy milk for use as storage media to preserve the viability of periodontal ligament cells: an in vitro study.

PubMed

Moura, Camilla Cristhian Gomes; Soares, Priscilla Barbosa Ferreira; de Paula Reis, Manuella Verdinelli; Fernandes Neto, Alfredo Júlio; Zanetta Barbosa, Darceny; Soares, Carlos José

2014-02-01

There is no consensus regarding the ability of coconut water and soy milk to maintain long-term cell viability. This study investigated the ability of pH-adjusted coconut water and soy milk to maintain the viability of periodontal ligament cells over a short and a longer period and compared these abilities with those of other solutions. Dog premolar teeth were extracted, dried for 30 min, and stored in the following media for 50 min or 24 h: long shelf-life whole milk (SWM), long shelf-life skim milk (SSM), Hank's Balanced Salt Solution (HBSS), soy milk (SM), and pH-adjusted coconut water (CW). The positive and two negative control groups corresponded to 0-min, 30-min (short-term), and 24-h (long-term) dry times, respectively. Cell viability was analyzed by trypan blue exclusion. Data were statistically analyzed using the Kruskal-Wallis test with post-analysis using the Dunn method. In the short-term experiment, the SSM resulted in significantly lower cell viability than SM and CW. At 24 h, SM and CW resulted in higher viability than HBSS and SSM and in comparable performance with the positive control group. Cell viability decreased over time, except in SM and CW. Soy milk and pH-adjusted coconut water showed promising results as storage solutions for avulsed teeth, preserving the viability for up to 24 h. © 2013 John Wiley & Sons A/S.

Role of cytoskeletal mechanics and cell membrane fluidity in the intracellular delivery of molecules mediated by laser-activated carbon nanoparticles.

PubMed

Holguin, Stefany Y; Anderson, Caleb F; Thadhani, Naresh N; Prausnitz, Mark R

2017-10-01

Exposure of cells and nanoparticles to near-infrared nanosecond pulsed laser light can lead to efficient intracellular delivery of molecules while maintaining high cell viability by a photoacoustic phenomenon known as transient nanoparticle energy transduction (TNET). Here, we examined the influence of cytoskeletal mechanics and plasma membrane fluidity on intracellular uptake of molecules and loss of cell viability due to TNET. We found that destabilization of actin filaments using latrunculin A led to greater uptake of molecules and less viability loss caused by TNET. Stabilization of actin filaments using jasplakinolide had no significant effect on uptake or viability loss caused by TNET. To study the role of plasma membrane fluidity, we increased fluidity by depletion of membrane cholesterol using methyl-β-cyclodextrin and decreased fluidity by enrichment of the membrane with cholesterol using water-soluble cholesterol. Neither of these membrane fluidity changes significantly altered cellular uptake or viability loss caused by TNET. We conclude that weakening mechanical integrity of the cytoskeleton can increase intracellular uptake and decrease loss of cell viability, while plasma membrane fluidity does not appear to play a significant role in uptake or viability loss caused by TNET. The positive effects of cytoskeletal weakening may be due to an enhanced ability of the cell to recover from the effects of TNET and maintain viability. Biotechnol. Bioeng. 2017;114: 2390-2399. © 2017 Wiley Periodicals, Inc. © 2017 Wiley Periodicals, Inc.

Investigation of Electrobiological Properties of Bioaerosols

NASA Astrophysics Data System (ADS)

Mainelis, G.; Yao, M.; An, H. R.

2004-05-01

Exposure to bioaerosols, especially to pathogenic or allergenic microorganisms, may cause a wide range of respiratory and other health disorders in occupational and general populations. One of bioaerosol characteristics - electric charge - can greatly influence their deposition in sampling lines and collection devices. The magnitude of electric charge carried by inhaled particles can have a significant effect on their deposition in the lung. In addition, electric charge may affect role of bioaerosols as ice and cloud condensation nuclei; charge (or electrical mobility) can control bioaerosol movement in electrical fields, such as created by power lines. Electrical charge is also important for the development of bioaerosol samplers that utilize electrostatics for particle collection - this technique has been shown to be more "gentle" collection method than traditionally used impactors and impingers. Our previous studies have shown that airborne environmental bacteria, such as Pseudomonas fluorescens and B. subtilis var. niger, have a net negative charge, with individual cells carrying as many as 10,000 elementary charge units, which sharply contrasted with low electrical charges carried by non-biological test particles. We have also found that magnitude and polarity of electrical charge can significantly affect viability of sensitive bacteria, such as P. fluorescens. In our continuing exploration of electrobiological properties of bioaerosols, we investigated application of electrostatic collection method for concurrent determination of total and viable bioaerosols, and also analyzed the effect of electrical fields on microbial viability. In our new bioaerosol collector, the biological particles are drawn into the sampler's electrical field and are concurrently deposited on an agar plate for determining viable microorganisms, and into a ELISA plate for determining total collected microorganisms. Experiments with B. subtilis var. niger and P. fluorescens vegetative cells have shown that on average 80 percent of airborne bacteria entering the sampler were removed from the air onto the plates when the sampler operated at 8 L/min and used collection voltage of -1,500V. From 15 to 25 percent of all bacteria entering the sampler were enumerated by the culture technique. Use of electrostatic analysis techniques may require application of strong electrical fields which could be damaging to biological particles. In our experiments, the airborne P. fluorescens bacteria were exposed to electric fields of 10kV/cm for 30 seconds, which did not result in viability reduction. In contrast, more than 90 percent of the P. fluorescens cells have been killed when the microorganisms were first deposited on filters and then exposed to positive electrical field of 15 kV/cm for at least 15 minutes. Electrical fields of 5 and 10 kV/cm also achieved similar effect when bacteria were exposed for 120 min. The exposure of bacteria to negative electrical fields resulted in even higher rates of inactivation. The B. subtilis var. niger bacteria proved to be hardier and 10 percent viability reduction was achieved with the use of 15kV/min for 2 hours. The obtained results demonstrate the importance of electrical charges and fields in behavior, collection and control of bioaerosols. The field studies will have to be performed to confirm laboratory findings.

Population viability impacts of habitat additions and subtractions: A simulation experiment with endangered kangaroo rats

EPA Science Inventory

Species viability is influenced by the quality, quantity and configuration of habitat. For species at risk, a principal challenge is to identify landscape configurations that, if realized, would improve a population’s viability or restoration potential. Critical habitat patche...

Identification, Description, and Perceived Viability of K-12 Consolidated Catholic School Systems

ERIC Educational Resources Information Center

Britt, Kenith C.

2011-01-01

Limited research has been conducted on Catholic school viability (James, Tichy, Collins, & Schwob, 2008; Lundy, 1999) and Catholic school systems (Goldschmidt, O'Keefe, & Walsh, 2004). But no research studies have investigated the viability of the consolidated Catholic school system (DeFiore, Convey, & Schuttloffel, 2009). This study investigates…

24 CFR 968.112 - Eligible costs.

Code of Federal Regulations, 2013 CFR

2013-04-01

...) Demonstration of viability. Except in the case of emergency work, a PHA shall only expend funds on a development... physical and social viability of the development at a reasonable cost (as defined in § 968.105), or for... for the long-term physical and social viability of the individual development. Development specific...

24 CFR 968.112 - Eligible costs.

Code of Federal Regulations, 2012 CFR

2012-04-01

...) Demonstration of viability. Except in the case of emergency work, a PHA shall only expend funds on a development... physical and social viability of the development at a reasonable cost (as defined in § 968.105), or for... for the long-term physical and social viability of the individual development. Development specific...

7 CFR 1703.131 - Approved purposes for a combination loan and grant.

Code of Federal Regulations, 2011 CFR

2011-01-01

... a cost which would not adversely affect the economic viability of the project; (e) Providing links... sources is not available at a cost that does not adversely impact the economic viability of the project as... impact the economic viability of the project, as determined by the Administrator. ...

7 CFR 1703.131 - Approved purposes for a combination loan and grant.

Code of Federal Regulations, 2010 CFR

2010-01-01

... a cost which would not adversely affect the economic viability of the project; (e) Providing links... sources is not available at a cost that does not adversely impact the economic viability of the project as... impact the economic viability of the project, as determined by the Administrator. ...

7 CFR 1703.131 - Approved purposes for a combination loan and grant.

Code of Federal Regulations, 2014 CFR

2014-01-01

... a cost which would not adversely affect the economic viability of the project; (e) Providing links... sources is not available at a cost that does not adversely impact the economic viability of the project as... impact the economic viability of the project, as determined by the Administrator. ...

24 CFR 968.112 - Eligible costs.

Code of Federal Regulations, 2011 CFR

2011-04-01

...) Demonstration of viability. Except in the case of emergency work, a PHA shall only expend funds on a development... physical and social viability of the development at a reasonable cost (as defined in § 968.105), or for... for the long-term physical and social viability of the individual development. Development specific...

7 CFR 1703.131 - Approved purposes for a combination loan and grant.

Code of Federal Regulations, 2012 CFR

2012-01-01

... a cost which would not adversely affect the economic viability of the project; (e) Providing links... sources is not available at a cost that does not adversely impact the economic viability of the project as... impact the economic viability of the project, as determined by the Administrator. ...

7 CFR 1703.131 - Approved purposes for a combination loan and grant.

Code of Federal Regulations, 2013 CFR

2013-01-01

... a cost which would not adversely affect the economic viability of the project; (e) Providing links... sources is not available at a cost that does not adversely impact the economic viability of the project as... impact the economic viability of the project, as determined by the Administrator. ...

Seed viability, germination, and radicle growth of dwarf mistletoe in California

Treesearch

Robert F. Scharpf

1970-01-01

Two species of dwarf mistletoe were studied: Arceuthobium abietinum (Engelm.) Hawksworth and Wiens and A. occidentale Engelm. Viability of fresh seeds was high and not significantly influenced by year of collection, where collected, or plant from which collected. Temperature affected viability most noticeably. It also significantly...

Assessing the Financial Viability of Academic Programmes

ERIC Educational Resources Information Center

Swift, Lynette

2012-01-01

This paper reviews and examines approaches to determining the financial viability of academic programmes as a critical component of assessing a programme's overall sustainability. Key to assessing the financial viability of a programme is understanding the teaching activities required to deliver the programme and the cost of those activities. A…

Financial Viability of Institutions. Issues in Post-Secondary Education.

ERIC Educational Resources Information Center

Jenny, Hans H.

Financial viability in postsecondary education is considered as part of the Postsecondary Education Core Design Project sponsored by the National Center for Education Statistics. Financial viability is defined within the scope of postsecondary education, and key policy issues at the national, state, and institutional levels are identified.…

A viability analysis for a stock/price model

NASA Astrophysics Data System (ADS)

Jerry, Chakib; Raissi, Nadia

2012-09-01

We examine the conditions for the sustainability of a stock/price system based on the use of a marine renewable resource. Instead of studying the environmental and economic interactions in terms of optimal control, we focus on the viability of the system. These viability/crisis situations are defined by a set of economic state constraints. This constraints combine a guaranteed consumption and a minimum income for fishermen. Using the mathematical concept of viability kernel, we reveal that with only economics constraints we guarantee a perennial stock/price system.

Enhanced viability of corneal epithelial cells for efficient transport/storage using a structurally modified calcium alginate hydrogel.

PubMed

Wright, Bernice; Cave, Richard A; Cook, Joseph P; Khutoryanskiy, Vitaliy V; Mi, Shengli; Chen, Bo; Leyland, Martin; Connon, Che J

2012-05-01

Therapeutic limbal epithelial stem cells could be managed more efficiently if clinically validated batches were transported for 'on-demand' use. In this study, corneal epithelial cell viability in calcium alginate hydrogels was examined under cell culture, ambient and chilled conditions for up to 7 days. Cell viability improved as gel internal pore size increased, and was further enhanced with modification of the gel from a mass to a thin disc. Ambient storage conditions were optimal for supporting cell viability in gel discs. Cell viability in gel discs was significantly enhanced with increases in pore size mediated by hydroxyethyl cellulose. Our novel methodology of controlling alginate gel shape and pore size together provides a more practical and economical alternative to established corneal tissue/cell storage methods.

Effects of Single P-Element Insertions on Bristle Number and Viability in Drosophila Melanogaster

PubMed Central

Lyman, R. F.; Lawrence, F.; Nuzhdin, S. V.; Mackay, TFC.

1996-01-01

Single P-element mutagenesis was used to construct 1094 lines with P[lArB] inserts on all three major chromosomes in an isogenic background previously free of P elements. The effects of insertions on bristle number and on viability were assessed by comparison to 392 control lines. The variance and effects of P-element inserts on bristle number and viability were larger than those inferred from spontaneous mutations. The distributions of effects on bristle number were symmetrical and highly leptokurtic, such that a few inserts with large effects caused most of the increase in variance. The distribution of effects on viability were negatively skewed and platykurtic. On average, the effects of P-element insertions on bristle number were partly recessive and on viability were completely recessive. P-element inserts with large effects on bristle number tended to have reduced viability, but the correlation between the absolute value of the effects on bristle number and on viability was not strong. Fifty P-element inserts tagging quantitative trait loci (QTLs) with large effects on bristle number were mapped cytogenetically. Two P-element-induced scabrous alleles and five extramacrochaetae alleles were generated. Single P-element mutagenesis is a powerful method for identifying QTLs at the level of genetic locus. PMID:8722781

Effects of single P-element insertions on bristle number and viability in Drosophila melanogaster.

PubMed

Lyman, R F; Lawrence, F; Nuzhdin, S V; Mackay, T F

1996-05-01

Single P-element mutagenesis was used to construct 1094 lines with P[lArB] inserts on all three major chromosomes in an isogenic background previously free of P elements. The effects of insertions on bristle number and on viability were assessed by comparison to 392 control lines. The variance and effects of P-element inserts on bristle number and viability were larger than those inferred from spontaneous mutations. The distributions of effects on bristle number were symmetrical and highly leptokurtic, such that a few inserts with large effects caused most of the increase in variance. The distribution of effects on viability were negatively skewed and platykurtic. On average, the effects of P-element insertions on bristle number were partly recessive and on viability were completely recessive. P-element inserts with large effects on bristle number tended to have reduced viability, but the correlation between the absolute value of the effects on bristle number and on viability was not strong. Fifty P-element inserts tagging quantitative trait loci (QTLs) with large effects on bristle number were mapped cytogenetically. Two P-element-induced scabrous alleles and five extramacrochaetae alleles were generated. Single P-element mutagenesis is a powerful method for identifying QTLs at the level of genetic locus.

The potential role of polyphenols in the modulation of skin cell viability by Aspalathus linearis and Cyclopia spp. herbal tea extracts in vitro.

PubMed

Magcwebeba, Tandeka Unathi; Riedel, Sylvia; Swanevelder, Sonja; Swart, Pieter; De Beer, Dalene; Joubert, Elizabeth; Andreas Gelderblom, Wentzel Christoffel

2016-11-01

The relationship between polyphenol constituents, antioxidant properties of aqueous and methanol extracts of green tea (Camellia sinensis), the herbal teas, rooibos (Aspalathus linearis) and honeybush (Cyclopia spp.), against skin cell viability was investigated in vitro. The effect of extracts, characterised in terms of polyphenol content and antioxidant properties, on cell viability of premalignant, normal and malignant skin cells was determined. Phenolic composition, particularly high levels of potent antioxidants, of rooibos and green tea methanol extracts was associated with a strong reduction in cell viability specifically targeting premalignant cells. In contrast, the aqueous extracts of Cyclopia spp. were more effective in reducing cell viability. This correlated with a relatively high flavanol/proanthocyanidin content and ABTS radical cation scavenging capacity. The major green tea flavanol (epigallocatechin gallate) and rooibos dihydrochalcone (aspalathin) exhibited differential effects against cell viability, while the major honeybush xanthone (mangiferin) and flavanone (hesperidin) lacked any effect presumably due to a cytoprotective effect. The underlying mechanisms against skin cell viability are likely to involve mitochondrial dysfunction resulting from polyphenol-iron interactions. The polyphenol constituents and antioxidant parameters of herbal tea extracts are useful tools to predict their activity against skin cell survival in vitro and potential chemopreventive effects in vivo. © 2016 Royal Pharmaceutical Society.

Longevity and viability of Taenia solium eggs in the digestive system of the beetle Ammophorus rubripes.

PubMed

Gomez-Puerta, Luis Antonio; Lopez-Urbina, Maria Teresa; Garcia, Hector Hugo; Gonzalez, Armando Emiliano

2014-03-01

The present study evaluated the capacity of Ammophorus rubripes beetles to carry Taenia solium eggs, in terms of duration and viability of eggs in their digestive system. One hundred beetles were distributed into five polyethylene boxes, and then they were infected with T. solium eggs. Gravid proglottids of T. solium were crushed and then mixed with cattle feces. One gram of this mixture was placed in each box for 24 hours, after which each group of beetles was transferred into a new clean box. Then, five beetles were dissected every three days. Time was strongly associated with viability (r=0.89; P<0.001) and the calculated time to cero viability is 36 days. The eggs in the intestinal system of each beetle were counted and tested for viability. Taenia solium eggs were present in the beetle's digestive system for up to 39 days (13th sampling day out of 20), gradually reducing in numbers and viability, which was 0 on day 36 post-infection. Egg viability was around 40% up to day 24 post-infection, with a median number of eggs of 11 per beetle at this time. Dung beetles may potentially contribute towards dispersing T. solium eggs in endemic areas.

LONGEVITY AND VIABILITY OF Taenia solium EGGS IN THE DIGESTIVE SYSTEM OF THE BEETLE Ammophorus rubripes

PubMed Central

Gomez-Puerta, Luis Antonio; Lopez-Urbina, Maria Teresa; Garcia, Hector Hugo; Gonzalez, Armando Emiliano

2015-01-01

The present study evaluated the capacity of Ammophorus rubripes beetles to carry Taenia solium eggs, in terms of duration and viability of eggs in their digestive system. One hundred beetles were distributed into five polyethylene boxes, and then they were infected with T. solium eggs. Gravid proglottids of T. solium were crushed and then mixed with cattle feces. One gram of this mixture was placed in each box for 24 hours, after which each group of beetles was transferred into a new clean box. Then, five beetles were dissected every three days. Time was strongly associated with viability (r=0.89; P<0.001) and the calculated time to cero viability is 36 days. The eggs in the intestinal system of each beetle were counted and tested for viability. Taenia solium eggs were present in the beetle’s digestive system for up to 39 days (13th sampling day out of 20), gradually reducing in numbers and viability, which was 0 on day 36 post-infection. Egg viability was around 40% up to day 24 post-infection, with a median number of eggs of 11 per beetle at this time. Dung beetles may potentially contribute towards dispersing T. solium eggs in endemic areas. PMID:24728368

Effects of steam autoclave treatment on Geobacillus stearothermophilus spores.

PubMed

Huesca-Espitia, L C; Suvira, M; Rosenbeck, K; Korza, G; Setlow, B; Li, W; Wang, S; Li, Y-Q; Setlow, P

2016-11-01

To determine the mechanism of autoclave killing of Geobacillus stearothermophilus spores used in biological indicators (BIs) for steam autoclave sterilization, and rates of loss of spore viability and a spore enzyme used in BIs. Spore viability, dipicolinic acid (DPA) release, nucleic acid staining, α-glucosidase activity, protein structure and mutagenesis were measured during autoclaving of G. stearothermophilus spores. Loss of DPA and increases in spore core nucleic acid staining were slower than loss of spore viability. Spore core α-glucosidase was also lost more slowly than spore viability, although soluble α-glucosidase in spore preparations was lost more rapidly. However, spores exposed to an effective autoclave sterilization lost all viability and α-glucosidase activity. Apparently killed autoclaved spores were not recovered by artificial germination in supportive media, much spore protein was denatured during autoclaving, and partially killed autoclave-treated spore preparations did not acquire mutations. These results indicate that autoclave-killed spores cannot be revived, spore killing by autoclaving is likely by protein damage, and spore core α-glucosidase activity is lost more slowly than spore viability. This work provides insight into the mechanism of autoclave killing of spores of an organism used in BIs, and that a spore enzyme in a BI is more stable to autoclaving than spore viability. © 2016 The Society for Applied Microbiology.

Combination of hyperthermia and photodynamic therapy on mesenchymal stem cell line treated with chloroaluminum phthalocyanine magnetic-nanoemulsion

NASA Astrophysics Data System (ADS)

de Paula, Leonardo B.; Primo, Fernando L.; Pinto, Marcelo R.; Morais, Paulo C.; Tedesco, Antonio C.

2015-04-01

The present study reports on the preparation and the cell viability assay of two nanoemulsions loaded with magnetic nanoparticle and chloroaluminum phthalocyanine. The preparations contain equal amount of chloroaluminum phthalocyanine (0.05 mg/mL) but different contents of magnetic nanoparticle (0.15×1013 or 1.50×1013 particle/mL). The human bone marrow mesenchymal stem cell line was used as the model to assess the cell viability and this type of cell can be used as a model to mimic cancer stem cells. The cell viability assays were performed in isolated as well as under combined magnetic hyperthermia and photodynamic therapy treatments. We found from the cell viability assay that under the hyperthermia treatment (1 MHz and 40 Oe magnetic field amplitude) the cell viability reduction was about 10%, regardless the magnetic nanoparticle content within the magnetic nanoparticle/chloroaluminum phthalocyanine formulation. However, cell viability reduction of about 50% and 60% were found while applying the photodynamic therapy treatment using the magnetic nanoparticle/chloroaluminum phthalocyanine formulation containing 0.15×1013 or 1.50×1013 magnetic particle/mL, respectively. Finally, an average reduction in cell viability of about 66% was found while combining the hyperthermia and photodynamic therapy treatments.

Comparison of reintroduction and enhancement effects on metapopulation viability

USGS Publications Warehouse

Halsey, Samniqueka J; Bell, Timothy J.; McEachern, A. Kathryn; Pavlovic, Noel B.

2015-01-01

Metapopulation viability depends upon a balance of extinction and colonization of local habitats by a species. Mechanisms that can affect this balance include physical characteristics related to natural processes (e.g. succession) as well as anthropogenic actions. Plant restorations can help to produce favorable metapopulation dynamics and consequently increase viability; however, to date no studies confirm this is true. Population viability analysis (PVA) allows for the use of empirical data to generate theoretical future projections in the form of median time to extinction and probability of extinction. In turn, PVAs can inform and aid the development of conservation, recovery, and management plans. Pitcher's thistle (Cirsium pitcheri) is a dune endemic that exhibited metapopulation dynamics. We projected viability of three natural and two restored populations with demographic data spanning 15–23 years to determine the degree the addition of reintroduced population affects metapopulation viability. The models were validated by comparing observed and projected abundances and adjusting parameters associated with demographic and environmental stochasticity to improve model performance. Our chosen model correctly predicted yearly population abundance for 60% of the population-years. Using that model, 50-year projections showed that the addition of reintroductions increases metapopulation viability. The reintroduction that simulated population performance in early-successional habitats had the maximum benefit. In situ enhancements of existing populations proved to be equally effective. This study shows that restorations can facilitate and improve metapopulation viability of species dependent on metapopulation dynamics for survival with long-term persistence of C. pitcheri in Indiana likely to depend on continued active management.

Combining biofilm matrix measurements with biomass and viability assays in susceptibility assessments of antimicrobials against Staphylococcus aureus biofilms.

PubMed

Skogman, Malena Elise; Vuorela, Pia Maarit; Fallarero, Adyary

2012-09-01

Despite that three types of assays (measuring biofilm viability, biomass, or matrix) are described to assess anti-biofilm activity, they are rarely used together. As infections can easily reappear if the matrix is not affected after antibiotic treatments, our goal was to explore the simultaneous effects of antibiotics on the viability, biomass and matrix of Staphylococcus aureus biofilms (ATCC 25923). Viability and biomass were quantified using resazurin and crystal violet staining sequentially in the same plate, while matrix staining was conducted with a wheat germ agglutinin-Alexa Fluor 488 fluorescent conjugate. Establishment of the detection limits and linearity ranges allowed concluding that all three methods were able to estimate biofilm formation in a similar fashion. In a susceptibility study with 18-h biofilms, two model compounds (penicillin G and ciprofloxacin) caused a reduction on the viability and biomass accompanied by an increase or not changed levels of the matrix, respectively. This response pattern was also proven for S. aureus Newman, S. epidermidis and E. coli biofilms. A classification of antibiotics based on five categories according to their effects on viability and matrix has been proposed earlier. Our data suggests a sixth group, represented by penicillin, causing decrease in bacterial viability but showing stimulatory effects on the matrix. Further, if effects on the matrix are not taken into account, the long-term chemotherapeutic effect of antibiotics can be jeopardized in spite of the positive effects on biofilms viability and biomass. Thus, measuring all these three endpoints simultaneously provide a more complete and accurate picture.

Microbial Community Structure of Subglacial Lake Whillans, West Antarctica

PubMed Central

Achberger, Amanda M.; Christner, Brent C.; Michaud, Alexander B.; Priscu, John C.; Skidmore, Mark L.; Vick-Majors, Trista J.; Adkins, W.

2016-01-01

Subglacial Lake Whillans (SLW) is located beneath ∼800 m of ice on the Whillans Ice Stream in West Antarctica and was sampled in January of 2013, providing the first opportunity to directly examine water and sediments from an Antarctic subglacial lake. To minimize the introduction of surface contaminants to SLW during its exploration, an access borehole was created using a microbiologically clean hot water drill designed to reduce the number and viability of microorganisms in the drilling water. Analysis of 16S rRNA genes (rDNA) amplified from samples of the drilling and borehole water allowed an evaluation of the efficacy of this approach and enabled a confident assessment of the SLW ecosystem inhabitants. Based on an analysis of 16S rDNA and rRNA (i.e., reverse-transcribed rRNA molecules) data, the SLW community was found to be bacterially dominated and compositionally distinct from the assemblages identified in the drill system. The abundance of bacteria (e.g., Candidatus Nitrotoga, Sideroxydans, Thiobacillus, and Albidiferax) and archaea (Candidatus Nitrosoarchaeum) related to chemolithoautotrophs was consistent with the oxidation of reduced iron, sulfur, and nitrogen compounds having important roles as pathways for primary production in this permanently dark ecosystem. Further, the prevalence of Methylobacter in surficial lake sediments combined with the detection of methanogenic taxa in the deepest sediment horizons analyzed (34–36 cm) supported the hypothesis that methane cycling occurs beneath the West Antarctic Ice Sheet. Large ratios of rRNA to rDNA were observed for several operational taxonomic units abundant in the water column and sediments (e.g., Albidiferax, Methylobacter, Candidatus Nitrotoga, Sideroxydans, and Smithella), suggesting a potentially active role for these taxa in the SLW ecosystem. Our findings are consistent with chemosynthetic microorganisms serving as the ecological foundation in this dark subsurface environment, providing new organic matter that sustains a microbial ecosystem beneath the West Antarctic Ice Sheet. PMID:27713727

Effects of dimethyl dicarbonate (DMDC) on the fermentation of litchi juice by Lactobacillus casei as an alternative of heat treatment.

PubMed

Yu, Yuanshan; Xiao, Gengsheng; Xu, Yujuan; Wu, Jijun; Wen, Jing

2014-05-01

This study investigated the effects of dimethyl dicarbonate (DMDC) on the fermentation of litchi juice by Lactobacillus casei as an alternative of heat treatment that may have undesirable effect on the juice. Quality attributes and products stability of both the fermented heat- and DMDC-treated litchi juice by L. casei were compared. It was found that residual indigenous microorganisms in both the heat- and DMDC-treated litchi juice cannot grow into dominant bacteria during further fermentation of litchi juice by L. casei. Compared with fermented heat-treated litchi juice, fermented DMDC-treated litchi juice showed a better color, flavor, and overall acceptance, and also retained more total phenolics and antioxidant capacity. The viability counts of L. casei in both the heat- and DMDC-treated litchi juice were more 8.0 lg CFU/mL after 4 wk of storage at 4 °C. Also, some quality attributes in both the fermented heat- and DMDC-treated litchi juices, including pH, total phenolics, ascorbic acid, antioxidant capacity, and so on, showed the tendency to slow decrease during storage at 4 °C, but the scores of overall acceptance showed no reduction after the storage of 4 wk at 4 °C. On the whole, the application of DMDC treatment could be an ideal alternative of heat treatment to ensure the microbial safety, consistent sensory, and nutritional quality of fermented litchi juice prior to fermentation. The pasteurization treatment is often recommended prior to fermentation of fruit juice by probiotics, as it would lead to a rapid inactivation and inhibition of spoilage and pathogenic bacteria, and ensure the fermented products with consistent sensory and nutritional quality. Dimethyl dicarbonate (DMDC) is a powerful antimicrobial agent, which was approved for use as a microbial control agent in juice beverages by FDA. This study provides a scientific basis for the application of DMDC prior to fermentation of litchi juice. © 2014 Institute of Food Technologists®

Comparison of human and southern sea otter (Enhydra lutris nereis) health risks for infection with protozoa in nearshore waters.

PubMed

Adell, A D; McBride, G; Wuertz, S; Conrad, P A; Smith, W A

2016-11-01

Cryptosporidium and Giardia spp. are waterborne, fecally-transmitted pathogens that cause economic loss due to gastroenteritis and beach closures. We applied quantitative microbial risk assessment (QMRA) to determine the health risks for humans and sea otters due to waterborne exposure of Cryptosporidium and Giardia spp. when swimming in three types of surface waters: river, stormwater and wastewater effluent during the wet and dry seasons in the central coast of California. This is the first application of QMRA to estimate both the probability of infection in Southern sea otters and the probability of illness in humans, using microbial source tracking (MST) as a variable. Children swimming close to stormwater discharges had an estimated Cryptosporidium-associated illness probability that exceeded the accepted U.S. EPA criteria (32 illnesses/1000 swimmers or 3.2%). Based on the assumption that sea otters are as susceptible as humans to Cryptosporidium infection, the infection probabilities were close to 2% and 16% when sea otters were swimming at the end of points of rivers and stormwater discharges, respectively. In the case of Giardia, infection probabilities of 11% and 23% were estimated for sea otters swimming at the end of point of wastewater discharges, assuming that sea otters are as susceptible as gerbils and humans, respectively. The results of this QMRA suggest that 1) humans and sea otters are at risk when swimming at outflow sites for rivers, stormwater and treated wastewater effluent; 2) reduced loads of viable protozoan cysts and oocysts in recreational water can lessen the probability of infection of humans and sea otters; and 3) the risk of infection of humans and sea otters can be reduced with the treatment of wastewater to decrease oocyst and cyst viability before effluent is released into the sea. Copyright © 2016 Elsevier Ltd. All rights reserved.

Do Long-Term Changes in Organic Matter Inputs to Forest Soils Affect Dissolved Organic Matter Chemistry and Export?

NASA Astrophysics Data System (ADS)

Lajtha, K.; Strid, A.; Lee, B. S.

2014-12-01

Dissolved organic matter (DOM) production and transport play an important role in regulating organic matter (OM) distribution through a soil profile and ultimately, OM stabilization or export to aquatic systems. The contributions of varying OM inputs to the quality and amount of DOM as it passes through a soil profile remain relatively unknown. The Detrital Input and Removal Treatment (DIRT) site at the H. J. Andrews Experimental Forest in Oregon has undergone 17 years of litter, wood and root input manipulations and allows us to guage shifts in DOM chemistry induced by long-term changes to aboveground and belowground OM additions and exclusions. Using fluorescence and UV spectroscopy to characterize fluorescent properties, extent of decomposition, and sources of DOM in streams and soil solutions collected with lysimeters and soil extractions, we have assessed the importance of fresh OM inputs to DOM chemistry. Soil extracts from DIRT plots had a higher fluorescence index (FI) than lysimeter solutions or stream water. A high FI in surface water is generally interpreted as indicative of a high proportion of microbially-derived DOM. However, we suspect that the high FI in soil extracts is due to a higher proportion of non-aromatic DOM from fresh soil that microorganisms consume in transit through the soil profile to lysimeters or to streams. High redox index (RI) values were observed in lysimeters from the April 2014 sampling compared with the November 2013 sampling. These RI values show evidence of more reducing conditions at the end of the rainy season in the spring compared to the onset of the rainy season in the fall. Lysimeter water collected in No Input, No Litter, and No Root treatments contained high proportions of protein, suggesting the absence of carbon inputs changes activities of the microbial community. Observed variations reflect the viability of using fluorescent properties to explore the terrestrial-aquatic interface.

Mild acid and alkali treated clay minerals enhance bioremediation of polycyclic aromatic hydrocarbons in long-term contaminated soil: A 14C-tracer study.

PubMed

Biswas, Bhabananda; Sarkar, Binoy; Rusmin, Ruhaida; Naidu, Ravi

2017-04-01

Bioremediation of polycyclic aromatic hydrocarbon (PAH)-contaminated soils requires a higher microbial viability and an increased PAH bioavailability. The clay/modified clay-modulated bacterial degradation could deliver a more efficient removal of PAHs in soils depending on the bioavailability of the compounds. In this study, we modified clay minerals (smectite and palygorskite) with mild acid (HCl) and alkali (NaOH) treatments (0.5-3 M), which increased the surface area and pore volume of the products, and removed the impurities without collapsing the crystalline structure of clay minerals. In soil incubation studies, supplements with the clay products increased bacterial growth in the order: 0.5 M HCl ≥ unmodified ≥ 0.5 M NaOH ≥ 3 M NaOH ≥ 3 M HCl for smectite, and 0.5 M HCl ≥ 3 M NaOH ≥ 0.5 M NaOH ≥ 3 M HCl ≥ unmodified for palygorskite. A 14 C-tracing study showed that the mild acid/alkali-treated clay products increased the PAH biodegradation (5-8%) in the order of 0.5 M HCl ≥ unmodified > 3 M NaOH ≥ 0.5 M NaOH for smectite, and 0.5 M HCl > 0.5 M NaOH ≥ unmodified ≥ 3 M NaOH for palygorskite. The biodegradation was correlated (r = 0.81) with the bioavailable fraction of PAHs and microbial growth as affected particularly by the 0.5 M HCl and 0.5 M NaOH-treated clay minerals. These results could be pivotal in developing a clay-modulated bioremediation technology for cleaning up PAH-contaminated soils and sediments in the field. Copyright © 2017 Elsevier Ltd. All rights reserved.

Cable in Boston; A Basic Viability Report.

ERIC Educational Resources Information Center

Hauben, Jan Ward; And Others

The viability of urban cable television (CATV) as an economic phenomenon is examined via a case study of its feasibility in Boston, a microcosm of general urban environment. To clarify cable's economics, a unitary concept of viability is used in which all local characteristics, cost assumptions, and growth estimates are structured dynamically as a…

Rapid assessment of corn seed viability using short wave infrared line-scan hyperspectral imaging and chemometrics

USDA-ARS?s Scientific Manuscript database

Knowledge of the viability status of seeds before sowing is important to farmers and seed suppliers. However, a myriad of factors can reduce viability of seeds or completely render seeds non-viable during pre- and post-harvest operations. Spectral imaging has shown potential for determining seed via...

7 CFR 1703.141 - Approved purposes for loans.

Code of Federal Regulations, 2010 CFR

2010-01-01

... from other sources at a cost which would not adversely affect the economic viability of the project; (e... other sources is not available at a cost which does not adversely impact the economic viability of the... that does not impact the economic viability of the project, as determined by the Administrator; (i) Any...

7 CFR 1703.141 - Approved purposes for loans.

Code of Federal Regulations, 2014 CFR

2014-01-01

... from other sources at a cost which would not adversely affect the economic viability of the project; (e... other sources is not available at a cost which does not adversely impact the economic viability of the... that does not impact the economic viability of the project, as determined by the Administrator; (i) Any...

7 CFR 1703.141 - Approved purposes for loans.

Code of Federal Regulations, 2013 CFR

2013-01-01

... from other sources at a cost which would not adversely affect the economic viability of the project; (e... other sources is not available at a cost which does not adversely impact the economic viability of the... that does not impact the economic viability of the project, as determined by the Administrator; (i) Any...

7 CFR 1703.141 - Approved purposes for loans.

Code of Federal Regulations, 2012 CFR

2012-01-01

... from other sources at a cost which would not adversely affect the economic viability of the project; (e... other sources is not available at a cost which does not adversely impact the economic viability of the... that does not impact the economic viability of the project, as determined by the Administrator; (i) Any...

Linking population viability, habitat suitability, and landscape simulation models for conservation planning

Treesearch

Michael A. Larson; Frank R., III Thompson; Joshua J. Millspaugh; William D. Dijak; Stephen R. Shifley

2004-01-01

Methods for habitat modeling based on landscape simulations and population viability modeling based on habitat quality are well developed, but no published study of which we are aware has effectively joined them in a single, comprehensive analysis. We demonstrate the application of a population viability model for ovenbirds (Seiurus aurocapillus)...

7 CFR 1703.141 - Approved purposes for loans.

Code of Federal Regulations, 2011 CFR

2011-01-01

... from other sources at a cost which would not adversely affect the economic viability of the project; (e... other sources is not available at a cost which does not adversely impact the economic viability of the... that does not impact the economic viability of the project, as determined by the Administrator; (i) Any...

Experiments with the Viability of Chicken Eggs

ERIC Educational Resources Information Center

Garigliano, Leonard J.

1975-01-01

Presents the results of an experiment designed to test two hypotheses: (1) a delay of two weeks at room temperature will have no effect on the viability of fertile chicken eggs and (2) refrigeration will have no effect on the viability of fertile chicken eggs. Experimenters were the author and two ninth-grade students. (PEB)

Shock Wave-Stimulated Periosteum for Cartilage Repair

DTIC Science & Technology

2013-12-01

were added to the Gtn-HPA prior to the gelation 6 process, at a cell density of 1×105 cells/ml. In the control groups, cells received no treatment...Mesenchymal Stem Cell Viability Viability test was performed 24 hours post- gelation using the Live/Dead assay. Viability/cytotoxicity kit was used (Molecular

NREL's Energy Storage and REopt Teams Awarded $525k from TCF to Study

Science.gov Websites

Commercial Viability of Optimal, Reliable Building-Integrated Energy Storage | News | NREL NREL's Energy Storage and REopt Teams Awarded $525k from TCF to Study Commercial Viability of Optimal Study Commercial Viability of Optimal, Reliable Building-Integrated Energy Storage November 14, 2017

Viability, Quality and Protein Content Associated with Sorghum Caryopses Infected with Grain Mold Fungi

USDA-ARS?s Scientific Manuscript database

Grain mold (GM) of sorghum is a yield-limiting disease that impacts caryopsis viability and quality. Several fungi, including Fusarium thapsinum (FT) and Curvularia lunata (CL), colonize the caryopsis during development. The viability of caryopses (including Sureno, Tx2911, SC170, BTx623, BTx631, an...

Viability, Advantages and Design Methodologies of M-Learning Delivery

ERIC Educational Resources Information Center

Zabel, Todd W.

2010-01-01

The purpose of this study was to examine the viability and principle design methodologies of Mobile Learning models in developing regions. Demographic and market studies were utilized to determine the viability of M-Learning delivery as well as best uses for such technologies and methods given socioeconomic and political conditions within the…

Population viability assessment of salmonids by using probabilistic networks

Treesearch

Danny C. Lee; Bruce E. Rieman

1997-01-01

Public agencies are being asked to quantitatively assess the impact of land management activities on sensitive populations of salmonids. To aid in these assessments, we developed a Bayesian viability assessment procedure (BayVAM) to help characterize land use risks to salmonids in the Pacific Northwest. This procedure incorporates a hybrid approach to viability...

24 CFR 954.306 - Rental housing: qualification as affordable housing and income targeting.

Code of Federal Regulations, 2010 CFR

2010-04-01

... adjustment is necessary to support the continued financial viability of the project and only by an amount that HUD determines is necessary to maintain continued financial viability of the project. HUD expects... income over time should help maintain the financial viability of a project within the qualifying rent...

45 CFR 1302.21 - Grantee shows legal status but not financial viability.

Code of Federal Regulations, 2010 CFR

2010-10-01

... legal status but not financial viability. (a) If a grantee shows legal status but impaired financial... approved application which restores the grantee's financial viability either by a reduction in the program... 45 Public Welfare 4 2010-10-01 2010-10-01 false Grantee shows legal status but not financial...

24 CFR 92.252 - Qualification as affordable housing: Rental housing.

Code of Federal Regulations, 2010 CFR

2010-04-01

... rents and in median income over time should be sufficient to maintain the financial viability of a..., only if HUD finds that an adjustment is necessary to support the continued financial viability of the project and only by an amount that HUD determines is necessary to maintain continued financial viability...

Hydrogen Supplementation of Preservation Solution Improves Viability of Osteochondral Grafts

PubMed Central

Yamada, Takuya; Onuma, Kenji; Kuzuno, Jun; Ujihira, Masanobu; Kurokawa, Ryosuke; Sakai, Rina; Takaso, Masashi

2014-01-01

Allogenic osteochondral tissue (OCT) is used for the treatment of large cartilage defects. Typically, OCTs collected during the disease-screening period are preserved at 4°C; however, the gradual reduction in cell viability during cold preservation adversely affects transplantation outcomes. Therefore, improved storage methods that maintain the cell viability of OCTs are needed to increase the availability of high-quality OCTs and improve treatment outcomes. Here, we evaluated whether long-term hydrogen delivery to preservation solution improved the viability of rat OCTs during cold preservation. Hydrogen-supplemented Dulbecco's Modified Eagles Medium (DMEM) and University of Wisconsin (UW) solution both significantly improved the cell viability of OCTs during preservation at 4°C for 21 days compared to nonsupplemented media. However, the long-term cold preservation of OCTs in DMEM containing hydrogen was associated with the most optimal maintenance of chondrocytes with respect to viability and morphology. Our findings demonstrate that OCTs preserved in DMEM supplemented with hydrogen are a promising material for the repair of large cartilage defects in the clinical setting. PMID:25506061

Assessment of bacterial endospore viability with fluorescent dyes.

PubMed

Laflamme, C; Lavigne, S; Ho, J; Duchaine, C

2004-01-01

To validate three fluorescence viability assays designed primarily for vegetative cells on pure Bacillus endospores. Purified fresh and gamma-irradiated Bacillus endospores (Bacillus cereus, B. coagulans and two strains of B. subtilis) were used. The viability assays were: 5-cyano-2,3-diotolyl tetrazolium chloride (CTC) to test respiratory activity and early germination, DiBAC4(3) and Live/Dead BacLight to measure membrane energization and permeabilization, respectively. Gamma irradiation treatment completely eliminated spore culturability and was used as negative control. The untreated spores showed respiratory activity after 1 h of incubation and this was characteristic of almost 100% of spores after 24 h. The membrane potential assessment gave no answer about spore viability. A lower proportion of untreated spores had permeabilized membrane compared with gamma-irradiated spores using Live/Dead BacLight (P < 0.02). It is possible to use CTC and Live/Dead BacLight to rapidly test endospore viability and evaluate the proportion of spores in a preparation that could not be recovered with plate count. This study shows that fluorescence tests could be applied to assess viability in potentially pathogenic Bacillus spore preparations within 1 h.

Economic viability of access broadband multiservice networks

NASA Astrophysics Data System (ADS)

Castelli, Francesco; Dammicco, Giacinto; Mocci, Ugo

1995-02-01

In this paper the economic viability of alternative architectures for optical access networks providing broad band services to different subscriber classes in a metropolitan environment, is investigated by a specific tool, NEVE (Network Economic Viability Evaluator), developed for broad band multiservice network planning, service evolutionary scenarios assessment, evaluation of tariff strategies and other actions taken at stimulating the demand growth. As the viability target can be achieved in different ways, different studies can be carried out by NEVE. In the paper some of them are discussed, particularly the ones addressed: to evaluate the impact on viability of alternative service scenarios; to determine the critical mass of broad band subscribers and the critical joint service adoption cost; to evaluate cross subsidiary policies among different subscriber classes and services; to perform sensitivity analysis with respect to variations of demand parameters and tariffs.

Coconut milk and probiotic milk as storage media to maintain periodontal ligament cell viability: an in vitro study.

PubMed

Saini, Divya; Gadicherla, Prahlad; Chandra, Prakash; Anandakrishna, Latha

2017-06-01

The viability of periodontal ligament (PDL) cells is a significant determinant of the long-term prognosis of replanted avulsed teeth. A storage medium is often required to maintain the viability of these cells during the extra-alveolar period. Many studies have been carried out to search for the most suitable storage medium for avulsed teeth, but an ideal solution has not yet been found. The purpose of the study was to compare and analyze the ability of coconut milk and probiotic milk to maintain PDL cell viability. In an in vitro setting, 69 caries free human premolars with normal periodontium that had been extracted for orthodontic purposes were randomly divided into two experimental groups on the basis of storage media used (i.e., coconut milk or probiotic milk) and a Hanks' balanced salt solution (HBSS) control group (23 samples per group). Immediately after extraction, the teeth were stored dry for 20 min and then immersed for 30 min in one of the storage media. The teeth were then subjected to collagenase-dispase assay and labeled with 0.5% trypan blue staining solution for determination of cell viability. The number of viable cells was counted under a light microscope and statistically analyzed using anova and post hoc Tukey test (P ≤ 0.05). Statistical analysis demonstrated there was a significant difference (P < 0.001) between coconut milk and probiotic milk as well as HBSS in maintaining cell viability. However, there was no significant difference between probiotic milk and HBSS in ability to maintain PDL cell viability (P > 0.05). Coconut milk may not be suitable as an interim transport media due to poor maintenance of cell viability. However, probiotic milk was able to maintain PDL cell viability as well as HBSS. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Viability of Staphylococcus xylosus isolated from artisanal sausages for application as starter cultures in meat products.

PubMed

Fiorentini, Angela Maria; Sawitzki, Maristela Cortez; Bertol, Teresinha Marisa; Sant'anna, Ernani S

2009-01-01

Viability of Staphylococcus xylosus isolated from artisanal sausages for application as starter cultures in meat products Viability of Staphylococcus xylosus strains AD1 and U5 isolated from natural fermented sausages was investigated as starter cultures in fermented sausages produced in the South Region of Brazil. The study demonstrated that the Staphylococcus xylosus strains AD1 and U5 showed significant growth during fermentation, stability over freeze-dried process, negative reaction for staphylococcal enterotoxins and viability for using as a single-strain culture or associated with lactic acid bacteria for production of fermented sausages.

Cell viability monitoring using Fano resonance in gold nanoslit array

NASA Astrophysics Data System (ADS)

Wu, Shu-Han; Hsieh, Shu-Yi; Lee, Kuang-Li; Weng, Ruei-Hung; Chiou, Arthur; Wei, Pei-Kuen

2013-09-01

Cell viability is a crucial issue in biological research. We present label-free monitoring of adhesion cells viability by gold nanoslits-based Fano resonance biosensors. Plastic multiple wells with gold nanoslits substrate were made using a thermal nanoimprint method. Adhesion cells in the wells were treated with doxorubicin for inducing cell death and compared with conventional colorimetric assay. The nanoslits method shows better respones of viability tests under low concentration and short interaction time due to its high surface sensitivies. The vinculin labelling indicates that the measured signals are in good agreement with the adhesion abilities of cells.

Viability of human fibroblasts in coconut water as a storage medium.

PubMed

Moreira-Neto, J J S; Gondim, J O; Raddi, M S G; Pansani, C A

2009-09-01

To evaluate the effectiveness of a new storage medium for avulsed teeth, coconut water, in maintaining the viability of human fibroblasts. Cell viability after different time periods was evaluated in the following storage media: coconut water, coconut water with sodium bicarbonate, milk, saline and still mineral water. Human fibroblasts were seeded in Eagle's minimal essential medium (EMEM) supplemented with 7.5% foetal calf serum. After trypsinisation, 100 microL of culture medium containing approximately 10(4) cells mL(-1) were collected and pipetted into the wells of 96-well plates, which were incubated overnight in 5% CO(2) and 95% air mixture at 37 degrees C. EMEM was then replaced by the storage media and the plates were incubated at 37 degrees C for 1, 2 and 4 h. Cell viability was determined using the neutral red assay. The proportions of viable cells after exposure to the storage media were analysed statistically by anova and the least significant difference (LSD) test (alpha = 5%). Milk had the greatest capacity to maintain cell viability (P < 0.05), followed by coconut water with sodium bicarbonate and saline. Coconut water was significantly worse at maintaining cell viability compared to milk, coconut water with sodium bicarbonate and saline. The smallest number of viable cells was observed for mineral water (P < 0.05). Coconut water was worse than milk in maintaining human fibroblast cell viability.

Soy milk as a storage medium to preserve human fibroblast cell viability: an in vitro study.

PubMed

Moura, Camilla Christian Gomes; Soares, Priscilla Barbosa Ferreira; Reis, Manuella Verdinelli de Paula; Fernandes Neto, Alfredo Júlio; Soares, Carlos José

2012-01-01

Soy milk (SM) is widely consumed worldwide as a substitute for cow milk. It is a source of vitamins, carbohydrates and sugars, but its capacity to preserve cell viability has not been evaluated. The purpose of the present study was to investigate the efficacy of SM to maintain the viability of human fibroblasts at short periods compared with different cow milks. Human mouth fibroblasts were cultured and stored in the following media at room temperature: 10% Dulbecco's Modified Eagle Medium (DMEM) (positive control group); long shelf-life ultra-high temperature whole cow milk (WM); long shelf-life ultra-high temperature skim cow milk (SKM); powdered cow milk (PM); and soy milk (SM). After 5, 15, 30 and 45 min, cell viability was analyzed using the MTT assay. Data were analyzed statistically by the Kruskal-Wallis test with post-analysis using the Dunn's method (α=0.05). SKM showed the lowest capacity to maintain cell viability in all analyzed times (p<0.05). At 30 and 45 min, the absorbance levels in control group (DMEM) and SM were significantly higher than in SKM (p<0.05). Cell viability decreased along the time (5-45 min). The results indicate that SM can be used as a more adequate storage medium for avulsed teeth. SKM was not as effective in preserving cell viability as the cell culture medium and SM.

What Are the Security Requirements for a Two-State Solution between Israel and Palestine?

DTIC Science & Technology

2011-03-01

Economic Viability of a Palestinian State...no. 2 (Spring, 1997), 215–229, http://www.jstor.org/stable/4329054. 103 Abed, The Economic Viability of a Palestinian State, 9. 58 might have...LIST OF REFERENCES Abed, George T. The Economic Viability of a Palestinian State. (Washington D.C.: Institute for Palestine Studies, 1990).

Viability and Virulence of Experimentally Stressed Nonculturable Salmonella typhimurium

PubMed Central

Caro, Audrey; Got, Patrice; Lesne, Jean; Binard, Sylvie; Baleux, Bernard

1999-01-01

Maintenance of pathogenicity of viable but nonculturable Salmonella typhimurium cells experimentally stressed with UV-C and seawater, was investigated relative to the viability level of the cellular population. Pathogenicity, tested in a mouse model, was lost concomitantly with culturability, whereas cell viability remained undamaged, as determined by respiratory activity and cytoplasmic membrane and genomic integrities. PMID:10388726

Scheduling viability tests for seeds in long-term storage based on a Bayesian Multi-Level Model

USDA-ARS?s Scientific Manuscript database

Genebank managers conduct viability tests on stored seeds so they can replace lots that have viability near a critical threshold, such as 50 or 85% germination. Currently, these tests are typically scheduled at uniform intervals; testing every 5 years is common. A manager needs to balance the cost...

45 CFR 1302.20 - Grantee to show both legal status and financial viability.

Code of Federal Regulations, 2010 CFR

2010-10-01

... both legal status and financial viability. (a) Upon the occurrence of a change in the legal condition... legal status and financial viability within 30 days after receiving the grantee's written submittal. (c... 45 Public Welfare 4 2010-10-01 2010-10-01 false Grantee to show both legal status and financial...

The effect of pressure and shear on tissue viability of human skin in relation to the development of pressure ulcers: a systematic review.

PubMed

Hoogendoorn, Iris; Reenalda, Jasper; Koopman, Bart F J M; Rietman, Johan S

2017-08-01

Pressure ulcers are a significant problem in health care, due to high costs and large impact on patients' life. In general, pressure ulcers develop as tissue viability decreases due to prolonged mechanical loading. The relation between load and tissue viability is highly influenced by individual characteristics. It is proposed that measurements of skin blood flow regulation could provide good assessment of the risk for pressure ulcer development, as skin blood flow is essential for tissue viability. . Therefore, the aim of this systematic review is to gain insight in the relation between mechanical load and the response of the skin and underlying tissue to this loading measured in-vivo with non-invasive techniques. A systematic literature search was performed to identify articles analysing the relation between mechanical load (pressure and/or shear) and tissue viability measured in-vivo. Two independent reviewers scored the methodological quality of the 22 included studies. Methodological information as well as tissue viability parameters during load application and after load removal were extracted from the included articles and used in a meta-analysis. Pressure results in a decrease in skin blood flow parameters, compared to baseline; showing a larger decrease with higher magnitudes of load. The steepness of the decrease is mostly dependent on the anatomical location. After load removal the magnitude of the post-reactive hyperaemic peak is related to the magnitude of pressure. Lastly, shear in addition to pressure, shows an additional negative effect, but the effect is less apparent than pressure on skin viability. Copyright © 2017 Tissue Viability Society. Published by Elsevier Ltd. All rights reserved.

Mps1 kinase regulates tumor cell viability via its novel role in mitochondria

PubMed Central

Zhang, X; Ling, Y; Guo, Y; Bai, Y; Shi, X; Gong, F; Tan, P; Zhang, Y; Wei, C; He, X; Ramirez, A; Liu, X; Cao, C; Zhong, H; Xu, Q; Ma, R Z

2016-01-01

Targeting mitotic kinase monopolar spindle 1 (Mps1) for tumor therapy has been investigated for many years. Although it was suggested that Mps1 regulates cell viability through its role in spindle assembly checkpoint (SAC), the underlying mechanism remains less defined. In an endeavor to reveal the role of high levels of mitotic kinase Mps1 in the development of colon cancer, we unexpectedly found the amount of Mps1 required for cell survival far exceeds that of maintaining SAC in aneuploid cell lines. This suggests that other functions of Mps1 besides SAC are also employed to maintain cell viability. Mps1 regulates cell viability independent of its role in cytokinesis as the genetic depletion of Mps1 spanning from metaphase to cytokinesis affects neither cytokinesis nor cell viability. Furthermore, we developed a single-cycle inhibition strategy that allows disruption of Mps1 function only in mitosis. Using this strategy, we found the functions of Mps1 in mitosis are vital for cell viability as short-term treatment of mitotic colon cancer cell lines with Mps1 inhibitors is sufficient to cause cell death. Interestingly, Mps1 inhibitors synergize with microtubule depolymerizing drug in promoting polyploidization but not in tumor cell growth inhibition. Finally, we found that Mps1 can be recruited to mitochondria by binding to voltage-dependent anion channel 1 (VDAC1) via its C-terminal fragment. This interaction is essential for cell viability as Mps1 mutant defective for interaction fails to main cell viability, causing the release of cytochrome c. Meanwhile, deprivation of VDAC1 can make tumor cells refractory to loss of Mps1-induced cell death. Collectively, we conclude that inhibition of the novel mitochondrial function Mps1 is sufficient to kill tumor cells. PMID:27383047

Mps1 kinase regulates tumor cell viability via its novel role in mitochondria.

PubMed

Zhang, X; Ling, Y; Guo, Y; Bai, Y; Shi, X; Gong, F; Tan, P; Zhang, Y; Wei, C; He, X; Ramirez, A; Liu, X; Cao, C; Zhong, H; Xu, Q; Ma, R Z

2016-07-07

Targeting mitotic kinase monopolar spindle 1 (Mps1) for tumor therapy has been investigated for many years. Although it was suggested that Mps1 regulates cell viability through its role in spindle assembly checkpoint (SAC), the underlying mechanism remains less defined. In an endeavor to reveal the role of high levels of mitotic kinase Mps1 in the development of colon cancer, we unexpectedly found the amount of Mps1 required for cell survival far exceeds that of maintaining SAC in aneuploid cell lines. This suggests that other functions of Mps1 besides SAC are also employed to maintain cell viability. Mps1 regulates cell viability independent of its role in cytokinesis as the genetic depletion of Mps1 spanning from metaphase to cytokinesis affects neither cytokinesis nor cell viability. Furthermore, we developed a single-cycle inhibition strategy that allows disruption of Mps1 function only in mitosis. Using this strategy, we found the functions of Mps1 in mitosis are vital for cell viability as short-term treatment of mitotic colon cancer cell lines with Mps1 inhibitors is sufficient to cause cell death. Interestingly, Mps1 inhibitors synergize with microtubule depolymerizing drug in promoting polyploidization but not in tumor cell growth inhibition. Finally, we found that Mps1 can be recruited to mitochondria by binding to voltage-dependent anion channel 1 (VDAC1) via its C-terminal fragment. This interaction is essential for cell viability as Mps1 mutant defective for interaction fails to main cell viability, causing the release of cytochrome c. Meanwhile, deprivation of VDAC1 can make tumor cells refractory to loss of Mps1-induced cell death. Collectively, we conclude that inhibition of the novel mitochondrial function Mps1 is sufficient to kill tumor cells.

The effect of simultaneous exposure of HEMn-DP and HEMn-LP melanocytes to nicotine and UV-radiation on the cell viability and melanogenesis

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

Delijewski, Marcin; Wrześniok, Dorota; Beberok, Ar

Nicotine is a main compound of tobacco plants and may affect more than a billion people all over the world that are permanently exposed to nicotine from cigarettes, various forms of smoking cessation therapies, electronic cigarettes or second-hand smoke. It is known that nicotine forms complexes with melanin what may lead to accumulation of this alkaloid in tissues of living organisms containing the pigment. This may affect the viability of cells and process of melanin biosynthesis that takes place in melanocytes. Although UV radiation is known to be a particular inductor of melanin biosynthesis, its simultaneous effect with nicotine onmore » this process as well as the viability of human cells containing melanin have not been assessed so far. The aim of this study was to examine the simultaneous impact of nicotine and UV radiation on viability and melanogenesis in cultured normal human melanocytes dark (HEMn-DP) and light (HEMn-LP) pigmented. Nicotine together with UV radiation induced concentration-dependent loss in melanocytes viability. The higher cell loss was observed in dark pigmented melanocytes in comparison to light pigmented cells. Simultaneous exposure of cells to nicotine and UV radiation also caused changes in melanization process in both tested cell lines. The data suggest that simultaneous exposure of melanocytes to nicotine and UV radiation up-regulates melanogenesis and affects cell viability. Observed processes are more pronounced in dark pigmented cells. - Highlights: • Nicotine and UVA induced concentration-dependent loss in melanocytes viability. • Nicotine and UVA modulated melanization process in melanocytes. • Changes in viability and melanization were more pronounced in dark pigmented cells.« less

Clinical evaluation of tuberculosis viability microscopy for assessing treatment response.

PubMed

Datta, Sumona; Sherman, Jonathan M; Bravard, Marjory A; Valencia, Teresa; Gilman, Robert H; Evans, Carlton A

2015-04-15

It is difficult to determine whether early tuberculosis treatment is effective in reducing the infectiousness of patients' sputum, because culture takes weeks and conventional acid-fast sputum microscopy and molecular tests cannot differentiate live from dead tuberculosis. To assess treatment response, sputum samples (n=124) from unselected patients (n=35) with sputum microscopy-positive tuberculosis were tested pretreatment and after 3, 6, and 9 days of empiric first-line therapy. Tuberculosis quantitative viability microscopy with fluorescein diacetate, quantitative culture, and acid-fast auramine microscopy were all performed in triplicate. Tuberculosis quantitative viability microscopy predicted quantitative culture results such that 76% of results agreed within ±1 logarithm (rS=0.85; P<.0001). In 31 patients with non-multidrug-resistant (MDR) tuberculosis, viability and quantitative culture results approximately halved (both 0.27 log reduction, P<.001) daily. For patients with non-MDR tuberculosis and available data, by treatment day 9 there was a >10-fold reduction in viability in 100% (24/24) of cases and quantitative culture in 95% (19/20) of cases. Four other patients subsequently found to have MDR tuberculosis had no significant changes in viability (P=.4) or quantitative culture (P=.6) results during early treatment. The change in viability and quantitative culture results during early treatment differed significantly between patients with non-MDR tuberculosis and those with MDR tuberculosis (both P<.001). Acid-fast microscopy results changed little during early treatment, and this change was similar for non-MDR tuberculosis vs MDR tuberculosis (P=.6). Tuberculosis quantitative viability microscopy is a simple test that within 1 hour predicted quantitative culture results that became available weeks later, rapidly indicating whether patients were responding to tuberculosis therapy. © The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America.

Assessment of myocardial viability: comparison of echocardiography versus cardiac magnetic resonance imaging in the current era.

PubMed

Tomlinson, David R; Becher, Harald; Selvanayagam, Joseph B

2008-06-01

Detecting viable myocardium, whether hibernating or stunned, is of clinical significance in patients with coronary artery disease and left ventricular dysfunction. Echocardiographic assessments of myocardial thickening and endocardial excursion during dobutamine infusion provide a highly specific marker for myocardial viability, but with relatively less sensitivity. The additional modalities of myocardial contrast echocardiography and tissue Doppler have recently been proposed to provide further, quantitative measures of myocardial viability assessment. Cardiac magnetic resonance (CMR) has become popular for the assessment of myocardial viability as it can assess cardiac function, volumes, myocardial scar, and perfusion with high-spatial resolution. Both 'delayed enhancement' CMR and dobutamine stress CMR have important roles in the assessment of patients with ischaemic cardiomyopathy. This article reviews the recent advances in both echocardiography and CMR for the clinical assessment of myocardial viability. It attempts to provide a pragmatic approach toward the patient-specific assessment of this important clinical problem.

A comparison of TO-PRO-1 iodide and 5-CFDA-AM staining methods for assessing viability of planktonic algae with epifluorescence microscopy.

PubMed

Gorokhova, Elena; Mattsson, Lisa; Sundström, Annica M

2012-06-01

Two fluorescent dyes, TO-PRO-1 iodide and 5-CFDA-AM, were evaluated for LIVE/DEAD assessment of unicellular marine algae Brachiomonas submarina and Tetraselmis suecica. Epifluorescence microscopy was used to estimate cell viability in predetermined mixtures of viable and non-viable algal cells and validated using microplate growth assay as reference measurements. On average, 5-CFDA-AM underestimated live cell abundance by ~25% compared with viability estimated by the growth assay, whereas TO-PRO-1 iodide provided accurate viability estimates. Furthermore, viability estimates based on staining with TO-PRO-1 iodide were not affected by a storage period of up to one month in -80°C, making the assay a good candidate for routine assessment of phytoplankton populations in field and laboratory studies. Copyright © 2012 Elsevier B.V. All rights reserved.

Exposure-related effects of formulated Pseudomonas fluorescens strain CL145A to glochidia from seven unionid mussel species

USGS Publications Warehouse

Luoma, James A.; Weber, Kerry L.; Severson, Todd J.; Schreier, Theresa M.; Mayer, Denise A.; Aloisi, Douglas B.; Eckert, Nathan L.

2015-01-01

Glochidia viability was reduced in two of the six species exposed to 50 mg/L SDP and in four of the six species exposed to 100 mg/L SDP when compared to untreated control groups at 6, 12, and 24 hours. Regardless of sample time, concentrations of 200 and 300 mg/L of SDP and 300 mg/L of heat-deactivated SDP (positive control) substantially reduced glochidia viability in all species except, L. higginsii. Glochidia viability was only reduced for L. cardium exposed to FDP at concentrations ≥ 200 mg/L. After 24 hours of FDP exposure, differences in glochidia viability were only detected in M. nervosa that were exposed to 300 mg/L of heat-deactivated SDP. However, given the low viability in the control group, the results for M. nervosa should be interpreted with caution.

Long term cryopreservation in 5% DMSO maintains unchanged CD34(+) cells viability and allows satisfactory hematological engraftment after peripheral blood stem cell transplantation.

PubMed

Abbruzzese, L; Agostini, F; Durante, C; Toffola, R T; Rupolo, M; Rossi, F M; Lleshi, A; Zanolin, S; Michieli, M; Mazzucato, M

2013-07-01

Peripheral blood stem cell cryopreservation is associated with cell damage and decreased viability. We evaluated the impact of up to 10 years of cryopreservation (5% DMSO) on viability of CD34(+) cells utilizing graft samples of consecutive patients (2002-2012) with different malignancies who underwent stem cell collection and transplantation. Viability of CD34(+) cells from oncohaematological patients measured after 5 weeks (97·2 ± 0·6%) or after 9-10 years of cryopreservation (95·9 ± 0·5%) was unaffected. Haemoglobin, granulocyte and platelet recovery after transplantation of long-term cryopreserved grafts occurred within 8-13 days. CD34(+) stem cells can be safely stored up to 9-10 years, without affecting cell viability and clinical effectiveness. © 2013 International Society of Blood Transfusion.

Doctors' perspectives on the viability of rural practice.

PubMed

Jones, J A; Humphreys, J S; Adena, M A

2004-01-01

Private practitioners play a vital role in meeting the health needs of rural communities. However, the prospect of operating a private practice business in rural Australia seems to be increasingly unattractive, because many communities are forced to recruit salaried or overseas-trained doctors. This study focuses on rural practices as businesses whose viability influences their attractiveness for the recruitment and retention of practitioners. The specific objectives are to ascertain which factors contribute to or threaten practice viability in rural areas, and whether they vary according to the degree of rurality or geographical remoteness. This study is based on data collected from a national study into the viability of rural general practice undertaken jointly by the Rural Doctors Association of Australia and Monash University School of Rural Health Bendigo. The Rural Remote and Metropolitan Area (RRMA) classification was used as the indicator of rurality. The study surveyed all general practitioners practising in rural or remote regions of Australia (RRMAs 3 to 7). Only practitioners with some financial interest in the practice were selected for this analysis. Free-text responses to the two questions 'What are the key factors contributing to the viability of your practice?' and 'What factors would put the viability of your practice at risk?' were analysed using qualitative content analysis. Factors were derived iteratively through higher-level aggregation of responses. Chi-square tests were used to make comparisons across the RRMA categories. The national survey achieved a response rate of 35% of the entire population of GPs practising in RRMA 3 to 7 regions. Of these, 1050 respondents were relevant to this analysis. Seven major factors were identified by practitioners as the main contributors to practice viability. 'Practice characteristics' was nominated by 59% of respondents, followed by 'Income' (31%), 'Personal circumstances', 'Workforce' and 'Community characteristics' (all approximately 23%), 'GP activities and workload' (16%) and 'Professional support' (12%). Eight main factors were identified by practitioners as threats to viability. 'Workforce' was nominated by 57% of respondents, followed by 'Financial' (44%), 'Medico-legal' (33%), 'Administration-political' (16%), 'Community characteristics' (15%), 'GP-practice characteristics' and 'Personal circumstances' (10%) and 'Family circumstances' (3%). Across RRMA 3 to 5 the order of the percentage of respondents identifying each factor was generally consistent, with significant differences in the magnitude of the percentages for three contributing factors and four risk factors. While respondent numbers in RRMA 6 and 7 communities were low, significance testing did reveal differences between them and the rural communities on two contributing and one risk factor. Practice viability is a major factor affecting the attractiveness of rural and remote practice for intending and existing GPs. Initiatives designed to contribute to viability will not be successful unless measures are also adopted to address perceived threats. This study highlights the systemic nature of the factors which contribute to and threaten practice viability. Although a primary component of practice viability is economic, with income from consultations being critical, the importance of the interrelationships between the main viability factors should not be underestimated. Clearly a multifaceted systemic response is required to overcome problems associated with rural workforce recruitment of future and burnout of current rural GPs.

In vitro radiolabel uptake viability assay for Onchocerca microfilariae

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

Callahan, H.L.; Wakeman, J.M.; Crouch, R.K.

1989-02-01

A radiolabel uptake viability assay for Onchocerca cervicalis using (/sup 3/H)2-deoxy-D-glucose in Hanks' balanced salt solution, pH 7.5, at 30 C is described and compared to the traditional visual motility assay. A correlation of r = 0.92 between the assays was found, with the radiolabel uptake method apparently a more sensitive indicator of microfilarial viability.

Poxvirus viability and signatures in historical relics.

PubMed

McCollum, Andrea M; Li, Yu; Wilkins, Kimberly; Karem, Kevin L; Davidson, Whitni B; Paddock, Christopher D; Reynolds, Mary G; Damon, Inger K

2014-02-01

Although it has been >30 years since the eradication of smallpox, the unearthing of well-preserved tissue material in which the virus may reside has called into question the viability of variola virus decades or centuries after its original occurrence. Experimental data to address the long-term stability and viability of the virus are limited. There are several instances of well-preserved corpses and tissues that have been examined for poxvirus viability and viral DNA. These historical specimens cause concern for potential exposures, and each situation should be approached cautiously and independently with the available information. Nevertheless, these specimens provide information on the history of a major disease and vaccination against it.

Viability of Staphylococcus xylosus isolated from artisanal sausages for application as starter cultures in meat products

PubMed Central

Fiorentini, Ângela Maria; Sawitzki, Maristela Cortez; Bertol, Teresinha Marisa; Sant’Anna, Ernani S.

2009-01-01

Viability of Staphylococcus xylosus isolated from artisanal sausages for application as starter cultures in meat products Viability of Staphylococcus xylosus strains AD1 and U5 isolated from natural fermented sausages was investigated as starter cultures in fermented sausages produced in the South Region of Brazil. The study demonstrated that the Staphylococcus xylosus strains AD1 and U5 showed significant growth during fermentation, stability over freeze-dried process, negative reaction for staphylococcal enterotoxins and viability for using as a single-strain culture or associated with lactic acid bacteria for production of fermented sausages. PMID:24031331

Assessment of cell concentration and viability of isolated hepatocytes using flow cytometry.

PubMed

Wigg, Alan J; Phillips, John W; Wheatland, Loretta; Berry, Michael N

2003-06-01

The assessment of cell concentration and viability of freshly isolated hepatocyte preparations has been traditionally performed using manual counting with a Neubauer counting chamber and staining for trypan blue exclusion. Despite the simple and rapid nature of this assessment, concerns about the accuracy of these methods exist. Simple flow cytometry techniques which determine cell concentration and viability are available yet surprisingly have not been extensively used or validated with isolated hepatocyte preparations. We therefore investigated the use of flow cytometry using TRUCOUNT Tubes and propidium iodide staining to measure cell concentration and viability of isolated rat hepatocytes in suspension. Analysis using TRUCOUNT Tubes provided more accurate and reproducible measurement of cell concentration than manual cell counting. Hepatocyte viability, assessed using propidium iodide, correlated more closely than did trypan blue exclusion with all indicators of hepatocyte integrity and function measured (lactate dehydrogenase leakage, cytochrome p450 content, cellular ATP concentration, ammonia and lactate removal, urea and albumin synthesis). We conclude that flow cytometry techniques can be used to measure cell concentration and viability of isolated hepatocyte preparations. The techniques are simple, rapid, and more accurate than manual cell counting and trypan blue staining and the results are not affected by protein-containing media.

Mechanism of H₂O₂-induced oxidative stress regulating viability and biocontrol ability of Rhodotorula glutinis.

PubMed

Chen, Jian; Li, Boqiang; Qin, Guozheng; Tian, Shiping

2015-01-16

The use of antagonistic yeasts to control postharvest pathogens is a promising alternative to fungicides. The effectiveness of the antagonists against fungal pathogens is greatly dependent on their viability, which is usually mediated by reactive oxygen species (ROS). Here, we investigated the effects of H₂O₂-induced oxidative stress on the viability and biocontrol efficacy of Rhodotorula glutinis and, using flow cytometric analysis, observed the changes of ROS accumulation and apoptosis in the yeast cells with or without H₂O₂ treatment. We found that the viability of R. glutinis decreased in a time- and dose-dependent manner under H₂O₂-induced oxidative stress. Compared to the control, yeast cells exposed to oxidative stress exhibited more accumulation of ROS and higher levels of protein oxidative damage, but showed lower efficacy for biocontrol of Penicillium expansum causing blue mold rot on peach fruit. The results indicate that apoptosis is a main cause of the cell viability loss in R. glutinis, which is attributed to ROS accumulation under oxidative stress. These findings offer a plausible explanation that oxidative stress affects biocontrol efficacy of R. glutinis via regulating its viability and cell apoptosis. Copyright © 2014 Elsevier B.V. All rights reserved.

The role of adrenergic activation on murine luteal cell viability and progesterone production.

PubMed

Wang, Jing; Tang, Min; Jiang, Huaide; Wu, Bing; Cai, Wei; Hu, Chuan; Bao, Riqiang; Dong, Qiming; Xiao, Li; Li, Gang; Zhang, Chunping

2016-09-15

Sympathetic innervations exist in mammalian CL. The action of catecholaminergic system on luteal cells has been the focus of a variety of studies. Norepinephrine (NE) increased progesterone secretion of cattle luteal cells by activating β-adrenoceptors. In this study, murine luteal cells were treated with NE and isoprenaline (ISO). We found that NE increased the viability of murine luteal cells and ISO decreased the viability of luteal cells. Both NE and ISO promoted the progesterone production. Nonselective β-adrenergic antagonist, propranolol reversed the effect of ISO on cell viability but did not reverse the effect of NE on cell viability. Propranolol blocked the influence of NE and ISO on progesterone production. These results reveal that the increase of luteal cell viability induced by NE is not dependent on β-adrenergic activation. α-Adrenergic activation possibly contributes to it. Both NE and ISO increased progesterone production through activating β-adrenergic receptor. Further study showed that CyclinD2 is involved in the increase of luteal cell induced by NE. 3β-Hydroxysteroid dehydrogenase, LHR, steroidogenic acute regulatory protein (StAR), and PGF2α contribute to the progesterone production induced by NE and ISO. Copyright © 2016 Elsevier Inc. All rights reserved.

Fluorescein diacetate for determination of cell viability in 3D fibroblast-collagen-GAG constructs.

PubMed

Powell, Heather M; Armour, Alexis D; Boyce, Steven T

2011-01-01

Quantification of cell viability and distribution within engineered tissues currently relies on representative histology, phenotypic assays, and destructive assays of viability. To evaluate uniformity of cell density throughout 3D collagen scaffolds prior to in vivo use, a nondestructive, field assessment of cell viability is advantageous. Here, we describe a field measure of cell viability in lyophilized collagen-glycosaminoglycan (C-GAG) scaffolds in vitro using fluorescein diacetate (FdA). Fibroblast-C-GAG constructs are stained 1 day after cellular inoculation using 0.04 mg/ml FdA followed by exposure to 366 nm UV light. Construct fluorescence quantified using Metamorph image analysis is correlated with inoculation density, MTT values, and histology of corresponding biopsies. Construct fluorescence correlates significantly with inoculation density (p  <  0.001) and MTT values (p  <  0.001) of biopsies collected immediately after FdA staining. No toxicity is detected in the constructs, as measured by MTT assay before and after the FdA assay at different time points; normal in vitro histology is demonstrated for the FdA-exposed constructs. In conclusion, measurement of intracellular fluorescence with FdA allows for the early, comprehensive measurement of cellular distributions and viability in engineered tissue.