Causative ehrlichial organisms in Potomac horse fever.

PubMed

Rikihisa, Y; Perry, B D

1985-09-01

An ehrlichia was consistently isolated from the peripheral blood leukocyte fraction of ponies that had been experimentally infected with Potomac horse fever by whole blood transfusion from naturally infected horses. The organism was propagated in a human histiocyte cell line for 3 to 5 weeks and then inoculated intravenously or intradermally into healthy adult ponies. Clinical signs of Potomac horse fever, which varied in the degree of severity, occurred 9 to 14 days post-inoculation in all of the ponies. One pony died 20 days post-inoculation. The ehrlichial organism was reisolated in the human histiocyte cell line from the blood leukocyte fraction of all of the experimental ponies on each day that samples were examined (days 9, 10, 11, 19, and 39). These organisms were identical to those originally detected in the wall of the intestine of ponies with clinically diagnosed Potomac horse fever when compared by light and electron microscopy and an immunofluorescence labeling technique. The immunofluorescent antibody titer became positive in a pony at 20 days postinjection. These results indicate that the ehrlichial organisms is the causative agent of Potomac horse fever.

Evaluation of antiviral efficacy of Chinese traditional medicine Babao Dan in rabbits infected with hepatitis E virus.

PubMed

Gong, Wanyun; Liu, Lin; Li, Manyu; Wang, Lin; Zhang, Mingyu; Luo, Zhengxin; Sridhar, Siddharth; Woo, Patrick C Y; Wang, Ling

2018-06-20

Hepatitis E virus (HEV) is a major cause of acute viral hepatitis. Patients with chronic hepatitis B superinfected with HEV may progress to liver failure. Babao Dan (BD) is a traditional Chinese medicine widely used as an auxiliary option for the treatment of chronic hepatitis and liver cancer in China. This study aimed to evaluate the effect of BD on the management of HEV infection in a rabbit model. Sixty-two specific-pathogen-free (SPF) rabbits were divided randomly into five groups and treated with BD or placebo for 2 weeks. All rabbits were inoculated intravenously with rabbit HEV after initial administration. Then, rabbits were administered BD or ribavirin or placebo at 2 weeks post-inoculation (wpi) until faecal virus shedding showed negative. The duration of faecal virus shedding and levels of HEV RNA in faeces were reduced, and anti-HEV antibodies were detected in all rabbits in groups treated with BD before or after inoculation. Ribavirin treatment rapidly cleared HEV infection in SPF rabbits, but anti-HEV antibodies remained negative in 50 % of rabbits treated with ribavirin. These results indicate that ribavirin treatment was more effective in clearing HEV infection, while administration of BD before or after inoculation was effective in clearing HEV infection. Further clinical studies are warranted.

Research on Dengue during World War II Revisited

PubMed Central

Snow, Grace E.; Haaland, Benjamin; Ooi, Eng Eong; Gubler, Duane J.

2014-01-01

Much of the basic clinical information about dengue infection comes from experimental human studies conducted in the 1920s and 1940s. Albert Sabin's original laboratory records from one such study were bequeathed to Duane J. Gubler. These records were reviewed and 150 experiments were included in our analyses. Persons were inoculated with dengue virus 1 (DENV-1) and DENV-2. Median fever duration was shorter in primary DENV-2 infections compared with DENV-1, although maximum temperature and severity of illness were comparable. At 1.5–9 months after primary infection, 20 persons were inoculated with the heterologous serotype. Only one person inoculated with a heterologous serotype at < 8 weeks showed development of a clinical infection with a maximum temperature of 38°C, and 7 (88%) of 8 persons inoculated with a heterologous serotype at 4–9 months post-primary infection showed development of fever. On average, persons had a shorter incubation period in secondary infection compared with primary infection. PMID:25311700

[Effects of hypoxia inducible factor-2α on promoting angiogenesis of residual hepatocellular carcinoma after high-intensity focused ultrasound ablation].

PubMed

Wu, Lun; Zhou, Wenbo; Zhou, Shiji; Liu, Chang'an; Li, Shengwei

2015-02-01

To investigate the dynamic features of angiogenesis in residual tumors after high intensity focused ultrasound (HIFU),and to determine the temporal effect and mechanism of hypoxia inducible factor-2 alpha (HIF-2a) in the angiogenic process of residual tumors. Xenograft tumors of HepG2 cells were generated by subcutaneously inoculating athymic BALB/c nu/nu mice with the hepatoma cells.About 30 days after inoculation,all mice (except in the control group) were treated by HIFU and assigned randomly to the following 7 groups according to various time intervals post-treatment:1st,3rd,5th day and 1st,2nd,3rd,4th week when the residual tumor tissues were obtained from the experimental groups.Protein levels of HIF-2a and vascular growth factor A (VEGF-A) were quantified by immunohistochemistry and western blotting,and mRNA levels were measured by (real-time quantitative) qPCR. Microvascular density (MVD) was calculated by counting the CD31-positive vascular endothelial cells identified by means of an immunohistochemical staining method. Compared with results from the control group,the protein and mRNA levels of HIF-2a expression reached the highest level in the experimental mice at the 2nd week (P=0.000 and P < 0.01 respectively),and were decreased thereafter(3rd week and 4th week, P=0.000 and P < 0.05).VEGF-A expression in the residual tumor tissues group that received HIFU was significantly decreased,compared with the control group,at all time points uPto 1 week (all P=0.000 and P < 0.01),but the levels increased compared to controls in the 2nd through 4th week (all P=0.000, P < 0.05). Similar results were obtained for MVD. HIFU treatment can inhibit angiogenesis in residual hepatoma tissues in the short-term (1 to 2 weeks post-treatment) in mice with hepatocellular carcinoma,but can promote angiogenesis overtime (2 to 4 weeks post-treatment); the angiogenic process may involve the HIF-2α/VEGFA pathway.

Sequential mesenteric arteriography in pony foals during repeated inoculations of Strongylus vulgaris and treatments with ivermectin.

PubMed

Holmes, R A; Klei, T R; McClure, J R; Turk, M A; Watters, J W; Chapman, M R

1990-04-01

Semiselective mesenteric arteriography was performed at regular intervals (inoculation weeks [IW] 0, 11, 18, and 24) in 9 of 10 pony foals raised to be free of parasites. Fifty infective larvae (L3) of Strongylus vulgaris were administered weekly for 4 weeks, then every 2 weeks through the 20th week. Three ponies were given ivermectin (oral paste, 0.2 mg/kg of body weight) treatment at IW 8, 16 and 24. Four ponies were inoculated, but did not receive ivermectin, and a third group of 2 ponies acted as uninoculated controls. Control ponies did not have gross or arteriographic lesions, whereas the inoculated untreated ponies had gross and progressive arteriographic lesions typical of verminous arteritis. Arteriographic lesions in the ivermectin-treated inoculated ponies were not as severe those in the untreated inoculated group, and there was either a partial resolution or a lack of progression of arteriographic lesions in all treated ponies. One untreated inoculated pony did not have progressive arterial lesions as did the 3 others in the group, and may develop resistance to the parasite.

Research on the influence of anaerobic stabilization of various dairy sewage sludge on biodegradation of polycyclic aromatic hydrocarbons PAHs with the use of effective microorganisms.

PubMed

Boruszko, Dariusz

2017-05-01

Sewage sludge was taken from a dairy WWTP belonging to Mlekovita Cooperative in Wysokie Mazowieckie. There were excess sludge, flotation sludge and a mixture of excess and flotation sludge from pre-treatment of dairy sewage. The initial content of 16 PAHs in excess sludge before fermentation was approximately 689µg·kg -1 in dry mass, whereas in post-flotation sludge (which constituted around 30% of raw sludge) it was approximately 95µg·kg -1 in dry mass. A mixture of excess and flotation sludge had the content of 497,7µg·kg -1 in dry mass. Through comparison of particular hydrocarbons content in raw sewage sludge to the total PAHs content, it was shown that tricyclic compounds, which constituted 46,3% of the PAHs sum (excess sludge), and tetracyclic compounds, which constituted 60,0% of the PAHs sum (flotation sludge), were the dominating fractions. In the sludge subjected to fermentation in reactors with mixed sludge and surplus activated sludge, the general trend of the course of changes in concentrations of PAHs was similar. Both in the sludge inoculated with EM and in that not inoculated with EM, a significant increase in the total PAHs contents was observed in the first fermentation phase (acidic fermentation) after 7 days of the process. Addition of EM into the sludge did not prevent the PAHs release, and therefore higher concentrations of PAHs sum were recorded during the hydrolysis stage than in sludge before fermentation. A decrease in the sum of PAHs was observed after 2 weeks of fermentation in relation to the quantity observed after 1 week of fermentation (except from post-flotation sludge). In the following weeks, there was further decrease in the concentration of the 16 PAHs sum in all sludge types. However, in sludge without EM inoculation, it was lower than in sludge with EM inoculation. The loss of the majority of tested hydrocarbons was reported in the final phase of fermentation. Copyright © 2017 Elsevier Inc. All rights reserved.

Modeling Powassan virus infection in Peromyscus leucopus, a natural host

PubMed Central

Meade-White, Kimberly; Saturday, Greg; Scott, Dana; Bloom, Marshall E.

2017-01-01

The tick-borne flavivirus, Powassan virus (POWV) causes life-threatening encephalitis in humans in North America and Europe. POWV is transmitted by ixodid tick vectors that feed on small to medium-sized mammals, such as Peromyscus leucopus mice, which may serve as either reservoir, bridge or amplification hosts. Intraperitoneal and intracranial inoculation of 4-week old Peromyscus leucopus mice with 103 PFU of POWV did not result in overt clinical signs of disease. However, following intracranial inoculation, infected mice seroconverted to POWV and histopathological examinations revealed that the mice uniformly developed mild lymphocytic perivascular cuffing and microgliosis in the brain and spinal cord from 5 to 15 days post infection (dpi), suggesting an early inflammatory response. In contrast, intracranial inoculation of 4-week old C57BL/6 and BALB/c mice was lethal by 5 dpi. Intraperitoneal inoculation was lethal in BALB/c mice, but 40% (2/5) of C57BL/6 mice survived. We concluded that Peromyscus leucopus mice infected i.c. with a lethal dose of POWV support a limited infection, restricted to the central nervous system and mount an antibody response to the virus. However, they fail to develop clinical signs of disease and are able to control the infection. These results suggest the involvement of restriction factors, and the mechanism by which Peromyscus leucopus mice restrict POWV infection remains under study. PMID:28141800

Modeling Powassan virus infection in Peromyscus leucopus, a natural host.

PubMed

Mlera, Luwanika; Meade-White, Kimberly; Saturday, Greg; Scott, Dana; Bloom, Marshall E

2017-01-01

The tick-borne flavivirus, Powassan virus (POWV) causes life-threatening encephalitis in humans in North America and Europe. POWV is transmitted by ixodid tick vectors that feed on small to medium-sized mammals, such as Peromyscus leucopus mice, which may serve as either reservoir, bridge or amplification hosts. Intraperitoneal and intracranial inoculation of 4-week old Peromyscus leucopus mice with 103 PFU of POWV did not result in overt clinical signs of disease. However, following intracranial inoculation, infected mice seroconverted to POWV and histopathological examinations revealed that the mice uniformly developed mild lymphocytic perivascular cuffing and microgliosis in the brain and spinal cord from 5 to 15 days post infection (dpi), suggesting an early inflammatory response. In contrast, intracranial inoculation of 4-week old C57BL/6 and BALB/c mice was lethal by 5 dpi. Intraperitoneal inoculation was lethal in BALB/c mice, but 40% (2/5) of C57BL/6 mice survived. We concluded that Peromyscus leucopus mice infected i.c. with a lethal dose of POWV support a limited infection, restricted to the central nervous system and mount an antibody response to the virus. However, they fail to develop clinical signs of disease and are able to control the infection. These results suggest the involvement of restriction factors, and the mechanism by which Peromyscus leucopus mice restrict POWV infection remains under study.

Nonencapsulated Streptococcus pneumoniae causes otitis media during single-species infection and during polymicrobial infection with nontypeable Haemophilus influenzae

PubMed Central

Murrah, Kyle A.; Pang, Bing; Richardson, Stephen; Perez, Antonia; Reimche, Jennifer; King, Lauren; Wren, John; Swords, W. Edward

2014-01-01

Streptococcus pneumoniae strains lacking capsular polysaccharide have been increasingly reported in carriage and disease contexts. Since most cases of otitis media involve more than one bacterial species, we aimed to determine the capacity of a nonencapsulated S. pneumoniae clinical isolate to induce disease in the context of a single-species infection and as a polymicrobial infection with nontypeable Haemophilus influenzae. Using the chinchilla model of otitis media, we found that nonencapsulated S. pneumoniae colonizes the nasopharynx following intranasal inoculation, but does not readily ascend into the middle ear. However, when we inoculated nonencapsulated S. pneumoniae directly into the middle ear, the bacteria persisted for two weeks post-inoculation and induced symptoms consistent with chronic otitis media. During coinfection with nontypeable H. influenzae, both species persisted for one week and induced polymicrobial otitis media. We also observed that nontypeable H. influenzae conferred passive protection from killing by amoxicillin upon S. pneumoniae from within polymicrobial biofilms in vitro. Therefore, based on these results, we conclude that nonencapsulated pneumococci are a potential causative agent of chronic/recurrent otitis media, and can also cause mutualistic infection with other opportunists, which could complicate treatment outcomes. PMID:26014114

Fate of Listeria monocytogenes on Fresh Apples under Different Storage Temperatures.

PubMed

Sheng, Lina; Edwards, Katheryn; Tsai, Hsieh-Chin; Hanrahan, Ines; Zhu, Mei-Jun

2017-01-01

Fresh apples are typically stored for up to 1 year commercially; different apple varieties require different storage temperatures to maintain their quality characteristics. There is sparse information available about Listeria monocytogenes survival on fresh apples under various storage temperatures. The objective of this study was to comprehensively evaluate the effect of storage temperature on apple fruit decay and L. monocytogenes survival. Unwaxed apple fruits of selected varieties (Fuji and Granny Smith) were dip inoculated in a three-strain L. monocytogenes cocktail to establish ∼3.5 and 6.0 Log 10 CFU/apple. Twenty-four hours post-inoculation, apples were subjected to 1, 4, 10, or 22°C storage for up to 3 months. Apples under the different storage treatments were sampled at 1-, 4-, 7- and 14-day for short-term storage under all four tested temperatures, and 2-, 4-, 8-, and 12-week for long-term storage at 1, 4, and 10°C. A set of uninoculated and unwaxed apples were simultaneously subjected to the previously mentioned storage temperatures and sampled biweekly for their total bacterial count (TPC) and yeasts/molds (Y/M) count. During the 2-week short-term storage, L. monocytogenes population on organic Granny Smith apples stored at 1, 4, or 10°C was reduced by 0.2-0.3 Log. When apples were stored at 22°C, there was a 0.5-1.2 Log 10 CFU/apple reduction 14-day post storage dependent on the initial inoculation level. During the 12-week cold storage under 1, 4, and 10°C, L. monocytogenes count on organic Granny Smith apples decreased by 0.5-1.5 Log 10 CFU/apple for both inoculation levels. L. monocytogenes had similar survival pattern on conventional Granny Smith and Fuji apples with 0.8-2.0 Log 10 CFU/apple reduction over a 3-month cold storage period. Interestingly, both TPC and Y/M count were stable regardless of apple variety or cultivation practice during the 12-week storage at all tested temperatures. In summary, while L. monocytogenes did not proliferate on apple surfaces during 12 weeks of refrigerated storage, only a limited reduction of L. monocytogenes was observed in this study. Therefore, the apple industry cannot rely on cold storage alone to control this pathogen. Additional interventions are needed to eradicate Listeria on fresh apples during long-term cold storage.

Immunologic and hematologic responses in ponies with experimentally induced Strongylus vulgaris infection.

PubMed

Bailey, M; Martin, S C; Lloyd, S

1989-08-01

Immunologic and hematologic responses were examined in 4 ponies with experimentally induced Strongylus vulgaris infection and in 5 helminth-free ponies. Two ponies were inoculated with 200 larvae and 2 were inoculated with 700 larvae of S vulgaris and then were reinoculated with the same numbers of larvae 34 weeks later. Initial response of the ponies inoculated with S vulgaris was S vulgaris antigen-induced lymphocyte response that developed 1.5 to 3 weeks after inoculation and did not persist. Development of antigen-reactive lymphocytes was followed sequentially by a biphasic complement-fixing antibody response, then biphasic eosinophilia. Antibody titer to S vulgaris antigen was higher in ponies inoculated with 700 larvae, compared with that in ponies given 200 larvae of S vulgaris. Also, the second peak in antibody titer and in absolute number of eosinophils was observed earlier in ponies inoculated with 700 larvae, compared with ponies inoculated with 200 S vulgaris larvae, and subsided before or from about 24 weeks after inoculation. The prepatent period for S vulgaris infection was 24 to 25 weeks. After reinoculation with S vulgaris, a degree of increased lymphocyte responsiveness was apparent but, by 17 weeks after reinoculation, only the primary peak in the absolute number of eosinophils indicated an anamnestic response. Essentially, antibody was not detectable after reinoculation.

Possible role of glial cells in the onset and progression of Lyme neuroborreliosis

PubMed Central

Ramesh, Geeta; Borda, Juan T; Gill, Amy; Ribka, Erin P; Morici, Lisa A; Mottram, Peter; Martin, Dale S; Jacobs, Mary B; Didier, Peter J; Philipp, Mario T

2009-01-01

Background Lyme neuroborreliosis (LNB) may present as meningitis, cranial neuropathy, acute radiculoneuropathy or, rarely, as encephalomyelitis. We hypothesized that glia, upon exposure to Borrelia burgdorferi, the Lyme disease agent, produce inflammatory mediators that promote the acute cellular infiltration of early LNB. This inflammatory context could potentiate glial and neuronal apoptosis. Methods We inoculated live B. burgdorferi into the cisterna magna of rhesus macaques and examined the inflammatory changes induced in the central nervous system (CNS), and dorsal root nerves and ganglia (DRG). Results ELISA of the cerebrospinal fluid (CSF) showed elevated IL-6, IL-8, CCL2, and CXCL13 as early as one week post-inoculation, accompanied by primarily lymphocytic and monocytic pleocytosis. In contrast, onset of the acquired immune response, evidenced by anti-B. burgdorferi C6 serum antibodies, was first detectable after 3 weeks post-inoculation. CSF cell pellets and CNS tissues were culture-positive for B. burgdorferi. Histopathology revealed signs of acute LNB: severe multifocal leptomeningitis, radiculitis, and DRG inflammatory lesions. Immunofluorescence staining and confocal microscopy detected B. burgdorferi antigen in the CNS and DRG. IL-6 was observed in astrocytes and neurons in the spinal cord, and in neurons in the DRG of infected animals. CCL2 and CXCL13 were found in microglia as well as in endothelial cells, macrophages and T cells. Importantly, the DRG of infected animals showed significant satellite cell and neuronal apoptosis. Conclusion Our results support the notion that innate responses of glia to B. burgdorferi initiate/mediate the inflammation seen in acute LNB, and show that neuronal apoptosis occurs in this context. PMID:19706181

Experimental giardiasis in goat kids.

PubMed

Koudela, B; Vítovec, J

1998-01-15

The clinical, pathological and parasitological features of giardiasis resulting from experimental inoculation with 3 x 10(6) Giardia cysts were studied in goat kids. All experimentally inoculated goat kids given Giardia cysts became infected. Three of the eight inoculated kids had decreased appetite, formless feces and become slightly depressed beginning 7 or 8 days post inoculation. The mean duration of the appearance of abnormal feces was 6 days. Irregular and intermitted cysts shedding started after prepatent periods of 6-10 days and lasted throughout this study (10 weeks). The evidence of high infectivity and fast transmission of Giardia were observed under standard zoohygienic conditions. The characteristics of intestinal lesions were similar to those found in other hosts infected with Giardia. The most severe lesions were seen in the duodenum and proximal jejunum, and consisted of moderate villus atrophy, villus blunting, crypt hyperplasia and inflammatory infiltration in the lamina propria. Scanning electron microscopy revealed ultrastructural alterations in the microvillus border of enterocytes. Mucosal smears and histological sections of the gall bladder displayed Giardia trophozoites and gall bladder epithelium hyperplasia together with bile ductular proliferation in the liver tissue in two kids.

Prior infection of pigs with a genotype 3 swine hepatitis E virus (HEV) protects against subsequent challenges with homologous and heterologous genotypes 3 and 4 human HEV

PubMed Central

Sanford, Brenton J.; Dryman, Barbara A.; Huang, Yao-Wei; Feagins, Alicia R.; LeRoith, Tanya; Meng, Xiang-Jin

2011-01-01

Hepatitis E virus (HEV) is an important human pathogen. At least four recognized and two putative genotypes of mammalian HEV have been reported: genotypes 1 and 2 are restricted to humans whereas genotypes 3 and 4 are zoonotic. The current experimental vaccines are all based on a single strain of HEV, even though multiple genotypes of HEV are co-circulating in some countries and thus an individual may be exposed to more than one genotype. Genotypes 3 and 4 swine HEV is widespread in pigs and known to infect humans. Therefore, it is important to know if prior infection with a genotype 3 swine HEV will confer protective immunity against subsequent exposure to genotypes 3 and 4 human and swine HEV. In this study, specific-pathogen-free pigs were divided into 4 groups of 6 each. Pigs in the three treatment groups were each inoculated with a genotype 3 swine HEV, and 12 weeks later, challenged with the same genotype 3 swine HEV, a genotype 3 human HEV, and a genotype 4 human HEV, respectively. The control group was inoculated and challenged with PBS buffer. Weekly sera from all pigs were tested for HEV RNA and IgG anti-HEV, and weekly fecal samples were also tested for HEV RNA. The pigs inoculated with swine HEV became infected as evidenced by fecal virus shedding and viremia, and the majority of pigs also developed IgG anti-HEV prior to challenge at 12 weeks post-inoculation. After challenge, viremia and fecal virus shedding of challenge viruses were not detected, suggesting that prior infection with a genotype 3 swine HEV prevented pigs from developing viremia and fecal virus shedding after challenges with homologous and heterologous genotypes 3 and 4 HEV. The results from this study have important implications for future development of an effective HEV vaccine. PMID:21536085

Genetically different isolates of Trypanosoma cruzi elicit different infection dynamics in raccoons (Procyon lotor) and Virginia opossums (Didelphis virginiana)

PubMed Central

Roellig, Dawn M.; Ellis, Angela E.; Yabsley, Michael J.

2009-01-01

Trypanosoma cruzi is a genetically and biologically diverse species. In the current study we determined T. cruzi infection dynamics in two common North American reservoirs, Virginia opossums (Didelphis virginiana) and raccoons (Procyon lotor). Based on previous molecular and culture data from naturally-exposed animals, we hypothesized that raccoons would have a longer patent period than opossums, and raccoons would be competent reservoirs for both genotypes T. cruzi I (TcI) and TcIIa, while opossums would only serve as hosts for TcI. Individuals (n = 2 or 3) of each species were inoculated with 1 × 106 culture-derived T. cruzi trypomastigotes of TcIIa (North American (NA) - raccoon), TcI (NA - opossum), TcIIb (South American - human), or both TcI and TcIIa. Parasitemias in opossums gradually increased and declined rapidly, whereas parasitemias peaked sooner in raccoons and they maintained relatively high parasitemia for 5 weeks. Raccoons became infected with all three T. cruzi strains, while opossums only became infected with TcI and TcIIb. Although opossums were susceptible to TcIIb, infection dynamics were dramatically different compared with TcI. Opossums inoculated with TcIIb seroconverted, but parasitemia duration was short and only detectable by PCR. In addition, raccoons seroconverted sooner (3–7 days post inoculation) than opossums (10 days post inoculation). These data suggest that infection dynamics of various T. cruzi strains can differ considerably in different wildlife hosts. PMID:19607833

Efficacy of enrofloxacin, florfenicol and amoxicillin against Ornithobacterium rhinotracheale and Escherichia coli O2:K1 dual infection in turkeys following APV priming.

PubMed

Marien, Maja; Decostere, Annemie; Duchateau, Luc; Chiers, Koen; Froyman, Robrecht; Nauwynck, Hans

2007-03-31

Experimental groups of 15 susceptible 3-week-old turkeys were inoculated oculonasally with avian metapneumovirus (APV) subtype A and susceptible Escherichia coli O2:K1 and Ornithobacterium rhinotracheale (ORT) bacteria, with a 3 days interval between viral and bacterial inoculation and approximately 8h between the two bacterial inoculations. The aims of the present study were to assess the efficacy of drinking-water administration of enrofloxacin for 3 and 5 days, amoxicillin for 5 days and florfenicol for 5 days for the treatment of the resulting respiratory disease, based on clinical and bacteriological examinations. Antimicrobial treatment started 1 day after dual bacterial inoculation. After infection, the birds were examined and scored for clinical signs daily, weighed at different times, and their tracheae swabbed daily. Five birds were euthanised and examined for macroscopic lesions at necropsy at 5 days post-bacterial inoculation (dpbi) and the remainder at 15dpbi. Samples of the turbinates, trachea, lungs, sinuses, air sacs, heart, pericardium and liver were collected for bacteriological examination. Recovery from respiratory disease caused by an APV/E. coli/ORT triple infection in 3-week-old turkey poults was overall most successful after enrofloxacin treatment, irrespective of treatment duration, followed by florfenicol treatment. Compared with the untreated group, clinical signs as well as ORT and E. coli multiplication in the respiratory tract were significantly reduced by both enrofloxacin treatments and the florfenicol treatment, with the enrofloxacin treatments showing significantly better reductions than the florfenicol treatment. Five-day treatment with amoxicillin, compared with the untreated group, did not cause a significant reduction in any of the aforementioned parameters.

Prior infection of pigs with a genotype 3 swine hepatitis E virus (HEV) protects against subsequent challenges with homologous and heterologous genotypes 3 and 4 human HEV.

PubMed

Sanford, Brenton J; Dryman, Barbara A; Huang, Yao-Wei; Feagins, Alicia R; Leroith, Tanya; Meng, Xiang-Jin

2011-07-01

Hepatitis E virus (HEV) is an important human pathogen. At least four recognized and two putative genotypes of mammalian HEV have been reported: genotypes 1 and 2 are restricted to humans whereas genotypes 3 and 4 are zoonotic. The current experimental vaccines are all based on a single strain of HEV, even though multiple genotypes of HEV are co-circulating in some countries and thus an individual may be exposed to more than one genotype. Genotypes 3 and 4 swine HEV is widespread in pigs and known to infect humans. Therefore, it is important to know if prior infection with a genotype 3 swine HEV will confer protective immunity against subsequent exposure to genotypes 3 and 4 human and swine HEV. In this study, specific-pathogen-free pigs were divided into 4 groups of 6 each. Pigs in the three treatment groups were each inoculated with a genotype 3 swine HEV, and 12 weeks later, challenged with the same genotype 3 swine HEV, a genotype 3 human HEV, and a genotype 4 human HEV, respectively. The control group was inoculated and challenged with PBS buffer. Weekly sera from all pigs were tested for HEV RNA and IgG anti-HEV, and weekly fecal samples were also tested for HEV RNA. The pigs inoculated with swine HEV became infected as evidenced by fecal virus shedding and viremia, and the majority of pigs also developed IgG anti-HEV prior to challenge at 12 weeks post-inoculation. After challenge, viremia was not detected and only two pigs challenged with swine HEV had 1-week fecal virus shedding, suggesting that prior infection with a genotype 3 swine HEV prevented pigs from developing viremia and fecal virus shedding after challenges with homologous and heterologous genotypes 3 and 4 HEV. The results from this study have important implications for future development of an effective HEV vaccine. Copyright © 2011 Elsevier B.V. All rights reserved.

Nonencapsulated Streptococcus pneumoniae causes otitis media during single-species infection and during polymicrobial infection with nontypeable Haemophilus influenzae.

PubMed

Murrah, Kyle A; Pang, Bing; Richardson, Stephen; Perez, Antonia; Reimche, Jennifer; King, Lauren; Wren, John; Swords, W Edward

2015-07-01

Streptococcus pneumoniae strains lacking capsular polysaccharide have been increasingly reported in carriage and disease contexts. Since most cases of otitis media involve more than one bacterial species, we aimed to determine the capacity of a nonencapsulated S. pneumoniae clinical isolate to induce disease in the context of a single-species infection and as a polymicrobial infection with nontypeable Haemophilus influenzae. Using the chinchilla model of otitis media, we found that nonencapsulated S. pneumoniae colonizes the nasopharynx following intranasal inoculation, but does not readily ascend into the middle ear. However, when we inoculated nonencapsulated S. pneumoniae directly into the middle ear, the bacteria persisted for two weeks post-inoculation and induced symptoms consistent with chronic otitis media. During coinfection with nontypeable H. influenzae, both species persisted for one week and induced polymicrobial otitis media. We also observed that nontypeable H. influenzae conferred passive protection from killing by amoxicillin upon S. pneumoniae from within polymicrobial biofilms in vitro. Therefore, based on these results, we conclude that nonencapsulated pneumococci are a potential causative agent of chronic/recurrent otitis media, and can also cause mutualistic infection with other opportunists, which could complicate treatment outcomes. © FEMS 2014. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Experimentally infected domestic ducks show efficient transmission of Indonesian H5N1 highly pathogenic avian influenza virus, but lack persistent viral shedding.

PubMed

Wibawa, Hendra; Bingham, John; Nuradji, Harimurti; Lowther, Sue; Payne, Jean; Harper, Jenni; Junaidi, Akhmad; Middleton, Deborah; Meers, Joanne

2014-01-01

Ducks are important maintenance hosts for avian influenza, including H5N1 highly pathogenic avian influenza viruses. A previous study indicated that persistence of H5N1 viruses in ducks after the development of humoral immunity may drive viral evolution following immune selection. As H5N1 HPAI is endemic in Indonesia, this mechanism may be important in understanding H5N1 evolution in that region. To determine the capability of domestic ducks to maintain prolonged shedding of Indonesian clade 2.1 H5N1 virus, two groups of Pekin ducks were inoculated through the eyes, nostrils and oropharynx and viral shedding and transmission investigated. Inoculated ducks (n = 15), which were mostly asymptomatic, shed infectious virus from the oral route from 1 to 8 days post inoculation, and from the cloacal route from 2-8 dpi. Viral ribonucleic acid was detected from 1-15 days post inoculation from the oral route and 1-24 days post inoculation from the cloacal route (cycle threshold <40). Most ducks seroconverted in a range of serological tests by 15 days post inoculation. Virus was efficiently transmitted during acute infection (5 inoculation-infected to all 5 contact ducks). However, no evidence for transmission, as determined by seroconversion and viral shedding, was found between an inoculation-infected group (n = 10) and contact ducks (n = 9) when the two groups only had contact after 10 days post inoculation. Clinical disease was more frequent and more severe in contact-infected (2 of 5) than inoculation-infected ducks (1 of 15). We conclude that Indonesian clade 2.1 H5N1 highly pathogenic avian influenza virus does not persist in individual ducks after acute infection.

Experimental infection of mice with tightly coiled spiral bacteria ("Candidatus Helicobacter suis") originating from the pig stomach.

PubMed

Park, J-H; Hong, J J; Park, J H

2003-01-01

Mice (n=34) were inoculated orally with a gastric homogenate from a pig infected with tightly coiled spiral bacteria (TCSB). In mice killed in pairs at 16 intervals up to 108 weeks post-inoculation (pi), TCSB were invariably found, mainly in the mucosal surface, gastric pits, intercellular spaces, cytoplasm of surface epithelial cells, and lumina of gastric glands. Histopathologically, infiltration of lymphocytes and plasma cells was seen from 8 weeks pi onwards, gradually increasing as infection progressed. From 64 weeks pi onwards, the formation of large follicles was observed in the lamina propria and submucosa, together with severe necrosis of surface epithelial cells. Glandular epithelial cells in the fundic mucosa were markedly dysplastic and intruded through the basement membrane into the submucosal layer. Common antigenicity between TCSB and Helicobacter pylori was demonstrated by Western blotting, ELISA, and immunohistochemistry. The sequence of the 16S rDNA fragment of 374 bp showed 100% homology with the 16S rRNA gene of "Candidatus Helicobacter suis". Experimental infection of the gastric mucosa of mice with TCSB was closely associated with chronic gastritis and dysplastic lesions.

Effects of repeated Strongylus vulgaris inoculations and concurrent ivermectin treatments on mesenteric arterial lesions in pony foals.

PubMed

Klei, T R; Turk, M A; McClure, J R; Holmes, R A; Dennis, V A; Chapman, M R

1990-04-01

Eight of 10 pony foals reared under helminth-free conditions were inoculated PO with 50 Strongylus vulgaris infective larvae/week for 4 weeks, at which time 1 foal died of acute verminous arteritis. Inoculation of 7 remaining foals continued at 2-week intervals for 20 weeks. Of the 7 foals, 3 were treated with ivermectin (0.2 mg/kg of body weight) in an oral paste formulation at experiment weeks 8, 16, 24; 4 foals were not treated. Two foals were not inoculated or treated and served as controls. After the first ivermectin treatment, ivermectin-treated foals had fewer days (12 +/- 2.9) with rectal temperatures greater than 38.6 C than did nontreated foals (23.3 +/- 3.8). Mean baseline rectal temperatures were 38 +/- 0.2 C. Adverse clinical reactions to ivermectin treatment were not observed in foals. Foals were euthanatized and necropsied 3 weeks after the last ivermectin treatment (week 24). Ivermectin was effective in reducing S vulgaris arterial larval and intestinal adult parasite numbers by 100% in 3 treated foals. Strongylus vulgaris arterial larvae and/or adults were recovered from all 4 nontreated inoculated foals. One nontreated inoculated foal lacked arterial larvae or active arterial lesions, indicating that protective resistance had developed in this individual. Marked gross and histopathologic lesions typical of chronic S vulgaris infection were observed in the 3 nontreated inoculated foals with arterial larvae. Repeated killing of intra-arterial S vulgaris fourth-stage larvae in ivermectin-treated foals did not exacerbate lesions associated with verminous arteritis or induce unique lesions associated with repeated destruction of arterial larvae.(ABSTRACT TRUNCATED AT 250 WORDS)

Beauveria bassiana and Metarhizium anisopliae endophytically colonize cassava roots following soil drench inoculation

PubMed Central

Greenfield, Melinda; Gómez-Jiménez, María I.; Ortiz, Viviana; Vega, Fernando E.; Kramer, Matthew; Parsa, Soroush

2016-01-01

We investigated the fungal entomopathogens Beauveria bassiana and Metarhizium anisopliae to determine if endophytic colonization could be achieved in cassava. An inoculation method based on drenching the soil around cassava stem cuttings using conidial suspensions resulted in endophytic colonization of cassava roots by both entomopathogens, though neither was found in the leaves or stems of the treated cassava plants. Both fungal entomopathogens were detected more often in the proximal end of the root than in the distal end. Colonization levels of B. bassiana were higher when plants were sampled at 7–9 days post-inoculation (84%) compared to 47–49 days post-inoculation (40%). In contrast, the colonization levels of M. anisopliae remained constant from 7–9 days post-inoculation (80%) to 47–49 days post-inoculation (80%), which suggests M. anisopliae is better able to persist in the soil, or as an endophyte in cassava roots over time. Differences in colonization success and plant growth were found among the fungal entomopathogen treatments. PMID:27103778

Experimental evidence supports the abscess theory of development of radicular cysts.

PubMed

Nair, P N R; Sundqvist, Göran; Sjögren, Ulf

2008-08-01

The objective of this study was to experimentally induce inflammatory cysts in an animal model so as to test the hypothesis that radicular cysts develop via the "abscess pathway." Twenty-eight perforated custom-made Teflon cages were surgically implanted into defined locations in the back of 7 Sprague Dawley rats. A week after the implantation of the cages, a known quantity of freshly grown, close allogeneic oral keratinocytes in phosphate buffer solution (PBS) was injected into each cage. One cage per animal was treated as the control that received only epithelial cells. The remaining 3 cages of each animal were trials. Seven days post epithelial cell inoculation; a suspension of 0.2 mL of Fusobacterium nucleatum (10(8) bacteria per mL) was injected into each of the 3 trial cages. Two, 12, and 24 weeks after the inoculation of the bacteria, the cages were taken out, and the tissue contents were fixed and processed by correlative light and transmission electron microscopy. Sixteen of the 21 trial cages could be processed and yielded results. Inoculations of epithelial cells followed 1 week later by F. nucleatum into tissue cages resulted in the development inflammatory cysts in 2 of the 16 cages. The 2 cages contained a total of 4 cystic sites. None of the control cages showed the presence of any cyst-like pathology. Inflammatory cysts were induced by initiating acute inflammatory foci (abscess/necrotic area) by bacterial injection that got enclosed by a proliferating epithelium. This finding provides strong experimental evidence in support of the "abscess theory" of development of radicular cysts.

Characterization of homologous and heterologous adaptive immune responses in porcine reproductive and respiratory syndrome virus infection

PubMed Central

2012-01-01

The present study characterized the homologous and heterologous immune response in type-I porcine reproductive and respiratory syndrome virus (PRRSV) infection. Two experiments were conducted: in experiment 1, eight pigs were inoculated with PRRSV strain 3262 and 84 days post-inoculation (dpi) they were challenged with either strain 3262 or strain 3267 and followed for the next 14 days (98 dpi). In experiment 2, eight pigs were inoculated with strain 3267 and challenged at 84 dpi as above. Clinical course, viremia, humoral response (neutralizing and non-neutralizing antibodies, NA) and virus-specific IFN-γ responses (ELISPOT) were evaluated all throughout the study. Serum levels of IL-1, IL-6, IL-8, TNF-α and TGF-β were determined (ELISA) after the second challenge. In experiment 1 primo-inoculation with strain 3262 induced viremia of ≤ 28 days, low titres of homologous NA but strong IFN-γ responses. In contrast, strain 3267 induced longer viremias (up to 56 days), higher NA titres (≤ 6 log2) and lower IFN-γ responses. Inoculation with 3267 produced higher serum IL-8 levels. After the re-challenge at 84 dpi, pigs in experiment 1 developed mostly a one week viremia regardless of the strain used. In experiment 2, neither the homologous nor the heterologous challenge resulted in detectable viremia although PRRSV was present in tonsils of some animals. Homologous re-inoculation with 3267 produced elevated TGF-β levels in serum for 7–14 days but this did not occur with the heterologous re-inoculation. In conclusion, inoculation with different PRRSV strains result in different virological and immunological outcomes and in different degrees of homologous and heterologous protection. PMID:22515169

Toxocara canis in experimentally infected silver and arctic foxes.

PubMed

Saeed, Isam; Taira, Kensuke; Kapel, Christian M O

2005-09-01

In two experiments, thirty-six farm foxes of two species were inoculated with various doses of infective Toxocara canis eggs or tissue larvae isolated from mice. In experiment I, six adult arctic foxes (Alopex lagopus; 11-month old) were each inoculated with 20,000 eggs and sacrificed 100, 220, or 300 days post infection (dpi), while ten silver fox cubs (Vulpes vulpes; 6-9-week old) were infected with varying doses of eggs (30-3000) and necropsied 120 dpi. In experiment II, two groups of five cubs and two groups of five adult silver foxes received both a primary inoculation and either one or two challenge inoculations: primary inoculation (day 0) with 400 embryonated eggs were administered to five cubs and five adults and another five cubs and five adults received 400 larvae. At 50 dpi, the first challenge inoculation (400 eggs) was inoculated in all animals. At 100 dpi, three animals from each group were necropsied. The remaining two animals in each group were received a second challenge inoculation of 400 tissue larvae on 100 dpi and were subsequently necropsied at 150 dpi. In both experiments, the highest numbers of larvae per gram (lpg) of tissue was found in the kidneys (100-300 dpi). In adult foxes receiving a high dose (20,000 eggs), increasing larval burdens were found in the kidneys over the course of the experiment (up to 300 dpi). The larval migration from the lungs to other tissues appeared to be dose-dependent with the highest larval burdens found in adult foxes. The faecal egg excretion, larval burden and intestinal worm burdens decreased from the first to the second challenge infection.

Transmission of West Nile virus by Culex quinquefasciatus say infected with Culex Flavivirus Izabal.

PubMed

Kent, Rebekah J; Crabtree, Mary B; Miller, Barry R

2010-05-04

The natural history and potential impact of mosquito-specific flaviviruses on the transmission efficiency of West Nile virus (WNV) is unknown. The objective of this study was to determine whether or not prior infection with Culex flavivirus (CxFV) Izabal altered the vector competence of Cx. quinquefasciatus Say for transmission of a co-circulating strain of West Nile virus (WNV) from Guatemala. CxFV-negative Culex quinquefasciatus and those infected with CxFV Izabal by intrathoracic inoculation were administered WNV-infectious blood meals. Infection, dissemination, and transmission of WNV were measured by plaque titration on Vero cells of individual mosquito bodies, legs, or saliva, respectively, two weeks following WNV exposure. Additional groups of Cx. quinquefasciatus were intrathoracically inoculated with WNV alone or WNV+CxFV Izabal simultaneously, and saliva collected nine days post inoculation. Growth of WNV in Aedes albopictus C6/36 cells or Cx. quinquefasciatus was not inhibited by prior infection with CxFV Izabal. There was no significant difference in the vector competence of Cx. quinquefasciatus for WNV between mosquitoes uninfected or infected with CxFV Izabal across multiple WNV blood meal titers and two colonies of Cx. quinquefasciatus (p>0.05). However, significantly more Cx. quinquefasciatus from Honduras that were co-inoculated simultaneously with both viruses transmitted WNV than those inoculated with WNV alone (p = 0.0014). Co-inoculated mosquitoes that transmitted WNV also contained CxFV in their saliva, whereas mosquitoes inoculated with CxFV alone did not contain virus in their saliva. In the sequential infection experiments, prior infection with CxFV Izabal had no significant impact on WNV replication, infection, dissemination, or transmission by Cx. quinquefasciatus, however WNV transmission was enhanced in the Honduras colony when mosquitoes were inoculated simultaneously with both viruses.

Experimentally Infected Domestic Ducks Show Efficient Transmission of Indonesian H5N1 Highly Pathogenic Avian Influenza Virus, but Lack Persistent Viral Shedding

PubMed Central

Wibawa, Hendra; Bingham, John; Nuradji, Harimurti; Lowther, Sue; Payne, Jean; Harper, Jenni; Junaidi, Akhmad; Middleton, Deborah; Meers, Joanne

2014-01-01

Ducks are important maintenance hosts for avian influenza, including H5N1 highly pathogenic avian influenza viruses. A previous study indicated that persistence of H5N1 viruses in ducks after the development of humoral immunity may drive viral evolution following immune selection. As H5N1 HPAI is endemic in Indonesia, this mechanism may be important in understanding H5N1 evolution in that region. To determine the capability of domestic ducks to maintain prolonged shedding of Indonesian clade 2.1 H5N1 virus, two groups of Pekin ducks were inoculated through the eyes, nostrils and oropharynx and viral shedding and transmission investigated. Inoculated ducks (n = 15), which were mostly asymptomatic, shed infectious virus from the oral route from 1 to 8 days post inoculation, and from the cloacal route from 2–8 dpi. Viral ribonucleic acid was detected from 1–15 days post inoculation from the oral route and 1–24 days post inoculation from the cloacal route (cycle threshold <40). Most ducks seroconverted in a range of serological tests by 15 days post inoculation. Virus was efficiently transmitted during acute infection (5 inoculation-infected to all 5 contact ducks). However, no evidence for transmission, as determined by seroconversion and viral shedding, was found between an inoculation-infected group (n = 10) and contact ducks (n = 9) when the two groups only had contact after 10 days post inoculation. Clinical disease was more frequent and more severe in contact-infected (2 of 5) than inoculation-infected ducks (1 of 15). We conclude that Indonesian clade 2.1 H5N1 highly pathogenic avian influenza virus does not persist in individual ducks after acute infection. PMID:24392085

Differential growth response of various crop species to arbuscular mycorrhizal inoculation.

PubMed

Eo, Ju-Kyeong; Eom, Ahn-Heum

2009-03-01

To investigate the growth response of various crop species to mycorrhizal inoculation, arbuscular mycorrhizal fungi were applied to Glycine max, Vigna angularis, Senna tora, Hordeum vulgare var. hexastichon. Zea mays, Sorghum bicolor, Allium tuberosum, Solanum melongena, and Capsicum annuum. The biomass of the inoculated crops was measured every two weeks for the 12-week growth period. By measuring biomass, we calculated the mycorrhizal responsiveness of the nine crop species. Among the nine crop species, four species showed a significant response to mycorrhizal inoculation. The shoot biomasses of V. angularis, C. annuum, A. tuberosum, and S. tora significantly increased with mycorrhizal inoculation.

Therapeutic use of a receptor mimic probiotic reduces intestinal Shiga toxin levels in a piglet model of hemolytic uremic syndrome

PubMed Central

2014-01-01

Background Hemolytic uremic syndrome (HUS) is a systemic and potentially fatal complication of gastroenteritis secondary to Shiga toxin-producing enterohemorrhagic Escherichia coli (EHEC) infection characterized by microangiopathic hemolytic anemia, thrombocytopenia, and acute renal damage. Shiga toxin (Stx), the toxin principle in HUS, is produced locally within the gut following EHEC colonization and is disseminated via the vasculature. Clinical development of HUS currently has no effective treatment and is a leading cause of renal failure in children. Novel post-exposure therapies are currently needed for HUS; therefore, the purpose of this study was to investigate the efficacy of a Stx receptor mimic probiotic in a porcine model of HUS. Edema disease, an infection of swine caused by host adapted Shiga toxin-producing Escherichia coli (STEC) and mediated by Shiga toxin 2e (Stx2e), shares many pathogenic similarities to HUS. In this study, three-week old piglets were inoculated with STEC and 24 hours later treated twice daily with a probiotic expressing an oligosaccharide receptor mimic for Stx2e to determine if the probiotic could reduce intestinal toxin levels. Methods Piglets were orally inoculated with 1010 CFU of STEC strain S1191 eight days after weaning. Beginning day 1 post-inoculation, piglets were treated orally twice daily with 5 × 1011 CFU of either the receptor mimic probiotic or a sham probiotic for 10 days. Intestinal Stx2e levels were assessed daily via Vero cell assay. The efficacy of the probiotic at reducing intestinal Stx2e, vascular lesions, and clinical disease was evaluated with repeated measures ANOVA and Fisher’s exact test as appropriate. Results The probiotic significantly reduced intestinal Stx2e, as reflected by decreased fecal toxin titers on days 3–8 post-inoculation (p < 0.01). Despite this reduction in intestinal toxin levels, however, the probiotic failed to reduce the incidence of vascular necrosis in target organs and had no effect on clinical disease. Conclusions The data suggest that post-exposure treatment with a Stx-binding probiotic is effective in reducing intestinal toxin burden. Future studies could target this approach for possible development of post-exposure interventions. PMID:24890228

Research on the influence of anaerobic stabilization of various dairy sewage sludge on biodegradation of polycyclic aromatic hydrocarbons PAHs with the use of effective microorganisms

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

Boruszko, Dariusz, E-mail: d.boruszko@pb.edu.pl

Sewage sludge was taken from a dairy WWTP belonging to Mlekovita Cooperative in Wysokie Mazowieckie. There were excess sludge, flotation sludge and a mixture of excess and flotation sludge from pre-treatment of dairy sewage. The initial content of 16 PAHs in excess sludge before fermentation was approximately 689 µg·kg{sup −1} in dry mass, whereas in post-flotation sludge (which constituted around 30% of raw sludge) it was approximately 95 µg·kg{sup −1} in dry mass. A mixture of excess and flotation sludge had the content of 497,7 µg·kg{sup −1} in dry mass. Through comparison of particular hydrocarbons content in raw sewage sludgemore » to the total PAHs content, it was shown that tricyclic compounds, which constituted 46,3% of the PAHs sum (excess sludge), and tetracyclic compounds, which constituted 60,0% of the PAHs sum (flotation sludge), were the dominating fractions. In the sludge subjected to fermentation in reactors with mixed sludge and surplus activated sludge, the general trend of the course of changes in concentrations of PAHs was similar. Both in the sludge inoculated with EM and in that not inoculated with EM, a significant increase in the total PAHs contents was observed in the first fermentation phase (acidic fermentation) after 7 days of the process. Addition of EM into the sludge did not prevent the PAHs release, and therefore higher concentrations of PAHs sum were recorded during the hydrolysis stage than in sludge before fermentation. A decrease in the sum of PAHs was observed after 2 weeks of fermentation in relation to the quantity observed after 1 week of fermentation (except from post-flotation sludge). In the following weeks, there was further decrease in the concentration of the 16 PAHs sum in all sludge types. However, in sludge without EM inoculation, it was lower than in sludge with EM inoculation. The loss of the majority of tested hydrocarbons was reported in the final phase of fermentation. - Highlights: • The influence of applying Effective Microorganisms on PAHs degradation in dairy sewage sludge during fermentation has been examined. • To study the relationship between quantitative parameters and to describe the correlation strength, Spearman's rank correlation coefficient was used. • In order to verify whether there was a statistically significant change, the Wilcoxon test was applied for individual observation pairs.« less

Semen variables of sheep (Ovis aries) experimentally infected with Toxoplasma gondii.

PubMed

Lopes, W D Z; Costa, A J; Souza, F A; Rodrigues, J D F; Costa, G H N; Soares, V E; Silva, G S

2009-04-01

The influence of Toxoplasma gondii on semen variables and sperm morphology of sheep was evaluated in eight reproductive males distributed into three experimental groups: GI, three sheep inoculated with 2.0x10(5) of P strain oocytes; GII, three sheep infected with 1.0x10(6) of RH strain tachyzoites and; GIII two control sheep. Clinical (rectal temperature, cardiac and respiratory frequencies), parasite and serology exams (IIF) were realized. Sperm variables (volume, motility, vigor and concentration) and semen morphology for each sheep were also evaluated. Thus, semen and blood collections were assessed on post-inoculation days (PIDs)-1,3,5,7,11,14 and weekly thereafter up to PID 70. Clinical alterations were observed (hypothermia and anorexia) in infected sheep from groups GI and GII. Parasitic outbreaks were detected in five sheep. All the infected sheep produced antibodies against T. gondii from PID 5 onwards, reaching a peak of 4096 and 8192 for group GI and GII sheep, respectively. Differences (P<0.05) were observed regarding the ejaculate volume between the inoculated groups (oocytes and tachyzoites) and control. Even though experimental toxoplasmic infection resulted in clinical symptomology in the inoculated sheep, the minimal alterations in sperm pathologies could not be directly attributed to T. gondii.

Response of dairy calves to vaccinia viruses that express foreign genes.

PubMed Central

Gillespie, J H; Geissinger, C; Scott, F W; Higgins, W P; Holmes, D F; Perkus, M; Mercer, S; Paoletti, E

1986-01-01

Repeated intradermal inoculations of calves with wild-type vaccinia virus and recombinant vaccinia viruses expressing human hepatitis B virus surface antigen and herpes simplex virus, type 1, glycoprotein D produced characteristic pox lesions at each site of injection. In some instances, calves were inoculated as many as five times at intervals from 4 to 7 weeks. The lesions invariably were more severe after the second inoculation. Subsequent inoculations produced a less severe area of redness, swelling, necrosis, and scab formation. No other signs of illness, such as an elevation in temperature, were noted in the calves. Vaccinia virus was isolated in low titers from scabs taken at various times after inoculation. No lesions were formed at the sites injected with tissue culture fluid and cellular debris at the same time that virus inoculations were made. Calf contact controls remained normal through the 8-week exposure in isolation units with calves inoculated twice with vaccinia virus. No neutralizing antibody to vaccinia virus was detected in the contact controls. In contrast, the virus-inoculated calves developed neutralizing antibody to vaccinia virus and to herpes simplex virus glycoprotein D in serum. In all cattle, a second inoculation significantly enhanced the neutralizing antibody response within 1 week, suggesting that an anamnestic response had occurred. No antibody to hepatitis B virus surface antigen was elicited in calves after repeated inoculations with vaccinia recombinants that express hepatitis B virus surface antigen and are known to elicit in rabbits antibodies reactive with hepatitis B virus surface antigen. Images PMID:3700615

Antitumour activity of cordycepin in mice.

PubMed

Yoshikawa, Noriko; Nakamura, Kazuki; Yamaguchi, Yu; Kagota, Satomi; Shinozuka, Kazumasa; Kunitomo, Masaru

2004-12-01

1. The antitumour effect of orally administered cordycepin, a component isolated from water extracts of Cordyceps sinensis, was examined in mice inoculated with B16 melanoma (B16-BL6) cells. 2. B16-BL6 (1 x 10(6)) cells were inoculated subcutaneously into the right footpad of mice. At 2 weeks after the cell inoculation, the enlarged primary tumour lump was weighed. Cordycepin (0, 5 and 15 mg/kg per day) was administered orally to the mice for 2 weeks from the date of tumour inoculation. Cordycepin (15 mg/kg per day) significantly reduced by 36% the wet weight of the primary tumour lump compared to that of the untreated control mice, without any loss of bodyweight or systemic toxicity. 3. Cordycepin (15 mg/kg per day) administered orally for 2 weeks inhibited the tumour enlargement in the right thigh inoculated with B16-BL6 cells premixed with extracellular matrix (Matrigel). 4. These results indicate that orally administered cordycepin inhibits melanoma cell growth in mice with no adverse effects.

An HPLC-MS characterization of the changes in sweet orange leaf metabolite profile following infection by the bacterial pathogen Candidatus Liberibacter asiaticus.

PubMed

Hijaz, Faraj M; Manthey, John A; Folimonova, Svetlana Y; Davis, Craig L; Jones, Shelley E; Reyes-De-Corcuera, José I

2013-01-01

Huanglongbing (HLB) presumably caused by Candidatus Liberibacter asiaticus (CLas) threatens the commercial U.S. citrus crop of an annual value of $3 billion. The earliest shift in metabolite profiles of leaves from greenhouse-grown sweet orange trees infected with Clas, and of healthy leaves, was characterized by HPLC-MS concurrently with PCR testing for the presence of Clas bacteria and observation of disease symptoms. Twenty, 8-month-old 'Valencia' and 'Hamlin' trees were grafted with budwood from PCR-positive HLB source trees. Five graft-inoculated trees of each variety and three control trees were sampled biweekly and analyzed by HPLC-MS and PCR. Thirteen weeks after inoculation, Clas was detected in newly growing flushes in 33% and 55% of the inoculated 'Hamlin' and 'Valencia' trees, respectively. Inoculated trees remained asymptomatic in the first 20 weeks, but developed symptoms 30 weeks after grafting. No significant differences in the leaf metabolite profiles were detected in Clas-infected trees 23 weeks after inoculation. However, 27 weeks after inoculation, differences in metabolite profiles between control leaves and those of Clas-infected trees were evident. Affected compounds were identified with authentic standards or structurally classified by their UV and mass spectra. Included among these compounds are flavonoid glycosides, polymethoxylated flavones, and hydroxycinnamates. Four structurally related hydroxycinnamate compounds increased more than 10-fold in leaves from 'Hamlin' and 'Valencia' sweet orange trees in response to Clas infection. Possible roles of these hydroxycinnamates as plant defense compounds against the Clas infection are discussed.

Influence of parainfluenza-1 respiratory tract viral infection on endothelin receptor-effector systems in mouse and rat tracheal smooth muscle.

PubMed Central

Knott, P. G.; Henry, P. J.; McWilliam, A. S.; Rigby, P. J.; Fernandes, L. B.; Goldie, R. G.

1996-01-01

1. In this study we have compared the effects of parainfluenza-1 respiratory tract viral infection on the density and function of ETA and ETB receptors in rat and mouse tracheal airway smooth muscle. 2. The bronchoconstrictor effect of inhaled methacholine was significantly enhanced in virus-infected rats, at both 4 and 12 days post-inoculation. That is, the concentration of methacholine causing an increase in resistance of 100% (PC100 methacholine) was significantly lower in virus-infected animals at both 4 and 12 days post-inoculation (n = 6-8; P < 0.05). 3. Total specific binding of [125I]-endothelin-1 and the relative proportions of ETA and ETB binding sites for [125I]-endothelin-1 were assessed in tracheal airway smooth muscle in parainfluenza-1-infected rats and mice at days 2, 4 and 12 post-inoculation using the ligands BQ-123 (1 microM; ETA receptor-selective) and sarafotoxin S6c (100 nM; ETB receptor-selective). Total specific binding in mice was significantly reduced at day 2 post-inoculation (n = 5; P < 0.05) but not at days 4 and 12 post-inoculation (n = 5). In control mice, the proportions of ETA and ETB binding sites were 53%:47% at day 2 and 43%:57% at day 4 and these were significantly altered by parainfluenza-1 infection such that, the ratios were 81%:19% at day 2 and 89%:11% at day 4 (P < 0.05). By day 12 post-inoculation, the proportion of ETA and ETB binding sites in tracheal smooth muscle from mice infected with parainfluenza-1 was not significantly different from control. In rat tracheal airway smooth muscle, neither total specific binding nor the ETA and ETB binding site ratio (64%:36%) were significantly altered in virus-inoculated rats at days 2, 4 or 12 post-inoculation (n = 5). 4. Parainfluenza-1 infection in mice had no effect on the sensitivity or maximal contractile effect of endothelin-1 in tracheal smooth muscle at days 2, 4 or 12 post-inoculation (n = 4). In contrast, contraction in response to the ETB receptor-selective agonist sarafotoxin S6c was attenuated by 39% at day 2 and by 93% at day 4 post-inoculation (P < 0.05). However, by day 12 post-inoculation, contractions to sarafotoxin S6c were not significantly different between control and virus-infected mice. In parainfluenza-1-infected rats, there were small but significant reductions in the sensitivity to carbachol, endothelin-1 and sarafotoxin S6c whilst the maximal responses to the highest concentrations of these agonists were not significantly altered by virus infection (n = 8). 5. BQ-123 (3 microM) had no significant effect on cumulative concentration-effect curves to endothelin-1 in tracheal preparations from control mice (n = 4) or parainfluenza-1-infected rats (n = 8). In contrast, in tissues taken from virus-infected mice at day 4 post-inoculation, BQ-123 caused a marked 9.6 fold rightward shift in the concentration-effect curve to endothelin-1 (n = 4). 6. In summary, we have demonstrated that parainfluenza-1 infection in mice transiently reduced the density of tracheal airway smooth muscle ETB receptors and this was reflected in reduced responsiveness to the ETB receptor-selective agonist sarafotoxin S6c. In contrast, whilst parainfluenza-1 infection in rats was associated with the pathological features and bronchial hyperresponsiveness common to respiratory tract viral infection, there was no selective down-regulation of ETB receptor expression or functional activity. The reasons for these species differences are not clear, but may relate to differences in the airway inflammatory response to parainfluenza-1 virus. PMID:8886411

Diverse mechanisms of plant resistance to cauliflower mosaic virus revealed by leaf skeleton hybridization.

PubMed

Melcher, U; Brannan, C M; Gardner, C O; Essenberg, R C

1992-01-01

Plants not hosts for cauliflower mosaic virus (CaMV) may prevent systemic CaMV infection by interfering with dissemination of infection through the plant or by preventing viral replication and maturation. Leaf skeleton hybridization allows distinction between these two barriers. The technique assesses the spatial distribution of CaMV in an inoculated leaf by hybridization of a skeleton of the leaf with a CaMV DNA probe. Leaves or leaflets of soybean, cucumber, peanut, tomato, lettuce, spinach, pepper, onion, wheat, maize and barley, inoculated with CaMV DNA or CaMV virions were processed for leaf skeleton hybridization either immediately after inoculation or two weeks thereafter. Autoradiographic images of soybean and cucumber skeletons had many dark spots suggesting that CaMV DNA replication and local spread had occurred. Images of onion leaf skeletons prepared two weeks after inoculation with CaMV DNA had fewer spots. To test whether these spots resulted from CaMV replication, DNA was extracted from inoculated onion leaves and analyzed by electrophoresis, blotting and hybridization. Molecules recovered two weeks after inoculation resembled those inoculated, indicating absence of replication. For the other species, we found no evidence of local spread of CaMV infections. Thus, many plant species resist systemic CaMV infection by preventing replication or local spread of CaMV, while others solely prevent systemic movement of infection.

Feed Supplementation with Red Seaweeds, Chondrus crispus and Sarcodiotheca gaudichaudii, Reduce Salmonella Enteritidis in Laying Hens

PubMed Central

Kulshreshtha, Garima; Rathgeber, Bruce; MacIsaac, Janice; Boulianne, Martine; Brigitte, Lehoux; Stratton, Glenn; Thomas, Nikhil A.; Critchley, Alan T.; Hafting, Jeff; Prithiviraj, Balakrishnan

2017-01-01

Salmonella Enteritidis is vertically transmitted to eggs from laying hens through infected ovaries and oviducts. S. Enteritidis can also penetrate the eggshell from contaminated feces. Reducing S. Enteritidis in laying hens is vital to provide safer eggs and minimize the spread of salmonellosis to humans. Antibiotics have been widely used to control bacterial diseases in broilers and laying hens. However, there is a major concern that the use of antibiotics leads to the development of antibiotic resistance and adverse effects on microbiota of the treated birds. Thus, there is an interest in developing alternatives to antibiotics, such as dietary prebiotics. In the present study, feed supplemented with the red seaweeds: Chondrus crispus (CC) or Sarcodiotheca gaudichaudii (SG), was offered to laying hens late in production to control S. Enteritidis. Diets contained one of the following; 2% or 4% Chondrus crispus (CC2, and CC4, respectively) or Sarcodiotheca gaudichaudii (SG2 and SG4, respectively). Chlortetracycline was used in the positive control diet. During week-4, 48 birds were orally challenged with 2 × 109 CFU/mL of S. Enteritidis. Eggs and fecal samples were collected 1, 3, 5, and 7 days’ post inoculation. Birds were euthanized and organs (ceca, ovary, liver, and spleen) were sampled and analyzed for the presence of S. Enteritidis, 7 days’ post inoculation. Results showed that seaweed reduced the negative effect on body weight and egg production in S. Enteritidis-challenged laying hens. Analysis of fecal samples showed that the antibiotic (CTC) reduced S. Enteritidis in the intestinal tract and fecal samples, 3 days’ post inoculation. Fecal samples from Chlortetracycline and CC4 supplemented birds tested negative for S. Enteritidis on days 5 and 7 post inoculation (lowest detection limit = 10-1). S. Enteritidis colonization in the ceca was also significantly reduced in birds fed CC (4%) and Chlortetracycline. Blood serum profiles revealed that there were no significant differences in serum aspartate aminotransferase (AST) and sodium. However, the level of serum immunoglobulin (IgA) was higher in the CC4 treatment. The relative abundance of Lactobacillus acidophilus was significantly higher in CC4 while, the abundance of the pathogenic bacteria, Clostridium perfringens and Salmonella Enteritidis were reduced compared to control. Results indicate that feed supplemented with 4% CC is effective in providing protection against Salmonella Enteritidis colonization in laying hens. PMID:28443073

Diversity and persistence of ectomycorrhizal fungi and their effect on nursery-inoculated Pinus pinaster in a post-fire plantation in Northern Portugal.

PubMed

Franco, Albina R; Sousa, Nadine R; Ramos, Miguel A; Oliveira, Rui S; Castro, Paula M L

2014-11-01

Ectomycorrhizal fungi (ECMF) play an important role in forest ecosystems, often mitigating stress factors and increasing seedling performance. The aim of this study was to investigate the effects of a nursery inoculation on Pinus pinaster growth and on the fungal communities established when reforesting burned areas. Inoculated P. pinaster saplings showed 1.5-fold higher stem height than the non-inoculated controls after a 5 year growth period, suggesting that fungal inoculation could potentiate tree growth in the field. Ordination analysis revealed the presence of different ECMF communities on both plots. Among the nursery-inoculated fungi, Laccaria sp., Rhizopogon sp., Suillus bovinus and Pisolithus sp. were detected on inoculated Pinus saplings on both sampling periods, indicating that they persisted after field establishment. Other fungi were also detected in the inoculated plants. Phialocephala sp. was found on the first assessment, while Terfezia sp. was detected on both sampling periods. Laccaria sp. and Rhizopogon sp. were identified in the control saplings, belonging however to different species than those found in the inoculated plot. Inocybe sp., Thelephora sp. and Paxillus involutus were present on both sampling periods in the non-inoculated plots. The results suggest that ECMF inoculation at nursery stage can benefit plant growth after transplantation to a post-fire site and that the inoculated fungi can persist in the field. This approach has great potential as a biotechnological tool to aid in the reforestation of burned areas.

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

PubMed Central

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

2006-01-01

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

Powassan virus infection in snowshoe hares (Lepus americanus).

PubMed

Zarnke, R L; Yuill, T M

1981-04-01

Sera from snowshoe hares (Lepus americanus) trapped near Rochester, Alberta, Canada were tested for Powassan virus antibody by the constant virus/serum dilution neutralization test. Of 1264 serum samples tested, 137 had an antibody titer of at least 1:4 for Powassan virus. Ten hares were inoculated with Powassan virus in the laboratory. Viremia lasted 4-5 days and ceased with the appearance of Powassan antibody in the serum. Neutralizing antibody reached a peak titer of 1:119 on day 15 post-inoculation and was still detectable 13 months post-inoculation.

Inoculation of hybrid poplar with the endophytic bacterium Enterobacter sp. 638 increases biomass but does not impact leaf level physiology

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

Rogers, A.; McDonald, K.; Muehlbauer, M. F.

Endophytic bacteria have been shown to provide several advantages to their host, including enhanced growth. Inoculating biofuel species with endophytic bacteria is therefore an attractive option to increase the productivity of biofuel feedstocks. Here, we investigated the effect of inoculating hard wood cuttings of Populus deltoides Bartr. x Populus. nigra L. clone OP367 with Enterobacter sp. 638. After 17 weeks, plants inoculated with Enterobacter sp. 638 had 55% greater total biomass than un-inoculated control plants. Study of gas exchange and fluorescence in developing and mature leaves over a diurnal cycle and over a 5 week measurement campaign revealed no effectsmore » of inoculation on photosynthesis, stomatal conductance, photosynthetic water use efficiency or the maximum and operating efficiency of photosystem II. However, plants inoculated with Enterobacter sp. 638 had a canopy that was 39% larger than control plants indicating that the enhanced growth was fueled by increased leaf area, not by improved physiology. Leaf nitrogen content was determined at two stages over the 5 week measurement period. No effect of Enterobacter sp. 638 on leaf nitrogen content was found indicating that the larger plants were acquiring sufficient nitrogen. Enterobacter sp. 638 lacks the genes for N{sub 2} fixation, therefore the increased availability of nitrogen likely resulted from enhanced nitrogen acquisition by the 84% larger root system. These data show that Enterobacter sp. 638 has the potential to dramatically increase productivity in poplar. If fully realized in the production environment, these results indicate that an increase in the environmental and economic viability of poplar as a biofuel feedstock is possible when inoculated with endophytic bacteria like Enterobacter sp. 638.« less

Parasite population dynamics in pigs infected with Trichuris suis and Oesophagostomum dentatum.

PubMed

Petersen, Heidi Huus; Andreasen, Annette; Kringel, Helene; Roepstorff, Allan; Thamsborg, Stig M

2014-01-17

The aim of the present study was to investigate the population dynamics and potential interactions between Trichuris suis and Oesophagostomum dentatum in experimentally co-infected pigs, by quantification of parasite parameters such as egg excretion, worm recovery and worm location. Forty-eight helminth naïve pigs were allocated into four groups. Group O was inoculated with 20 O. dentatum L3/kg/day and Group T with 10 T. suis eggs/kg/day. Group OT was inoculated with both 20 O. dentatum L3/kg/day and 10 T. suis eggs/kg/day, while Group C was kept as an uninfected control group. All inoculations were trickle infections administered twice weekly and were continued until slaughter. Faecal samples were collected from the rectum of all pigs at day 0, and twice weekly from 2 to 9 weeks post first infection (wpi). Six pigs from each group were necropsied 5 wpi and the remaining 6 pigs from each group were necropsied 10 wpi. The faecal egg counts (FEC) and total worm burdens of O. dentatum were dramatically influenced by the presence of T. suis, with significantly lower mean FECs and worm burdens at 5 and 10 wpi compared to single infected pigs. Furthermore, in the presence of T. suis we found that O. dentatum was located more posteriorly in the gut. The changes in the Trichuris population were less prominent, but faecal egg counts, worm counts 5 wpi (57% recovered vs. 39%) and the proportion of infected animals at 10 wpi were higher in Group OT compared to Group T. The location of T. suis was unaffected by the presence of O. dentatum. These results indicate an antagonistic interaction between T. suis and O. dentatum which is dominated by T. suis. Copyright © 2013 Elsevier B.V. All rights reserved.

Evaluation of the impact of quorum sensing transcriptional regulator SdiA on long-term persistence and fecal shedding of Escherichia coli O157:H7 in weaned calves.

PubMed

Sharma, V K; Bearson, S M D

2013-04-01

Escherichia coli O157:H7 (O157) colonization of bovine intestine is mediated through the locus of enterocyte effacement (LEE)-encoded type III secretion system and secreted virulence proteins that promote colonization of the recto-anal junction (RAJ) of the large intestine of cattle. The quorum sensing transcriptional regulator SdiA, a homolog of LuxR, has been shown in vitro to repress LEE strongly when overexpressed from a multi-copy recombinant plasmid or when its activity is enhanced by the binding of N-acyl-L-homoserine lactones (AHLs), the quorum sensing signals that are detected by SdiA. Since LEE has been shown to be essential for colonization and persistence of O157 in bovine intestine, we examined whether a mutation in sdiA, which normally represses LEE in vitro, would also exert negative effect on colonization and long-term persistence of O157 in weaned calves. Ten-week old weaned calves (n = 4/group) were inoculated orally with 10(10) cfu of either the wild-type or sdiA mutant strain. Initial fecal shedding of the sdiA mutant and the wild-type strain were similar in magnitude and declined during the first 2 weeks post-inoculation. The sdiA mutant was detected in feces of only one of the four calves at low levels (≥10(2) cfu/g feces) from days 19 - 27 post-inoculation, whereas, the fecal shedding of the wild-type strain persisted at approximately 4-logs in all four calves from days 19 - 27. We also confirmed that SdiA represses ler, which encodes a positive transcriptional regulator of LEE, in response to AHLs, and reduces adherence of O157 to HEp-2 cells. In conclusion, this study demonstrates that although in vitro the sdiA gene represses LEE and LEE-mediated adherence to cultured cells, the presence of sdiA is necessary for colonization of bovine large intestine that in turn promotes persistent fecal shedding of O157 by these animals. Published by Elsevier Ltd.

Bio-preservation of ground beef meat by Enterococcus faecalis CECT7121.

PubMed

Sparo, M D; Confalonieri, A; Urbizu, L; Ceci, M; Bruni, S F Sánchez

2013-01-01

Meat and particularly ground beef is frequently associated with Food Poisoning episodes and breeches in Food Safety. The main goal of this research was to evaluate the bactericide effect of the probiotic Enterococcus faecalis CECT7121, against different pathogens as: Escherichia coli O157:H7, Staphylococcus aureus, Clostridium perfringens and Listeria monocytogenes, inoculated in ground beef meat. Three studies were performed to evaluate the inhibition of E. faecalis CECT7121 on ground beef meat samples inoculated with pathogens: Study I: Samples (100 g meat) were inoculated with pathogens (10(3) CFU/g)) and E. faecalis CECT7121 (10(4) CFU/g) simultaneously. Study II: Samples were inoculated with E. faecalis CECT7121 24 h before the pathogens. Study III: E. faecalis CECT7121were inoculated 24 h after pathogens. The viable counts were performed at 0, 24, 48 and 72 h post-inoculation. The simultaneous inoculation of E. faecalis CECT7121 with E. coli O157:H7 strains resulted in the absence of viable counts of bacteria at 72 h post-treatment. However, when the probiotic was added 24 h before and 24 h after the pathogen E. coli O157:H7, viable cells were not detected at 24 h and 48 h post-treatment, respectively. Consistently, neither S. aureus nor Cl. perfringens viable bacteria were detected at 48 h in whole assays when inoculated with E. faecalis CECT7121. The same trend than described before was obtained after applying the 3 models assayed for L. monocytogenes. The current assays demonstrated the bactericide activity of E. faecalis CECT7121 strain on bacterial pathogens in ground beef meat.

Standardization and validation of real time PCR assays for the diagnosis of histoplasmosis using three molecular targets in an animal model.

PubMed

López, Luisa F; Muñoz, César O; Cáceres, Diego H; Tobón, Ángela M; Loparev, Vladimir; Clay, Oliver; Chiller, Tom; Litvintseva, Anastasia; Gade, Lalitha; González, Ángel; Gómez, Beatriz L

2017-01-01

Histoplasmosis is considered one of the most important endemic and systemic mycoses worldwide. Until now few molecular techniques have been developed for its diagnosis. The aim of this study was to develop and evaluate three real time PCR (qPCR) protocols for different protein-coding genes (100-kDa, H and M antigens) using an animal model. Fresh and formalin-fixed and paraffin-embedded (FFPE) lung tissues from BALB/c mice inoculated i.n. with 2.5x106 Histoplasma capsulatum yeast or PBS were obtained at 1, 2, 3, 4, 8, 12 and 16 weeks post-infection. A collection of DNA from cultures representing different clades of H. capsulatum (30 strains) and other medically relevant pathogens (36 strains of related fungi and Mycobacterium tuberculosis) were used to analyze sensitivity and specificity. Analytical sensitivity and specificity were 100% when DNAs from the different strains were tested. The highest fungal burden occurred at first week post-infection and complete fungal clearance was observed after the third week; similar results were obtained when the presence of H. capsulatum yeast cells was demonstrated in histopathological analysis. In the first week post-infection, all fresh and FFPE lung tissues from H. capsulatum-infected animals were positive for the qPCR protocols tested except for the M antigen protocol, which gave variable results when fresh lung tissue samples were analyzed. In the second week, all qPCR protocols showed variable results for both fresh and FFPE tissues. Samples from the infected mice at the remaining times post-infection and uninfected mice (controls) were negative for all protocols. Good agreement was observed between CFUs, histopathological analysis and qPCR results for the 100-kDa and H antigen protocols. We successfully standardized and validated three qPCR assays for detecting H. capsulatum DNA in fresh and FFPE tissues, and conclude that the 100-kDa and H antigen molecular assays are promising tests for diagnosing this mycosis.

Green tea extract inhibits proliferation of uterine leiomyoma cells in vitro and in nude mice

PubMed Central

ZHANG, Dong; AL-HENDY, Mohamed; RICHARD-DAVIS, Gloria; MONTGOMERY-RICE, Valerie; SHARAN, Chakradhari; RAJARATNAM, Veera; KHURANA, Anjali; AL-HENDY, Ayman

2010-01-01

Objective Investigate the effect of epigallocatechin gallate (EGCG), on rat leiomyoma (ELT3) cells in vitro and in nude mice model. Study Design ELT3 cells were treated with various concentrations of EGCG. Cell proliferation, PCNA and Cdk4 protein levels were evaluated. ELT3 cells were inoculated subcutaneously in female athymic nude mice. Animals were fed 1.25mg EGCG (in drinking water)/mouse/day. Tumors were collected and evaluated at 4 and 8 weeks post-treatment. Results Inhibitory effect of EGCG (200 μM) on ELT3 cells was observed after 24 h treatment (p<0.05). At ≥50μM, EGCG significantly decreased PCNA and Cdk4 protein levels (p<0.05). In vivo, EGCG treatment dramatically reduced the volume and weight of tumors at 4 and 8 weeks post-treatment (p<0.05). The PCNA and Cdk4 protein levels were significantly reduced in EGCG treated group (p<0.05). Conclusion EGCG effectively inhibits the proliferation and induce apoptosis in rat ELT3 uterine leiomyoma cells in vitro and in vivo. PMID:20074693

Respiratory syncytial virus increases lung cellular bioenergetics in neonatal C57BL/6 mice

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

Alsuwaidi, Ahmed R., E-mail: alsuwaidia@uaeu.ac.ae; Albawardi, Alia, E-mail: alia.albawardi@uaeu.ac.ae; Almarzooqi, Saeeda, E-mail: saeeda.almarzooqi@uaeu.ac.ae

2014-04-15

We have previously reported that lung cellular bioenergetics (cellular respiration and ATP) increased in 4–10 week-old BALB/c mice infected with respiratory syncytial virus (RSV). This study examined the kinetics and changes in cellular bioenergetics in ≤2-week-old C57BL/6 mice following RSV infection. Mice (5–14 days old) were inoculated intranasally with RSV and the lungs were examined on days 1–10 post-infection. Histopathology and electron microscopy revealed preserved pneumocyte architectures and organelles. Increased lung cellular bioenergetics was noted from days 1–10 post-infection. Cellular GSH remained unchanged. These results indicate that the increased lung cellular respiration (measured by mitochondrial O{sub 2} consumption) and ATPmore » following RSV infection is independent of either age or genetic background of the host. - Highlights: • RSV infection increases lung cellular respiration and ATP in neonatal C57BL/6 mice. • Increased lung cellular bioenergetics is a biomarker of RSV infection. • Lung cellular glutathione remains unchanged in RSV infection.« less

Assessing the impact of egg sweating on Salmonella Enteritidis penetration into shell eggs.

PubMed

Gradl, Janet A; Curtis, Patricia A; Jones, Deana R; Anderson, Kenneth E

2017-07-01

Salmonella Enteritidis (SE) prevalence in eggs is a major concern to the egg industry. Some research has shown that egg sweating can increase Salmonella penetration into egg contents when refrigerated eggs are moved to a warmer temperature. This occurs when eggs are tempered before wash, to minimize thermal cracks. The effect of egg sweating on SE penetration into shell eggs over a 6 week storage period at 4°C was assessed. A 2 × 2 factorial of SE inoculation and egg sweating was utilized. Treatments included (SES) nalidixic acid (NA)-resistant SE inoculated and sweated, (SENS) NA-resistant SE inoculated and not sweated, (NSES) buffered peptone water (BPW) inoculated and sweated, and (NSENS) BPW inoculated and not sweated. Eggs were inoculated with 108 SE. Eggs formed condensation for approximately 17 min in a 32°C incubator. Shell rinse, shell emulsion, and egg contents were sampled then enumerated and assessed for prevalence of SE over a 6 wk storage period at 4°C. After wk 1, the SENS shell rinse had higher SE counts (0.32 log10 CFU/mL) than the other 3 treatments, where no SE was enumerated. A significant week by treatment interaction was found for the shell rinse SE detection (P < 0.05). In subsequent weeks, no SE counts were obtained from the egg shell rinse, shell emulsion, or egg contents. The SENS shell rinses had significantly higher SE prevalence than the SES rinses in weeks 1 (100% vs. 34.3%), 2 (57.6% vs. 22.2%), and 3 (38.2% vs. 11.1%) (P < 0.05). In samples from weeks 4, 5, and 6, there was no difference in SE prevalence between SES and SENS. Egg sweating did not increase SE penetration into the shell emulsion across treatment or week (P < 0.05). The decreasing trend of SE prevalence obtained over the study period indicate that refrigeration is effective at inhibiting SE growth. These results indicate that egg sweating occurring under common US egg handling practices is not harmful to egg safety. © 2017 Poultry Science Association Inc.

Survival of introduced phosphate-solubilizing bacteria (PSB) and their impact on microbial community structure during the phytoextraction of Cd-contaminated soil.

PubMed

Jeong, Seulki; Moon, Hee Sun; Shin, Doyun; Nam, Kyoungphile

2013-12-15

This study was conducted to investigate whether or not phosphate-solubilizing bacteria (PSB) as a kind of plant growth promoting rhizobacteria enhance the uptake of Cd by plants. In addition, the effect of PSB augmentation during phytoextraction on the microbial community of indigenous soil bacteria was also studied. In the initial Cd-contaminated soil, the major phyla were Proteobacteria (35%), Actinobacteria (38%) and Firmicutes (8%). While Proteobacteria were dominant at the second and sixth week (41 and 54%, respectively) in inoculated soil, Firmicutes (mainly belonging to the Bacilli class-61%), dramatically increased in the eight-week soil. For the uninoculated soil, the proportion of α-Proteobacteria increased after eight weeks (32%). Interestingly, Actinobacteria class, which was originally present in the soil (37%), seemed to disappear during phytoremediation, irrespective of whether PSB was inoculated or not. Cluster analysis and Principal Component Analysis revealed that the microbial community of eight-week inoculated soil was completely separated from the other soil samples, due to the dramatic increase of Bacillus aryabhattai. These findings revealed that it took at least eight weeks for the inoculated Bacillus sp. to functionally adapt to the introduced soil, against competition with indigenous microorganisms in soil. An ecological understanding of interaction among augmented bacteria, plant and indigenous soil bacteria is needed, for proper management of phytoextraction. Copyright © 2013 Elsevier B.V. All rights reserved.

Thymic Dendritic Cells Are Primary Targets for the Oncogenic Virus SL3-3

PubMed Central

Uittenbogaart, Christel H.; Law, Wendy; Leenen, Pieter J. M.; Bristol, Gregory; van Ewijk, Willem; Hays, Esther F.

1998-01-01

The murine retrovirus SL3-3 causes malignant transformation of thymocytes and thymic lymphoma in mice of the AKR and NFS strains when they are inoculated neonatally. The objective of the present study was to identify the primary target cells for the virus in the thymuses of these mice. Immunohistochemical studies of the thymus after neonatal inoculation of the SL3-3 virus showed that cells expressing the viral envelope glycoprotein (gp70+ cells) were first seen at 2 weeks of age. These virus-expressing cells were found in the cortex and at the corticomedullary junction in both mouse strains. The gp70+ cells had the morphology and immunophenotype of dendritic cells. They lacked macrophage-specific antigens. Cell separation studies showed that bright gp70+ cells were detected in a fraction enriched for dendritic cells. At 3 weeks of age, macrophages also expressed gp70. At that time, both gp70+ dendritic cells and macrophages were found at the corticomedullary junction and in foci in the thymic cortex. At no time during this 3-week period was the virus expressed in cortical and medullary epithelial cells or in thymic lymphoid cells. Infectious cell center assays indicated that cells expressing infectious virus were present in small numbers at 2 weeks after inoculation but increased at 5 weeks of age by several orders of magnitude, indicating virus spread to the thymic lymphoid cells. Thus, at 2 weeks after neonatal inoculation of SL3-3, thymic dendritic cells are the first cells to express the virus. At 3 weeks of age, macrophages also express the virus. In subsequent weeks, the virus spreads to the thymocytes. This pathway of virus expression in the thymus allows the inevitable provirus integration in a thymocyte that results in a clonal lymphoma. PMID:9811752

Co-infection of turkeys with Escherichia coli (O78) and H6N1 avian influenza virus.

PubMed

Umar, Sajid; Delverdier, Maxence; Delpont, Mattias; Belkasmi, Sakhia F Z; Teillaud, Angélique; Bleuart, Céline; Pardo, Isabelle; El Houadfi, Mohammed; Guérin, Jean-Luc; Ducatez, Mariette F

2018-06-01

Respiratory diseases are responsible for major economic losses in poultry farms. While in most cases a single pathogen is not alone responsible for the clinical outcome, the impact of co-infections is not well known, especially in turkeys. The purpose of this study was to assess the possible synergism between Escherichia coli (O78) and low pathogenic avian influenza virus (LPAIV, H6N1), in the turkey model. Four-week-old commercial turkeys were inoculated with either H6N1, O78 or both agents simultaneously or three days apart. We have established an experimental infection model of turkeys using aerosolization that better mimics field infections. Birds were observed clinically and swabbed on a daily basis. Necropsies were performed at 4 and 14 days post single or dual inoculation and followed by histological and immunohistochemical analyses. Combined LPAIV/E. coli infections resulted in more severe clinical signs, were associated with higher mortality and respiratory organ lesions (mucous or fibrinous exudative material in lungs and air sacs), in comparison with the groups given single infections (P < 0.05). The time interval or the sequence between H6N1 and E. coli inoculation (none or three days) did not have a significant effect on the outcome of the dual infection and disease although slightly greater (P > 0.05) respiratory signs were observed in turkeys of the E. coli followed by H6N1 inoculated group. Microscopic lesions and immunohistochemical staining supported clinical and macroscopic findings. Efficient virus and bacteria replication was observed in all inoculated groups. E. coli and H6N1 thus exercise an additive or synergistic pathogenic effect in the reproduction of respiratory disease.

Susceptibility of calves to porcine circovirus-2 (PCV2).

PubMed

Halami, Mohammad Y; Freick, Markus; Shehata, Awad A; Müller, Hermann; Vahlenkamp, Thomas W

2014-09-17

Circoviruses are known to infect pigs and birds and cause severe diseases with various clinical signs. Porcine circovirus-2 (PCV2), associated with severe economic losses, was detected in rodents, mosquitoes, cattle, and in calves affected with bovine neonatal pancytopenia (BNP). However, molecular and serological investigations on circovirus infections in cattle revealed inconsistent results. The aim of the study was to investigate the susceptibility and immune response of calves to experimental PCV2 inoculation. Animals were either intravenously inoculated with tissue-culture grown PCV2, with bone marrow from PCV2 positive and negative calves or immunized with a commercial inactivated PCV2 vaccine. The results showed that the animals inoculated with tissue-culture grown PCV2 and with PCV2 positive bone marrow displayed clinical signs including lymph node swelling, reddening of oral and ocular mucosa, and diarrhoea 7-18 days post inoculation (p.i.). PCV2-specific antibodies were detected in the tissue-culture grown PCV2-infected animals and in the PCV2-immunized animals from day 11 and 7 p.i. onwards, respectively, but were absent in both bone marrow inoculated groups. PCV2 was detected by real-time quantitative PCR only in blood samples of the tissue-culture grown PCV2-infected animals and in various tissues (e.g. spleen, lymph nodes, thymus), with high copy numbers in blood between day 4 (5.16log10 genomic copy number/ml) and 46 (5.33log10 genomic copy number/ml) p.i. In conclusion, the seroconversion and the detection of PCV2 in lymphoid tissues for more than five weeks p.i. revealed that host susceptibility of PCV2 is not solely restricted to pigs. Copyright © 2014 Elsevier B.V. All rights reserved.

Pneumococcal meningitis: development of a new animal model

PubMed Central

Wei, Benjamin P.C.; Shepherd, Robert K.; Robins-Browne, Roy M.; Clark, Graeme M.; O’Leary, Stephen J.

2007-01-01

Hypothesis The rat is a suitable animal to establish a model for the study of pneumococcal meningitis post cochlear implantation Background There has been an increase in the number of cases of cochlear implant-related meningitis. The most common organism identified was Streptococcus pneumoniae. Whether cochlear implantation increases the risk of pneumococcal meningitis in healthy subjects without other risk factors remains to be determined. Previous animal studies do not focus on the pathogenesis and risk of pneumococcal meningitis post implantation and are based on relatively small animal numbers, making it difficult to assess the cause and effect relationship. There is, therefore, a need to develop a new animal model allowing direct examination of the pathogenesis of meningitis in the presence of a cochlear implant. Methods Eighteen non-implanted rats were infected with 1× 106 and 1 × 108 colony forming units (CFU) of a clinical isolate of S. pneumoniae via three different inoculation routes (middle ear, inner ear and intraperitoneal) to examine for evidence of meningitis over 24 hours. Six implanted rats were infected with the highest amount of bacteria possible for each route of inoculation (4 × 1010 CFU intraperitoneal, 3 × 108CFU middle ear, 1 × 106 CFU inner ear) to examine for evidence of meningitis with the presence of an implant. Histological pattern of cochlear infections for each of the three different inoculating routes were examined. Results Pneumococcal meningitis was evident in all 6 implanted animals for each of the three different routes of inoculation. Once in the inner ear, bacteria were found to enter the central nervous system either via the cochlear aqueduct or canaliculi perforantes of osseous spiral lamina, reaching the perineural and perivascular space then the internal acoustic meatus. The rate, extent and pattern of infection within the cochleae depended on the route of inoculation. Finally, there was no evidence of pneumococcal meningitis observed in 18 non-implanted rats inoculated at a lower concentration of S. pneumoniae when observed for 24 hours post-inoculation. Conclusion Meningitis in implanted rats following inoculation with a clinical isolate of S. pneumoniae is possible via all three potential routes of infection via the upper respiratory tract. The lack of meningitis observed in the 18 non-implanted rats suggests longer post-inoculation monitoring periods are required to ensure whether or not meningitis will develop. Based on this work we have developed a new animal model that will allow quantitative risk assessment of meningitis post cochlear implantation, and the assessment of the efficacy of potential interventional strategies in future studies. PMID:16936571

Experimental Infection of Pig-Tailed Macaques (Macaca nemestrina) with Mycoplasma genitalium.

PubMed

Wood, Gwendolyn E; Patton, Dorothy L; Cummings, Peter K; Iverson-Cabral, Stefanie L; Totten, Patricia A

2017-02-01

Mycoplasma genitalium is an underappreciated cause of human reproductive tract disease, characterized by persistent, often asymptomatic, infection. Building on our previous experiments using a single female pig-tailed macaque as a model for M. genitalium infection (G. E. Wood, S. L. Iverson-Cabral, D. L. Patton, P. K. Cummings, Y. T. Cosgrove Sweeney, and P. A. Totten, Infect Immun 81:2938-2951, 2013, https://doi.org/10.1128/IAI.01322-12), we cervically inoculated eight additional animals, two of which were simultaneously inoculated in salpingeal tissue autotransplanted into abdominal pockets. Viable M. genitalium persisted in the lower genital tract for 8 weeks in three animals, 4 weeks in two, and 1 week in one; two primates resisted infection. In both animals inoculated in salpingeal pockets, viable M. genitalium was recovered for 2 weeks. Recovery of viable M. genitalium from lower genital tract specimens was improved by diluting the specimen in broth and by Vero cell coculture. Ascension to upper reproductive tract tissues was not detected, even among three persistently infected animals. M. genitalium-specific serum antibodies targeting the immunodominant MgpB and MgpC proteins appeared within 1 week in three animals inoculated both cervically and in salpingeal pockets and in one of three persistently infected animals inoculated only in the cervix. M. genitalium-specific IgG, but not IgA, was detected in cervical secretions of serum antibody-positive animals, predominantly against MgpB and MgpC, but was insufficient to clear M. genitalium lower tract infection. Our findings further support female pig-tailed macaques as a model of M. genitalium infection, persistence, and immune evasion. Copyright © 2017 American Society for Microbiology.

Experimental inoculation study indicates swine as a potential host for Hendra virus

PubMed Central

Li, Mingyi; Embury-Hyatt, Carissa; Weingartl, Hana M.

2010-01-01

Hendra virus (HeV) is a zoonotic virus from the family Paramyxoviridae causing fatal disease in humans and horses. Five-week-old Landrace pigs and 5-month-old Gottingen minipigs were inoculated with approximately 107 plaque forming units per animal. In addition to fever and depression exhibited in all infected pigs, one of the two Landrace pigs developed respiratory signs at 5 days post-inoculation (dpi) and one of the Gottingen minipigs developed respiratory signs at 5 dpi and mild neurological signs at 7 dpi. Virus was detected in all infected pigs at 2–5 dpi from oral, nasal, and rectal swabs and at 3–5 dpi from ocular swabs by real-time RT-PCR targeting the HeV M gene. Virus titers in nasal swab samples were as high as 104.6 TCID50/mL. The viral RNA was mainly distributed in tissues from respiratory and lymphoid systems at an early stage of infection and the presence of virus was confirmed by virus isolation. Pathological changes and immunohistochemical staining for viral antigen were consistent with the tissue distribution of the virus. This new finding indicates that pigs are susceptible to HeV infections and could potentially play a role as an intermediate host in transmission to humans. PMID:20167195

Potential of nisin-incorporated sodium caseinate films to control Listeria in artificially contaminated cheese.

PubMed

Cao-Hoang, Lan; Chaine, Aline; Grégoire, Lydie; Waché, Yves

2010-10-01

A sodium caseinate film containing nisin (1000 IU/cm(2)) was produced and used to control Listeria innocua in an artificially contaminated cheese. Mini red Babybel cheese was chosen as a model semi-soft cheese. L. innocua was both surface- and in-depth inoculated to investigate the effectiveness of the antimicrobial film as a function of the distance from the surface in contact with the film. The presence of the active film resulted in a 1.1 log CFU/g reduction in L. innocua counts in surface-inoculated cheese samples after one week of storage at 4 degrees C as compared to control samples. With regard to in-depth inoculated cheese samples, antimicrobial efficiency was found to be dependent on the distance from the surface in contact with the active films to the cheese matrix. The inactivation rates obtained were 1.1, 0.9 and 0.25 log CFU/g for distances from the contact surface of 1 mm, 2 mm and 3 mm, respectively. Our study demonstrates the potential application of sodium caseinate films containing nisin as a promising method to overcome problems associated with post-process contamination, thereby extending the shelf life and possibly enhancing the microbial safety of cheeses. 2010 Elsevier Ltd. All rights reserved.

Calcium phosphate coupled Newcastle disease vaccine elicits humoral and cell mediated immune responses in chickens.

PubMed

Koppad, Sanganagouda; Raj, G Dhinakar; Gopinath, V P; Kirubaharan, J John; Thangavelu, A; Thiagarajan, V

2011-12-01

Calcium phosphate (CaP) particles were coupled with inactivated Newcastle disease virus (NDV) vaccine. The surface morphology of CaP particles coupled to NDV was found to be spherical, smooth and with a tendency to agglomerate. The mean (± SE) size of CaP particles was found 557.44 ± 18.62 nm. The mean percent encapsulation efficiency of CaP particles coupled to NDV assessed based on total protein content and haemagglutination (HA) activity in eluate was found to be 10.72 ± 0.89 and 12.50 ± 2.09, respectively. The humoral and cell mediated immune responses induced by CaP coupled NDV vaccine were assessed in comparison to a commercial live vaccine (RDV 'F'). CaP coupled NDV vaccine elicited prolonged haemagglutination inhibition (HI) and enzyme linked immunosorbent assay (ELISA) titres in the serum even at fourth and fifth week post-vaccination (PV), unlike RDV 'F' inoculated chickens whose titres declined to insignificant levels by this time. CaP coupled NDV vaccine could stimulate HI antibodies in tracheal washings and tears from second and first week PV, respectively. IgA ELISA antibodies were also seen in tracheal washings of these birds from third week PV and in tears from second week PV. CaP coupled NDV vaccine elicited cell mediated immune responses (CMI) from two to four weeks PV. The stimulation indices obtained after stimulation with specific antigen was not significantly different between CaP coupled antigen and live NDV virus except on first week PV. However, CaP coupled antigen did not cause suppression of lympo proliferation as indicated by statistically similar responses to mitogen, concanavalin A between the two groups. Overall, CaP coupled NDV vaccine elicited stronger and prolonged immune responses in comparison to the commercial live vaccine. No increase in the serum calcium and phosphorous levels were seen in CaP coupled NDV vaccine inoculated chickens. Copyright © 2010 Elsevier Ltd. All rights reserved.

EFFECTS OF ELECTROMAGNETICALLY SIGNALIZED MEDIA ON HOST-PATHOGEN INTERACTION.

PubMed

D'Hallewin, G; Venditti, T; Cubaiu, L; Ladu, G; Renati, P

2014-01-01

Up to date, limited data are available about electromagnetic phase signaling effects on host-pathogen interactions during the postharvest of horticultural commodities. Inspired by the last striking works on water physics, quantum signaling through phase transfer and its impact on biological and histological structures, we studied the effect of different electromagnetic signals on pome blue mold (Penicillium expansum) pathogenesis. Tags with different electromagnetic-signals (EmS) were used to generate 3 Coherent Electro Dynamic (CED) environments. Artificially wounded 'Coscia' pears, placed onto 3 EmS tags (QF, QA and QR), were employed for the in vivo experiment. Whereas, a set of wounded-fruit placed onto an un-electromagnetic-signalized tag (QN) or kept without tag were used as blank or control, respectively. Inoculation was performed 2 or 24 h post-wounding with P. expansum conidia. The same tags placed under Petri dishes containing dot-inoculated PDA served for the in vitro experiment. Both experiments performed at 25 degrees C endured 7 days. The percentage of infected wounds was calculated and the radial growth measured in vitro. Concerning the in vivo experiment, 100% of control and blank fruit inoculated 2 h post-wounding was infected after 5 days, while, 97% after 7 days, when inoculation occurred 24 h post-wounding. Compared to control and blank, the pathogenesis in fruit placed on the EmS tags resulted inhibited, and when fruit was inoculated 2 h post-wounding, the infection degree on QF, QA and QR tags resulted 19, 52 and 64%, respectively. The degree for the same EmS tags was significantly lower when fruit was inoculated 24 h post-wounding (9, 32 and 42%, respectively). The in vitro experiment evidenced a notable inhibition of the radial growth by all EmS tags in comparison to control and blank (51 mm), while the QF tag provided the greatest inhibition (12 mm).

Two different genotypes of H1N2 swine influenza virus isolated in northern China and their pathogenicity in animals.

PubMed

Yang, Huanliang; Chen, Yan; Qiao, Chuanling; Xu, Chuantian; Yan, Minghua; Xin, Xiaoguang; Bu, Zhigao; Chen, Hualan

2015-02-25

During 2006 and 2007, two swine-origin triple-reassortant influenza A (H1N2) viruses were isolated from pigs in northern China, and the antigenic characteristics of the hemagglutinin protein of the viruses were examined. Genotyping and phylogenetic analyses demonstrated different emergence patterns for the two H1N2 viruses, Sw/Hebei/10/06 and Sw/Tianjin/1/07. Sequences for the other genes encoding the internal proteins were compared with the existing data to determine their origins and establish the likely mechanisms of genetic reassortment. Sw/Hebei/10/06 is an Sw/Indiana/9K035/99-like virus, whereas Sw/Tianjin/1/07 represents a new H1N2 genotype with surface genes of classic swine and human origin and internal genes originating from the Eurasian avian-like swine H1N1 virus. Six-week-old female BALB/c mice infected with the Sw/HeB/10/06 and Sw/TJ/1/07 viruses showed an average weight loss of 12.8% and 8.1%, respectively. Healthy six-week-old pigs were inoculated intranasally with either the Sw/HeB/10/06 or Sw/TJ/1/07 virus. No considerable changes in the clinical presentation were observed post-inoculation in any of the virus-inoculated groups, and the viruses effectively replicated in the nasal cavity and lung tissue. Based on the results, it is possible that the new genotype of the swine H1N2 virus that emerged in China may become widespread in the swine population and pose a potential threat to public health. Copyright © 2014 Elsevier B.V. All rights reserved.

Effectiveness of oxfendazole against early and later 4th-stage Strongylus vulgaris in ponies.

PubMed

Slocombe, J O; McCraw, B M; Pennock, P; Ducharme, N G; Baird, J D

1986-03-01

Twenty pony foals (reared worm free), 6.5 to 10 weeks of age, were inoculated with Strongylus vulgaris and allocated to 5 groups, each with 4 foals. One week after inoculation, 1 group of 4 foals was given oxfendazole (OFZ) at a dosage rate of 10 mg/kg of body weight, another group was given 2 such treatments 48 hours apart, and a 3rd group was given a placebo. All treatments were administered by stomach tube. Three weeks later, foals were euthanatized and necropsied in a test for efficacy against early 4th-stage larvae. Oxfendazole was 80% and 94.9% effective against early 4th-stage S vulgaris with 1 and 2 doses, respectively. A 4th group of 4 foals was given 2 treatments of OFZ, 48 hours apart, about 8 weeks after inoculation, and a 5th group was given a placebo. These foals were euthanatized and necropsied 5 weeks after treatment in a test for efficacy against later 4th-stage larvae. Two doses of OFZ were 96.6% effective against later 4th-stage larvae.

Type of tomatoes and water rinse affect efficacy of acid washes against salmonella enterica inoculated on stem scar areas of tomatoes and on product quality

USDA-ARS?s Scientific Manuscript database

The study was conducted to evaluate the effects of post-treatment rinsing with water on the inactivation efficacy of acid treatments against Salmonella inoculated onto stem scar areas of two types of tomatoes. In addition, impact on fruit quality was investigated during 21 days post-treatment storag...

Construction of an infectious cDNA clone of avian hepatitis E virus (avian HEV) recovered from a clinically healthy chicken in the United States and characterization of its pathogenicity in specific-pathogen-free chickens.

PubMed

Kwon, Hyuk Moo; LeRoith, Tanya; Pudupakam, R S; Pierson, F William; Huang, Yao-Wei; Dryman, Barbara A; Meng, Xiang-Jin

2011-01-27

A genetically distinct strain of avian hepatitis E virus (avian HEV-VA strain) was isolated from a healthy chicken in Virginia, and thus it is important to characterize and compare its pathogenicity with the prototype strain (avian HEV-prototype) isolated from a diseased chicken. Here we first constructed an infectious clone of the avian HEV-VA strain. Capped RNA transcripts from the avian HEV-VA clone were replication-competent after transfection of LMH chicken liver cells. Chickens inoculated intrahepatically with RNA transcripts of avian HEV-VA clone developed active infection as evidenced by fecal virus shedding, viremia, and seroconversion. To characterize the pathogenicity, RNA transcripts of both avian HEV-VA and avian HEV-prototype clones were intrahepatically inoculated into the livers of chickens. Avian HEV RNA was detected in feces, serum and bile samples from 10/10 avian HEV-VA-inoculated and 9/9 avian HEV-prototype-inoculated chickens although seroconversion occurred only in some chickens during the experimental period. The histopathological lesion scores were lower for avian HEV-VA group than avian HEV-prototype group in the liver at 3 and 5 weeks post-inoculation (wpi) and in the spleen at 3 wpi, although the differences were not statistically significant. The liver/body weight ratio, indicative of liver enlargement, of both avian HEV-VA and avian HEV-prototype groups were significantly higher than that of the control group at 5 wpi. Overall, the avian HEV-VA strain still induces histological liver lesions even though it was isolated from a healthy chicken. The results also showed that intrahepatic inoculation of chickens with RNA transcripts of avian HEV infectious clone may serve as an alternative for live virus in animal pathogenicity studies. Copyright © 2010 Elsevier B.V. All rights reserved.

Effect of housing arrangement on fecal-oral transmission of avian hepatitis E virus in chicken flocks.

PubMed

Liu, Baoyuan; Sun, Yani; Chen, Yiyang; Du, Taofeng; Nan, Yuchen; Wang, Xinjie; Li, Huixia; Huang, Baicheng; Zhang, Gaiping; Zhou, En-Min; Zhao, Qin

2017-09-07

Avian hepatitis E virus (HEV) infection is common in chicken flocks in China, as currently no measures exist to prevent the spread of the disease. In this study, we analyzed the effect of caged versus cage-free housing arrangements on avian HEV transmission. First, 127 serum and 110 clinical fecal samples were collected from 4 chicken flocks including the two arrangements in Shaanxi Province, China and tested for HEV antibodies and/or virus. Concurrently, 36 specific-pathogen-free chickens were divided equally into four experimental living arrangement groups, designated cage-free (Inoculated), caged (Inoculated), cage-free (Negative) and caged (Negative) groups. In caged groups, three cages contained 3 chickens each. Three chickens each from cage-free (Inoculated) and caged (Inoculated) groups (one chicken of each cage) were inoculated by cutaneous ulnar vein with the same dose of avian HEV, respectively. The cage-free (Negative) and caged (Negative) groups served as negative control. Serum and fecal samples were collected at 1 to 7 weeks post-inoculation (wpi) and liver lesions were scored at 7 wpi. The results of serology showed that the avian HEV infection rate (54.10%) of the cage-free chickens was significantly higher than the one (12.12%) for caged chickens (P < 0.05). Also, the rate of detection of avian HEV RNA in the clinical fecal samples was significantly higher in the cage-free (22.80%, 13/57) than caged birds (5.66%, 3/53). Moreover, under experimental conditions, the infected number of uninoculated cage-free chickens (6) was significantly higher than the one for the uninoculated caged birds (2), as evidenced by seroconversion, fecal virus shedding, viremia and gross and microscopic liver lesions. These results suggest that reduction of contact with feces as seen in the caged arrangement of housing chickens can reduce avian HEV transmission. This study provides insights for prevention and control of avian HEV infection in chicken flocks.

Growth, cadmium uptake and accumulation of maize (Zea mays L.) under the effects of arbuscular mycorrhizal fungi.

PubMed

Liu, Lingzhi; Gong, Zongqiang; Zhang, Yulong; Li, Peijun

2014-12-01

The effects of three arbuscular mycorrhizal fungi isolates on Cd uptake and accumulation by maize (Zea mays L.) were investigated in a planted pot experiment. Plants were inoculated with Glomus intraradices, Glomus constrictum and Glomus mosseae at three different Cd concentrations. The results showed that root colonization increased with Cd addition during a 6-week growth period, however, the fungal density on roots decreased after 9-week growth in the treatments with G. constrictum and G. mosseae isolates. The percentage of mycorrhizal colonization by the three arbuscular mycorrhizal fungi isolates ranged from 22.7 to 72.3%. Arbuscular mycorrhizal fungi inoculations decreased maize biomass especially during the first 6-week growth before Cd addition, and this inhibitory effect was less significant with Cd addition and growth time. Cd concentrations and uptake in maize plants increased with arbuscular mycorrhizal fungi colonization at low Cd concentration (0.02 mM): nonetheless, it decreased at high Cd concentration (0.20 mM) after 6-week growth period. Inoculation with G. constrictum isolates enhanced the root Cd concentrations and uptake, but G. mosseae isolates showed the opposite results at high Cd concentration level after 9 week growth period, as compared to non-mycorrhizal plants. In conclusion, maize plants inoculated with arbuscular mycorrhizal fungi were less sensitive to Cd stress than uninoculated plants. G. constrictum isolates enhanced Cd phytostabilization and G. mosseae isolates reduced Cd uptake in maize (Z. mays L.).

Screenhouse and field persistence of nonpathogenic endophytic Fusarium oxysporum in Musa tissue culture plants.

PubMed

Paparu, Pamela; Dubois, Thomas; Gold, Clifford S; Niere, Björn; Adipala, Ekwamu; Coyne, Daniel

2008-04-01

Two major biotic constraints to highland cooking banana (Musa spp., genome group AAA-EA) production in Uganda are the banana weevil Cosmopolites sordidus and the burrowing nematode Radopholus similis. Endophytic Fusarium oxysporum strains inoculated into tissue culture banana plantlets have shown control of the banana weevil and the nematode. We conducted screenhouse and field experiments to investigate persistence in the roots and rhizome of two endophytic Fusarium oxysporum strains, V2w2 and III4w1, inoculated into tissue-culture banana plantlets of highland cooking banana cultivars Kibuzi and Nabusa. Re-isolation of F. oxysporum showed that endophyte colonization decreased faster from the rhizomes than from the roots of inoculated plants, both in the screenhouse and in the field. Whereas rhizome colonization by F. oxysporum decreased in the screenhouse (4-16 weeks after inoculation), root colonization did not. However, in the field (17-33 weeks after inoculation), a decrease was observed in both rhizome and root colonization. The results show a better persistence in the roots than rhizomes of endophytic F. oxysporum strains V2w2 and III4w1.

Leishmania-derived trimannose modulates inflammatory response to significantly reduce Leishmania (L.) major-induced lesions.

PubMed

Grinnage-Pulley, Tara L; Kabotso, Daniel E K; Rintelmann, Chelsea L; Roychoudhury, Rajarshi; Schaut, Robert G; Toepp, Angela J; Gibson-Corley, Katherine N; Parrish, Molly; Pohl, Nicola L B; Petersen, Christine A

2017-10-23

Leishmania lipophosphoglycan (LPG) is a key virulence factor, initiating inflammation resulting in cutaneous lesions. LPG is capped by various oligosaccharides. How these glycans are recognized and how they alter the course of Leishmania infection is poorly understood. Previous studies synthesized α-1,2-trimannose cap sugars on latex beads demonstrated that C57BL/6 mice co-inoculated with L. major and trimannose-coated beads produced significantly higher levels of IL-12 p40 and other pro-inflammatory, type 1 cytokines compared L. major infection alone within the first 48 h of infection. However, as L. major infection typically progress over weeks to months, the role of trimannose in altering disease progression over the course of infection was unknown. Wild-type mice were inoculated with either trimannose or carrier (uncoated) beads, infected with L. major alone, co-inoculated with carrier beads and L. major, or co-inoculated with trimannose beads and L. major Trimannose treatment of L. major- infected mice decreased parasite load and significantly decreased lesion size at 14 days post infection (pi) compared to non-treated, infected mice. Infected, trimannose-treated mice had decreased IL-12p40 and IL-10 secretion and increased IFN-γ at 14 days pi. Mice lacking the ability to detect trimannose, mannose-receptor deleted mice (MR -/- ), when treated with trimannose beads and infected with L. major did not have decreased lesion size. Leishmania -derived trimannose represents a novel immunomodulator that provides early type 1-skewed cytokine production to control parasite load and alter the course of cutaneous leishmaniasis. Copyright © 2017 American Society for Microbiology.

Cross protective immune responses in nursing piglets infected with a US spike-insertion deletion porcine epidemic diarrhea virus strain and challenged with an original US PEDV strain.

PubMed

Annamalai, Thavamathi; Lin, Chun-Ming; Gao, Xiang; Liu, Xinsheng; Lu, Zhongyan; Saif, Linda J; Wang, Qiuhong

2017-10-06

We investigated cross-protective immunity of a US spike-insertion deletion porcine epidemic diarrhea virus (PEDV) Iowa106 (S-INDEL) strain against the original US PEDV (PC21A) strain in nursing piglets. Piglets were inoculated orally with S-INDEL, PC21A or mock. At 20-29 days post-inoculation (dpi), all pigs were challenged with the PC21A strain. The S-INDEL-inoculated pigs had lower ileal IgA antibody secreting cells, serum IgA and neutralizing antibody titers compared with PC21A-inoculated pigs. No pigs in the PC21A-group developed diarrhea, whereas 81 and 100% of pigs in the S-INDEL and mock-groups had diarrhea post challenge, respectively. S-INDEL induced partial protective immunity against the original US PEDV strain.

Coexistence of Helicobacter pylori spiral and coccoid forms in experimental mice

PubMed Central

Hua, Jiesong; Ho, Bow; Zheng, Pengyuan; Yeoh, Khay Guan; Ng, Han Chong; Lim, Seng Gee

1998-01-01

AIM: To infect mice with Helicobacter pylori and detect immune response against two form of H. pylori. METHODS: An isolate of H. pylori obtained from a patient with gastric cancer was used to infect mice. Fifty mice were divided into eight groups. Two groups served as negative control without any inoculation and internal negative control with 0.5 M NaHCO3 and brain heart infusion (HBI), respectively. Mice in each experimental group were first inoculated with 0.5 M NaHCO3 and then H. pylori suspension for 3 times at a 2-d interval. Mice from controls and infectious groups were sacrificed at a weekly interval postinfection. Gastric samples were trimmed, inoculated onto chocolate blood agar and then incujbated in microaerophilic atmosphere at 37¡æ for 14 d. Sera were examined for immunoglobulins against H. pylori spiral and coccoid antigens by ELISA. RESULTS: After inoculation H. pylori was isolated in one mouse from one week postinfection. No H. pylori was detected in control mice. However, urease test was positive in 50% (5/10) control mice, 70% (7/10) mice inoculated with NaHCO3 and BHI and 77% (23/30) mice infected with H. pylori. The systemic immune responses of the mice to H. pylori strain were determined by ELISA. The mice showed immune responses to both H. pylori spiral and coccoid antigens one week after infection with H. pylori. The peak mean absorbances of antibodies against spiral and coccoid forms were four weeks postinfection which showed 6 and 18 times higher than that of negative control group respectively (P < 0.01). CONCLUSION: Spiral and coccoid forms of H. pylori coexist in experimental mice studied. PMID:11819350

Effects of gamma radiation and azathioprine on Brucella abortus infection in BALB/c mice

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

Elzer, P.H.; Rowe, G.E.; Enright, F.M.

Sublethal irradiation of BALB/c mice 4 hours prior to inoculation with 5 {times} 10(4) virulent Brucella abortus, caused significant (P less than 0.01) reductions in bacterial numbers in comparison with numbers in unirradiated controls. Numbers of brucellae in the spleen were significantly lower by 5 days after inoculation and decreased thereafter, so that at 2 and 3 weeks after inoculation, there were up to 1,000-fold fewer organisms in the spleen of irradiated mice. The number of brucellae in the spleen increased in irradiated mice thereafter. The course of events in the liver was similar, but developed more slowly, and peakmore » differences in bacterial numbers were about 1 log less. These phenomena were not attributable to differences in implantation of brucellae in the liver or spleen, nor to an abnormal distribution of organisms in other organs of irradiated mice. Irradiation of mice during the plateau phase of infection also resulted in significant (P less than 0.05) reductions in bacterial counts in the spleen during the succeeding 4 weeks. Macrophage activation in the spleen, measured by a Listeria monocytogenes-killing assay, was significantly (P less than 0.01) increased by irradiation alone at 1 week after inoculation and at that time was significantly (P less than 0.01) greater in B abortus-infected, irradiated mice than in B abortus-infected controls. Histologic, cytologic, and immunologic studies revealed that the decrease in numbers of organisms between 1 and 2 weeks after inoculation in irradiated mice occurred at a time when their immune response to B abortus was suppressed and when numbers of neutrophils and monocytes infiltrating the spleen were significantly (P less than 0.01) diminished.« less

Uptake and Persistence of Homologous and Heterologous Zooxanthellae in the Temperate Sea Anemone Cereus pedunculatus (Pennant).

PubMed

Davy, S K; Lucas, I A N; Turner, J R

1997-04-01

The uptake and persistence of symbiotic dinoflagellates (zooxanthellae) were measured in the temperate sea anemone Cereus pedunculatus (Pennant). Aposymbiotic specimens of C. pedunculatus were inoculated with zooxanthellae freshly isolated from a range of temperate and subtropical Anthozoa. Each inoculate consisted of zooxanthellae from a single host species and was either homologous (zooxanthellae from a host of the same species as the one being inoculated) or heterologous (from a host of a different species than the one being inoculated). The densities of zooxanthellae in host tissues were determined at regular intervals. C. pedunculatus took up homologous and heterologous zooxanthellae to similar degrees, except for zooxanthellae from the temperate Anthopleura ballii, which were taken up to a lesser extent. The densities of all zooxanthellae declined between 4 hours and 4 days after uptake, indicating that zooxanthellae were expelled, digested, or both during this period. The densities of all zooxanthellae increased between 2 and 8 weeks after inoculation, indicating zooxanthella growth. Over the entire 8-week period after uptake, densities of homologous zooxanthellae were always greater than those of heterologous zooxanthellae. Between 8 and 36 weeks after infection, densities of homologous zooxanthellae declined markedly and densities of some heterologous zooxanthellae increased further, resulting in homologous and heterologous zooxanthella densities being the same at 36 weeks. These densities were the same as those in naturally infected C. pedunculatus of similar size. The results suggest that zooxanthellae from a range of host species and environments can establish symbioses with C. pedunculatus and that, over long periods under laboratory conditions, heterologous zooxanthellae may populate C. pedunculatus to the same extent as homologous zooxanthellae.

Immunization against East Coast Fever in field cattle with low infectivity Theileria parva stabilate--preliminary assessment.

PubMed

Mbassa, G K; Kweka, L E; Dulla, P N

1998-05-01

Two Theileria parva sporozoite stabilates stored at -196 degrees C, then at -70 degrees C for six weeks (stabilate 1) and more than six months (stabilate 2) were inoculated into four eight-month old male calves, 1 and 2 (stabilate 1), and 3 and 4 (stabilate 2). Calves 1 and 2 developed pyrexia, enlargement of lymph nodes, and the former died of East Coast Fever. Calves 3 and 4 showed slight enlargement of lymph nodes without fever. Lymph node smears from all calves (from day 10 to 20 post-inoculation) showed lymphoblasts, phagocytic macrophages, and schizonts. Piroplasms were detected in erythrocytes in blood smears from calves 1 and 2 but not in calves 3 and 4. Calves 2, 3 and 4 recovered without any treatment while calf 1 died of East Coast Fever on day 20. Serum samples from recovered calves taken on day 30 of the experiment were positive for antischizont antibodies to T. parva at 1:640 dilution, but pre-inoculation serum samples were negative. Stabilate 2 was used to immunize 64 Boran, Friesian, Ayrshire and crosses with Zebu cattle in four herds with 25% reduction of oxytetracycline dose. All the animals except one calf recovered without any severe reactions. The latter died of disease other than ECF after the monitoring period was over (day 24). Day 30 post-inoculation serum samples were positive for T. parva antischizont antibodies. A follow-up of the remaining animals for over one year revealed no further ECF incidences in these herds. This experiment shows the loss of infectivity of the vaccine stored at temperatures higher than -196 degrees C. dependent on the duration. However, despite the lack of clinical signs in calves 3 and 4, there was cellular response and antibody production, and the stabilate for vaccine against East Coast fever can thus be stored prior to use at higher than -196 degrees C and still maintain capability to produce antibodies in field cattle, eliminating the use of oxytetracycline and monitoring. The vaccine will be cheaper and easier to use and the requirement for liquid nitrogen in the field reduced and the scale of application of the vaccine widened.

Effects of atmospheric ammonia on young pigs experimentally infected with Bordetella bronchiseptica

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

Drummond, J.G.; Curtis, S.E.; Meyer, R.C.

1981-06-01

Effects of atmospheric ammonia on performance and respiratory tract health of young pigs experimentally infected with Bordetella bronchiseptica were studied. Treatments were: (1) control, (2) Bordetella inoculation (approx 10(9) bacteria/naris) alone, (3) Bordetella inoculation plus exposure to atmospheric ammonia at 34.7 mg/m3 (50 ppm), and (4) Bordetella inoculation plus exposure to atmospheric ammonia at 69.4 mg/m3 (100 ppm). Pigs weighted 8.01 kg (av) at start of treatment. Body weight and feed disappearance were measured weekly. After 4 weeks, all pigs were killed and examined grossly, and appropriate specimens were obtained for histopathologic examination. Regression models were fitted to growth, feedmore » disappearance, and gain-to-feed data. The growth model indicated that Bordetella-inoculated pigs gained 26% less body weight than did controls, regardless of atmospheric ammonia concentration. Bordetella inoculation, regardless of ammonia exposure, reduced feed disappearance 12% below the control rate. Treatment difference was not noted in gain/feed data. Shrunken turbinates were observed in Bordetella-inoculated pigs. Shrinkage also appeared to be related directly to ammonia concentration. Rhinitis was confirmed histopathologically, and its severity was related with atmospheric ammonia concentration, but no difference was seen in the osseous core of the turbinates.« less

Expression profiles of defence related cDNAs in oil palm (Elaeis guineensis Jacq.) inoculated with mycorrhizae and Trichoderma harzianum Rifai T32.

PubMed

Tan, Yung-Chie; Wong, Mui-Yun; Ho, Chai-Ling

2015-11-01

Basal stem rot is one of the major diseases of oil palm (Elaies guineensis Jacq.) caused by pathogenic Ganoderma species. Trichoderma and mycorrhizae were proposed to be able to reduce the disease severity. However, their roles in improving oil palm defence system by possibly inducing defence-related genes in the host are not well characterized. To better understand that, transcript profiles of eleven putative defence-related cDNAs in the roots of oil palm inoculated with Trichoderma harzianum T32 and mycorrhizae at different time points were studied. Transcripts encoding putative Bowman-Birk protease inhibitor (EgBBI2) and defensin (EgDFS) increased more than 2 fold in mycorrhizae-treated roots at 6 weeks post inoculation (wpi) compared to those in controls. Transcripts encoding putative dehydrin (EgDHN), glycine-rich RNA binding protein (EgGRRBP), isoflavone reductase (EgIFR), type 2 ribosome inactivating protein (EgT2RIP), and EgDFS increased in the oil palm roots treated with T. harzianum at 6 and/or 12 wpi compared to those in the controls. Some of these genes were also expressed in oil palm roots treated with Ganoderma boninense. This study provides an insight of some defence-related genes induced by Trichoderma and mycorrhizae, and their roles as potential agents to boost the plant defence system. Copyright © 2015 Elsevier Masson SAS. All rights reserved.

Melon Resistance to Cucurbit yellow stunting disorder virus Is Characterized by Reduced Virus Accumulation.

PubMed

Marco, Cristina F; Aguilar, Juan M; Abad, Jesús; Gómez-Guillamón, María Luisa; Aranda, Miguel A

2003-07-01

ABSTRACT The pattern of accumulation of Cucurbit yellow stunting disorder virus (CYSDV; genus Crinivirus, family Closteroviridae) RNA has been analyzed in several cucurbit accessions. In susceptible accessions of melon (Cucumis melo), cucumber (Cucumis sativus), marrow (Cucurbita maxima), and squash (Cucurbita pepo), CYSDV RNA accumulation peaked during the first to second week postinoculation in the first to third leaf above the inoculated one; younger leaves showed very low or undetectable levels of CYSDV. Three melon accessions previously shown to remain asymptomatic after CYSDV inoculation under natural conditions were also assayed for their susceptibility to CYSDV. Hybridization and reverse transcription-polymerase chain reaction (RT-PCR) analysis of noninoculated leaves showed that only one of these, C-105, remained virus-free for up to 6 weeks after whitefly inoculation. In this accession, very low CYSDV levels were detected by RT-PCR in whitefly-inoculated leaves, and therefore, multiplication or spread of CYSDV in C-105 plants appeared to remain restricted to the inoculated leaves. When C-105 plants were graft inoculated, CYSDV RNA could be detected in phloem tissues, but the systemic colonization of C-105 by CYSDV upon graft inoculation seemed to be seriously impeded. Additionally, in situ hybridization experiments showed that, after C-105 graft inoculation, only a portion of the vascular bundles in petioles and stems were colonized by CYSDV and virus could not be found in leaf veins. RT-PCR experiments using primers to specifically detect negative-sense CYSDV RNA were carried out and showed that CYSDV replication took place in graft-inoculated C-105 scions. Therefore, the resistance mechanism may involve a restriction of the virus movement in the vascular system of the plants and/or prevention of high levels of virus accumulation.

An evaluation of the wilt-causing bacterium Ralstonia solanacearum as a potential biological control agent for the alien Kahili ginger (Hedychium gardnerianum) in Hawaiian forests

USGS Publications Warehouse

1999-01-01

Kahili ginger (Hedychium gardnerianum) is an invasive weed in tropical forests in Hawaii and elsewhere. Bacterial wilt caused by the ginger strain of Ralstonia(=Pseudomonas) solanacearum systemically infects edible ginger (Zingiber officinale) and ornamental gingers (Hedychium spp.), causing wilt in infected plants. The suitability of R. solanacearum as a biological control agent for kahili ginger was investigated by inoculating seedlings and rooted cuttings of native forest plants, ornamental ginger, and solanaceous species to confirm host specificity. Inoculation via stem injection or root wounding with a bacterial–water suspension was followed by observation for 8 weeks. Inoculations on H. gardnerianum were then carried out in ohia-lehua (Metrosideros polymorpha) wet forests of Hawaii Volcanoes National Park to determine the bacterium's efficacy in the field. No native forest or solanaceous species developed wilt or other symptoms during the study. The bacterium caused limited infection near the inoculation site on H. coronarium, Z. zerumbet, Heliconia latispatha, and Musa sapientum. However, infection did not become systemic in any of these species, and normal growth resumed following appearance of initial symptoms. All inoculated H. gardnerianum plants developed irreversible chlorosis and severe wilting 3–4 weeks following inoculation. Systemic infection also caused death and decay of rhizomes. Most plants were completely dead 16–20 weeks following inoculation. The destructiveness of the ginger strain of R. solanacearum to edible ginger has raised questions regarding its use for biological control. However, because locations of kahili ginger infestations are often remote, the risk of contaminating edible ginger plantings is unlikely. The ability of this bacterium to cause severe disease in H. gardnerianum in the field, together with its lack of virulence in other ginger species, contributes to its potential as a biological control agent.

Standardization and validation of real time PCR assays for the diagnosis of histoplasmosis using three molecular targets in an animal model

PubMed Central

López, Luisa F.; Muñoz, César O.; Cáceres, Diego H.; Tobón, Ángela M.; Loparev, Vladimir; Clay, Oliver; Chiller, Tom; Litvintseva, Anastasia; Gade, Lalitha; González, Ángel

2017-01-01

Histoplasmosis is considered one of the most important endemic and systemic mycoses worldwide. Until now few molecular techniques have been developed for its diagnosis. The aim of this study was to develop and evaluate three real time PCR (qPCR) protocols for different protein-coding genes (100-kDa, H and M antigens) using an animal model. Fresh and formalin-fixed and paraffin-embedded (FFPE) lung tissues from BALB/c mice inoculated i.n. with 2.5x106 Histoplasma capsulatum yeast or PBS were obtained at 1, 2, 3, 4, 8, 12 and 16 weeks post-infection. A collection of DNA from cultures representing different clades of H. capsulatum (30 strains) and other medically relevant pathogens (36 strains of related fungi and Mycobacterium tuberculosis) were used to analyze sensitivity and specificity. Analytical sensitivity and specificity were 100% when DNAs from the different strains were tested. The highest fungal burden occurred at first week post-infection and complete fungal clearance was observed after the third week; similar results were obtained when the presence of H. capsulatum yeast cells was demonstrated in histopathological analysis. In the first week post-infection, all fresh and FFPE lung tissues from H. capsulatum-infected animals were positive for the qPCR protocols tested except for the M antigen protocol, which gave variable results when fresh lung tissue samples were analyzed. In the second week, all qPCR protocols showed variable results for both fresh and FFPE tissues. Samples from the infected mice at the remaining times post-infection and uninfected mice (controls) were negative for all protocols. Good agreement was observed between CFUs, histopathological analysis and qPCR results for the 100-kDa and H antigen protocols. We successfully standardized and validated three qPCR assays for detecting H. capsulatum DNA in fresh and FFPE tissues, and conclude that the 100-kDa and H antigen molecular assays are promising tests for diagnosing this mycosis. PMID:29287097

Derivation of host and pathogen genotypes in the fusiform rust pathosystem on slash pine using a complimentary genetics model and diallel data

Treesearch

H.E. Stelzel; Robert L. Doudrick; Thomas L. Kubisiak

1997-01-01

Seedlings from 20, full-sib families five-parent slash pine diallel were inoculated using two, single urediniospore-derived cultures of the fusiform rust fungus on two different dates during the 1994 growing season. Presence or absence of fusiform rust galls was recorded for each inoculated seedling at nine months post-inoculation and percent infection levels for each...

Urban inoculation and the decline of smallpox mortality in eighteenth-century cities-a reply to Razzell.

PubMed

Davenport, Romola J; Boulton, Jeremy; Schwarz, Leonard

2016-02-01

Smallpox was probably the single most lethal disease in eighteenth-century Britain but was reduced to a minor cause of death by the mid-nineteenth century due to vaccination programmes post-1798. While the success of vaccination is unquestionable, it remains disputed to what extent the prophylactic precursor of vaccination, inoculation, reduced smallpox mortality in the eighteenth century. Smallpox was most lethal in urban populations, but most researchers have judged inoculation to have been unpopular in large towns. Recently, however, Razzell argued that inoculation significantly reduced smallpox mortality of adults and older children in London in the last third of the eighteenth century. This article uses demographic evidence from London and Manchester to confirm previous findings of a sudden fall in adult smallpox mortality and a rise in the importance of smallpox in early childhood c . 1770. The nature of these changes is consistent with an increase in smallpox transmission in London and Manchester after 1770 and indicates that smallpox inoculation was insufficient to reduce smallpox mortality in large towns. It remains unclear whether inoculation could have operated to enhance smallpox transmission or whether changes in the properties of the smallpox virus drove the intensification of smallpox mortality among young children post-1770.

Competition between influenza A virus subtypes through heterosubtypic immunity modulates re-infection and antibody dynamics in the mallard duck.

PubMed

Latorre-Margalef, Neus; Brown, Justin D; Fojtik, Alinde; Poulson, Rebecca L; Carter, Deborah; Franca, Monique; Stallknecht, David E

2017-06-01

Our overall hypothesis is that host population immunity directed at multiple antigens will influence the prevalence, diversity and evolution of influenza A virus (IAV) in avian populations where the vast subtype diversity is maintained. To investigate how initial infection influences the outcome of later infections with homologous or heterologous IAV subtypes and how viruses interact through host immune responses, we carried out experimental infections in mallard ducks (Anas platyrhynchos). Mallards were pre-challenged with an H3N8 low-pathogenic IAV and were divided into six groups. At five weeks post H3N8 inoculation, each group was challenged with a different IAV subtype (H4N5, H10N7, H6N2, H12N5) or the same H3N8. Two additional pre-challenged groups were inoculated with the homologous H3N8 virus at weeks 11 and 15 after pre-challenge to evaluate the duration of protection. The results showed that mallards were still resistant to re-infection after 15 weeks. There was a significant reduction in shedding for all pre-challenged groups compared to controls and the outcome of the heterologous challenges varied according to hemagglutinin (HA) phylogenetic relatedness between the viruses used. There was a boost in the H3 antibody titer after re-infection with H4N5, which is consistent with original antigenic sin or antigenic seniority and suggest a putative strategy of virus evasion. These results imply competition between related subtypes that could regulate IAV subtype population dynamics in nature. Collectively, we provide new insights into within-host IAV complex interactions as drivers of IAV antigenic diversity that could allow the circulation of multiple subtypes in wild ducks.

Competition between influenza A virus subtypes through heterosubtypic immunity modulates re-infection and antibody dynamics in the mallard duck

PubMed Central

Brown, Justin D.; Carter, Deborah; Franca, Monique; Stallknecht, David E.

2017-01-01

Our overall hypothesis is that host population immunity directed at multiple antigens will influence the prevalence, diversity and evolution of influenza A virus (IAV) in avian populations where the vast subtype diversity is maintained. To investigate how initial infection influences the outcome of later infections with homologous or heterologous IAV subtypes and how viruses interact through host immune responses, we carried out experimental infections in mallard ducks (Anas platyrhynchos). Mallards were pre-challenged with an H3N8 low-pathogenic IAV and were divided into six groups. At five weeks post H3N8 inoculation, each group was challenged with a different IAV subtype (H4N5, H10N7, H6N2, H12N5) or the same H3N8. Two additional pre-challenged groups were inoculated with the homologous H3N8 virus at weeks 11 and 15 after pre-challenge to evaluate the duration of protection. The results showed that mallards were still resistant to re-infection after 15 weeks. There was a significant reduction in shedding for all pre-challenged groups compared to controls and the outcome of the heterologous challenges varied according to hemagglutinin (HA) phylogenetic relatedness between the viruses used. There was a boost in the H3 antibody titer after re-infection with H4N5, which is consistent with original antigenic sin or antigenic seniority and suggest a putative strategy of virus evasion. These results imply competition between related subtypes that could regulate IAV subtype population dynamics in nature. Collectively, we provide new insights into within-host IAV complex interactions as drivers of IAV antigenic diversity that could allow the circulation of multiple subtypes in wild ducks. PMID:28640898

Porcine circovirus type 2 (PCV2) infection decreases the efficacy of an attenuated classical swine fever virus (CSFV) vaccine

PubMed Central

2011-01-01

The Lapinized Philippines Coronel (LPC) vaccine, an attenuated strain of classical swine fever virus (CSFV), is an important tool for the prevention and control of CSFV infection and is widely and routinely used in most CSF endemic areas, including Taiwan. The aim of this study was to investigate whether PCV2 infection affects the efficacy of the LPC vaccine. Eighteen 6-week-old, cesarean-derived and colostrum-deprived (CDCD), crossbred pigs were randomly assigned to four groups. A total of 105.3 TCID50 of PCV2 was experimentally inoculated into pigs through both intranasal and intramuscular routes at 0 days post-inoculation (dpi) followed by LPC vaccination 12 days later. All the animals were challenged with wild-type CSFV (ALD stain) at 27 dpi and euthanized at 45 dpi. Following CSFV challenge, the LPC-vaccinated pigs pre-inoculated with PCV2 showed transient fever, viremia, and viral shedding in the saliva and feces. The number of IgM+, CD4+CD8-CD25+, CD4+CD8+CD25+, and CD4-CD8+CD25+ lymphocyte subsets and the level of neutralizing antibodies against CSFV were significantly higher in the animals with LPC vaccination alone than in the pigs with PCV2 inoculation/LPC vaccination. In addition, PCV2-derived inhibition of the CSFV-specific cell proliferative response of peripheral blood mononuclear cells (PBMCs) was demonstrated in an ex vivo experiment. These findings indicate that PCV2 infection decreases the efficacy of the LPC vaccine. This PCV2-derived interference may not only allow the invasion of wild-type CSFV in pig farms but also increases the difficulty of CSF prevention and control in CSF endemic areas. PMID:22129109

Composition of Gut Microbiota Influences Resistance of Newly Hatched Chickens to Salmonella Enteritidis Infection

PubMed Central

Varmuzova, Karolina; Kubasova, Tereza; Davidova-Gerzova, Lenka; Sisak, Frantisek; Havlickova, Hana; Sebkova, Alena; Faldynova, Marcela; Rychlik, Ivan

2016-01-01

Since poultry is a very common source of non-typhoid Salmonella for humans, different interventions aimed at decreasing the prevalence of Salmonella in chickens are understood as an effective measure for decreasing the incidence of human salmonellosis. One such intervention is the use of probiotic or competitive exclusion products. In this study we tested whether microbiota from donor hens of different age will equally protect chickens against Salmonella Enteritidis infection. Newly hatched chickens were therefore orally inoculated with cecal extracts from 1-, 3-, 16-, 28-, and 42-week-old donors and 7 days later, the chickens were infected with S. Enteritidis. The experiment was terminated 4 days later. In the second experiment, groups of newly hatched chickens were inoculated with cecal extracts of 35-week-old hens either on day 1 of life followed by S. Enteritidis infection on day 2 or were infected with S. Enteritidis infection on day 1 followed by therapeutic administration of the cecal extract on day 2 or were inoculated on day 1 of life with a mixture of the cecal extract and S. Enteritidis. This experiment was terminated when the chickens were 5 days old. Both Salmonella culture and chicken gene expression confirmed that inoculation of newly hatched chickens with microbiota from 3-week-old or older chickens protected them against S. Enteritidis challenge. On the other hand, microbiota from 1-week-old donors failed to protect chickens against S. Enteritidis challenge. Microbiota from 35-week-old hens protected chickens even 24 h after administration. However, simultaneous or therapeutic microbiota administration failed to protect chickens against S. Enteritidis infection. Gut microbiota can be used as a preventive measure against S. Enteritidis infection but its composition and early administration is critical for its efficacy. PMID:27379083

Effects of the inoculations using bacteria producing ACC deaminase on ethylene metabolism and growth of wheat grown under different soil water contents.

PubMed

Zhang, Guozhuang; Sun, Yonglin; Sheng, Hao; Li, Haichao; Liu, Xiping

2018-04-01

Crop growth and productivity are often impacted by the increased ethylene content induced by adverse environmental conditions such drought. Inoculations with bacteria producing ACC deaminase is considered as a potential biological approach to improve the growth and tolerance of stressed plants by lowering endogenous ethylene level. In this study, germinated wheat seeds were inoculated using three species of the rhizobacteria, which were isolated from the rhizosphere of wheat growing in dryland, and sown in pots. After three weeks, wheat seedlings were exposed to non-limiting water condition, medium drought and severe drought, respectively, for six weeks. The results showed that, irrespective of rhizobacterial inoculations, decreased soil water contents stimulated wheat ethylene metabolism, which was reflected by the significantly increased activity of ACC synthetase and ACC oxidase, besides an increased content of ACC both in the roots and leaves, and an enhanced capacity of leaves to release ethylene, concomitant with a significant decline in shoot and roots biomass. The inoculations of all three rhizobacterial species under each water condition reduced ACC content in wheat leaves, but effects of the inoculations on ACC synthase and ACC oxidase activity in the leaves and roots, ACC content in the roots, the capacity of leaves to release ethylene, and wheat growth varied with water conditions and bacterial species. Hence, both soil water conditions and rhizobacterial inoculations acted on all the processes of ethylene metabolism, with the former being dominant. The inoculations under non-limiting water condition and medium drought promoted shoot and root growth of wheat plants. Copyright © 2018 Elsevier Masson SAS. All rights reserved.

Inoculation effects on root-colonizing arbuscular mycorrhizal fungal communities spread beyond directly inoculated plants

PubMed Central

Krak, Karol; Vosátka, Miroslav; Püschel, David; Štorchová, Helena

2017-01-01

Inoculation with arbuscular mycorrhizal fungi (AMF) may improve plant performance at disturbed sites, but inoculation may also suppress root colonization by native AMF and decrease the diversity of the root-colonizing AMF community. This has been shown for the roots of directly inoculated plants, but little is known about the stability of inoculation effects, and to which degree the inoculant and the inoculation-induced changes in AMF community composition spread into newly emerging seedlings that were not in direct contact with the introduced propagules. We addressed this topic in a greenhouse experiment based on the soil and native AMF community of a post-mining site. Plants were cultivated in compartmented pots with substrate containing the native AMF community, where AMF extraradical mycelium radiating from directly inoculated plants was allowed to inoculate neighboring plants. The abundances of the inoculated isolate and of native AMF taxa were monitored in the roots of the directly inoculated plants and the neighboring plants by quantitative real-time PCR. As expected, inoculation suppressed root colonization of the directly inoculated plants by other AMF taxa of the native AMF community and also by native genotypes of the same species as used for inoculation. In the neighboring plants, high abundance of the inoculant and the suppression of native AMF were maintained. Thus, we demonstrate that inoculation effects on native AMF propagate into plants that were not in direct contact with the introduced inoculum, and are therefore likely to persist at the site of inoculation. PMID:28738069

Inoculation effects on root-colonizing arbuscular mycorrhizal fungal communities spread beyond directly inoculated plants.

PubMed

Janoušková, Martina; Krak, Karol; Vosátka, Miroslav; Püschel, David; Štorchová, Helena

2017-01-01

Inoculation with arbuscular mycorrhizal fungi (AMF) may improve plant performance at disturbed sites, but inoculation may also suppress root colonization by native AMF and decrease the diversity of the root-colonizing AMF community. This has been shown for the roots of directly inoculated plants, but little is known about the stability of inoculation effects, and to which degree the inoculant and the inoculation-induced changes in AMF community composition spread into newly emerging seedlings that were not in direct contact with the introduced propagules. We addressed this topic in a greenhouse experiment based on the soil and native AMF community of a post-mining site. Plants were cultivated in compartmented pots with substrate containing the native AMF community, where AMF extraradical mycelium radiating from directly inoculated plants was allowed to inoculate neighboring plants. The abundances of the inoculated isolate and of native AMF taxa were monitored in the roots of the directly inoculated plants and the neighboring plants by quantitative real-time PCR. As expected, inoculation suppressed root colonization of the directly inoculated plants by other AMF taxa of the native AMF community and also by native genotypes of the same species as used for inoculation. In the neighboring plants, high abundance of the inoculant and the suppression of native AMF were maintained. Thus, we demonstrate that inoculation effects on native AMF propagate into plants that were not in direct contact with the introduced inoculum, and are therefore likely to persist at the site of inoculation.

Comparative pathogenesis in specific-pathogen-free chickens of two strains of avian hepatitis E virus recovered from a chicken with Hepatitis-Splenomegaly syndrome and from a clinically healthy chicken.

PubMed

Billam, P; LeRoith, T; Pudupakam, R S; Pierson, F W; Duncan, R B; Meng, X J

2009-11-18

Avian hepatitis E virus (avian HEV) is the primary causative agent of Hepatitis-Splenomegaly (HS) syndrome in chickens. Recently, a genetically unique strain of avian HEV, designated avian HEV-VA, was recovered from healthy chickens in Virginia. The objective of this study was to experimentally compare the pathogenicity of the prototype strain recovered from a chicken with HS syndrome and the avian HEV-VA strain in specific-pathogen-free chickens. An infectious stock of the avian HEV-VA strain was first generated and its infectivity titer determined in chickens. For the comparative pathogenesis study, 54 chickens of 6-week-old were assigned to 3 groups of 18 chickens each. The group 1 chickens were each intravenously inoculated with 5x10(2.5) 50% chicken infectious dose of the prototype strain. The group 2 received the same dose of the avian HEV-VA strain, and the group 3 served as negative controls. Six chickens from each group were necropsied at 2, 3 and 4 weeks post-inoculation (wpi). Most chickens in both inoculated groups seroconverted by 3wpi, and the mean anti-avian HEV antibody titers were higher for the prototype strain group than the avian HEV-VA strain group. There was no significant difference in the patterns of viremia and fecal virus shedding. Blood analyte profiles did not differ between treatment groups except for serum creatine phosphokinase levels which were higher for prototype avian HEV group than avian HEV-VA group. The hepatic lesion score was higher for the prototype strain group than the other two groups. The results indicated that the avian HEV-VA strain is only slightly attenuated compared to the prototype strain, suggesting that the full spectrum of HS syndrome is likely associated with other co-factors.

Persistence of fecal shedding of Salmonella Enteritidis by experimentally infected laying hens housed in conventional or enriched cages.

PubMed

Gast, Richard K; Guraya, Rupa; Jones, Deana R; Anderson, Kenneth E

2015-07-01

Salmonella Enteritidis can be deposited inside eggs laid by infected hens, so the prevalence of this pathogen in commercial egg-producing flocks is an important risk factor for human illness. Opportunities for the introduction, transmission, and persistence of salmonellae in poultry are potentially influenced by flock housing and management systems. Animal welfare concerns have spurred the development of alternatives to traditional cage-based housing. However, the consequences of poultry housing systems for food safety have not been fully resolved by prior research. The present study assessed the effects of two different housing systems (conventional cages and colony cages enriched with perching and nesting areas) on the persistence of fecal shedding of Salmonella Enteritidis by groups of experimentally infected laying hens. In each of two trials, 136 hens were distributed among cages of both housing systems and orally inoculated with doses of 10(8) cfu of Salmonella Enteritidis (phage type 13a in one trial and phage type 4 in the other). At weekly intervals, samples of voided feces were collected from beneath each cage and cultured to detect Salmonella Enteritidis. Fecal shedding of Salmonella Enteritidis was detected for up to 8 wk post-inoculation by hens housed in enriched colony cages and 10 wk by hens housed in conventional cages. For both trials combined, the frequency of positive fecal cultures was significantly (P < 0.05) greater for conventional cages than for enriched colony cages at 1 wk (84.7 vs. 71.5%), 2 wk (54.2 vs. 31.3%), 3 wk (21.5 vs. 7.6%), and 4 wk (9.7 vs. 2.8%) post-inoculation. These results demonstrate that the susceptibility of hens to intestinal colonization by Salmonella Enteritidis can differ between conventional and enriched cage-based production systems, although this effect does not necessarily translate into a corresponding difference in the longer-term persistence of fecal shedding. © 2015 Poultry Science Association Inc.

Leaf elemental analysis in mycorrhizal post oak seedlings

NASA Astrophysics Data System (ADS)

Boling, B. C.; Naab, F. U.; Smith, D.; Duggan, J. L.; McDaniel, F. D.

2006-09-01

Growth and element assimilation was investigated in the leaves of post oak seedlings exposed to four different treatment combinations of fertilization and ectomycorrhizal inoculation. Element concentration was analyzed via particle-induced X-ray emission spectrometry (PIXE). PIXE detected 10 of the 13 essential macro and micronutrients: P, S, Mg, Ca, K, Cu, Zn, Mn, Fe and Cl. Mean growth and dry weight was significantly different across the treatment groups as well as the mean concentration of Mg, Al, S, K, Ca, Fe, Cu and Zn. The data suggest that fertilization rather than mycorrhizal inoculation had a stronger influence on nutrient uptake. This study is the first to analyze element concentration in post oak and to investigate the potential benefits of mycorrhizal symbiosis in post oak seedlings in terms of nutrient uptake.

The Role of Attachment and Cognitive Inhibition in Children's Memory and Suggestibility for a Stressful Event.

ERIC Educational Resources Information Center

Alexander, Kristen Weede; Goodman, Gail S.; Scharf, Jennifer M.; Edelstein, Robin S.; Quas, Jodi A.; Shaver, Phillip R.

2002-01-01

This study interviewed 51 children, ages 3 to 7, about an inoculation after an approximate 2-week delay. Responses indicated a relationship between parents' attachment avoidance and children's distress during the inoculation. Parental attachment anxiety and the interaction between parental avoidance and children's stress predicted children's…

Interactions of Heterodera daverti, H. goldeni and H. zeae with Meloidogyne incognita on rice

USDA-ARS?s Scientific Manuscript database

The interactions of the cyst nematodes Heterodera daverti, H. goldeni and H. zeae with the root-knot nematode Meloidogyne incognita on rice (Oryza sativa) cultivars Giza 178 and Sakha 101 were studied in the greenhouse. Inoculation with H. goldeni alone or one week before inoculation with M. incogni...

The effect of plant growth-promoting rhizobacteria on asparagus seedlings and germinating seeds subjected to water stress under greenhouse conditions.

PubMed

Liddycoat, Scott M; Greenberg, Bruce M; Wolyn, David J

2009-04-01

Plant growth-promoting rhizobacteria (PGPR) can have positive effects on vigour and productivity, especially under stress conditions. In asparagus (Asparagus officinalis L.) field culture, seeds are planted in high-density nurseries, and 1-year-old crowns are transplanted to production fields. Performance can be negatively affected by water stress, transplant shock, and disease pressure on wounded roots. PGPR inoculation has the potential to alleviate some of the stresses incurred in the production system. In this study, the effects of PGPR (Pseudomonas spp.) treatment were determined on 3-week-old greenhouse-grown seedlings and germinating seeds of 2 asparagus cultivars. The pots were irrigated to a predetermined level that resulted in optimum growth or the plants were subjected to drought or flooding stress for 8 weeks. The cultivars responded differently to PGPR: single inoculations of seedlings enhanced growth of 'Guelph Millennium' under optimum conditions and 'Jersey Giant' seedlings under drought stress. Seed inoculations with PGPR resulted in a positive response only for 'Guelph Millennium', for which both single or multiple inoculations enhanced plant growth under drought stress.

Molecular Characterization of the Onset and Progression of Colitis in Inoculated Interleukin-10 Gene-Deficient Mice: A Role for PPARα

PubMed Central

Knoch, Bianca; Barnett, Matthew P. G.; Cooney, Janine; McNabb, Warren C.; Barraclough, Diane; Laing, William; Zhu, Shuotun; Park, Zaneta A.; MacLean, Paul; Knowles, Scott O.; Roy, Nicole C.

2010-01-01

The interleukin-10 gene-deficient (Il10 −/−) mouse is a model of human inflammatory bowel disease and Ppara has been identified as one of the key genes involved in regulation of colitis in the bacterially inoculated Il10 −/− model. The aims were to (1) characterize colitis onset and progression using a histopathological, transcriptomic, and proteomic approach and (2) investigate links between PPARα and IL10 using gene network analysis. Bacterial inoculation resulted in severe colitis in Il10 −/− mice from 10 to 12 weeks of age. Innate and adaptive immune responses showed differences in gene expression relating to colitis severity. Actin cytoskeleton dynamics, innate immunity, and apoptosis-linked gene and protein expression data suggested a delayed remodeling process in 12-week-old Il10 −/− mice. Gene expression changes in 12-week-old Il10 −/− mice were related to PPARα signaling likely to control colitis, but how PPARα activation might regulate intestinal IL10 production remains to be determined. PMID:20671959

Sensitivity of Pseudomonas fluorescens to gamma irradiation following surface inoculations on romaine lettuce and baby spinach

USDA-ARS?s Scientific Manuscript database

Irradiation of fresh fruits and vegetables is a post-harvest intervention measure often used to inactivate pathogenic food-borne microbes. We evaluated the sensitivity of Pseudomonas fluorescens strains (2-79, Q8R1, Q287) to gamma irradiation following surface inoculations on romaine lettuce and spi...

Biochemical Changes in Tissues during Infectious Illness: Metabolic Consequences of Interactions between Infectious Illness and Forced Exercise.

DTIC Science & Technology

1980-09-01

with the inoculation of S. typhimurium at 24 hrs post infection. Fasting ketosis was dramatically altered by the inoculation. Both plasma...as were both fasting and exercise associated ketosis . During fasting plasma Phydroxybutyrate increased 20-fold in the controls but only 13-fold in

Potentially Deceptive Health Nutrition-Related Advertising Claims: The Role of Inoculation in Conferring Resistance

ERIC Educational Resources Information Center

Mason, Alicia M.; Miller, Claude H.

2016-01-01

Objective: This study sought to examine the efficacy of inoculation message treatments to facilitate resistance to health nutrition-related (HNR) commercial food advertising claims. Design: Data were collected across three phases extending across a 5-week period conducted over two semesters at a Midwest US university. A 2 × 3 between-subjects…

Association mapping analysis of the response to Macrophomina phaseolina in the Andean Diversity Panel

USDA-ARS?s Scientific Manuscript database

The Andean Diversity Panel was evaluated for response to inoculation with charcoal rot in a field trial in Isabela, Puerto Rico in 2012. The trial was inoculated with backpack sprayers at flowering and a visual evaluation was conducted three weeks later. There was a broad range of response, with abo...

Immunological alterations during the clinical and recovery phases of experimental swine dysentery.

PubMed

Jonasson, Robert; Andersson, Märit; Råsbäck, Therese; Johannisson, Anders; Jensen-Waern, Marianne

2006-07-01

The aim of this study was to examine changes in the systemic immune response during the incubation period and following the onset of clinical swine dysentery, including the recovery period. Ten healthy conventional pigs were inoculated with Brachyspira hyodysenteriae. Blood was sampled at pre-inoculation, at days 4 and 14 post-inoculation, during the first 4 days with clinical signs of dysentery and at days 1, 3, 7, 11 and 15 of the recovery period. Eight pigs developed haemorrhagic diarrhoea. Flow-cytometric analyses of lymphocyte subpopulations showed that all animals, including the two that remained healthy, had an increase in CD8alpha+ CD4- cells and gammadelta T cells at days 4 and 14 post-inoculation. In addition, an increase in CD4+ CD8alpha+ cells and CD8alpha+ CD8beta+ cells was observed at days 4 and 14 post-inoculation in animals that developed dysentery. During clinical signs of dysentery, the acute-phase protein serum amyloid A was increased. There was a two- to threefold increase in both neutrophils and monocytes during signs of dysentery and at the beginning of the recovery period. The numbers of CD8alpha+ CD8beta- CD4-, CD45RA- lymphocytes also increased during the dysentery period. Circulating CD21+ cells and CD21+ CD45RA- cells decreased at the end of the incubation period, during signs of dysentery and at the beginning of the recovery period. The dysentery-affected animals developed antibodies to B. hyodysenteriae-specific antigens (approximately 16 kDa and approximately 30 kDa) from the first day of recovery, and gammadelta T cells showed an increase during the recovery period. In comparison with pre-inoculation, increased numbers of monocytes, neutrophils, CD8alpha+ CD8beta- CD4- lymphocytes and CD45RA- lymphocytes were observed during clinical dysentery. Increased numbers of neutrophils, gammadelta T cells and specific antibodies were seen during the recovery period.

Evaluation of thermophilic fungal consortium for organic municipal solid waste composting.

PubMed

Awasthi, Mukesh Kumar; Pandey, Akhilesh Kumar; Khan, Jamaluddin; Bundela, Pushpendra Singh; Wong, Jonathan W C; Selvam, Ammaiyappan

2014-09-01

Influence of fungal consortium and different turning frequency on composting of organic fraction of municipal solid waste (OFMSW) was investigated to produce compost with higher agronomic value. Four piles of OFMSW were prepared: three piles were inoculated with fungal consortium containing 5l each spore suspensions of Trichoderma viride, Aspergillus niger and Aspergillus flavus and with a turning frequency of weekly (Pile 1), twice a week (Pile 2) and daily (Pile 3), while Pile 4 with weekly turning and without fungal inoculation served as control. The fungal consortium with weekly (Pile 1) turning frequency significantly affected temperature, pH, TOC, TKN, C/N ratio and germination index. High degradation of organic matter and early maturity was observed in Pile 1. Results indicate that fungal consortium with weekly turning frequency of open windrows were more cost-effective in comparison with other technologies for efficient composting and yield safe end products. Copyright © 2014 Elsevier Ltd. All rights reserved.

Potential for horizontal transmission of Salmonella and Campylobacter among caged and cage free laying hens

USDA-ARS?s Scientific Manuscript database

Nine hens were inoculated orally and intravaginally with a marker strain of Salmonella and Campylobacter at 56 wk-of-age and housed in individual cages in isolation. Challenged hens were comingled with non-challenged hens 2 wk post-inoculation, at a ratio of 1 challenged hen per 4 non-challenged he...

[Herpes simplex virus-mediated RNA interference targeting vesicular glutamate transporter 3 attenuates tactile allodynia in mice].

PubMed

Liu, Jie-Qiong; Li, Chen-Hong; Luo, Qiong; Yin, Ping-Ping; Lei, Tao; Luo, Fang

2016-11-20

To construct a replication-deficient herpes simplex virus (HSV-1) for delivering a short hairpin RNA (shRNA) targeting vesicular glutamate transporter 3 (VGLUT3) and observe its effect in alleviating allodynia in mice. The recombinant HSV-1 vector carrying the shRNA targeting Vglut3 (HSV-1-shvglut3) was constructed and inoculated in the sciatic nerve in a mouse model of mechanical allodynia to test its analgesia effect. Mechanical allodynia and heat hypersensitivity of the mice were tested by von Frey filaments and Hargreaves' test, respectively. VGLUT3 expression in the dorsal root ganglion (DRG) was evaluated by immunohistochemistry and Western blotting. Following inoculation in the sciatic nerve, the HSV vector HSV-1-shvglut3 was retrogradely transported to the DRG. Mechanical withdraw thresholds of the mouse models receiving HSV-1-shvglut3 inoculation were reversed to nearly the baseline level, and VGLUT3 expression in the DRG was down-regulated 2 weeks after vector inoculation. The analgesic effect lasted for over 2 weeks in these mice without obvious systematic side effects or changes in heat hypersensitivity threshold. Vglut3 in the DRG is a promising therapeutic target for alleviating mechanical allodynia, and HSV-1 vector-mediated RNA interference is safe and efficient for inducing long-lasting analgesia after peripheral inoculation of the vector.

Boron neutron capture therapy (BNCT) for liver metastasis in an experimental model: dose–response at five-week follow-up based on retrospective dose assessment in individual rats

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

Emiliano C. C. Pozzi; Veronica A. Trivilin; Lucas L. Colombo

Boron neutron capture therapy (BNCT) was proposed for untreatable colorectal liver metastases. Employing an experimental model of liver metastases in rats, we recently demonstrated that BNCT mediated by boronophenylalanine (BPA-BNCT) at 13 Gy prescribed to tumor is therapeutically useful at 3-week follow-up. The aim of the present study was to evaluate dose–response at 5-week follow-up, based on retrospective dose assessment in individual rats. BDIX rats were inoculated with syngeneic colon cancer cells DHD/K12/TRb. Tumor-bearing animals were divided into three groups: BPA-BNCT (n = 19), Beam only (n = 8) and Sham (n = 7) (matched manipulation, no treatment). For eachmore » rat, neutron flux was measured in situ and boron content was measured in a pre-irradiation blood sample for retrospective individual dose assessment. For statistical analysis (ANOVA), individual data for the BPA-BNCT group were pooled according to absorbed tumor dose, BPA-BNCT I: 4.5–8.9 Gy and BPA-BNCT II: 9.2–16 Gy. At 5 weeks post-irradiation, the tumor surface area post-treatment/pre-treatment ratio was 12.2 +/- 6.6 for Sham, 7.8 +/- 4.1 for Beam only, 4.4 +/- 5.6 for BPA-BNCT I and 0.45 +/- 0.20 for BPA-BNCT II; tumor nodule weight was 750 +/- 480 mg for Sham, 960 +/- 620 mg for Beam only, 380 +/- 720 mg for BPA-BNCT I and 7.3 +/- 5.9 mg for BPA-BNCT II. The BPA-BNCT II group exhibited statistically significant tumor control with no contributory liver toxicity. Potential threshold doses for tumor response and significant tumor control were established at 6.1 and 9.2 Gy, respectively.« less

Effect of the South African asinine-94 strain of equine arteritis virus (EAV) in pregnant donkey mares and duration of maternal immunity in foals.

PubMed

Paweska, J T

1997-06-01

Clinical, virological and serological responses were investigated in five pregnant donkey mares after experimental exposure to the South African asinine-94 strain of equine arteritis virus (EAV), and the duration of maternal immunity to EAV was studied in their foals. In four intranasally inoculated mares, fever with maximum rectal temperatures of 39.1-40.7 degrees C was recorded 2-11 d after challenge. All the inoculated mares developed mild depression, and a serous ocular and nasal discharge; in three mares mild conjuctivitis was observed. The virus was recovered from the nasopharynx and from buffy-coat samples of all the mares 3-10 d, and 2-18 d post inoculation (p.i.), respectively. Seroconversion to EAV was detected on days 8-10 p.i. Peak serum-virus-neutralizing antibody titres of log10 1.8-2.4, and IgG ELISA OD values of 0.85-2.15 were recorded 2-3 weeks p.i. The in-contact (p.c.) control mare developed fever on days 15-19 post exposure, and showed mild clinical signs of equine viral arteritis similar to those observed in the inoculated mares. Seroconversion to EAV was detected in the p.c. mare on day 20 post exposure, and virus was isolated from nasal swabs and blood samples collected at the time of the febrile response and 1-3 d afterwards. None of the mares aborted. After they had given normal birth 45-128 d p.i. or after p.c. exposure, no virus could be isolated from their placentas. The concentration of EAV-neutralizing antibody in colostrum was two to eight times higher than in serum samples collected at the time of parturition. All the foals born to infected mares were clinically normal at the time of birth and throughout the subsequent 1-2 months of observation. No EAV was recovered from the buffy-coat fraction of blood samples collected at birth nor from those collected on days 1, 2 and 7 after birth. Also, no virus-serum-neutralizing or IgG ELISA antibody to EAV was detected in sera collected immediately after birth before the foals started nursing. The colostrum-derived maternal antibodies against EAV gradually declined and could not be detected by either the VN test or ELISA for 2-3 months after birth. This study demonstrates that the asinine-94 strain of EAV does not cause abortion in pregnant donkey mares. Furthermore, no carrier state could be demonstrated in foals born to mares infected at the time of pregnancy.

Experimental palatal candidosis and saliva flow in monkeys.

PubMed

Olsen, I; Haanaes, H R

1977-01-01

Maxillary acrylic plates, inoculated with Candida albicans, were inserted for 3 weeks in 10 monkeys (Cercopithecus aethiops) (Series I), and reinserted in five of the animals 8 weeks after removal (Series II). To suppress saliva flow oxyphencyclimine was injected intramuscularly (0.125 mg/kg) thrice daily for 3 weeks in six monkeys of Series I, while four controls received no drug. In Series II the oxyphencyclimine dose was doubled in three animals, and two controls were sham-treated with sodium chloride. Mean saliva flow was reduced to 58% after 1 week and to 63% after 3 weeks with the low dose of oxyphencyclimine. The values with the high dose were 56% and 64%, respectively. After 1 week thrush had developed beneath the plates of all monkeys. The patches were more extensive and regressed slower with oxyphencyclimine. Enlarged lesions were seen with the double dose. In Series I intraepithelial invasion by hyphae was detected more frequently and longer after inoculation in the oxyphencyclimine group. Such invasion was not found in biopsies from Series II. It is likely that saliva offers some protection against yeasts colonizing the fitting size of a denture.

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

PubMed Central

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

2014-01-01

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

Immunization with a Borrelia burgdorferi BB0172-Derived Peptide Protects Mice against Lyme Disease

PubMed Central

Small, Christina M.; Ajithdoss, Dharani K.; Rodrigues Hoffmann, Aline; Mwangi, Waithaka; Esteve-Gassent, Maria D.

2014-01-01

Lyme disease is the most prevalent arthropod borne disease in the US and it is caused by the bacterial spirochete Borrelia burgdorferi (Bb), which is acquired through the bite of an infected Ixodes tick. Vaccine development efforts focused on the von Willebrand factor A domain of the borrelial protein BB0172 from which four peptides (A, B, C and D) were synthesized and conjugated to Keyhole Limpet Hemocyanin, formulated in Titer Max® adjuvant and used to immunize C3H/HeN mice subcutaneously at days 0, 14 and 21. Sera were collected to evaluate antibody responses and some mice were sacrificed for histopathology to evaluate vaccine safety. Twenty-eight days post-priming, protection was evaluated by needle inoculation of half the mice in each group with 103 Bb/mouse, whereas the rest were challenged with 105Bb/mouse. Eight weeks post-priming, another four groups of similarly immunized mice were challenged using infected ticks. In both experiments, twenty-one days post-challenge, the mice were sacrificed to determine antibody responses, bacterial burdens and conduct histopathology. Results showed that only mice immunized with peptide B were protected against challenge with Bb. In addition, compared to the other the treatment groups, peptide B-immunized mice showed very limited inflammation in the heart and joint tissues. Peptide B-specific antibody titers peaked at 8 weeks post-priming and surprisingly, the anti-peptide B antibodies did not cross-react with Bb lysates. These findings strongly suggest that peptide B is a promising candidate for the development of a new DIVA vaccine (Differentiate between Infected and Vaccinated Animals) for protection against Lyme disease. PMID:24505447

Responses of Wound-Inoculated Seedlings of Pinus elliottii var. elliottii and Pinus taeda to Mycelial Cultures Derived from Multople and Singel Basidiospores of Cronartium quercuum f. sp. fusiforme

Treesearch

T. Miller; K.P. Gramacho; R.A. Schmidt; H.V. Amerson; E.G. Kuhlman

1998-01-01

In 1991, a series of experiments was initiated to examine the effectiveness and research value of inoculating 6-week-old seedlings of slash (Pinus elliottii var. elliottii) and loblolly pine (P. raeda) with suspensions of basidiospores of Cronartium quercuum f. sp. fusiforme...

Atypical patterns of neural infection produced in mice by drug-resistant strains of herpes simplex virus.

PubMed

Field, H J; Anderson, J R; Wildy, P

1982-03-01

Mice inoculated intracerebrally (i.c.) with a mutant strain of HSV were found to develop cataracts 1 to 2 months after inoculation. Cataract formation was subsequently shown to follow an acute retinitis which commenced within 1 week of inoculation. The mutant had been selected for high resistance to the nucleoside analogue acyclovir and has been shown previously to be defective in the induction of thymidine kinase and also to express an altered DNA polymerase. The LD50 for mice inoculated i.c. was greater than 10(5) p.f.u. compared with approx 7 p.f.u. for the parental strain. Studies of virus replication following i.c. inoculation with a sublethal dose of the mutant revealed that only small amounts of infectious virus were produced in the brain, but during a period from 6 to 12 days after inoculation vigorous replication occurred in retinal tissue, producing very high titres of virus.

Pathogenicity of the Korean H5N8 highly pathogenic avian influenza virus in commercial domestic poultry species.

PubMed

Lee, Dong-Hun; Kwon, Jung-Hoon; Noh, Jin-Yong; Park, Jae-Keun; Yuk, Seong-Su; Erdene-Ochir, Tseren-Ochir; Lee, Joong-Bok; Park, Seung-Yong; Choi, In-Soo; Lee, Sang-Won; Song, Chang-Seon

2016-01-01

In 2014, the highly pathogenic avian influenza (HPAI) virus H5N8 triggered outbreaks in wild birds and poultry farms in South Korea. In the present study, we investigated the pathogenicity of the H5N8 HPAI virus, belonging to the clade 2.3.4.4, in different species of poultry. For this, we examined clinical signs and viral shedding levels following intranasal inoculation of the virus in 3-week-old commercial layer chickens and quails, 10-week-old Korean native chickens, and 8-week-old Muscovy ducks. Intranasal inoculation with 10(6.0) viruses at 50% egg-infective dose resulted in 100% mortality in the layer chickens (8/8) and quails (4/4), but 60% and 0% deaths in the Korean native chickens (3/5) and Muscovy ducks (0/4), respectively. In addition, transmission of the inoculated virus to contact-exposed birds was evident in all the species used in this study. Based on our results, we conclude that the H5N8 HPAI virus has lower pathogenicity and transmissibility in poultry species compared with previously reported H5N1 HPAI viruses.

Oral keratinocytes support non-replicative infection and transfer of harbored HIV-1 to permissive cells.

PubMed

Vacharaksa, Anjalee; Asrani, Anil C; Gebhard, Kristin H; Fasching, Claudine E; Giacaman, Rodrigo A; Janoff, Edward N; Ross, Karen F; Herzberg, Mark C

2008-07-17

Oral keratinocytes on the mucosal surface are frequently exposed to HIV-1 through contact with infected sexual partners or nursing mothers. To determine the plausibility that oral keratinocytes are primary targets of HIV-1, we tested the hypothesis that HIV-1 infects oral keratinocytes in a restricted manner. To study the fate of HIV-1, immortalized oral keratinocytes (OKF6/TERT-2; TERT-2 cells) were characterized for the fate of HIV-specific RNA and DNA. At 6 h post inoculation with X4 or R5-tropic HIV-1, HIV-1gag RNA was detected maximally within TERT-2 cells. Reverse transcriptase activity in TERT-2 cells was confirmed by VSV-G-mediated infection with HIV-NL4-3Deltaenv-EGFP. AZT inhibited EGFP expression in a dose-dependent manner, suggesting that viral replication can be supported if receptors are bypassed. Within 3 h post inoculation, integrated HIV-1 DNA was detected in TERT-2 cell nuclei and persisted after subculture. Multiply spliced and unspliced HIV-1 mRNAs were not detectable up to 72 h post inoculation, suggesting that HIV replication may abort and that infection is non-productive. Within 48 h post inoculation, however, virus harbored by CD4 negative TERT-2 cells trans infected co-cultured peripheral blood mononuclear cells (PBMCs) or MOLT4 cells (CD4+ CCR5+) by direct cell-to-cell transfer or by releasing low levels of infectious virions. Primary tonsil epithelial cells also trans infected HIV-1 to permissive cells in a donor-specific manner. Oral keratinocytes appear, therefore, to support stable non-replicative integration, while harboring and transmitting infectious X4- or R5-tropic HIV-1 to permissive cells for up to 48 h.

Development and application of a quantitative PCR assay to study equine herpesvirus 5 invasion and replication in equine tissues in vitro and in vivo.

PubMed

Zarski, Lila M; High, Emily A; Nelli, Rahul K; Bolin, Steven R; Williams, Kurt J; Hussey, Gisela

2017-10-01

Equine herpesvirus 5 (EHV-5) infection is associated with pulmonary fibrosis in horses, but further studies on EHV-5 persistence in equine cells are needed to fully understand viral and host contributions to disease pathogenesis. Our aim was to develop a quantitative PCR (qPCR) assay to measure EHV-5 viral copy number in equine cell cultures, blood lymphocytes, and nasal swabs of horses. Furthermore, we used a recently developed equine primary respiratory cell culture system to study EHV-5 pathogenesis at the respiratory tract. PCR primers and a probe were designed to target gene E11 of the EHV-5 genome. Sensitivity and repeatability were established, and specificity was verified by testing multiple isolates of EHV-5, as well as DNA from other equine herpesviruses. Four-week old fully differentiated (mature), newly seeded (immature) primary equine respiratory epithelial cell (ERECs), and equine dermal cell cultures were inoculated with EHV-5 and the cells and supernatants collected daily for 14days. Blood lymphocytes and nasal swabs were collected from horses experimentally infected with equine herpesvirus 1 (EHV-1). The qPCR assay detected EHV-5 at stable concentrations throughout 14days in inoculated mature EREC and equine dermal cell cultures (peaking at 202 and 5861 viral genomes per 10 6 cellular β actin, respectively). EHV-5 copies detected in the immature EREC cultures increased over 14days and reached levels greater than 10,000 viral genomes per 10 6 cellular β actin. Moreover, EHV-5 was detected in the lymphocytes of 76% of horses and in the nasal swabs of 84% of horses experimentally infected with EHV-1 pre-inoculation with EHV-1. Post-inoculation with EHV-1, EHV-5 was detected in lymphocytes of 52% of horses while EHV-5 levels in nasal swabs were not significantly different from pre-inoculation levels. In conclusion, qPCR was a reliable technique to investigate viral load in in vivo and in vitro samples, and EHV-5 replication in equine epithelial cells may be influenced by cellular stages of differentiation. Copyright © 2017 Elsevier B.V. All rights reserved.

Association between nasal shedding and fever that influenza A (H3N2) induces in dogs.

PubMed

Song, Daesub; Moon, Hyoungjoon; Jung, Kwonil; Yeom, Minjoo; Kim, Hyekwon; Han, Sangyoon; An, Dongjun; Oh, Jinsik; Kim, Jongman; Park, Bongkyun; Kang, Bokyu

2011-01-05

Avian origin canine influenza virus was reported in Korea. The dog to dog contact transmission of the avian origin canine influenza virus (CIV) H3N2 and CIV H3N8 was shown by experimental contact transmission. This study was focused on viral excretion and fever in order to elucidate the epidemiological associations which might be helpful to control the disease transmissions in CIV outbreak in dogs. An influenza seronegative 10-week-old Beagle dog was experimentally inoculated with the canine influenza virus A/canine/01/2007, subtype H3N2. Eight hours after inoculation, the infected dog was cohoused with seven uninfected Beagle dogs. Clinical signs including fever were recorded for 14 days post inoculation. The infected dog and four of seven contact dogs in the study showed clinical signs (sneezing, nasal discharge and coughing) during the study. Viral shedding occurred in all of the animals tested and began on 1 to 6 DPI in dogs with clinical signs. Elevated body temperatures above 39.5 °C (geometric mean temperature of 39.86 °C ± 0.49) were observed in all symptomatic dogs. The mean viral titer during fever was 2.99 log EID₅₀/ml, which was significantly higher than the viral titer detected in the non fever. The data show that contact dogs with a canine influenza infected dog shed different levels of virus in their nasal excretions and demonstrate that clinical signs, including fever, significantly correlate with the viral shedding.

Efficacy of an accelerated hydrogen peroxide disinfectant to inactivate porcine epidemic diarrhea virus in swine feces on metal surfaces.

PubMed

Holtkamp, Derald J; Myers, Jacqueline; Thomas, Paul R; Karriker, Locke A; Ramirez, Alejandro; Zhang, Jianqiang; Wang, Chong

2017-04-01

In May of 2013, porcine epidemic diarrhea virus (PEDV) was detected in swine for the first time in North America. It spread rapidly, in part due to contaminated livestock trailers. The objective of this study was to test the efficacy of an accelerated hydrogen peroxide disinfectant for inactivating PEDV in the presence of feces on metal surfaces, such as those found in livestock trailers. Three-week-old barrows were inoculated intragastrically with 5 mL of PEDV-negative feces for the negative control, 5 mL of untreated PEDV-positive feces for the positive control, and 5 mL or 10 mL of PEDV-positive feces that was subjected to treatment with a 1:16 or 1:32 concentrations of accelerated hydrogen peroxide disinfectant for a contact time of 30 min at 20°C. These pigs served as a bioassay to determine the infectivity of virus following treatment. Rectal swabs collected from the inoculated pigs on days 3 and 7 post-inoculation were tested by using PEDV-specific real-time reverse transcription polymerase chain reaction and the proportion of pigs in each group that became infected with PEDV was assessed. None of the pigs used for the bioassay in the 4 treatment groups and the negative control group became infected with PEDV, which was significantly different from the positive control group ( P < 0.05) in which all pigs were infected. The results suggest that the application of the accelerated hydrogen peroxide under these conditions was sufficient to inactivate the virus in feces found on metal surfaces.

Effect of vaccination against pneumonia on the survival of bighorn sheep (Ovis canadensis) commingled with carrier animals.

PubMed

Raghavan, Bindu; Bavananthasivam, Jegarubee; Kugadas, Abirami; Haldorson, Gary J; Srikumaran, Subramaniam

2017-05-01

Leukotoxin producing (lkt+) members of Pasteurellaceae, particularly Mannheimia haemolytica and Bibersteinia trehalosi are important pathogens of pneumonia in bighorn sheep (BHS; Ovis canadensis), causing fatal disease. Predisposing or concurrent infection with Mycoplasma ovipneumoniae enhances the severity of the disease, resulting in increased morbidity and mortality. Several studies have investigated the effectiveness of vaccines against lkt+ members of Pasteurellaceae in preventing fatal pneumonia in BHS. In all of these studies, however, vaccinated animals were challenged experimentally, by direct inoculation of the pathogens, rather than by natural challenge. Moreover, none has investigated the efficacy of the vaccines under conditions of concurrent infection with M. ovipneumoniae. We immunized three bighorn rams and one pregnant ewe with an experimental multivalent vaccine along with a commercial vaccine. The immunized animals were then commingled with two bighorn ewes known to be carriers of lkt+ members of Pasteurellaceae, to simulate natural infection or disease transmission. All vaccinated animals remained healthy. We then inoculated the two carrier ewes with nasal washings from domestic sheep containing M. ovipneumoniae. Within a week, all animals developed mild to moderate signs of pneumonia. While the rams died within two-three months post-inoculation (p.i.), the vaccinated ewe and her lamb died five and eight months p.i., respectively. Taken together, these results suggest that vaccination of BHS against lkt+ members of Pasteurellaceae alone can protect them from natural challenge by these pathogens. However, it may not be adequate to protect them against pneumonia compounded by concurrent infection with M. ovipneumoniae. Copyright © 2017 Elsevier B.V. All rights reserved.

Efficacy of an accelerated hydrogen peroxide disinfectant to inactivate porcine epidemic diarrhea virus in swine feces on metal surfaces

PubMed Central

Holtkamp, Derald J.; Myers, Jacqueline; Thomas, Paul R.; Karriker, Locke A.; Ramirez, Alejandro; Zhang, Jianqiang; Wang, Chong

2017-01-01

In May of 2013, porcine epidemic diarrhea virus (PEDV) was detected in swine for the first time in North America. It spread rapidly, in part due to contaminated livestock trailers. The objective of this study was to test the efficacy of an accelerated hydrogen peroxide disinfectant for inactivating PEDV in the presence of feces on metal surfaces, such as those found in livestock trailers. Three-week-old barrows were inoculated intragastrically with 5 mL of PEDV-negative feces for the negative control, 5 mL of untreated PEDV-positive feces for the positive control, and 5 mL or 10 mL of PEDV-positive feces that was subjected to treatment with a 1:16 or 1:32 concentrations of accelerated hydrogen peroxide disinfectant for a contact time of 30 min at 20°C. These pigs served as a bioassay to determine the infectivity of virus following treatment. Rectal swabs collected from the inoculated pigs on days 3 and 7 post-inoculation were tested by using PEDV-specific real-time reverse transcription polymerase chain reaction and the proportion of pigs in each group that became infected with PEDV was assessed. None of the pigs used for the bioassay in the 4 treatment groups and the negative control group became infected with PEDV, which was significantly different from the positive control group (P < 0.05) in which all pigs were infected. The results suggest that the application of the accelerated hydrogen peroxide under these conditions was sufficient to inactivate the virus in feces found on metal surfaces. PMID:28408777

Effects of host age on susceptibility to infection and immune-gene expression in honey bee queens (Apis mellifera) inoculated with Nosema ceranae

USDA-ARS?s Scientific Manuscript database

Nosema ceranae is a microsporidium parasite infecting honey bees worldwide. All colony members including workers, drones and queens can become infected. In this study, we inoculated queens of age 1, 6 and 12 days post adult emergence, with N. ceranae spores of different doses and allowed them to age...

Suppression of bacterial infection in rice by treatment with a sulfated peptide.

PubMed

Wei, Tong; Chern, Mawsheng; Liu, Furong; Ronald, Pamela C

2016-12-01

The rice XA21 receptor kinase confers robust resistance to bacterial blight disease caused by Xanthomonas oryzae pv. oryzae (Xoo). A tyrosine-sulfated peptide from Xoo, called RaxX, triggers XA21-mediated immune responses, including the production of ethylene and reactive oxygen species and the induction of defence gene expression. It has not been tested previously whether these responses confer effective resistance to Xoo. Here, we describe a newly established post-inoculation treatment assay that facilitates investigations into the effect of the sulfated RaxX peptide in planta. In this assay, rice plants were inoculated with a virulent strain of Xoo and then treated with the RaxX peptide 2 days after inoculation. We found that post-inoculation treatment of XA21 plants with the sulfated RaxX peptide suppresses the development of Xoo infection in XA21 rice plants. The treated plants display restricted lesion development and reduced bacterial growth. Our findings demonstrate that exogenous application of sulfated RaxX activates XA21-mediated immunity in planta, and provides a potential strategy for the control of bacterial disease in the field. © 2016 BSPP and John Wiley & Sons Ltd.

In vitro and in vivo antivirus activity of an anti-programmed death-ligand 1 (PD-L1) rat-bovine chimeric antibody against bovine leukemia virus infection.

PubMed

Nishimori, Asami; Konnai, Satoru; Okagawa, Tomohiro; Maekawa, Naoya; Ikebuchi, Ryoyo; Goto, Shinya; Sajiki, Yamato; Suzuki, Yasuhiko; Kohara, Junko; Ogasawara, Satoshi; Kato, Yukinari; Murata, Shiro; Ohashi, Kazuhiko

2017-01-01

Programmed death-1 (PD-1), an immunoinhibitory receptor on T cells, is known to be involved in immune evasion through its binding to PD-ligand 1 (PD-L1) in many chronic diseases. We previously found that PD-L1 expression was upregulated in cattle infected with bovine leukemia virus (BLV) and that an antibody that blocked the PD-1/PD-L1 interaction reactivated T-cell function in vitro. Therefore, this study assessed its antivirus activities in vivo. First, we inoculated the anti-bovine PD-L1 rat monoclonal antibody 4G12 into a BLV-infected cow. However, this did not induce T-cell proliferation or reduction of BLV provirus loads during the test period, and only bound to circulating IgM+ B cells until one week post-inoculation. We hypothesized that this lack of in vivo effects was due to its lower stability in cattle and so established an anti-PD-L1 rat-bovine chimeric antibody (Boch4G12). Boch4G12 was able to bind specifically with bovine PD-L1, interrupt the PD-1/PD-L1 interaction, and activate the immune response in both healthy and BLV-infected cattle in vitro. Therefore, we experimentally infected a healthy calf with BLV and inoculated it intravenously with 1 mg/kg of Boch4G12 once it reached the aleukemic (AL) stage. Cultivation of peripheral blood mononuclear cells (PBMCs) isolated from the tested calf indicated that the proliferation of CD4+ T cells was increased by Boch4G12 inoculation, while BLV provirus loads were significantly reduced, clearly demonstrating that this treatment induced antivirus activities. Therefore, further studies using a large number of animals are required to support its efficacy for clinical application.

In vitro and in vivo antivirus activity of an anti-programmed death-ligand 1 (PD-L1) rat-bovine chimeric antibody against bovine leukemia virus infection

PubMed Central

Nishimori, Asami; Okagawa, Tomohiro; Maekawa, Naoya; Ikebuchi, Ryoyo; Goto, Shinya; Sajiki, Yamato; Suzuki, Yasuhiko; Kohara, Junko; Ogasawara, Satoshi; Kato, Yukinari; Murata, Shiro; Ohashi, Kazuhiko

2017-01-01

Programmed death-1 (PD-1), an immunoinhibitory receptor on T cells, is known to be involved in immune evasion through its binding to PD-ligand 1 (PD-L1) in many chronic diseases. We previously found that PD-L1 expression was upregulated in cattle infected with bovine leukemia virus (BLV) and that an antibody that blocked the PD-1/PD-L1 interaction reactivated T-cell function in vitro. Therefore, this study assessed its antivirus activities in vivo. First, we inoculated the anti-bovine PD-L1 rat monoclonal antibody 4G12 into a BLV-infected cow. However, this did not induce T-cell proliferation or reduction of BLV provirus loads during the test period, and only bound to circulating IgM+ B cells until one week post-inoculation. We hypothesized that this lack of in vivo effects was due to its lower stability in cattle and so established an anti-PD-L1 rat-bovine chimeric antibody (Boch4G12). Boch4G12 was able to bind specifically with bovine PD-L1, interrupt the PD-1/PD-L1 interaction, and activate the immune response in both healthy and BLV-infected cattle in vitro. Therefore, we experimentally infected a healthy calf with BLV and inoculated it intravenously with 1 mg/kg of Boch4G12 once it reached the aleukemic (AL) stage. Cultivation of peripheral blood mononuclear cells (PBMCs) isolated from the tested calf indicated that the proliferation of CD4+ T cells was increased by Boch4G12 inoculation, while BLV provirus loads were significantly reduced, clearly demonstrating that this treatment induced antivirus activities. Therefore, further studies using a large number of animals are required to support its efficacy for clinical application. PMID:28445479

Optimizing the efficacy of Paecilomyces lilacinus (strain 251) for the control of root-knot nematodes.

PubMed

Kiewnick, S; Sikora, R A

2004-01-01

The egg pathogenic fungus Paecilomyces lilacinus (strain 251) is a biocontrol fungus with a potential range of activity to control the worldwide most important plant parasitic nematodes. This biological nematicide may be an useful tool in an integrated approach to control mainly sedentary nematodes. Greenhouse experiments were conducted with the root-knot nematodes Meloidogyne incognita and M. hapla on tomato. P. lilacinus, formulated as WG (BIOACT WG), was incorporated into soil inoculated with root-knot nematode eggs prior to transplanting the susceptible tomato cultivar "Hellfrucht". Furthermore, soil treatments were combined with seedling treatments 24 hours before transplanting and a soil drench 2 weeks after planting, respectively. Seedling and post planting treatment was also combined with a soil treatment at planting. All single or combination treatments tested decreased the gall index and the number of egg masses compared to the untreated control 12 weeks after planting. However, the combination of the seedling treatment with a pre- or at-planting application of P. lilacinus was necessary to achieve higher levels of control. Additional post plant drenching resulted in only a slight increase In efficacy. To the feasibility of this modified application system for the control of root-knot nematodes, a yield experiment was conducted with M. hapla and the susceptible cultivar "Gnom F1 Hybrid". It could be demonstrated that the above mentioned combination of pre-planting application plus the seedling and one post plant drench gave the best control and resulted in a significant fruit yield increase in concurrence with a decrease in number of galls per root.

Serologic response of domestic ferrets (Mustela putorius furo) to canine distemper and rabies virus vaccines.

PubMed

Hoover, J P; Baldwin, C A; Rupprecht, C E

1989-01-15

Nine unrelated 12-week-old naive domestic ferrets (Mustela putorius furo) were used to evaluate the serologic responses to commercial canine distemper virus (CDV) and rabies virus (RV) vaccines. Five of the ferrets (group 1) were inoculated 3 times at 2-week intervals with a multivalent modified-live virus vaccine of canine cell-line origin, containing CDV and an inactivated RV vaccine. Four of the ferrets (group 2) were inoculated once with the multivalent modified-live virus vaccine containing CDV and were not inoculated with the RV vaccine. Both group-1 and group-2 ferrets seroconverted to the CDV component of the vaccine. Group-1 ferrets also seroconverted after RV vaccination and maintained serum antibody titers to both CDV and RV for at least 7 months. Domestic ferret sera were found to have IgG epitopes similar to sera of domestic dogs and cats. Domestic ferret sera did not contain antibodies to feline coronavirus or FeLV antigens.

Post-Natal Persistent Infection With Classical Swine Fever Virus in Wild Boar: A Strategy for Viral Maintenance?

PubMed

Cabezón, O; Colom-Cadena, A; Muñoz-González, S; Pérez-Simó, M; Bohórquez, J A; Rosell, R; Marco, I; Domingo, M; Lavín, S; Ganges, L

2017-04-01

In this study, fifteen wild boar piglets were intranasally inoculated <10 h after birth with the moderately virulent classical swine fever virus (CSFV) strain Catalonia 01. At 5 days post-inoculation, seven other animals within 48 h of birth were put in contact with them. Viral replication and innate and specific immune responses were evaluated. Of the inoculated animals, 46.67% remained post-natally persistently infected and were apparently healthy with neither humoral nor cellular immunological responses specific to CSFV and with high viral loads in their blood, organs and body secretions. Moreover, the present data extend the time period to 48 h after birth when a moderately virulent CSFV strain could lead to post-natal persistent infection given the generation of persistently infected wild boars in the contact group (33.33%). The innate immune response to the virus, as measured by type I IFN-α in serum, was mostly not impaired in the persistently infected wild boars. Interestingly, a decrease and lack of IFN-γ-producing cells against CSFV and PHA was observed. In endemic countries where wild swine species are increasing and low and moderate virulence CSFV strains are prevalent, the possible generation of this form of disease cannot be ruled out. © 2015 Blackwell Verlag GmbH.

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

PubMed

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

2014-06-01

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

Transient and Prolonged Response of Chicken Cecum Mucosa to Colonization with Different Gut Microbiota

PubMed Central

Volf, Jiri; Polansky, Ondrej; Varmuzova, Karolina; Gerzova, Lenka; Sekelova, Zuzana; Faldynova, Marcela; Babak, Vladimir; Medvecky, Matej; Smith, Adrian L.; Kaspers, Bernd; Velge, Philippe; Rychlik, Ivan

2016-01-01

In this study we determined protein and gene expression in the caeca of newly hatched chickens inoculated with cecal contents sourced from hens of different ages. Over 250 proteins exhibited modified expression levels in response to microbiota inoculation. The most significant inductions were observed for ISG12-2, OASL, ES1, LYG2, DMBT1-L, CDD, ANGPTL6, B2M, CUZD1, IgM and Ig lambda chain. Of these, ISG12-2, ES1 and both immunoglobulins were expressed at lower levels in germ-free chickens compared to conventional chickens. In contrast, CELA2A, BRT-2, ALDH1A1, ADH1C, AKR1B1L, HEXB, ALDH2, ALDOB, CALB1 and TTR were expressed at lower levels following inoculation of microbiota. When chicks were given microbiota preparations from different age donors, the recipients mounted differential responses to the inoculation which also differed from the response profile in naturally colonised birds. For example, B2M, CUZD1 and CELA2A responded differently to the inoculation with microbiota of 4- or 40-week-old hens. The increased or decreased gene expression could be recorded 6 weeks after the inoculation of newly hatched chickens. To characterise the proteins that may directly interact with the microbiota we characterised chicken proteins that co-purified with the microbiota and identified a range of host proteins including CDD, ANGPTL6, DMBT1-L, MEP1A and Ig lambda. We propose that induction of ISG12-2 results in reduced apoptosis of host cells exposed to the colonizing commensal microbiota and that CDD, ANGPTL6, DMBT1-L, MEP1A and Ig lambda reduce contact of luminal microbiota with the gut epithelium thereby reducing the inflammatory response. PMID:27685470

Effect of diethylcarbamazine on Strongylus vulgaris infection in ponies.

PubMed

Hofing, G L; Bennett, D G

1982-02-01

Shetland ponies (n = 4) were given diethylcarbamazine orally at a dose level of 22 mg/kg/day for 1 week before they were inoculated with 800 third-stage larvae of Strongylus vulgaris. Treatment was continued for 86 (1 pony) or 200 days (3 ponies) after the inoculation. As compared with the changes seen in a similarly inoculated group of ponies (group 2) which were not treated, diethylcarbamazine did not prevent the clinical or pathologic changes due to the migrating larvae. Fewer adult parasites were recovered at necropsy from treated ponies than from nontreated (group 2) ponies, even when treatment was discontinued 86 days after inoculation. Treatment appeared to have a detrimental effect on 4th-stage larvae either in the arteries or their intestinal wall, but not until after arterial lesions resulted.

Evaluation of hha and hha sepB mutant strains of Escherichia coli O157:H7 as bacterins for reducing E. coli O157:H7 shedding in cattle.

PubMed

Sharma, Vijay K; Dean-Nystrom, Evelyn A; Casey, Thomas A

2011-07-12

Escherichia coli O157:H7 colonizes cattle intestines by using the locus of enterocyte effacement (LEE)-encoded proteins. The induction of systemic immune response against LEE-encoded proteins, therefore, will prove effective in reducing E. coli O157:H7 colonization in cattle. The previous studies have demonstrated that a hha (encodes for a hemolysin expression modulating protein) deletion enhances expression of LEE-encoded proteins and a sepB (encodes an ATPase required for the secretion of LEE-encoded proteins) deletion results in intracellular accumulation of LEE proteins. In this study, we demonstrate the efficacy of the hha and hha sepB deletion mutants as bacterins for reducing fecal shedding of E. coli O157:H7 in experimentally inoculated weaned calves. The weaned calves were injected intramuscularly with the bacterins containing 10(9) heat-killed cells of the hha(+) wild-type or hha or hha sepB isogenic mutants, and boosted with the same doses 2- and 4-weeks later. The evaluation of the immune response two weeks after the last booster immunization revealed that the calves vaccinated with the hha mutant bacterin had higher antibody titers against LEE proteins compared to the titers for these antibodies in the calves vaccinated with the hha sepB mutant or hha(+) wild-type bacterins. Following oral inoculations with 10(10) CFU of the wild-type E. coli O157:H7, the greater numbers of calves in the group vaccinated with the hha or hha sepB mutant bacterins stopped shedding the inoculum strain within a few days after the inoculations compared to the group of calves vaccinated with the hha(+) wild-type bacterin or PBS sham vaccine. Thus, the use of bacterins prepared from the hha and hha sepB mutants for reducing colonization of E. coli O157:H7 in cattle could represent a potentially important pre-harvest strategy to enhance post-harvest safety of bovine food products, water and produce. Copyright © 2011 Elsevier Ltd. All rights reserved.

The role of virus dose in experimental bovine leukemia virus infection in sheep.

PubMed

Stirtzinger, T; Valli, V E; Miller, J M

1988-04-01

Twenty-four, six month old lambs were assembled into four groups of five animals each and one group of four animals. All groups were inoculated with lymphocytes from a single donor lamb infected with bovine leukemia virus. The inoculum varied from 250 to 250,000 lymphocytes, in tenfold increments. Animals were exposed by intradermal injection in the neck region immediately anterior to the left shoulder joint. All groups were monitored at 0, 3, 7 and 12 weeks after inoculation using the following procedures: a. Syncytia induction assay for detection of bovine leukemia virus in peripheral blood lymphocytes. b. Agar gel immunodiffusion against the gp51 antigen of bovine leukemia virus for the detection of antibovine leukemia virus gp51 antibody. c. Lymphocyte stimulation test for the assessment of cell-mediated immunity using mitogen, nonfractionated bovine leukemia virus antigen, and partially purified bovine lymphoma tumor-associated antigen for the in vitro activation of lymphocytes from bovine leukemia virus-inoculated and sham-inoculated, control animals. d. Routine hematological techniques for the assessment of total leukocyte and lymphocyte counts. The median infectious dose for lymphocytes from the single bovine leukemia virus-infected donor used in this study was determined to be 2000 cells. The syncytia induction assay detected more infected individuals (13/23) at an earlier time than did the agar gel immunodiffusion assay (10/23). Using either serological or virus isolation techniques, infected animals were first detected at three weeks postinoculation in the group receiving the high-dose inoculum and at seven weeks postinoculation in groups receiving low- or medium-dose inocula.(ABSTRACT TRUNCATED AT 250 WORDS)

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

PubMed

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

2018-02-01

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

At-hatch administration of probiotic to chickens can introduce beneficial changes in gut microbiota.

PubMed

Baldwin, Stephen; Hughes, Robert J; Hao Van, Thi Thu; Moore, Robert J; Stanley, Dragana

2018-01-01

Recent advances in culture-free microbiological techniques bring new understanding of the role of intestinal microbiota in heath and performance. Intestinal microbial communities in chickens assume a near-stable state within the week which leaves a very small window for permanent microbiota remodelling. It is the first colonisers that determine the fate of microbial community in humans and birds alike, and after the microbiota has matured there are very small odds for permanent modification as stable community resists change. In this study we inoculated broiler chicks immediately post hatch, with 3 species of Lactobacillus, identified by sequencing of 16S rRNA and pheS genes as L. ingluviei, L. agilis and L. reuteri. The strains were isolated from the gut of healthy chickens as reproducibly persistent Lactobacillus strains among multiple flocks. Birds inoculated with the probiotic mix reached significantly higher weight by 28 days of age. Although each strain was able to colonise when administered alone, administering the probiotic mix at-hatch resulted in colonisation by only L. ingluviei. High initial abundance of L. ingluviei was slowly reducing, however, the effects of at-hatch administration of the Lactobacillus mix on modifying microbiota development and structure remained persistent. There was a tendency of promotion of beneficial and reduction in pathogenic taxa in the probiotic administered group.

Laboratory-acquired vaccinia virus infection in a recently immunized person--Massachusetts, 2013.

PubMed

Hsu, Christopher H; Farland, Julien; Winters, Thomas; Gunn, Julia; Caron, Donna; Evans, Jennifer; Osadebe, Lynda; Bethune, Leon; McCollum, Andrea M; Patel, Nishi; Wilkins, Kimberly; Davidson, Whitni; Petersen, Brett; Barry, M Anita

2015-05-01

On November 26, 2013, the CDC poxvirus laboratory was notified by the Boston Public Health Commission (BPHC) of an inadvertent inoculation of a recently vaccinated (ACAM2000 smallpox vaccine) laboratory worker with wild type vaccinia virus (VACV) Western Reserve. A joint investigation by CDC and BPHC confirmed orthopoxvirus infection in the worker, who had reported a needle stick in his thumb while inoculating a mouse with VACV. He experienced a non-tender, red rash on his arm, diagnosed at a local emergency department as cellulitis. He subsequently developed a necrotic lesion on his thumb, diagnosed as VACV infection. Three weeks after the injury, the thumb lesion was surgically debrided and at 2 months post-injury, the skin lesion had resolved. The investigation confirmed that the infection was the first reported VACV infection in the United States in a laboratory worker vaccinated according to the Advisory Committee on Immunization Practices (ACIP) recommendations. The incident prompted the academic institution to outline biosafety measures for working with biologic agents, such as biosafety training of laboratory personnel, vaccination (if appropriate), and steps in incident reporting. Though vaccination has been shown to be an effective measure in protecting personnel in the laboratory setting, this case report underscores the importance of proper safety measures and incident reporting.

Age and cellular context influence rectal prolapse formation in mice with caecal wall colorectal cancer xenografts.

PubMed

Tommelein, Joke; Gremonprez, Félix; Verset, Laurine; De Vlieghere, Elly; Wagemans, Glenn; Gespach, Christian; Boterberg, Tom; Demetter, Pieter; Ceelen, Wim; Bracke, Marc; De Wever, Olivier

2016-11-15

In patients with rectal prolapse is the prevalence of colorectal cancer increased, suggesting that a colorectal tumor may induce rectal prolapse. Establishment of tumor xenografts in immunodeficient mice after orthotopic inoculations of human colorectal cancer cells into the caecal wall is a widely used approach for the study of human colorectal cancer progression and preclinical evaluation of therapeutics. Remarkably, 70% of young mice carrying a COLO320DM caecal tumor showed symptoms of intussusception of the large bowel associated with intestinal lumen obstruction and rectal prolapse. The quantity of the COLO320DM bioluminescent signal of the first three weeks post-inoculation predicts prolapse in young mice. Rectal prolapse was not observed in adult mice carrying a COLO320DM caecal tumor or young mice carrying a HT29 caecal tumor. In contrast to HT29 tumors, which showed local invasion and metastasis, COLO320DM tumors demonstrated a non-invasive tumor with pushing borders without presence of metastasis. In conclusion, rectal prolapse can be linked to a non-invasive, space-occupying COLO320DM tumor in the gastrointestinal tract of young immunodeficient mice. These data reveal a model that can clarify the association of patients showing rectal prolapse with colorectal cancer.

Effect of Field Inoculation with Sinorhizobium meliloti L33 on the Composition of Bacterial Communities in Rhizospheres of a Target Plant (Medicago sativa) and a Non-Target Plant (Chenopodium album)—Linking of 16S rRNA Gene-Based Single-Strand Conformation Polymorphism Community Profiles to the Diversity of Cultivated Bacteria

PubMed Central

Schwieger, Frank; Tebbe, Christoph C.

2000-01-01

Fourteen weeks after field release of luciferase gene-tagged Sinorhizobium meliloti L33 in field plots seeded with Medicago sativa, we found that the inoculant also occurred in bulk soil from noninoculated control plots. In rhizospheres of M. sativa plants, S. meliloti L33 could be detected in noninoculated plots 12 weeks after inoculation, indicating that growth in the rhizosphere preceded spread into bulk soil. To determine whether inoculation affected bacterial diversity, 1,119 bacteria were isolated from the rhizospheres of M. sativa and Chenopodium album, which was the dominant weed in the field plots. Amplified ribosomal DNA restriction analysis (ARDRA) revealed plant-specific fragment size frequencies. Dominant ARDRA groups were identified by 16S rRNA gene nucleotide sequencing. Database comparisons indicated that the rhizospheres contained members of the Proteobacteria (α, β, and γ subgroups), members of the Cytophaga-Flavobacterium group, and gram-positive bacteria with high G+C DNA contents. The levels of many groups were affected by the plant species and, in the case of M. sativa, by inoculation. The most abundant isolates were related to Variovorax sp., Arthrobacter ramosus, and Acinetobacter calcoaceticus. In the rhizosphere of M. sativa, inoculation reduced the numbers of cells of A. calcoaceticus and members of the genus Pseudomonas and increased the number of rhizobia. Cultivation-independent PCR–single-strand conformation polymorphism (SSCP) profiles of a 16S rRNA gene region confirmed the existence of plant-specific rhizosphere communities and the effect of the inoculant. All dominant ARDRA groups except Variovorax species could be detected. On the other hand, the SSCP profiles revealed products which could not be assigned to the dominant cultured isolates, indicating that the bacterial diversity was greater than the diversity suggested by cultivation. PMID:10919821

A dry-inoculation method for nut kernels.

PubMed

Blessington, Tyann; Theofel, Christopher G; Harris, Linda J

2013-04-01

A dry-inoculation method for almonds and walnuts was developed to eliminate the need for the postinoculation drying required for wet-inoculation methods. The survival of Salmonella enterica Enteritidis PT 30 on wet- and dry-inoculated almond and walnut kernels stored under ambient conditions (average: 23 °C; 41 or 47% RH) was then compared over 14 weeks. For wet inoculation, an aqueous Salmonella preparation was added directly to almond or walnut kernels, which were then dried under ambient conditions (3 or 7 days, respectively) to initial nut moisture levels. For the dry inoculation, liquid inoculum was mixed with sterilized sand and dried for 24 h at 40 °C. The dried inoculated sand was mixed with kernels, and the sand was removed by shaking the mixture in a sterile sieve. Mixing procedures to optimize the bacterial transfer from sand to kernel were evaluated; in general, similar levels were achieved on walnuts (4.8-5.2 log CFU/g) and almonds (4.2-5.1 log CFU/g). The decline of Salmonella Enteritidis populations was similar during ambient storage (98 days) for both wet-and dry-inoculation methods for both almonds and walnuts. The dry-inoculation method mimics some of the suspected routes of contamination for tree nuts and may be appropriate for some postharvest challenge studies. Copyright © 2012 Elsevier Ltd. All rights reserved.

Topical Administration Is a Promising Inoculating Route versus Intramuscular Inoculation for the Nanoparticle-Carried DNA Vaccine to Prevent Corneal Infections.

PubMed

Hu, Kai; Malla, Tejsu; Zhai, Yujia; Dong, Lili; Tang, Ru

2015-01-01

To evaluate the comparative effect of topical versus intramuscular administration of nanoparticle-carried DNA vaccine in preventing corneal herpes simplex virus type 1 (HSV-1) infection. Nanoparticle [polyethylenimine (PEI)-Fe3O4]-carried DNA vaccine (PEI-Fe3O4-pRSC-gD-IL-21) or DNA vaccine (pRSC-gD-IL-21) alone were topically versus intramuscularly inoculated into one eye each of mice on days 0, 14 and 28. Three weeks after the final immunization, the specific immune responses and clinical degrees of primary herpes simplex keratitis were evaluated. Topical inoculation of nanoparticle-carried DNA vaccine induced mice to generate similar levels of specific HSV-1-neutralizing antibody, IFN-γ and IL-4 in serum and specific killing (cytotoxicity) and proliferative activities of the splenic lymphocytes, but a significantly higher level of secretory IgA in tears compared to those of intramuscular inoculation. More importantly, the mice inoculated topically showed a significantly decreased herpes simplex keratitis severity than the mice inoculated intramuscularly after HSV-1 challenge on the corneas of the mice. Topical inoculation of nanoparticle-carried DNA vaccine elicits a stronger specific local immune response and more effectively inhibits herpes simplex keratitis as compared to intramuscular inoculation in an HSV-1 ocular challenge mouse model. Thus, topical administration may be a promising inoculating route for the nanoparticle-carried DNA vaccine to prevent corneal infections. © 2015 S. Karger AG, Basel.

In vivo whole animal body imaging reveals colonization of Chlamydia muridarum to the lower genital tract at early stages of infection.

PubMed

Gupta, Rishein; Wali, Shradha; Yu, Jieh-Juen; Chambers, James P; Zhong, Guangming; Murthy, Ashlesh K; Bakar, Sazaly Abu; Guentzel, M N; Arulanandam, Bernard P

2014-10-01

The leading cause of sexually transmitted bacterial infection is Chlamydia trachomatis. The aim of this study is to investigate the early events in colonization of this bacterium within the murine genital tract. An in vivo animal body imaging technology was used to track fluorophore labeled C. muridarum elementary bodies (EBs) inoculated intravaginally in C57BL/6 mice during the first 24 h of infection. Ascension of viable EBs was observed (1) to be localized to the lower regions of the murine genital tract within the first 24 h post challenge and (2) was dose independent during this early exposure period. Molecular detection revealed enhanced bacterial load in lower regions of the genital tract with increasing bacterial load in the upper region beginning 12 h post inoculation. This study provides additional insight into chlamydial colonization in the murine genital tract during the first 12-24 h following inoculation.

Infection of some cayenne pepper varieties (Capsicum frustescens L.) by Tobacco mosaic virus at different growth stages

NASA Astrophysics Data System (ADS)

Damiri, N.; Sofita, I. S.; Effend, T. A.; Rahim, S. E.

2017-09-01

This research aimed to study the infection of three varieties of cayenne pepper (Capsicum frustescens L.) by Tobacco Mosaic Virus when they were inoculated at 2, 4, 6, 8 and 10 weeks old after planting. This experiment was conducted in a green house, at the Plant pests and diseases department, Agriculture Faculty, Sriwijaya University, Indralaya, South Sumatra Indonesia from March to October 2014. The study was arranged in factorial completely randomized design with three replicates. First factor was varieties of cayenne pepper namely green, white and small. Second factor was growth stage. Results of the study showed that TMV inoculated at different growth stages of three cayenne pepper varieties affected the incubation period of TMV symptom, time for flowering and productions. The infection of TMV on various ages affected the disease severity on cayenne pepper variety. The highest disease severity was taking place on small cayenne pepper variety that was inoculated at the early stages of age namely 2 weeks after planting. Inoculation of TMV at younger stages of all Cayenne peppers varieties caused a significant reduction in the number of fruits and its weights. TMV has caused a reduction of more than 50% in weight of cayenne pepper fruits regardless of the variety.

Ability of herpes simplex virus vectors to boost immune responses to DNA vectors and to protect against challenge by simian immunodeficiency virus

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

Kaur, Amitinder; Sanford, Hannah B.; Garry, Deirdre

2007-01-20

The immunogenicity and protective capacity of replication-defective herpes simplex virus (HSV) vector-based vaccines were examined in rhesus macaques. Three macaques were inoculated with recombinant HSV vectors expressing Gag, Env, and a Tat-Rev-Nef fusion protein of simian immunodeficiency virus (SIV). Three other macaques were primed with recombinant DNA vectors expressing Gag, Env, and a Pol-Tat-Nef-Vif fusion protein prior to boosting with the HSV vectors. Robust anti-Gag and anti-Env cellular responses were detected in all six macaques. Following intravenous challenge with wild-type, cloned SIV239, peak and 12-week plasma viremia levels were significantly lower in vaccinated compared to control macaques. Plasma SIV RNAmore » in vaccinated macaques was inversely correlated with anti-Rev ELISPOT responses on the day of challenge (P value < 0.05), anti-Tat ELISPOT responses at 2 weeks post challenge (P value < 0.05) and peak neutralizing antibody titers pre-challenge (P value 0.06). These findings support continued study of recombinant herpesviruses as a vaccine approach for AIDS.« less

T helper 1 background protects against airway hyperresponsiveness and inflammation in guinea pigs with persistent respiratory syncytial virus infection.

PubMed

Sutton, Troy C; Tayyari, Farnoosh; Khan, M Aatif; Manson, Heather E; Hegele, Richard G

2007-05-01

A family history of allergy has been implicated in children who develop post-bronchiolitis wheezing and asthma. In a guinea pig model of respiratory syncytial virus (RSV) lung infection, we evaluated the role of host Th1 background (either genetic or induced) on the development of a persistent infection, nonspecific airway hyperresponsiveness (AHR) and airway inflammation. Allergy resistant/T helper 1 (Th1)-skewed strain 2 guinea pigs (STR2) and cytosine phosphate guanine oligodeoxynucleotides (CpG-ODN) (Th1 stimuli) pretreated Cam Hartley guinea pigs (CH) were inoculated with RSV and compared with virus-inoculated allergy-susceptible/Th2-skewed CHs and to sham-inoculated STR2 and CH, 60 d post-inoculation. We measured titers of intrapulmonary RSV, lung interferon (IFN)-gamma and interleukin (IL)-5 mRNA expression, AHR and airway T cells and eosinophils. All virus-inoculated groups of animals showed evidence of persistent RSV lung infection; however, Th2-skewed guinea pigs had virus-associated AHR and significantly greater levels of airway T cells and eosinophils. In conclusion, RSV can establish persistent infection of the guinea pig lung regardless of host Th1/Th2 background; however; a host Th1 background limits the extent of virus-associated AHR and airway inflammation. Heterogeneity in virus-host interactions may be relevant to understanding why some children hospitalized for RSV bronchiolitis go on to develop recurrent wheezing/asthma symptoms.

Effect of Low Dose of Fumonisins on Pig Health: Immune Status, Intestinal Microbiota and Sensitivity to Salmonella

PubMed Central

Burel, Christine; Tanguy, Mael; Guerre, Philippe; Boilletot, Eric; Cariolet, Roland; Queguiner, Marilyne; Postollec, Gilbert; Pinton, Philippe; Salvat, Gilles; Oswald, Isabelle P.; Fravalo, Philippe

2013-01-01

The objective of this study was to measure the effects of chronic exposure to fumonisins via the ingestion of feed containing naturally contaminated corn in growing pigs infected or not with Salmonella spp. This exposure to a moderate dietary concentration of fumonisins (11.8 ppm) was sufficient to induce a biological effect in pigs (Sa/So ratio), but no mortality or pathology was observed over 63 days of exposure. No mortality or related clinical signs, even in cases of inoculation with Salmonella (5 × 104 CFU), were observed either. Fumonisins, at these concentrations, did not affect the ability of lymphocytes to proliferate in the presence of mitogens, but after seven days post-inoculation they led to inhibition of the ability of specific Salmonella lymphocytes to proliferate following exposure to a specific Salmonella antigen. However, the ingestion of fumonisins had no impact on Salmonella translocation or seroconversion in inoculated pigs. The inoculation of Salmonella did not affect faecal microbiota profiles, but exposure to moderate concentrations of fumonisins transiently affected the digestive microbiota balance. In cases of co-infection with fumonisins and Salmonella, the microbiota profiles were rapidly and clearly modified as early as 48 h post-Salmonella inoculation. Therefore under these experimental conditions, exposure to an average concentration of fumonisins in naturally contaminated feed had no effect on pig health but did affect the digestive microbiota balance, with Salmonella exposure amplifying this phenomenon. PMID:23612754

Latency and Persistence of 'Candidatus Liberibacter asiaticus' in Its Psyllid Vector, Diaphorina citri (Hemiptera: Liviidae).

PubMed

Canale, Maria Cristina; Tomaseto, Arthur Fernando; Haddad, Marineia de Lara; Della Coletta-Filho, Helvécio; Lopes, João Roberto Spotti

2017-03-01

Although 'Candidatus Liberibacter asiaticus' (Las) is a major pathogen associated with citrus huanglongbing (HLB), some characteristics of transmission by the psyllid vector Diaphorina citri are not fully understood. We examined the latent period and persistence of transmission of Las by D. citri in a series of experiments at 25°C, in which third-instar psyllid nymphs and 1-week-old adults were confined on infected citrus for an acquisition access period (AAP), and submitted to sequential inoculation access periods (IAPs) on healthy citrus seedlings. The median latent period (LP 50 , i.e., acquisition time after which 50% of the individuals can inoculate) of 16.8 and 17.8 days for psyllids that acquired Las as nymphs and adults, respectively, was determined by transferring single individuals in 48-h IAPs. Inoculation events were intermittent and randomly distributed over the IAPs, but were more frequent after acquisition by nymphs. A minimum latent period of 7 to 10 days was observed by transferring groups of 10 psyllids in 48-h IAPs, after a 96-h AAP by nymphs. Psyllids transmitted for up to 5 weeks, when submitted to sequential 1-week IAPs after a 14-day AAP as nymphs. The long latent period and persistence of transmission are indirect evidences of circulative propagation of Las in D. citri.

Experimental Inoculation of Egyptian Fruit Bats (Rousettus aegyptiacus) with Ebola Virus

PubMed Central

Paweska, Janusz T.; Storm, Nadia; Grobbelaar, Antoinette A.; Markotter, Wanda; Kemp, Alan; Jansen van Vuren, Petrus

2016-01-01

Colonized Egyptian fruit bats (Rousettus aegyptiacus), originating in South Africa, were inoculated subcutaneously with Ebola virus (EBOV). No overt signs of morbidity, mortality, or gross lesions were noted. Bats seroconverted by Day 10–16 post inoculation (p.i.), with the highest mean anti-EBOV IgG level on Day 28 p.i. EBOV RNA was detected in blood from one bat. In 16 other tissues tested, viral RNA distribution was limited and at very low levels. No seroconversion could be demonstrated in any of the control bats up to 28 days after in-contact exposure to subcutaneously-inoculated bats. The control bats were subsequently inoculated intraperitoneally, and intramuscularly with the same dose of EBOV. No mortality, morbidity or gross pathology was observed in these bats. Kinetics of immune response was similar to that in subcutaneously-inoculated bats. Viral RNA was more widely disseminated to multiple tissues and detectable in a higher proportion of individuals, but consistently at very low levels. Irrespective of the route of inoculation, no virus was isolated from tissues which tested positive for EBOV RNA. Viral RNA was not detected in oral, nasal, ocular, vaginal, penile and rectal swabs from any of the experimental groups. PMID:26805873

Preempting Performance Challenges: The Effects of Inoculation Messaging on Attacks to Task Self-Efficacy

PubMed Central

Jackson, Ben; Compton, Josh; Whiddett, Ryan; Anthony, David R.; Dimmock, James A.

2015-01-01

Although inoculation messages have been shown to be effective for inducing resistance to counter-attitudinal attacks, researchers have devoted relatively little attention toward studying the way in which inoculation theory principles might support challenges to psychological phenomena other than attitudes (e.g., self-efficacy). Prior to completing a physical (i.e., balance) task, undergraduates (N = 127, Mage = 19.20, SD = 2.16) were randomly assigned to receive either a control or inoculation message, and reported their confidence in their ability regarding the upcoming task. During the task, a confederate provided standardized negative feedback to all participants regarding their performance, and following the completion of the task, participants again reported their self-efficacy along with measures assessing in-task processes. Findings supported the viability of efficacy inoculation; controlling for pre-task self-efficacy, task performance, and relevant psycho-social variables (e.g., resilience, self-confidence robustness), participants in the inoculation condition reported greater confidence in their ability (i.e., task self-efficacy) than those in the control condition at post-task. Relative to those in the inoculation condition, participants in the control condition also experienced greater concentration disruption and self-presentation concerns during the task. PMID:25898287

Recombinant Measles AIK-C Vaccine Strain Expressing the prM-E Antigen of Japanese Encephalitis Virus.

PubMed

Higuchi, Akira; Toriniwa, Hiroko; Komiya, Tomoyoshi; Nakayama, Tetsuo

2016-01-01

An inactivated Japanese encephalitis virus (JEV) vaccine, which induces neutralizing antibodies, has been used for many years in Japan. In the present study, the JEV prM-E protein gene was cloned, inserted at the P/M junction of measles AIK-C cDNA, and an infectious virus was recovered. The JEV E protein was expressed in B95a cells infected with the recombinant virus. Cotton rats were inoculated with recombinant virus. Measles PA antibodies were detected three weeks after immunization. Neutralizing antibodies against JEV developed one week after inoculation, and EIA antibodies were detected three weeks after immunization. The measles AIK-C-based recombinant virus simultaneously induced measles and JEV immune responses, and may be a candidate for infant vaccines. Therefore, the present strategy of recombinant viruses based on a measles vaccine vector would be applicable to the platform for vaccine development.

Amblyomma maculatum Feeding Augments Rickettsia parkeri Infection in a Rhesus Macaque Model: A Pilot Study

PubMed Central

Banajee, Kaikhushroo H.; Embers, Monica E.; Langohr, Ingeborg M.; Doyle, Lara A.; Hasenkampf, Nicole R.; Macaluso, Kevin R.

2015-01-01

Rickettsia parkeri is an emerging eschar-causing human pathogen in the spotted fever group of Rickettsia and is transmitted by the Gulf coast tick, Amblyomma maculatum. Tick saliva has been shown to alter both the cellular and humoral components of the innate and adaptive immune systems. However, the effect of this immunomodulation on Rickettsia transmission and pathology in an immunocompetent vertebrate host has not been fully examined. We hypothesize that, by modifying the host immune response, tick feeding enhances infection and pathology of pathogenic spotted fever group Rickettsia sp. In order to assess this interaction in vivo, a pilot study was conducted using five rhesus macaques that were divided into three groups. One group was intradermally inoculated with low passage R. parkeri (Portsmouth strain) alone (n = 2) and another group was inoculated during infestation by adult, R. parkeri-free A. maculatum (n = 2). The final macaque was infested with ticks alone (tick feeding control group). Blood, lymph node and skin biopsies were collected at several time points post-inoculation/infestation to assess pathology and quantify rickettsial DNA. As opposed to the tick-only animal, all Rickettsia-inoculated macaques developed inflammatory leukograms, elevated C-reactive protein concentrations, and elevated TH1 (interferon-γ, interleukin-15) and acute phase inflammatory cytokines (interleukin-6) post-inoculation, with greater neutrophilia and interleukin-6 concentrations in the tick plus R. parkeri group. While eschars formed at all R. parkeri inoculation sites, larger and slower healing eschars were observed in the tick feeding plus R. parkeri group. Furthermore, dissemination of R. parkeri to draining lymph nodes early in infection and increased persistence at the inoculation site were observed in the tick plus R. parkeri group. This study indicates that rhesus macaques can be used to model R. parkeri rickettsiosis, and suggests that immunomodulatory factors introduced during tick feeding may enhance the pathogenicity of spotted fever group Rickettsia. PMID:26244337

[Use the Markov-decision tree model to optimize vaccination strategies of hepatitis E among women aged 15 to 49].

PubMed

Chen, Z M; Ji, S B; Shi, X L; Zhao, Y Y; Zhang, X F; Jin, H

2017-02-10

Objective: To evaluate the cost-utility of different hepatitis E vaccination strategies in women aged 15 to 49. Methods: The Markov-decision tree model was constructed to evaluate the cost-utility of three hepatitis E virus vaccination strategies. Parameters of the models were estimated on the basis of published studies and experience of experts. Both methods on sensitivity and threshold analysis were used to evaluate the uncertainties of the model. Results: Compared with non-vaccination group, strategy on post-screening vaccination with rate as 100%, could save 0.10 quality-adjusted life years per capital in the women from the societal perspectives. After implementation of screening program and with the vaccination rate reaching 100%, the incremental cost utility ratio (ICUR) of vaccination appeared as 5 651.89 and 6 385.33 Yuan/QALY, respectively. Vaccination post to the implementation of a screening program, the result showed better benefit than the vaccination rate of 100%. Results from the sensitivity analysis showed that both the cost of hepatitis E vaccine and the inoculation compliance rate presented significant effects. If the cost were lower than 191.56 Yuan (RMB) or the inoculation compliance rate lower than 0.23, the vaccination rate of 100% strategy was better than the post-screening vaccination strategy, otherwise the post-screening vaccination strategy appeared the optimal strategy. Conclusion: Post-screening vaccination for women aged 15 to 49 from social perspectives seemed the optimal one but it had to depend on the change of vaccine cost and the rate of inoculation compliance.

Anticoccidial effect of Piper sarmentosum extracts in experimental coccidiosis in broiler chickens.

PubMed

Wang, Dingfa; Zhou, Luli; Li, Wei; Zhou, Hanlin; Hou, Guanyu

2016-06-01

To study the anticoccidial effect of Piper sarmentosum extracts (PSE) in experimental broiler coccidiosis, 270 one-day-old Wenchang broiler chickens were randomly assigned to six groups, each with three replicates (n = 15). The six groups were blank control group (BC), negative control group (NC), positive control group (PC), and another three PSE addition groups. Chickens in three control groups were fed a basal diet without PSE supplementation. Chickens in the three PSE addition groups were fed a basal diet supplemented with PSE at 100 (T100), 200 (T200), and 300 (T300) mg/kg of feed, respectively. At 15 days of age, chickens in group NC, PC, and three PSE addition groups were challenged with an oral dose of 1 × 10(5) Eimeria tenella oocysts each chick. Chickens in group PC were fed with diclazuril solution in water for 5 days after 48 h with oocysts inoculation. The results showed that PSE and diclazuril improved growth performance and significantly (P < 0.05) decreased oocysts per gram in inoculated broiler chickens. PSE and diclazuril significantly (P < 0.05) decreased nitric oxide at 6 and 9 days post-inoculation relative to the NC group, respectively. At 6 and 9 days post-inoculation, PSE supplementation at 200 mg/kg in the diet increased concentration of interleukin 2 (IL-2) and interferon gamma (IFN-γ) (P < 0.05). PSE supplementation at 200 mg/kg in the diet significantly (P < 0.05) increased mRNA expressions of IFN-γ and IL-2 in the cecum of chickens at 9 days post-inoculation relative to the BC and NC group. The current results showed the anticoccidial properties, and beneficial effect on intestinal mucosa damage of PSE in broiler chickens that had been challenged by coccidiosis.

Q Fever in Pregnant Goats: Pathogenesis and Excretion of Coxiella burnetii

PubMed Central

Roest, Hendrik-Jan; van Gelderen, Betty; Dinkla, Annemieke; Frangoulidis, Dimitrios; van Zijderveld, Fred; Rebel, Johanna; van Keulen, Lucien

2012-01-01

Coxiella burnetii is an intracellular bacterial pathogen that causes Q fever. Infected pregnant goats are a major source of human infection. However, the tissue dissemination and excretion pathway of the pathogen in goats are still poorly understood. To better understand Q fever pathogenesis, we inoculated groups of pregnant goats via the intranasal route with a recent Dutch outbreak C. burnetii isolate. Tissue dissemination and excretion of the pathogen were followed for up to 95 days after parturition. Goats were successfully infected via the intranasal route. PCR and immunohistochemistry showed strong tropism of C. burnetii towards the placenta at two to four weeks after inoculation. Bacterial replication seemed to occur predominantly in the trophoblasts of the placenta and not in other organs of goats and kids. The amount of C. burnetii DNA in the organs of goats and kids increased towards parturition. After parturition it decreased to undetectable levels: after 81 days post-parturition in goats and after 28 days post-parturition in kids. Infected goats gave birth to live or dead kids. High numbers of C. burnetii were excreted during abortion, but also during parturition of liveborn kids. C. burnetii was not detected in faeces or vaginal mucus before parturition. Our results are the first to demonstrate that pregnant goats can be infected via the intranasal route. C. burnetii has a strong tropism for the trophoblasts of the placenta and is not excreted before parturition; pathogen excretion occurs during birth of dead as well as healthy animals. Besides abortions, normal deliveries in C. burnetii-infected goats should be considered as a major zoonotic risk for Q fever in humans. PMID:23152826

Q fever in pregnant goats: pathogenesis and excretion of Coxiella burnetii.

PubMed

Roest, Hendrik-Jan; van Gelderen, Betty; Dinkla, Annemieke; Frangoulidis, Dimitrios; van Zijderveld, Fred; Rebel, Johanna; van Keulen, Lucien

2012-01-01

Coxiella burnetii is an intracellular bacterial pathogen that causes Q fever. Infected pregnant goats are a major source of human infection. However, the tissue dissemination and excretion pathway of the pathogen in goats are still poorly understood. To better understand Q fever pathogenesis, we inoculated groups of pregnant goats via the intranasal route with a recent Dutch outbreak C. burnetii isolate. Tissue dissemination and excretion of the pathogen were followed for up to 95 days after parturition. Goats were successfully infected via the intranasal route. PCR and immunohistochemistry showed strong tropism of C. burnetii towards the placenta at two to four weeks after inoculation. Bacterial replication seemed to occur predominantly in the trophoblasts of the placenta and not in other organs of goats and kids. The amount of C. burnetii DNA in the organs of goats and kids increased towards parturition. After parturition it decreased to undetectable levels: after 81 days post-parturition in goats and after 28 days post-parturition in kids. Infected goats gave birth to live or dead kids. High numbers of C. burnetii were excreted during abortion, but also during parturition of liveborn kids. C. burnetii was not detected in faeces or vaginal mucus before parturition. Our results are the first to demonstrate that pregnant goats can be infected via the intranasal route. C. burnetii has a strong tropism for the trophoblasts of the placenta and is not excreted before parturition; pathogen excretion occurs during birth of dead as well as healthy animals. Besides abortions, normal deliveries in C. burnetii-infected goats should be considered as a major zoonotic risk for Q fever in humans.

[Amyloidosis in infected Didelphis marsupialis].

PubMed

Roa, Diana Milena; Sarmiento, Ladys; Rodríguez, Gerzaín

2002-09-01

A male opossum, Didelphis marsupialis, captured in Teruel (Huila), Colombia, was inoculated intraperitoneally with 1 x 10(6) promastigotes of Leishmania chagasi (MHOM/CO/84/CL044B). The animal died 5 weeks after inoculation. Autopsy revealed signs of visceral leishmaniasis along with amastigote parasite form in Kupffer cells and spleen macrophages. Amyloid deposits in liver and spleen were demonstrated by histological staining and electron microscopy. The rapid death was considered a consequence of a secondary, reactive amyloidosis.

Passive serum therapy with polyclonal antibodies against Mycobacterium tuberculosis protects against post-chemotherapy relapse of tuberculosis infection in SCID mice.

PubMed

Guirado, Evelyn; Amat, Isabel; Gil, Olga; Díaz, Jorge; Arcos, Virginia; Caceres, Neus; Ausina, Vicenç; Cardona, Pere-Joan

2006-04-01

We investigated the protective role of immune-sera against reactivation of Mycobacterium tuberculosis infection in SCID mice and found that passive immunization with sera obtained from mice treated with detoxified M. tuberculosis extracts (delivered in liposomes in a composition known as RUTI) exerted significant protection. Our SCID mouse model consisted of aerosol infection by M. tuberculosis, followed by 3 to 8weeks of chemotherapy with isoniazid+rifampicin (INH+RIF) (25 and 10mg/kg, respectively). After infection and antibiotic administration, two groups of mice were treated for up to 10weeks with intraperitoneal passive immunization using hyperimmune serum (HS) obtained from mice infected with M. tuberculosis, treated with chemotherapy (INH+RIF) for 8weeks and inoculated with RUTI (HS group) or with normal serum (CT group). Significant differences were found between HS and CT groups in the number of bacilli in the lungs (3.68+/-2.02 vs. 5.72+/-1.41log(10) c.f.u.), extent of pulmonary granulomatomous infiltration (10.33+/-0.67 vs. 31.2+/-1.77%), and percentage of animals without pulmonary abscesses (16.7% vs. 45.5%). These data strongly suggest a protective role of specific antibodies against lung dissemination of M. tuberculosis infection.

Experimentally induced Fasciola hepatica infection in white-tailed deer. II. Pathological features.

PubMed Central

Presidente, P J; McCraw, B M; Lumsden, J H

1975-01-01

Six white-tailed deer (Odocoileus virginianus) and six sheep were inoculated with metacercariae of Fasciola hepatica. Two animals of each species were given 100, 500 or 2500 metacercariae. One animal in each inocluated group was killed and examined at six weeks postinoculation and the remainder at 15 weeks postinoculation. At six weeks postinoculation the parietal surface of the livers from inoculated deer was covered with gray fibrous plaques and rust colored patches. Fibroplasia with mononuclear cell infiltration characterized Glisson's capsule on the parietal surface. Granulomas were found in the hepatic parenchyma and on the dorsal surface of the lung. Fresh and healing tracks were occasionally found in the liver. In the sheep fibrinous exudate and numerous subcapsular tracks were found on both surfaces of the liver. Inflammatory changes in portal areas and numerous fresh and healing tracks in the hepatic parenchyma were prominent features. At 15 weeks postinoculation inflammatory changes in Glisson's capsule of inoculated deer were less marked than at six weeks but portal fibrosis and hyperplasia of bile duct epithelium were more advanced. A zone of hemorrhage surrounded ducts that contained mature F. hepatica in one deer. The livers from the sheep were rough, pitted and covered with fibrous tags and adhesions to the diaphragm and greater omentum were common. Hemorrhagic tracks were common in the sheep given 500 and 2500 metacercariae. Portal fibrosis and hyperplasia of bile duct epithelium were seen in the sheep (100 metacercariae) that harbored mature F. hepatica. Images Fig. 1. Fig. 2. Fig. 3. Fig. 4. Fig. 5. Fig. 6. Fig. 7. Fig. 8. Fig. 9. Fig. 10. Fig. 11. Fig. 12. PMID:1125834

Oronasal and intramuscular vaccination of swine with a modified live porcine parvovirus vaccine: multiplication and transmission of the vaccine virus.

PubMed

Paul, P S; Mengeling, W L

1984-12-01

An attenuated strain NADL-2 of porcine parvovirus (PPV) has been used at the 54th cell culture passage as a modified live-virus (MLV) vaccine. The present study was conducted to determine the minimum immunizing dose of MLV, the extent of MLV multiplication in swine tissues, and its transmission from swine administered MLV oronasally or intramuscularly. Immune response to MLV was dose dependent and swine responded to as little as 10(2) median cell-culture infective doses (CCID50). A 10(5) CCID50 of MLV, the largest dose given, induced the best immune response and was used in subsequent experiments. Route of MLV administration also was found to be important. The MLV replicated in tissues of swine after IM inoculation; however, viral antigen in tissues was less, as measured by immunofluorescence, and serum hemagglutination-inhibition titers for PPV were lower in MLV-inoculated swine than we have previously observed in virulent PPV-inoculated swine. In contrast, oronasal inoculation with MLV did not consistently result in infection of pigs; only 5 of 23 swine had virologic and/or serologic evidence of infection. Virus transmission studies indicated that MLV is shed in feces, but shedding occurs later than that in virulent-PPV-inoculated swine and is inconsistent. Delayed transmission of MLV was observed in contact pigs, which were seronegative at 2 weeks, but became seropositive at 4 weeks--indicating that perhaps a virus population capable of infecting pigs by oronasal route was selected by passage through the pig.(ABSTRACT TRUNCATED AT 250 WORDS)

Arbuscular mycorrhizal wheat inoculation promotes alkane and polycyclic aromatic hydrocarbon biodegradation: Microcosm experiment on aged-contaminated soil.

PubMed

Ingrid, Lenoir; Lounès-Hadj Sahraoui, Anissa; Frédéric, Laruelle; Yolande, Dalpé; Joël, Fontaine

2016-06-01

Very few studies reported the potential of arbuscular mycorrhizal symbiosis to dissipate hydrocarbons in aged polluted soils. The present work aims to study the efficiency of arbuscular mycorrhizal colonized wheat plants in the dissipation of alkanes and polycyclic aromatic hydrocarbons (PAHs). Our results demonstrated that the inoculation of wheat with Rhizophagus irregularis allowed a better dissipation of PAHs and alkanes after 16 weeks of culture by comparison to non-inoculated condition. These dissipations observed in the inoculated soil resulted from several processes: (i) a light adsorption on roots (0.5% for PAHs), (ii) a bioaccumulation in roots (5.7% for PAHs and 6.6% for alkanes), (iii) a transfer in shoots (0.4 for PAHs and 0.5% for alkanes) and mainly a biodegradation. Whereas PAHs and alkanes degradation rates were respectively estimated to 12 and 47% with non-inoculated wheat, their degradation rates reached 18 and 48% with inoculated wheat. The mycorrhizal inoculation induced an increase of Gram-positive and Gram-negative bacteria by 56 and 37% compared to the non-inoculated wheat. Moreover, an increase of peroxidase activity was assessed in mycorrhizal roots. Taken together, our findings suggested that mycorrhization led to a better hydrocarbon biodegradation in the aged-contaminated soil thanks to a stimulation of telluric bacteria and hydrocarbon metabolization in mycorrhizal roots. Copyright © 2016 Elsevier Ltd. All rights reserved.

The fungistatic and fungicidal effects of volatiles from metathoracic glands of soybean-attacking stink bugs (Heteroptera: Pentatomidae) on the entomopathogen Beauveria bassiana.

PubMed

Lopes, Rogério B; Laumann, Raul A; Blassioli-Moraes, Maria C; Borges, Miguel; Faria, Marcos

2015-11-01

This study was initially designed to evaluate the differential susceptibility of three soybean-attacking pentatomids to the entomopathogenic fungus Beauveria bassiana in standardized bioassays. Euschistus heros (Eh) was shown to be significantly less susceptible than Chinavia ubica (Cu), whereas Dichelops melacanthus (Dm) adults were highly susceptible to fungal infections. A deeper look at the mechanisms involved in the possible role of volatiles from metathoracic glands on fungal infections was undertaken, and gland extracts from Nezara viridula (Nv), a species known for its resilience to fungal infections, were also included in the assays. Atmospheres with volatiles from pentatomids with very low-susceptibility to B. bassiana infections (Eh and Nv) had a significant effect on speed of germination as shown in counts performed up to 22h post-inoculation, by which time 0.1 (control), 0.6 (Dm), 17.9 (Cu), 32.6 (Eh), and 43.4% (Nv) of conidia had not germinated. The fungistatic (inhibitory) and fungicidal (lethal) effects of Eh and Nv volatile-rich atmospheres were subsequently quantified in Petri dishes with either PDA or PDA medium amended with carbendazim, which allowed germination rates to be determined at 18 and 48h post-inoculation, respectively. As opposed to control, Eh volatile-rich atmosphere had a clear fungistatic effect, since germination rate was only 27.4% within 18h, but reached 99.4% at 48h post-inoculation. For Nv volatile-rich atmospheres, only 15.1% of conidia germinated within 18h, and by 48h post-inoculation, approx. 18% of conidia were unviable (neither germ tubes nor intumescence), whereas in the control treatment rates were >99% at both reading times. Therefore, the gaseous phase of defensive secretions from fungus-resilient pentatomids possess a strong inhibitory effect and may display a less pronounced lethal effect on fungal germination, as was the case for Nv. Copyright © 2015 Elsevier Inc. All rights reserved.

Effect of PKC412, an inhibitor of protein kinase C, on spontaneous metastatic model mice.

PubMed

Nakamura, Kazuki; Yoshikawa, Noriko; Yamaguchi, Yu; Kagota, Satomi; Shinozuka, Kazumasa; Kunitomo, Masaru

2003-01-01

We investigated the anti-metastatic effect of PKC412, a selective inhibitor of protein kinase C (PKC), on a spontaneous metastatic mouse model, which was prepared by inoculation with B16-BL6 mouse melanoma cells into the footpad of the right hind leg. At two weeks after inoculation, the primary tumor was amputated completely. PKC412 (200 mg/kg) administered orally for four weeks after the tumor inoculation, significantly prolonged survival compared with the control. Furthermore, to elucidate the mechanism of the anti-metastatic effect of PKC412, we examined the growth rate of B16-BL6 cells premixed with Matrigel in vivo and the invasiveness of B16-BL6 cells using a chemo-invasion chamber in vitro. PKC412 significantly reduced the growth rate of cells in vivo (100 and 200 mg/kg) and the invading cells in vitro (10, 30 and 100 nM) in a dose-dependent manner. Thus, PKC412 exerts an anti-metastatic action through inhibition of the invasiveness of melanoma cells in the extracellular matrix.

Effect of feed restriction on performance and postprandial nutrient metabolism in pigs co-infected with Mycoplasma hyopneumoniae and swine influenza virus.

PubMed

Le Floc'h, Nathalie; Deblanc, Céline; Cariolet, Roland; Gautier-Bouchardon, Anne V; Merlot, Elodie; Simon, Gaëlle

2014-01-01

As nutritional status and inflammation are strongly connected, feeding and nutritional strategies could be effective to improve the ability of pigs to cope with disease. The aims of this study were to investigate the impact of a feed restriction on the ability of pigs to resist and be tolerant to a coinfection with Mycoplasma hyopneumoniae (Mhp) and the European H1N1 swine influenza virus, and the consequences for nutrient metabolism, with a focus on amino acids. Two groups of specific pathogen-free pigs were inoculated with Mhp and H1N1 21 days apart. One group was fed ad libitum, the other group was subjected to a two-week 40% feed restriction starting one week before H1N1 infection. The two respective mock control groups were included. Three days post-H1N1 infection, 200 g of feed was given to pigs previously fasted overnight and serial blood samples were taken over 4 hours to measure plasma nutrient concentrations. Throughout the study, clinical signs were observed and pathogens were detected in nasal swabs and lung tissues. Feed-restricted pigs presented shorter hyperthermia and a positive mean weight gain over the 3 days post-H1N1 infection whereas animals fed ad libitum lost weight. Both infection and feed restriction reduced postprandial glucose concentrations, indicating changes in glucose metabolism. Post-prandial plasma concentrations of the essential amino acids histidine, arginine and threonine were lower in co-infected pigs suggesting a greater use of those amino acids for metabolic purposes associated with the immune response. Altogether, these results indicate that modifying feeding practices could help to prepare animals to overcome an influenza infection. Connections with metabolism changes are discussed.

Effect of Feed Restriction on Performance and Postprandial Nutrient Metabolism in Pigs Co-Infected with Mycoplasma hyopneumoniae and Swine Influenza Virus

PubMed Central

Cariolet, Roland; Gautier-Bouchardon, Anne V.; Merlot, Elodie; Simon, Gaëlle

2014-01-01

As nutritional status and inflammation are strongly connected, feeding and nutritional strategies could be effective to improve the ability of pigs to cope with disease. The aims of this study were to investigate the impact of a feed restriction on the ability of pigs to resist and be tolerant to a coinfection with Mycoplasma hyopneumoniae (Mhp) and the European H1N1 swine influenza virus, and the consequences for nutrient metabolism, with a focus on amino acids. Two groups of specific pathogen-free pigs were inoculated with Mhp and H1N1 21 days apart. One group was fed ad libitum, the other group was subjected to a two-week 40% feed restriction starting one week before H1N1 infection. The two respective mock control groups were included. Three days post-H1N1 infection, 200 g of feed was given to pigs previously fasted overnight and serial blood samples were taken over 4 hours to measure plasma nutrient concentrations. Throughout the study, clinical signs were observed and pathogens were detected in nasal swabs and lung tissues. Feed-restricted pigs presented shorter hyperthermia and a positive mean weight gain over the 3 days post-H1N1 infection whereas animals fed ad libitum lost weight. Both infection and feed restriction reduced postprandial glucose concentrations, indicating changes in glucose metabolism. Post-prandial plasma concentrations of the essential amino acids histidine, arginine and threonine were lower in co-infected pigs suggesting a greater use of those amino acids for metabolic purposes associated with the immune response. Altogether, these results indicate that modifying feeding practices could help to prepare animals to overcome an influenza infection. Connections with metabolism changes are discussed. PMID:25101681

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

PubMed

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

2001-05-01

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

Root ethylene signalling is involved in Miscanthus sinensis growth promotion by the bacterial endophyte Herbaspirillum frisingense GSF30T

PubMed Central

Ludewig, Uwe

2013-01-01

The bacterial endophyte Herbaspirillum frisingense GSF30T is a colonizer of several grasses grown in temperate climates, including the highly nitrogen-efficient perennial energy grass Miscanthus. Inoculation of Miscanthus sinensis seedlings with H. frisingense promoted root and shoot growth but had only a minor impact on nutrient concentrations. The bacterium affected the root architecture and increased fine-root structures. Although H. frisingense has the genetic requirements to fix nitrogen, only minor changes in nitrogen concentrations were observed. Herbaspirillum agglomerates were identified primarily in the root apoplast but also in the shoots. The short-term (3h) and long-term (3 weeks) transcriptomic responses of the plant to bacterial inoculation revealed that H. frisingense induced rapid changes in plant hormone signalling, most prominent in jasmonate signalling. Ethylene signalling pathways were also affected and persisted after 3 weeks in the root. Growth stimulation of the root by the ethylene precursor 1-aminocyclopropane 1-carboxylic acid was dose dependent and was affected by H. frisingense inoculation. Minor changes in the proteome were identified after 3 weeks. This study suggests that H. frisingense improves plant growth by modulating plant hormone signalling pathways and provides a framework to understand the beneficial effects of diazotrophic plant-growth-promoting bacteria, such as H. frisingense, on the biomass grass Miscanthus. PMID:24043849

Detection of bluetongue virus in the blood of inoculated calves: comparison of virus isolation, PCR assay, and in vitro feeding of Culicoides variipennis.

PubMed

MacLachlan, N J; Nunamaker, R A; Katz, J B; Sawyer, M M; Akita, G Y; Osburn, B I; Tabachnick, W J

1994-01-01

The interval after infection when bluetongue virus (BTV) was present in the blood of calves inoculated with BTV serotype 10 (BTV 10) was evaluated by virus isolation (VI) in embryonated chicken eggs (ECE), BTV-specific polymerase chain reaction (PCR), and in vitro blood feeding of vector Culicoides variipennis (C.v.) sonorensis. BTV nucleic acid was detected by PCR in blood cells for 16 to 20 weeks after infection whereas infectious virus was detected by VI in ECE for 2 to 8 weeks. BTV was detected in calf blood by in vitro feeding of C.v. sonorensis for only 0 to 2 weeks after inoculation of calves with BTV 10. Selected bloods which were positive by PCR analysis but not by VI in ECE were not infectious for sheep. The data are consistent with the hypothesis that prolonged viremia in BTV-infected cattle results from association of the virus with blood cells, especially erythrocytes. The fact that calf blood that contained viral nucleic acid as determined by PCR analysis, but not infectious virus as determined by VI in ECE, was not infectious for either the insect vector or sheep suggests that cattle whose blood contains BTV nucleic acid but not infectious virus are unimportant to the epidemiology of BTV infection.

Critical evaluation of post-consumption food waste composting employing thermophilic bacterial consortium.

PubMed

Awasthi, Mukesh Kumar; Selvam, Ammaiyappan; Lai, Ka Man; Wong, Jonathan W C

2017-12-01

Effect of single-function (oil degrading) and multi-functional bacterial consortium with zeolite as additive for post-consumption food waste (PCFW) composting was investigated through assessing the oil content reduction in a computer controlled 20-L composter. Three treatments of PCFWs combined with 10% zeolite were developed: Treatment-2 and Treatment-3 were inoculated with multi-functional (BC-1) and oil degrading bacterial consortium (BC-2), respectively, while T-1 was without bacterial inoculation and served as control. Results revealed that BC-2 inoculated treatment (T-3) was superior to control treatment and marginally better than T-2 in terms of oil degradation. The reduction of oil content was >97.8% in T-3 and 92.27% in T-2, while total organic matter degradation was marginally higher in T-2 (42.95%) than T-3 (41.67%). Other parameters of compost maturity including germination test indicated that T-2 was marginally better than T-3 and significantly enhanced the oily PCFW decomposition and shortened the composting period by 20days. Copyright © 2017 Elsevier Ltd. All rights reserved.

Synovial fluid cytology in experimental acute canine monocytic ehrlichiosis (Ehrlichia canis).

PubMed

Theodorou, Konstantina; Leontides, Leonidas; Siarkou, Victoria I; Petanides, Theodoros; Tsafas, Konstantinos; Harrus, Shimon; Mylonakis, Mathios E

2015-05-15

Evidence-based information of a cause-and-effect relationship between Ehrlichia canis infection and polyarthritis in naturally- or experimentally-infected dogs is currently lacking. The aim of this prospective study was to investigate whether synovial fluid cytological evidence of arthritis could be documented in dogs with acute monocytic ehrlichiosis. Direct synovial fluid cytology smears from eight Beagle dogs experimentally infected with E. canis were examined prior to, and on 21, 35 and 63 days post-inoculation. The cytological variables assessed included cellularity, percentages of mononuclear cells and neutrophils, macrophage reactivity and evidence of E. canis morulae. The median cellularity and percentages of mononuclear cells and neutrophils prior to inoculation did not differ when compared to post-inoculation cytological evaluation. Increased cellularity, E. canis morulae or cytological evidence of arthritis or macrophage reactivity were not observed throughout the course of the study. In the present study, no cytological evidence of arthritis was found in dogs with experimental acute canine monocytic ehrlichiosis, suggesting that E. canis infection should be considered a rather uncommon cause of arthritis in dogs. Copyright © 2015 Elsevier B.V. All rights reserved.

Boron neutron capture therapy (BNCT) for liver metastasis in an experimental model: dose–response at five-week follow-up based on retrospective dose assessment in individual rats.

PubMed

Pozzi, Emiliano C C; Trivillin, Verónica A; Colombo, Lucas L; Monti Hughes, Andrea; Thorp, Silvia I; Cardoso, Jorge E; Garabalino, Marcela A; Molinari, Ana J; Heber, Elisa M; Curotto, Paula; Miller, Marcelo; Itoiz, Maria E; Aromando, Romina F; Nigg, David W; Schwint, Amanda E

2013-11-01

Boron neutron capture therapy (BNCT) was proposed for untreatable colorectal liver metastases. Employing an experimental model of liver metastases in rats, we recently demonstrated that BNCT mediated by boronophenylalanine (BPA-BNCT) at 13 Gy prescribed to tumor is therapeutically useful at 3-week follow-up. The aim of the present study was to evaluate dose–response at 5-week follow-up, based on retrospective dose assessment in individual rats. BDIX rats were inoculated with syngeneic colon cancer cells DHD/K12/TRb. Tumor-bearing animals were divided into three groups: BPA-BNCT (n = 19), Beam only (n = 8) and Sham (n = 7) (matched manipulation, no treatment). For each rat, neutron flux was measured in situ and boron content was measured in a pre-irradiation blood sample for retrospective individual dose assessment. For statistical analysis (ANOVA), individual data for the BPA-BNCT group were pooled according to absorbed tumor dose, BPA-BNCT I: 4.5–8.9 Gy and BPA-BNCT II: 9.2–16 Gy. At 5 weeks post-irradiation, the tumor surface area post-treatment/pre-treatment ratio was 12.2 ± 6.6 for Sham, 7.8 ± 4.1 for Beam only, 4.4 ± 5.6 for BPA-BNCT I and 0.45 ± 0.20 for BPA-BNCT II; tumor nodule weight was 750 ± 480 mg for Sham, 960 ± 620 mg for Beam only, 380 ± 720 mg for BPA-BNCT I and 7.3 ± 5.9 mg for BPA-BNCT II. The BPA-BNCT II group exhibited statistically significant tumor control with no contributory liver toxicity. Potential threshold doses for tumor response and significant tumor control were established at 6.1 and 9.2 Gy, respectively.

Enhancement of growth and salt tolerance of red pepper seedlings (Capsicum annuum L.) by regulating stress ethylene synthesis with halotolerant bacteria containing 1-aminocyclopropane-1-carboxylic acid deaminase activity.

PubMed

Siddikee, Md Ashaduzzaman; Glick, Bernard R; Chauhan, Puneet S; Yim, Woo jong; Sa, Tongmin

2011-04-01

Three 1-aminocyclopropane-1-carboxylic acid (ACC) deaminase-producing halotolerant bacteria were isolated from West Coast soil of Yellow Sea, Incheon, South Korea and evaluated for their efficiency in improving red pepper plant growth under salt stress. The strains RS16, RS656 and RS111 were identified by 16S rRNA gene sequencing as Brevibacterium iodinum, Bacillus licheniformis and Zhihengliuela alba, respectively. Two hour exposure of 100, 150 and 200 mM NaCl stress on 8 day old red pepper seedlings caused 44, 64 and 74% increase ethylene production, while at 150 mM NaCl stress, inoculation of B. licheniformis RS656, Z. alba RS111, and Br. iodinum RS16 reduces ethylene production by 44, 53 and 57%, respectively. Similarly, 3 week old red pepper plants were subjected to salt stress for two weeks and approximately ∼50% reduction in growth recorded at 150 mM NaCl stress compared to negative control whereas bacteria inoculation significantly increase the growth compared to positive control. Salt stress also caused 1.3-fold reduction in the root/shoot dry weight ratio compared to the absence of salt while bacteria inoculation retained the biomass allocation similar to control plants. The salt tolerance index (ratio of biomass of salt stressed to non-stressed plant) was also significantly increased in inoculated plants compared to non-inoculated. Increase nutrient uptakes under salt stress by red pepper further evident that bacteria inoculation ameliorates salt stress effect. In summary, this study indicates that the use of ACC deaminase-producing halotolerant bacteria mitigates the salt stress by reducing salt stress-induced ethylene production on growth of red pepper plants. Copyright © 2011 Elsevier Masson SAS. All rights reserved.

Genetic bottlenecks during systemic movement of Cucumber mosaic virus vary in different host plants

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

Ali, Akhtar; Roossinck, Marilyn J., E-mail: mroossinck@noble.or

2010-09-01

Genetic bottlenecks are stochastic events that narrow variation in a population. We compared bottlenecks during the systemic infection of Cucumber mosaic virus (CMV) in four host plants. We mechanically inoculated an artificial population of twelve CMV mutants to young leaves of tomato, pepper, Nicotiana benthamiana, and squash. The inoculated leaves and primary and secondary systemically infected leaves were sampled at 2, 10, and 15 days post-inoculation. All twelve mutants were detected in all of the inoculated leaves. The number of mutants recovered from the systemically infected leaves of all host species was reduced significantly, indicating bottlenecks in systemic movement. Themore » recovery frequencies of a few of the mutants were significantly different in each host probably due to host-specific selective forces. These results have implications for the differences in virus population variation that is seen in different host plants.« less

Host innate inflammatory factors and staphylococcal protein A influence the duration of human Staphylococcus aureus nasal carriage

PubMed Central

Cole, Amy L.; Muthukrishnan, Gowrishankar; Chong, Christine; Beavis, Ashley; Eade, Colleen R.; Wood, Matthew P.; Deichen, Michael G.; Cole, Alexander M.

2016-01-01

Human Staphylococcus aureus (SA) nasal carriage provides a reservoir for the dissemination of infectious strains; however, factors regulating the establishment and persistence of nasal colonization are mostly unknown. We measured carriage duration and nasal fluid inflammatory markers after nasally inoculating healthy participants with their previously isolated SA strains. Ten out of 15 studies resulted in rapid clearance (9±6 days) that corresponded with upregulated chemokines, growth factors, and predominantly Th1-type cytokines, but not IL-17. Nasal SA persistence corresponded with elevated baseline levels of MIP-1β, IL-1β, and IL-6, no induction of inflammatory factors post-inoculation, and decreased IL-1RA:IL-1β ratio. SA-expressed staphylococcal protein A (SpA) levels correlated positively with carriage duration. Competitive inoculation studies revealed that isogenic SpA knockout (ΔSpA) strains were cleared faster than wild-type only in participants with upregulated inflammatory markers post-inoculation. The remaining participants did not mount an inflammatory response and did not clear either strain. ΔSpA strains demonstrated lower growth rates in carrier nasal fluids and lower survival rates when incubated with neutrophils. Collectively, the presented studies identify innate immune effectors that cooperatively modulate nasal carriage duration, and confirm SpA as a bacterial co-determinant of SA nasal carriage. PMID:26838052

Comparison of a modified shell vial culture procedure with conventional mouse inoculation for rabies virus isolation.

PubMed

Ribas Antúnez, María de los Angeles; Girón, Blanca; Monsalvez, Iraima; Morier, Luis; Acosta, Gretel; Tejero, Yahisel; Cordero, Yanislet; Piedra, Dainelyd

2013-04-01

Rabies is a neurotropic disease that is often lethal. The early diagnosis of rabies infection is important and requires methods that allow for the isolation of the virus from animals and humans. The present study compared a modified shell vial (MSV) procedure using 24-well tissue culture plates with the mouse inoculation test (MIT), which is considered the gold standard for rabies virus isolation. Thirty brain samples (25 positive and 5 negative by the fluorescent antibody test) obtained from different animal species at the National Institute of Hygiene Rafael Rangel in Caracas, Venezuela, were studied by the MIT and MSV assays. Nine samples (36%) were positive at 24 h, 10 (40%) were positive at 48 h and six (24%) were positive at 72 h by the MSV assay. With the MIT assay, 76% were positive at six days post inoculation and 12% were positive at 12 and 18 days post inoculation. One sample that was negative according to the MSV assay was positive with MIT on the 12th day. The MSV procedure exhibited a sensitivity of 96.2%, a specificity of 100%, a positive predictive value of 100% and a negative predictive value 80%. This procedure allowed for rapid rabies virus detection. MIT can be employed as an alternative method in laboratories without tissue culture facilities.

Comparative pathogenesis of H3N2 canine influenza virus in beagle dogs challenged by intranasal and intratracheal inoculation.

PubMed

Luo, Jie; Lu, Gang; Ye, Shaotang; Ou, Jiajun; Fu, Cheng; Zhang, Xin; Wang, Xiangbin; Huang, Ji; Wu, Peixin; Xu, Haibin; Wu, Liyan; Li, Shoujun

2018-05-31

As important companion animals, dogs may serve as intermediate hosts for transmitting influenza virus to humans. However, knowledge regarding H3N2 canine influenza virus (CIV) pathogenicity is not comprehensive, which directly affects the animal models of pathogenicity in H3N2 CIV vaccine research. Here, to assess H3N2 CIV pathogenicity, we utilized 30 ten-week-old purpose-bred beagles intratracheally or intranasally inoculated with 10 6 50 % egg-infectious dose. Intratracheal inoculation was more virulent to dogs than intranasal inoculation as shown by lung pathology score, histopathological changes, clinical symptoms, and body temperature. More intense virus replication was observed in the upper and lower respiratory tracts by intratracheal than intranasal inoculation according to nasal swabs, various organ virus titers, and antigen expression. These results may enhance the H3N2 CIV infection model, providing a more complete experimental basis for studying intrinsic H3N2 CIV pathogenic mechanism, and also serving a reference role for CIV prevention and treatment. Copyright © 2018. Published by Elsevier B.V.

Q fever in pregnant goats: humoral and cellular immune responses

PubMed Central

2013-01-01

Q fever is a zoonosis caused by the intracellular bacterium Coxiella burnetii. Both humoral and cellular immunity are important in the host defence against intracellular bacteria. Little is known about the immune response to C. burnetii infections in domestic ruminants even though these species are the major source of Q fever in humans. To investigate the goat’s immune response we inoculated groups of pregnant goats via inhalation with a Dutch outbreak isolate of C. burnetii. All animals were successfully infected. Phase 1 and Phase 2 IgM- and IgG-specific antibodies were measured. Cellular immune responses were investigated by interferon-gamma, enzyme-linked immunosorbent spot test (IFN-γ Elispot), lymphocyte proliferation test (LPT) and systemic cytokines. After two weeks post inoculation (wpi), a strong anti-C. burnetii Phase 2 IgM and IgG antibody response was observed while the increase in IgM anti-Phase 1 antibodies was less pronounced. IgG anti-Phase 1 antibodies started to rise at 6 wpi. Cellular immune responses were observed after parturition. Our results demonstrated humoral and cellular immune responses to C. burnetii infection in pregnant goats. Cell-mediated immune responses did not differ enough to distinguish between Coxiella-infected and non-infected pregnant animals, whereas a strong-phase specific antibody response is detected after 2 wpi. This humoral immune response may be useful in the early detection of C. burnetii-infected pregnant goats. PMID:23915213

Cutaneous leishmaniasis in the dorsal skin of hamsters: a useful model for the screening of antileishmanial drugs.

PubMed

Robledo, Sara M; Carrillo, Lina M; Daza, Alejandro; Restrepo, Adriana M; Muñoz, Diana L; Tobón, Jairo; Murillo, Javier D; López, Anderson; Ríos, Carolina; Mesa, Carol V; Upegui, Yulieth A; Valencia-Tobón, Alejandro; Mondragón-Shem, Karina; Rodríguez, Berardo; Vélez, Iván D

2012-04-21

Traditionally, hamsters are experimentally inoculated in the snout or the footpad. However in these sites an ulcer not always occurs, measurement of lesion size is a hard procedure and animals show difficulty to eat, breathe and move because of the lesion. In order to optimize the hamster model for cutaneous leishmaniasis, young adult male and female golden hamsters (Mesocricetus auratus) were injected intradermally at the dorsal skin with 1 to 1.5 x l0(7) promastigotes of Leishmania species and progression of subsequent lesions were evaluated for up to 16 weeks post infection. The golden hamster was selected because it is considered the adequate bio-model to evaluate drugs against Leishmania as they are susceptible to infection by different species. Cutaneous infection of hamsters results in chronic but controlled lesions, and a clinical evolution with signs similar to those observed in humans. Therefore, the establishment of the extent of infection by measuring the size of the lesion according to the area of indurations and ulcers is feasible. This approach has proven its versatility and easy management during inoculation, follow up and characterization of typical lesions (ulcers), application of treatments through different ways and obtaining of clinical samples after different treatments. By using this method the quality of animal life regarding locomotion, search for food and water, play and social activities is also preserved.

Evaluation of physicochemical properties and antioxidant activities of kombucha "Tea Fungus" during extended periods of fermentation.

PubMed

Amarasinghe, Hashani; Weerakkody, Nimsha S; Waisundara, Viduranga Y

2018-05-01

Kombucha fermentation is traditionally carried out by inoculating a previously grown tea fungal mat into a freshly prepared tea broth and incubating under aerobic conditions for 7-10 days. In this study, four kombucha beverages were prepared by placing the tea fungal mats in sugared Sri Lankan black tea at varying concentrations for a period of 8 weeks. The antioxidant activities, physicochemical, and qualitative properties were monitored prior to the commencement of the fermentation process, one day after the inoculation with the microorganisms and subsequently on a weekly basis. All samples displayed a statistically significant decrease ( p  <   .05) in the antioxidant activity at the end of 8 weeks, which was indicative of the decreasing functional properties of the beverage. The physicochemical properties indicated increased acidity and turbidity, which might decrease consumer appeal of the fermented beverage. Further studies are necessary to test the accumulation of organic acids, nucleic acids, and toxicity of kombucha on human organs following the extended period of fermentation.

Periapical inflammation subsequent to coronal inoculation of dog teeth root filled with resilon/epiphany in 1 or 2 treatment sessions with chlorhexidine medication.

PubMed

Santos, João M; Palma, Paulo J; Ramos, João C; Cabrita, António S; Friedman, Shimon

2014-06-01

Therapeutic methods that inhibit microbial ingress into filled root canals are desirable. This in vivo study assessed the inhibition of periapical inflammation subsequent to coronal inoculation in canals medicated with 2% chlorhexidine gel and filled with Resilon/Epiphany (Pentron Clinical Technologies, Wallingford, CT). Six Beagle dogs each had 10 two-rooted premolars treated. In group 1 (n = 36 roots), 1 root/tooth had the canal conditioned with Primer Epiphany, filled with Epiphany sealer and Resilon core in 1 session, and coronally sealed with PhotacFil. In group 2 (n = 36 roots), the second root/tooth had the canal medicated with 2% chlorhexidine gel for 1 week and then filled and coronally sealed as in group 1. After 3 weeks, canals were exposed to the oral environment for 7 days, inoculated with isologous plaque, and coronally sealed. Negative controls treated as groups 1 and 2 remained sealed. Positive controls had canals unfilled and exposed. Seven months after inoculation, dogs were euthanized; jaw blocks processed for histologic examination; and periapical inflammation (PI) recorded as none, mild, or severe. In groups 1 and 2, severe PI occurred in 5 of 65 roots (8%) and mild PI in 18 of 65 roots (28%) with a significantly higher (P = .031) PI incidence in group 2 than in group 1. Negative controls had only mild PI in 9 of 29 roots (31%). Roots medicated with 2% chlorhexidine gel had mild PI significantly more (P = .009) than roots filled in 1 session (more than 2-fold). Intracanal medication with 2% chlorhexidine gel and root filling with Resilon/Epiphany did not effectively inhibit apical periodontitis subsequent to coronal inoculation. Copyright © 2014 American Association of Endodontists. All rights reserved.

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

PubMed Central

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

2016-01-01

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

Biochemical responses and oxidative stress in Francisella tularensis infection: a European brown hare model

PubMed Central

2011-01-01

Background The aim of the present study was to investigate biochemical and oxidative stress responses to experimental F. tularensis infection in European brown hares, an important source of human tularemia infections. Methods For these purposes we compared the development of an array of biochemical parameters measured in blood plasma using standard procedures of dry chemistry as well as electrochemical devices following a subcutaneous infection with a wild Francisella tularensis subsp. holarctica strain (a single dose of 2.6 × 109 CFU pro toto). Results Subcutaneous inoculation of a single dose with 2.6 × 109 colony forming units of a wild F. tularensis strain pro toto resulted in the death of two out of five hares. Plasma chemistry profiles were examined on days 2 to 35 post-infection. When compared to controls, the total protein, urea, lactate dehydrogenase, aspartate aminotransferase and alanine aminotransferase were increased, while albumin, glucose and amylase were decreased. Both uric and ascorbic acids and glutathione dropped on day 2 and then increased significantly on days 6 to 12 and 6 to 14 post-inoculation, respectively. There was a two-fold increase in lipid peroxidation on days 4 to 8 post-inoculation. Conclusions Contrary to all expectations, the present study demonstrates that the European brown hare shows relatively low susceptibility to tularemia. Therefore, the circumstances of tularemia in hares under natural conditions should be further studied. PMID:21232117

Virus excretion and antibody dynamics in goats inoculated with a field isolate of peste des petits ruminants virus.

PubMed

Liu, W; Wu, X; Wang, Z; Bao, J; Li, L; Zhao, Y; Li, J

2013-11-01

A field isolate of peste des petits ruminants virus (PPRV) from an outbreak in Tibet, China, was inoculated into goats to investigate the dynamics of virus excretion and antibody production. Further, animals received PPRV vaccine strain Nigeria 75/1. Ocular, nasal and oral samples were tested for the presence of virus antigen by one-step real-time qualitative RT-PCR (qRT-PCR); competitive ELISA (c-ELISA) was used for the measurement of specific antibodies against PPRV. Virus particles could be detected as early as day 3 post-inoculation (pi) and virus excretion lasted for up to day 26 pi. All four goats inoculated with the PPRV field isolate were seropositive as early as day 10 pi. In animals inoculated with the vaccine strain, antibody was detected at day 14 pi, and levels of neutralizing antibodies remained above the protection threshold level (1 : 8) for 8 months. Both virus particles and neutralizing antibodies were detected earlier in goats challenged with the field isolate than in those receiving the vaccine strain. © 2013 Blackwell Verlag GmbH.

Moxidectin steady state prior to inoculation protects cats from subsequent, repeated infection with Dirofilaria immitis.

PubMed

Little, Susan E; Hostetler, Joe A; Thomas, Jennifer E; Bailey, Keith L; Barrett, Anne W; Gruntmeir, Kaylynn; Gruntmeir, Jeff; Starkey, Lindsay A; Basel, Chris; Blagburn, Byron L

2015-02-18

Infection of cats with Dirofilaria immitis causes seroconversion on antibody tests and pulmonary pathology, often without subsequent development of adult heartworms. Consistent administration of topical 10% imidacloprid-1% moxidectin has been shown to result in sustained plasma levels of moxidectin in cats after three to five treatments, a pharmacokinetic behavior known as "steady state". To evaluate the ability of moxidectin at "steady state" to protect cats from subsequent infection with D. immitis, cats (n = 10) were treated with the labeled dose of topical 10% imidacloprid-1% moxidectin for four monthly treatments. Each cat was inoculated with 25 third-stage larvae of D. immitis 7, 14, 21, and 28 days after the last treatment; non-treated cats (n = 9) were inoculated on the same days, serving as infection controls. Blood samples were collected from each cat from 1 month prior to treatment until 7 months after the final inoculation and tested for antibody to, and antigen and microfilaria of, D. immitis. Measurement of serum levels of moxidectin confirmed steady state in treated cats. Cats treated with topical 10% imidacloprid-1% moxidectin prior to trickle inoculation of D. immitis L3 larvae throughout the 28 day post-treatment period remained negative on antibody and antigen tests throughout the study and did not develop gross or histologic lesions characteristic of heartworm infection. A majority of non-treated cats tested antibody positive by 3-4 months post infection (6/9) and, after heat treatment, tested antigen positive by 6-7 months post-infection (5/9). Histologic lesions characteristic of D. immitis infection, including intimal and medial thickening of the pulmonary artery, were present in every cat with D. immitis antibodies (6/6), although adult D. immitis were confirmed in only 5/6 antibody-positive cats at necropsy. Microfilariae were not detected at any time. Taken together, these data indicate that prior treatment with 10% imidacloprid-1% moxidectin protected cats from subsequent infection with D. immitis for 28 days, preventing both formation of a detectable antibody response and development of pulmonary lesions by either immature stages of D. immitis or young adult heartworms.

Expression profiles of differentially regulated genes during the early stages of apple flower infection with Erwinia amylovora

PubMed Central

Sarowar, Sujon; Zhao, Youfu; Soria-Guerra, Ruth Elena; Ali, Shahjahan; Zheng, Danman; Wang, Dongping; Korban, Schuyler S.

2011-01-01

To identify genes involved in the response to the fire blight pathogen Erwinia amylovora in apple (Malus×domestica), expression profiles were investigated using an apple oligo (70-mer) array representing 40, 000 genes. Blossoms of a fire blight-susceptible apple cultivar Gala were collected from trees growing in the orchard, placed on a tray in the laboratory, and spray-inoculated with a suspension of E. amylovora at a concentration of 108 cfu ml−1. Uninoculated detached flowers served as controls at each time point. Expression profiles were captured at three different time points post-inoculation at 2, 8, and 24 h, together with those at 0 h (uninoculated). A total of about 3500 genes were found to be significantly modulated in response to at least one of the three time points. Among those, a total of 770, 855, and 1002 genes were up-regulated, by 2-fold, at 2, 8, and 24 h following inoculation, respectively; while, 748, 1024, and 1455 genes were down-regulated, by 2-fold, at 2, 8, and 24 h following inoculation, respectively. Over the three time points post-inoculation, 365 genes were commonly up-regulated and 374 genes were commonly down-regulated. Both sets of genes were classified based on their functional categories. The majority of up-regulated genes were involved in metabolism, signal transduction, signalling, transport, and stress response. A number of transcripts encoding proteins/enzymes known to be up-regulated under particular biotic and abiotic stress were also up-regulated following E. amylovora treatment. Those up- or down-regulated genes encode transcription factors, signaling components, defense-related, transporter, and metabolism, all of which have been associated with disease responses in Arabidopsis and rice, suggesting similar response pathways are involved in apple blossoms. PMID:21725032

Wild-type and mutant AvrA- Salmonella induce broadly similar immune pathways in the chicken ceca with key differences in signaling intermediates and inflammation.

PubMed

Arsenault, Ryan J; Genovese, Kenneth J; He, Haiqi; Wu, Huixia; Neish, Andrew S; Kogut, Michael H

2016-02-01

Salmonella enterica serovar Typhimurium (ST) is a serious infectious disease throughout the world, and a major reservoir for Salmonella is chicken. Chicken infected with Salmonella do not develop clinical disease, this may be the result of important host interactions with key virulence proteins. To study this, we inoculated chicken with mutant Salmonella Typhimurium that lacked the virulence protein AvrA (AvrA(-)). AvrA is referred to as an avirulence factor, as it moderates the host immune response. The lack of the AvrA virulence gene in ST resulted in reduced weight gain, enhanced persistence and greater extraintestinal organ invasion in chickens, as compared to wild-type (WT) ST. Kinome analysis was performed on inoculated cecal tissue. The majority of the signal transduction pathways induced by AvrA(-) and WT ST were similar; however, we observed alterations in innate immune system signaling. In addition, a leukocyte migration pathway was altered by AvrA(-) ST that may allow greater gut barrier permeability and invasion by the mutant. Cytokine expression did not appear significantly altered at 7 d post-inoculation; at 14 d post-inoculation, there was an observed increase in the expression of anti-inflammatory IL-10 in the WT inoculated ceca. This study is the first to describe mutant AvrA(-) ST infection of chicken and provides further insight into the Salmonella responses observed in chicken relative to other species such as humans and cattle. Published by Oxford University Press on behalf of Poultry Science Association 2015. This work is written by (a) US Government employee(s) and is in the public domain in the US.

Surface ultrastructural studies on the germination, penetration and conidial development of Aspergillus flavus Link:Fries infecting silkworm, Bombyx mori Linn.

PubMed

Kumar, Vineet; Singh, G P; Babu, A M

2004-01-01

Aspergillosis is a common disease of the silkworm Bombyx mori Linn., caused by an insect mycopathogen Aspergillus flavus Link:Fries. The present study reveals the germination, penetration and conidial development of A. flavus on the larval integument of B. mori under SEM. Four different strains (NB18, KA, NB4D2 and NB7) of B. mori was surface inoculated with ca. of 1 x 10(6) conidia/ml. Each conidium germinated on the cuticle approximately 6 h after inoculation, forming a humpy or suctorial appressoria within 24 h. The hyphae which entered into haemocoel 2 day post-inoculation, grew and multiplied extensively, forming a mycelial complex, causing death of the host larva in about 4-5 days. This occurred with minimal breakdown of the internal tissues. Death of the host was followed by ramification of the fungus through the mesodermal and epidermal tissues, leading to larval mummification about 5-6 days after inoculation. Extensive fungal growths on the entire larval body followed, consisting of aerial hyphae, which developed branched conidiophores. The aerial hyphae with abundant conidiophores formed a confluent yellowish green fungal mat over the entire larval body in 6-7 days of post-inoculation. The tip of each emerging conidiophores gradually dilated and developed to become a bulbous head known as the vesicle. A large number of conidiogenous cells were produced over the entire surface of vesicle, which later developed into finger-like projections termed as sterigmata or phialides. The phialides matured within 2 days after the aerial hyphae emerged as evidenced by chains of conidia at their tips. The conidia were globose with externally roughened walls. The life cycle of the fungus on B. mori was completed in six to seven days.

High Incubation Temperature and Threonine Dietary Level Improve Ileum Response Against Post-Hatch Salmonella Enteritidis Inoculation in Broiler Chicks

PubMed Central

de Oliveira, Heraldo Bezerra; Campos, Danila Barreiro; Guerra, Ricardo Romão; Costa, Fernando Guilherme Perazzo

2015-01-01

This study assessed the effect of both embryonic thermal manipulation and dietary threonine level on the response of broilers inoculated with Salmonella Enteritidis, considering bacterial counts in the cecal contents, intestinal morphology, mucin and heat shock protein 70 gene expression, body weight and weight gain. Thermal manipulation was used from 11 days of incubation until hatch, defining three treatments: standard (37.7°C), continuous high temperature (38.7°C) and continuous low temperature (36.7°C). After hatch, chicks were distributed according to a 3x2+1 factorial arrangement (three temperatures and two threonine levels and one sham-inoculated control). At two days of age, all chicks were inoculated with Salmonella Enteritidis, except for the sham-inoculated control group. There was no interaction between the factors on any analyses. High temperature during incubation was able to reduce colonization by Salmonella Enteritidis in the first days, reducing both Salmonella counts and the number of positive birds. It also increased mucin expression and decreased Hsp70 expression compared with other inoculated groups. High temperature during incubation and high threonine level act independently to reduce the negative effects associated to Salmonella Enteritidis infection on intestinal morphology and performance, with results similar to sham-inoculated birds. The findings open new perspectives for practical strategies towards the pre-harvest Salmonella control in the poultry industry. PMID:26131553

Biofumigation on Post-Harvest Diseases of Fruits Using a New Volatile-Producing Fungus of Ceratocystis fimbriata

PubMed Central

Li, Qian; Wu, Lei; Hao, Jianjun; Luo, Laixin; Cao, Yongsong; Li, Jianqiang

2015-01-01

A variety of volatile organic compounds (VOCs) produced by Ceratocystis fimbriata have strong bioactivity against a wide range of fungi, bacteria and oomycetes. Mycelial growth, conidial production, and spore germination of fungi and oomycetes were significantly inhibited after exposure to cultures of C. fimbriata, and colony formation of bacteria was also inhibited. Two post-harvest diseases, peach brown rot caused by Monilinia fructicola and citrus green mold caused by Penicillium digitatum, were controlled during a 4-day storage by enclosing wound-inoculated fruits with 10 standard diameter Petri plate cultures of C. fimbriata in a 15 L box. The fruits were freshly inoculated at onset of storage and the cultures of C. fimbriata were 6 days old. Percentage of control was 92 and 97%, respectively. After exposure to C. fimbriata VOCs, severely misshapen hyphae and conidia of these two post-harvest pathogens were observed by scanning electron microscopy, and their pathogenicity was lost or greatly reduced. PMID:26147922

Healthy rabbits are susceptible to Epstein-Barr virus infection and infected cells proliferate in immunosuppressed animals.

PubMed

Khan, Gulfaraz; Ahmed, Waqar; Philip, Pretty S; Ali, Mahmoud H; Adem, Abdu

2015-02-18

Epstein-Barr virus (EBV) is an oncogenic virus implicated in the pathogenesis of several human malignancies. However, due to the lack of a suitable animal model, a number of fundamental questions pertaining to the biology of EBV remain poorly understood. Here, we explore the potential of rabbits as a model for EBV infection and investigate the impact of immunosuppression on viral proliferation and gene expression. Six healthy New Zealand white rabbits were inoculated intravenously with EBV and blood samples collected prior to infection and for 7 weeks post-infection. Three weeks after the last blood collection, animals were immunosuppressed with daily intramuscular injections of cyclosporin A at doses of 20 mg/kg for 15 days and blood collected twice a week from each rabbit. The animals were subsequently sacrificed and tissues from all major organs were collected for subsequent analysis. Following intravenous inoculation, all 6 rabbits seroconverted with raised IgG and IgM titres to EBV, but viral DNA in peripheral blood mononuclear cells (PBMCs) could only be detected intermittently. Following immunosuppression however, EBV DNA could be readily detected in PBMCs from all 4 rabbits that survived the treatment. Quantitative PCR indicated an increase in EBV viral load in PBMCs as the duration of immunosuppression increased. At autopsy, splenomegaly was seen in 3/4 rabbits, but spleens from all 4 rabbit were EBV PCR positive. EBER-in situ hybridization and immunoshistochemistry revealed the presence of a large number of EBER-positive and LMP-1 positive lymphoblasts in the spleens of 3/4 rabbits. To a lesser extent, EBER-positive cells were also seen in the portal tract regions of the liver of these rabbits. Western blotting indicated that EBNA-1 and EBNA-2 were also expressed in the liver and spleen of infected animals. EBV can infect healthy rabbits and the infected cells proliferate when the animals are immunocompromised. The infected cells expressed several EBV-latent gene products which are probably driving the proliferation, reminiscent of what is seen in immunocompromised individuals. Further work is required to explore the potential of rabbits as an animal model for studying EBV biology and tumorigenesis.

In vivo monitoring of transfected DNA, gene expression kinetics, and cellular immune responses in mice immunized with a human NIS gene-expressing plasmid.

PubMed

Son, Hye-Youn; Jeon, Yong-Hyun; Chung, June-Key; Kim, Chul-Woo

2016-12-01

In assessing the effectiveness of DNA vaccines, it is important to monitor: (1) the kinetics of target gene expression in vivo; and (2) the movement of cells that become transfected with the plasmid DNA used in the immunization of a subject. In this study, we used, as a visual imaging marker, expression of the transfected human sodium/iodide symporter (hNIS) gene, which enhances intracellular radio-pertechnetate (TcO4-) accumulation. After intradermal (i.d.) and systemic injection of mice with pcDNA-hNIS and radioactive Technetium-99m (Tc-99m), respectively, whole-body images were obtained by nuclear scintigraphy. The migration of mice cells transfected with the hNIS gene was monitored over a 2-week period by gamma-radioactivity counting of isolated cell populations and was demonstrated in peripheral lymphoid tissues, especially in the draining lymph nodes (dLNs). Beginning at 24 h after DNA inoculation and continuing for the 2-week monitoring period, hNIS-expressing cells were observed specifically in the T-cell-rich zones of the paracortical area of the dLNs. Over the same time period, high levels of INF-γ-secreting CD8 T-cells were found in the dLNs of the pcDNA-hNIS immunized mice. Tumor growth was also significantly retarded in the mice that received hNIS DNA immunization followed by inoculation with CT26 colorectal adenocarcinoma cells that had been transfected with the rat NIS gene (rNIS), which is 93% homologous to the hNIS gene. In conclusion, mouse cells transfected with hNIS DNA after i.d. immunization were found to traffic to the dLNs, and hNIS gene expression in these cells continued for at least 2 weeks post immunization. Furthermore, sequential presentation of NIS DNA to T-cells by migratory antigen presenting cells could induce NIS DNA-specific Th1 immune responses and thus retard the growth of NIS-expressing tumors. © The Author(s) 2016.

Lactobacillus gasseri K7 modulates the blood cell transcriptome of conventional mice infected with Escherichia coli O157:H7.

PubMed

Sagaya, F M; Hacin, B; Tompa, G; Ihan, A; Špela, Š; Černe, M; Hurrell, R F; Matijašić, B B; Rogelj, I; Vergères, G

2014-05-01

As the immune cells underlying the intestinal barrier sense luminal microbial signals, blood cell transcriptomics may identify subclinical changes triggered by gut bacteria that may otherwise not be detected. We have therefore investigated how Lactobacillus gasseri K7 and enterohemorrhagic Escherichia coli O157:H7 modulate the blood cell transcriptome of mice possessing an intact microbiota. We have analysed the transcriptome of five groups of C57BL/6J mice: (i) control, (ii) inoculated with a single dose of E. coli, (iii) inoculated during 2 weeks with Lact. gasseri, (iv) co-inoculated with E. coli and Lact. gasseri, (v) inoculated with Lact. gasseri prior to E. coli infection. The transcriptome could distinguish between the five treatment groups. Gene characteristics of bacterial infection, in particular inflammation, were upregulated in the mice inoculated with E. coli. Lact. gasseri had only mild effects on the transcriptome but modified the gene expression induced by E. coli. The transcriptome differentiates mice inoculated orally with E. coli, Lact. gasseri and combinations of these two strains. These results suggest that the blood cell transcriptome can be used as a source of biomarkers to monitor the impact of probiotics in subclinical models of infectious disease. © 2014 The Society for Applied Microbiology.

Evaluation the virulence of Mycobacterium bovis isolated from milk samples through histopathological study in laboratory animals.

PubMed

Al-Saqur, I M; Al-Thwani, A N; Al-Attar, I M; Al-Mashhadani, M S

2016-12-01

Mycobacterium bovis has a broad host range, and it is the principal agent responsible for tuberculosis (TB) in bovine, domestic and wild mammals. M. bovis also infects human, causing zoonotic TB through ingestion, inhalation and, less frequently by contact with mucous membranes and broken skin. Zoonotic TB was formerly an endemic disease, usually transmitted to man by consumption of raw cow's milk. It is indistinguishable clinically or pathologically from TB caused by M. tuberculosis. The aims of this study were, to isolate and identified M. bovis from raw milk samples by different methods, and evaluate the virulence of M. bovis in laboratory animals (Rabbit). To conduct the study, ninety three cow's milk samples were collected from farms around Baghdad governorate. The decontamination of milk samples was firstly carried out, then samples were subjected to routine tests which include, direct smear for Ziehl Neelsen acid fast stain, culture, each sample was cultured on Lowenstein Jensen media with Sodium pyruvite (All cultures incubated on 37°C for 4-10weeks with continuous observation), and biochemical testes as Nitrate reduction test, Niacin paper strip test and pyrazinamidase test, were employed to diagnose and identified the bacteria. Beside molecular assay was used to confirm the identification of the isolates by Polymerase Chain Reaction (PCR) using specific primers for M. bovis. The virulence of these isolates were investigated through inoculate it in group of laboratory animals consist of 8 rabbit in addition to other group of 4 animals as control (inoculate with Phosphate Buffer Saline). The animals were scarified after 6weeks of inoculation, post- mortem examination was carried out, smears were taken from lesions, and tissue samples were collected from lymph nodes and different organs. The results revealed five isolates of M. bovis in direct smear by acid fast Ziehl-Neelsen stain, while eight isolates observed by culture, the colonies appeared with characteristic feature of cream color, rough, and with irregular edge. The molecular assay using PCR technique confirmed the diagnosis of eight positive isolates in smears and culture. The virulence of these isolates were investigated through the pathological effects appeared in inoculated rabbit which showed lesions scattered mainly in lymph nodes and different organs as lung, liver, spleen and kidney when compared with control group which were naive. Beside the infiltration of mononuclear cells in the internal organs particularly in the lungs. The result of histopathological examination clarified the virulence of M. bovis isolates, and its impact on tissue and organs of the rabbit. Our study conclude the presence of M. bovis isolates in milk in high percentage pause important source of tuberculosis infection for human being. Copyright © 2016.

Rapid CD4(+) T-cell loss induced by human immunodeficiency virus type 1(NC) in uninfected and previously infected chimpanzees.

PubMed

Novembre, F J; de Rosayro, J; Nidtha, S; O'Neil, S P; Gibson, T R; Evans-Strickfaden, T; Hart, C E; McClure, H M

2001-02-01

To investigate the pathogenicity of a virus originating in a chimpanzee with AIDS (C499), two chimpanzees were inoculated with a plasma-derived isolate termed human immunodeficiency virus type 1(NC) (HIV-1(NC)). A previously uninfected chimpanzee, C534, experienced rapid peripheral CD4(+) T-cell loss to fewer than 26 cells/microl by 14 weeks after infection. CD4(+) T-cell depletion was associated with high plasma HIV-1 loads but a low virus burden in the peripheral lymph node. The second chimpanzee, C459, infected 13 years previously with HIV-1(LAV), experienced a more protracted course of peripheral CD4(+) T-cell loss after HIV-1(NC) inoculation, resulting in fewer than 200 cells/microl by 96 weeks postinoculation. The quantities of viral RNA in the plasma and peripheral lymph node from C459 were below the lower limits of detection prior to inoculation with HIV-1(NC) but were significantly and persistently increased after superinfection, with HIV-1(NC) representing the predominant viral genotype. These results show that viruses derived from C499 are more pathogenic for chimpanzees than any other HIV-1 isolates described to date.

Exploring virulence and immunogenicity in the emerging pathogen Sporothrix brasiliensis

PubMed Central

Della Terra, Paula Portella; Fernandes, Geisa Ferreira; Nishikaku, Angela Satie; Burger, Eva; de Camargo, Zoilo Pires

2017-01-01

Sporotrichosis is a polymorphic chronic infection of humans and animals classically acquired after traumatic inoculation with soil and plant material contaminated with Sporothrix spp. propagules. An alternative and successful route of transmission is bites and scratches from diseased cats, through which Sporothrix yeasts are inoculated into mammalian tissue. The development of a murine model of subcutaneous sporotrichosis mimicking the alternative route of transmission is essential to understanding disease pathogenesis and the development of novel therapeutic strategies. To explore the impact of horizontal transmission in animals (e.g., cat-cat) and zoonotic transmission on Sporothrix fitness, the left hind footpads of BALB/c mice were inoculated with 5×106 yeasts (n = 11 S. brasiliensis, n = 2 S. schenckii, or n = 1 S. globosa). Twenty days post-infection, our model reproduced both the pathophysiology and symptomology of sporotrichosis with suppurating subcutaneous nodules that progressed proximally along lymphatic channels. Across the main pathogenic members of the S. schenckii clade, S. brasiliensis was usually more virulent than S. schenckii and S. globosa. However, the virulence in S. brasiliensis was strain-dependent, and we demonstrated that highly virulent isolates disseminate from the left hind footpad to the liver, spleen, kidneys, lungs, heart, and brain of infected animals, inducing significant and chronic weight loss (losing up to 15% of their body weight). The weight loss correlated with host death between 2 and 16 weeks post-infection. Histopathological features included necrosis, suppurative inflammation, and polymorphonuclear and mononuclear inflammatory infiltrates. Immunoblot using specific antisera and homologous exoantigen investigated the humoral response. Antigenic profiles were isolate-specific, supporting the hypothesis that different Sporothrix species can elicit a heterogeneous humoral response over time, but cross reaction was observed between S. brasiliensis and S. schenckii proteomes. Despite great diversity in the immunoblot profiles, antibodies were mainly derived against 3-carboxymuconate cyclase, a glycoprotein oscillating between 60 and 70 kDa (gp60-gp70) and a 100-kDa molecule in nearly 100% of the assays. Thus, our data broaden the current view of virulence and immunogenicity in the Sporothrix-sporotrichosis system, substantially expanding the possibilities for comparative genomic with isolates bearing divergent virulence traits and helping uncover the molecular mechanisms and evolutionary pressures underpinning the emergence of Sporothrix virulence. PMID:28854184

Exploring virulence and immunogenicity in the emerging pathogen Sporothrix brasiliensis.

PubMed

Della Terra, Paula Portella; Rodrigues, Anderson Messias; Fernandes, Geisa Ferreira; Nishikaku, Angela Satie; Burger, Eva; de Camargo, Zoilo Pires

2017-08-01

Sporotrichosis is a polymorphic chronic infection of humans and animals classically acquired after traumatic inoculation with soil and plant material contaminated with Sporothrix spp. propagules. An alternative and successful route of transmission is bites and scratches from diseased cats, through which Sporothrix yeasts are inoculated into mammalian tissue. The development of a murine model of subcutaneous sporotrichosis mimicking the alternative route of transmission is essential to understanding disease pathogenesis and the development of novel therapeutic strategies. To explore the impact of horizontal transmission in animals (e.g., cat-cat) and zoonotic transmission on Sporothrix fitness, the left hind footpads of BALB/c mice were inoculated with 5×106 yeasts (n = 11 S. brasiliensis, n = 2 S. schenckii, or n = 1 S. globosa). Twenty days post-infection, our model reproduced both the pathophysiology and symptomology of sporotrichosis with suppurating subcutaneous nodules that progressed proximally along lymphatic channels. Across the main pathogenic members of the S. schenckii clade, S. brasiliensis was usually more virulent than S. schenckii and S. globosa. However, the virulence in S. brasiliensis was strain-dependent, and we demonstrated that highly virulent isolates disseminate from the left hind footpad to the liver, spleen, kidneys, lungs, heart, and brain of infected animals, inducing significant and chronic weight loss (losing up to 15% of their body weight). The weight loss correlated with host death between 2 and 16 weeks post-infection. Histopathological features included necrosis, suppurative inflammation, and polymorphonuclear and mononuclear inflammatory infiltrates. Immunoblot using specific antisera and homologous exoantigen investigated the humoral response. Antigenic profiles were isolate-specific, supporting the hypothesis that different Sporothrix species can elicit a heterogeneous humoral response over time, but cross reaction was observed between S. brasiliensis and S. schenckii proteomes. Despite great diversity in the immunoblot profiles, antibodies were mainly derived against 3-carboxymuconate cyclase, a glycoprotein oscillating between 60 and 70 kDa (gp60-gp70) and a 100-kDa molecule in nearly 100% of the assays. Thus, our data broaden the current view of virulence and immunogenicity in the Sporothrix-sporotrichosis system, substantially expanding the possibilities for comparative genomic with isolates bearing divergent virulence traits and helping uncover the molecular mechanisms and evolutionary pressures underpinning the emergence of Sporothrix virulence.

Transfer of adult Strongylus vulgaris via stomach tube.

PubMed

Hofing, G L; Bennett, D G

1983-10-01

Patent infections with Strongylus vulgaris were established in 6 of 8 helminth-free ponies given 41 to 101 adult worms via nasogastric tube. The parasites were removed from the cecum and ventral colon and transferred within 1 to 2 hours of the death of the donor horses. Eggs were found in the feces of the recipients in 2 or 3 days; egg counts reached maximum, 28 eggs per gram of feces, at 4 weeks after ponies were inoculated. In 6 ponies euthanatized 3 to 7 weeks after parasitic transfers were done, 28% of the inoculated worms were found alive at necropsy. A 7th pony was maintained as a donor for establishing infections for chemotherapy trials and, although never passing more than 6 eggs per gram of feces, shed infective larvae over a period of 2 years.

Decreased egg production in laying hens associated with infection with genotype 3 avian hepatitis E virus strain from China.

PubMed

Zhao, Qin; Liu, Baoyuan; Sun, Yani; Du, Taofeng; Chen, Yiyang; Wang, Xinjie; Li, Huixia; Nan, Yuchen; Zhang, Gaiping; Zhou, En-Min

2017-05-01

To determine the relationship between decreased egg production and avian HEV infection, thirty healthy 23-week-old Hy-Line Variety Brown layer hens were randomly divided into 3 groups with 10 hens per group. Next, a genotype 3 avian HEV strain from China was used to inoculate laying hens via oronasal or intravenous routes using a 50% chicken infectious dose of 500. All hens were necropsied at 14 weeks postinoculation (wpi). Fecal virus shedding, viremia, seroconversion, serum alanine aminotransferase (ALT) increases and liver lesions showed that after intravenous (i.v.) and oronasal inoculation, the laying hens were successfully infected. Compared with the uninoculated group, the i.v. and oronasally inoculated groups exhibited egg production decreases at 1wpi and 2wpi, reaching peak production at 3wpi and 8wpi, respectively. In both groups, decreased production was evident for 12 weeks and overall decreases ranged from 10% to 30%. In addition, in the 7 field layer farms exhibiting decreased egg production, vaccination regimens had been completed against Newcastle disease, infectious bronchitis, avian influenza H9N2 and H5N1 and egg drop syndrome virus. However, circulating avian HEV was confirmed on these farms using tests to detect avian HEV IgG antibodies and RNA. Therefore, the experimental and field data indicate that avian HEV infection acting alone could account for observed decreases in egg production in laying hens. Copyright © 2017 Elsevier B.V. All rights reserved.

Plasmodium falciparum incidence relative to entomologic inoculation rates at a site proposed for testing malaria vaccines in western Kenya.

PubMed

Beier, J C; Oster, C N; Onyango, F K; Bales, J D; Sherwood, J A; Perkins, P V; Chumo, D K; Koech, D V; Whitmire, R E; Roberts, C R

1994-05-01

Relationships between Plasmodium falciparum incidence and entomologic inoculation rates (EIRs) were determined for a 21-month period in Saradidi, western Kenya, in preparation for malaria vaccine field trials. Children, ranging in age from six months to six years and treated to clear malaria parasites, were monitored daily for up to 12 weeks to detect new malaria infections. Overall, new P. falciparum infections were detected in 77% of 809 children. The percentage of children that developed infections per two-week period averaged 34.7%, ranging from 7.3% to 90.9%. Transmission by vector populations was detected in 86.4% (38 of 44) of the two-week periods, with daily EIRs averaging 0.75 infective bites per person. Periods of intense transmission during April to August, and from November to January, coincided with seasonal rains. Relationships between daily malaria attack rates and EIRs indicated that an average of only 7.5% (1 in 13) of the sporozoite inoculations produced new infections in children. Regression analysis demonstrated that EIRs accounted for 74% of the variation in attack rates. One of the components of the EIR, the human-biting rate, alone accounted for 68% of the variation in attack rates. Thus, measurements of either the EIR or the human-biting rate can be used to predict corresponding attack rates in children. These baseline epidemiologic studies indicate that the intense transmission patterns of P. falciparum in Saradidi will provide excellent conditions for evaluating malaria vaccine efficacy.

Survival of Salmonella enterica serovar infantis on and within stored table eggs.

PubMed

Lublin, Avishai; Maler, Ilana; Mechani, Sara; Pinto, Riky; Sela-Saldinger, Shlomo

2015-02-01

Contaminated table eggs are considered a primary source of foodborne salmonellosis globally. Recently, a single clone of Salmonella enterica serovar Infantis emerged in Israel and became the predominant serovar isolated in poultry. This clone is currently the most prevalent strain in poultry and is the leading cause of salmonellosis in humans. Because little is known regarding the potential transmission of this strain from contaminated eggs to humans, the objective of this study was to evaluate the ability of Salmonella Infantis to survive on the eggshell or within the egg during cold storage or at room temperature. Salmonella cells (5.7 log CFU per egg) were inoculated on the surface of 120 intact eggs or injected into the egg yolk (3.7 log CFU per egg) of another 120 eggs. Half of the eggs were stored at 5.5 ± 0.3°C and half at room temperature (25.5 ± 0.1°C) for up to 10 weeks. At both temperatures, the number of Salmonella cells on the shell declined by 2 log up to 4 weeks and remained constant thereafter. Yolk-inoculated Salmonella counts at cold storage declined by 1 log up to 4 weeks and remained constant, while room-temperature storage supported the growth of the pathogen to a level of 8 log CFU/ml of total egg content, as early as 4 weeks postinoculation. Examination of egg content following surface inoculation revealed the presence of Salmonella in a portion of the eggs at both temperatures up to 10 weeks, suggesting that this strain can also penetrate through the shell and survive within the egg. These findings imply that Salmonella enterica serovar Infantis is capable of survival both on the exterior and interior of table eggs and even multiply inside the egg at room temperature. Our findings support the need for prompt refrigeration to prevent Salmonella multiplication during storage of eggs at room temperature.

Investigation of a cutaneous delayed hypersensitivity response as a means of detecting Salmonella dublin infection in cattle.

PubMed

Aitken, M M; Hall, G A; Jones, P W

1978-05-01

Delayed hypersensitivity reactions developed 48 to 96 h after intradermal injection of killed Salmonella dublin in 25 of 28 cattle which had been inoculated intravenously, and in five of 10 cattle which had been inoculated orally with S dublin 24 to 493 days previously. Control animals showed no delayed hypersensitivity reactions. Persistence of infection in five of the intravenously inoculated and in four of the orally inoculated animals was confirmed by isolation of S dublin from the carcases at necropsy one week after skin testing. Failure to isolate the organism from the carcases of 21 animals which had reacted positively to the intradermal test did not eliminate the possibility of their being carriers of S dublin. Skin testing was concluded to be a reliable means of identifying animals which had been, and possibly still were, infected systemically with S dublin. However recovered animals might be falsely identified as infected. Repeated testing gave misleading results.

Colonization of onions by endophytic fungi and their impacts on the biology of Thrips tabaci.

PubMed

Muvea, Alexander M; Meyhöfer, Rainer; Subramanian, Sevgan; Poehling, Hans-Michael; Ekesi, Sunday; Maniania, Nguya K

2014-01-01

Endophytic fungi, which live within host plant tissues without causing any visible symptom of infection, are important mutualists that mediate plant-herbivore interactions. Thrips tabaci (Lindeman) is one of the key pests of onion, Allium cepa L., an economically important agricultural crop cultivated worldwide. However, information on endophyte colonization of onions, and their impacts on the biology of thrips feeding on them, is lacking. We tested the colonization of onion plants by selected fungal endophyte isolates using two inoculation methods. The effects of inoculated endophytes on T. tabaci infesting onion were also examined. Seven fungal endophytes used in our study were able to colonize onion plants either by the seed or seedling inoculation methods. Seed inoculation resulted in 1.47 times higher mean percentage post-inoculation recovery of all the endophytes tested as compared to seedling inoculation. Fewer thrips were observed on plants inoculated with Clonostachys rosea ICIPE 707, Trichoderma asperellum M2RT4, Trichoderma atroviride ICIPE 710, Trichoderma harzianum 709, Hypocrea lixii F3ST1 and Fusarium sp. ICIPE 712 isolates as compared to those inoculated with Fusarium sp. ICIPE 717 and the control treatments. Onion plants colonized by C. rosea ICIPE 707, T. asperellum M2RT4, T. atroviride ICIPE 710 and H. lixii F3ST1 had significantly lower feeding punctures as compared to the other treatments. Among the isolates tested, the lowest numbers of eggs were laid by T. tabaci on H. lixii F3ST1 and C. rosea ICIPE 707 inoculated plants. These results extend the knowledge on colonization of onions by fungal endophytes and their effects on Thrips tabaci.

Colonization of Onions by Endophytic Fungi and Their Impacts on the Biology of Thrips tabaci

PubMed Central

Muvea, Alexander M.; Meyhöfer, Rainer; Subramanian, Sevgan; Poehling, Hans-Michael; Ekesi, Sunday; Maniania, Nguya K.

2014-01-01

Endophytic fungi, which live within host plant tissues without causing any visible symptom of infection, are important mutualists that mediate plant–herbivore interactions. Thrips tabaci (Lindeman) is one of the key pests of onion, Allium cepa L., an economically important agricultural crop cultivated worldwide. However, information on endophyte colonization of onions, and their impacts on the biology of thrips feeding on them, is lacking. We tested the colonization of onion plants by selected fungal endophyte isolates using two inoculation methods. The effects of inoculated endophytes on T. tabaci infesting onion were also examined. Seven fungal endophytes used in our study were able to colonize onion plants either by the seed or seedling inoculation methods. Seed inoculation resulted in 1.47 times higher mean percentage post-inoculation recovery of all the endophytes tested as compared to seedling inoculation. Fewer thrips were observed on plants inoculated with Clonostachys rosea ICIPE 707, Trichoderma asperellum M2RT4, Trichoderma atroviride ICIPE 710, Trichoderma harzianum 709, Hypocrea lixii F3ST1 and Fusarium sp. ICIPE 712 isolates as compared to those inoculated with Fusarium sp. ICIPE 717 and the control treatments. Onion plants colonized by C. rosea ICIPE 707, T. asperellum M2RT4, T. atroviride ICIPE 710 and H. lixii F3ST1 had significantly lower feeding punctures as compared to the other treatments. Among the isolates tested, the lowest numbers of eggs were laid by T. tabaci on H. lixii F3ST1 and C. rosea ICIPE 707 inoculated plants. These results extend the knowledge on colonization of onions by fungal endophytes and their effects on Thrips tabaci. PMID:25254657

Susceptibility of Pigs to Zoonotic Hepatitis E Virus Genotype 3 Isolated from a Wild Boar.

PubMed

Thiry, D; Rose, N; Mauroy, A; Paboeuf, F; Dams, L; Roels, S; Pavio, N; Thiry, E

2017-10-01

In Europe, zoonotic hepatitis E virus (HEV) genotype 3 strains mainly circulate in humans, swine and wild boar. The aim of this study was to investigate the potential transmission of a wild boar originating HEV strain (WbHEV) to swine by intravenous or oral inoculation and to study the consequences of infection of a WbHEV strain, a WbHEV strain previously passaged in a pig and a swine HEV strain after oral inoculation. Firstly, an intravenous infection was performed for which five piglets were divided into two groups with three pigs inoculated with a WbHEV field strain and two pigs inoculated with a HEV-negative swine liver homogenate. All pigs were necropsied 8, 9 and 10 days post-inoculation. Secondly, an oral infection of 56 days was performed on 12 piglets divided into four groups inoculated with a WbHEV strain, a WbHEV strain previously passaged in swine, a swine HEV strain or a HEV-negative swine liver homogenate. After intravenous inoculation, HEV RNA was detected in serum, bile, liver, spleen, duodenum, jejunum, colon, lung, gastro-hepatic lymph nodes and faeces in all infected piglets. After oral inoculation, HEV RNA was detected in serum, bile, liver, gastro-hepatic lymph nodes and faeces. Most of HEV-inoculated pigs became seropositive at day 15. This study provides experimental evidence of early viral spread throughout the organism after intravenous infection with a WbHEV strain and supports the notion that such a zoonotic strain could be transmitted via the natural faecal-oral route of infection between wild boar and pigs but also between pigs. © 2016 Blackwell Verlag GmbH.

Effect of age of cook-in-bag delicatessen meats formulated with lactate-diacetate on the behavior of Listeria monocytogenes contamination introduced when opening the packages during storage.

PubMed

Geornaras, Ifigenia; Toczko, Darren; Sofos, John N

2013-07-01

This study evaluated the potential effect of age of cook-in-bag ham and turkey breast delicatessen meats formulated with lactate-diacetate on survival and/or growth of Listeria monocytogenes introduced after opening of packages and slicing of product. Commercially prepared cured ham and turkey breast products formulated with potassium lactate and sodium diacetate were stored at 1.7°C unsliced, in their original cook-in-bags, and without postlethality exposure. On days 5, 90, 120, and 180 of storage, product slices (10.2 by 7.6 cm) were surface inoculated (1 to 2 log CFU/cm²) with a 10-strain mixture of L. monocytogenes, vacuum packaged (seven slices per bag), and stored at 4°C for up to 13 weeks. Inoculated levels of L. monocytogenes on both products were 1.4 to 1.5 log CFU/cm². Irrespective of product age at slicing and inoculation, after 13 weeks of vacuum-packaged storage (4°C), pathogen counts on product slices were 1.5 to 2.3 (ham) and 2.3 to 2.5 (turkey) log CFU/cm². Overall, the results of the study showed that the age of the cook-in-bag products prior to slicing and inoculation with the pathogen did not (P ≥ 0.05) affect the behavior of L. monocytogenes during vacuum-packaged storage (4°C, up to 13 weeks) of ham and turkey slices. Mean counts of lactic acid bacteria and yeasts and molds, when detected, did not exceed approximately 1 and 2 log CFU/cm², respectively, among all stored samples. Findings of the study will be useful to the meat industry and risk assessors in their efforts to control L. monocytogenes in ready-to-eat meat products.

Wheat differential gene expression induced by different races of Puccinia triticina.

PubMed

Neugebauer, Kerri A; Bruce, Myron; Todd, Tim; Trick, Harold N; Fellers, John P

2018-01-01

Puccinia triticina, the causal agent of wheat leaf rust, causes significant losses in wheat yield and quality each year worldwide. During leaf rust infection, the host plant recognizes numerous molecules, some of which trigger host defenses. Although P. triticina reproduces clonally, there is still variation within the population due to a high mutation frequency, host specificity, and environmental adaptation. This study explores how wheat responds on a gene expression level to different P. triticina races. Six P. triticina races were inoculated onto a susceptible wheat variety and samples were taken at six days post inoculation, just prior to pustule eruption. RNA sequence data identified 63 wheat genes differentially expressed between the six races. A time course, conducted over the first seven days post inoculation, was used to examine the expression pattern of 63 genes during infection. Forty-seven wheat genes were verified to have differential expression. Three common expression patterns were identified. In addition, two genes were associated with race specific gene expression. Differential expression of an ER molecular chaperone gene was associated with races from two different P. triticina lineages. Also, differential expression in an alanine glyoxylate aminotransferase gene was associated with races with virulence shifts for leaf rust resistance genes.

Rhizobial symbiosis effect on the growth, metal uptake, and antioxidant responses of Medicago lupulina under copper stress.

PubMed

Kong, Zhaoyu; Mohamad, Osama Abdalla; Deng, Zhenshan; Liu, Xiaodong; Glick, Bernard R; Wei, Gehong

2015-08-01

The effects of rhizobial symbiosis on the growth, metal uptake, and antioxidant responses of Medicago lupulina in the presence of 200 mg kg(-1) Cu(2+) throughout different stages of symbiosis development were studied. The symbiosis with Sinorhizobium meliloti CCNWSX0020 induced an increase in plant growth and nitrogen content irrespective of the presence of Cu(2+). The total amount of Cu uptake of inoculated plants significantly increased by 34.0 and 120.4% in shoots and roots, respectively, compared with non-inoculated plants. However, although the rhizobial symbiosis promoted Cu accumulation both in shoots and roots, the increase in roots was much higher than in shoots, thus decreasing the translocation factor and helping Cu phytostabilization. The rate of lipid peroxidation was significantly decreased in both shoots and roots of inoculated vs. non-inoculated plants when measured either 8, 13, or 18 days post-inoculation. In comparison with non-inoculated plants, the activities of superoxide dismutase and ascorbate peroxidase of shoots of inoculated plants exposed to excess Cu were significantly elevated at different stages of symbiosis development; similar increases occurred in the activities of superoxide dismutase, catalase, and glutathione reductase of inoculated roots. The symbiosis with S. meliloti CCNWSX0020 also upregulated the corresponding genes involved in antioxidant responses in the plants treated with excess Cu. The results indicated that the rhizobial symbiosis with S. meliloti CCNWSX0020 not only enhanced plant growth and metal uptake but also improved the responses of plant antioxidant defense to excess Cu stress.

Effect of enrofloxacin on Haemophilus parasuis infection, disease and immune response.

PubMed

Macedo, Nubia; Cheeran, Maxim C J; Rovira, Albert; Holtcamp, Andrew; Torremorell, Montserrat

2017-02-01

Haemophilus parasuis, the causative agent of Glasser's disease, is a pathogen that colonizes the upper respiratory tract (URT) of pigs, invades the bloodstream and causes polyserositis. Because antimicrobials are highly effective against H. parasuis, we hypothesized that they could have a detrimental effect on the establishment of an immune response if given at the time of URT colonization. In this study, we characterized clinical outcomes and antibody and IFN-γ responses to H. parasuis in pigs treated with enrofloxacin before or after low dose inoculation with a pathogenic H. parasuis strain. Pigs that were only inoculated with the agent (EXP group) and pigs that were treated with enrofloxacin and then inoculated (ABT/EXP group) developed signs of disease starting at 4days post inoculation (DPI), presented a significant increase in serum IgG and were protected against a subsequent homologous challenge. In contrast, pigs treated with antibiotic after inoculation (EXP/ABT group) neither showed signs of disease nor seroconverted (IgG) after low dose inoculation. EXP/ABT pigs as well as naïve control pigs [enrofloxacin only (ABT) and challenge only (CHA)] were susceptible to challenge. Variable levels of antibodies in bronchioalveolar fluid and IFN-γ in peripheral blood mononuclear cells were observed after H. parasuis inoculation, but were not associated with protection. In summary, only pigs treated before low dose H. parasuis inoculation seroconverted and were protected against subsequent challenge. Results from this study can help determine timing of antimicrobial use and contribute to our current understanding of judicious antibiotic use. Copyright © 2016 Elsevier B.V. All rights reserved.

Enhancement of Thiamine Biosynthesis in Oil Palm Seedlings by Colonization of Endophytic Fungus Hendersonia toruloidea

PubMed Central

Kamarudin, Amirah N.; Lai, Kok S.; Lamasudin, Dhilia U.; Idris, Abu S.; Balia Yusof, Zetty N.

2017-01-01

Thiamine, or vitamin B1 plays an indispensable role as a cofactor in crucial metabolic reactions including glycolysis, pentose phosphate pathway and the tricarboxylic acid cycle in all living organisms. Thiamine has been shown to play a role in plant adaptation toward biotic and abiotic stresses. The modulation of thiamine biosynthetic genes in oil palm seedlings was evaluated in response to root colonization by endophytic Hendersonia toruloidea. Seven-month-old oil palm seedlings were inoculated with H. toruloidea and microscopic analyses were performed to visualize the localization of endophytic H. toruloidea in oil palm roots. Transmission electron microscopy confirmed that H. toruloidea colonized cortical cells. The expression of thiamine biosynthetic genes and accumulation of total thiamine in oil palm seedlings were also evaluated. Quantitative real-time PCR was performed to measure transcript abundances of four key thiamine biosynthesis genes (THI4, THIC, TH1, and TPK) on days 1, 7, 15, and 30 in response to H. toruloidea colonization. The results showed an increase of up to 12-fold in the expression of all gene transcripts on day 1 post-inoculation. On days 7, 15, and 30 post-inoculation, the relative expression levels of these genes were shown to be downregulated. Thiamine accumulation was observed on day 7 post-colonization and subsequently decreased until day 30. This work provides the first evidence for the enhancement of thiamine biosynthesis by endophytic colonization in oil palm seedlings. PMID:29089959

Enhancement of Thiamine Biosynthesis in Oil Palm Seedlings by Colonization of Endophytic Fungus Hendersonia toruloidea.

PubMed

Kamarudin, Amirah N; Lai, Kok S; Lamasudin, Dhilia U; Idris, Abu S; Balia Yusof, Zetty N

2017-01-01

Thiamine, or vitamin B1 plays an indispensable role as a cofactor in crucial metabolic reactions including glycolysis, pentose phosphate pathway and the tricarboxylic acid cycle in all living organisms. Thiamine has been shown to play a role in plant adaptation toward biotic and abiotic stresses. The modulation of thiamine biosynthetic genes in oil palm seedlings was evaluated in response to root colonization by endophytic Hendersonia toruloidea . Seven-month-old oil palm seedlings were inoculated with H. toruloidea and microscopic analyses were performed to visualize the localization of endophytic H. toruloidea in oil palm roots. Transmission electron microscopy confirmed that H. toruloidea colonized cortical cells. The expression of thiamine biosynthetic genes and accumulation of total thiamine in oil palm seedlings were also evaluated. Quantitative real-time PCR was performed to measure transcript abundances of four key thiamine biosynthesis genes ( THI4 , THIC , TH1 , and TPK ) on days 1, 7, 15, and 30 in response to H. toruloidea colonization. The results showed an increase of up to 12-fold in the expression of all gene transcripts on day 1 post-inoculation. On days 7, 15, and 30 post-inoculation, the relative expression levels of these genes were shown to be downregulated. Thiamine accumulation was observed on day 7 post-colonization and subsequently decreased until day 30. This work provides the first evidence for the enhancement of thiamine biosynthesis by endophytic colonization in oil palm seedlings.

Percutaneous Implants with Porous Titanium Dermal Barriers: An In Vivo Evaluation of Infection Risk

PubMed Central

Isackson, Dorthyann; McGill, Lawrence D.; Bachus, Kent N.

2010-01-01

Osseointegrated percutaneous implants are a promising prosthetic alternative for a subset of amputees. However, as with all percutaneous implants, they have an increased risk of infection since they breach the skin barrier. Theoretically, host tissues could attach to the metal implant creating a barrier to infection. When compared with smooth surfaces, it is hypothesized that porous surfaces improve the attachment of the host tissues to the implant, and decrease the infection risk. In this study, 4 titanium implants, manufactured with a percutaneous post and a subcutaneous disk, were placed subcutaneously on the dorsum of eight New Zealand White rabbits. Beginning at four weeks post-op, the implants were inoculated weekly with 108 CFU Staphylococcus aureus until signs of clinical infection presented. While we were unable to detect a difference in the incidence of infection of the porous metal implants, smooth surface (no porous coating) percutaneous and subcutaneous components had a 7-fold increased risk of infection compared to the implants with a porous coating on one or both components. The porous coated implants displayed excellent tissue ingrowth into the porous structures; whereas, the smooth implants were surrounded with a thick, organized fibrotic capsule that was separated from the implant surface. This study suggests that porous coated metal percutaneous implants are at a significantly lower risk of infection when compared to smooth metal implants. The smooth surface percutaneous implants were inadequate in allowing a long-term seal to develop with the soft tissue, thus increasing vulnerability to the migration of infecting microorganisms. PMID:21145778

Improved hatchability and efficient protection after in ovo vaccination with live-attenuated H7N2 and H9N2 avian influenza viruses

PubMed Central

2011-01-01

Mass in ovo vaccination with live attenuated viruses is widely used in the poultry industry to protect against various infectious diseases. The worldwide outbreaks of low pathogenic and highly pathogenic avian influenza highlight the pressing need for the development of similar mass vaccination strategies against avian influenza viruses. We have previously shown that a genetically modified live attenuated avian influenza virus (LAIV) was amenable for in ovo vaccination and provided optimal protection against H5 HPAI viruses. However, in ovo vaccination against other subtypes resulted in poor hatchability and, therefore, seemed impractical. In this study, we modified the H7 and H9 hemagglutinin (HA) proteins by substituting the amino acids at the cleavage site for those found in the H6 HA subtype. We found that with this modification, a single dose in ovo vaccination of 18-day old eggs provided complete protection against homologous challenge with low pathogenic virus in ≥70% of chickens at 2 or 6 weeks post-hatching. Further, inoculation of 19-day old egg embryos with 106 EID50 of LAIVs improved hatchability to ≥90% (equivalent to unvaccinated controls) with similar levels of protection. Our findings indicate that the strategy of modifying the HA cleavage site combined with the LAIV backbone could be used for in ovo vaccination against avian influenza. Importantly, with protection conferred as early as 2 weeks post-hatching, with this strategy birds would be protected prior to or at the time of delivery to a farm or commercial operation. PMID:21255403

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

PubMed

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

2011-01-01

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

Propagation of an Avirulent Turkey Hemorrhagic Enteritis Virus Isolate in Chickens.

PubMed

Gerber, Priscilla F; Hossain, Mohammad F; Reynolds, Paul; Hoang, Phuong; Burgess, Susan K; Renz, Katrin; McMillan, Mary; Katz, Margaret E; Walkden-Brown, Stephen W

2018-03-01

A series of studies were undertaken to optimize the propagation of hemorrhagic enteritis virus (HEV) in specific-pathogen-free (SPF) chickens. A total of 562 SPF chickens were orally inoculated with an Australian avirulent HEV isolate of turkey origin at 9, 14, 21, or 28 days of age with 5, 6, 7, or 8 log  10 genomic copies (GC), while 102 chickens served as uninfected controls. No clinical signs were observed in infected chickens. There was an inoculum-dose-dependent increase in the relative spleen and liver weight ( P < 0.01). Relative spleen weight 7 days post-HEV inoculation was up to 85% higher in chickens that were inoculated with 6 to 7 GC compared with controls, with no further increase at higher doses. Relative liver weight increased up to 14% in chickens inoculated with 6 GC, with no further increase. Birds inoculated with a 7 GC dose had a higher frequency of HEV DNA-positive birds (77% to 86%) than birds inoculated with lower doses (33% to 59%; P < 0.01). The most efficient dose for live passage propagation was 7 GC inoculated in 9-to-14-day-old birds, yielding an infection rate of 81%. Livers and spleens from infected birds at all doses were processed to produce a putative vaccine with a final GC recovery in the vaccine material of 8.6 GC/bird. In summary, HEV of turkey origin can be readily propagated in SPF chickens, and conditions to maximize viral retrieval were established.

Crimean-Congo haemorrhagic fever virus replication in adult Hyalomma truncatum and Amblyomma variegatum ticks.

PubMed

Gonzalez, J P; Cornet, J P; Wilson, M L; Camicas, J L

1991-01-01

The kinetics of the replication of the Crimean-Congo haemorrhagic fever virus (CCHFV) was studied in intra-anally inoculated adult Hyalomma truncatum and Amblyomma variegatum ticks. The virus was re-isolated by suckling mouse inoculation and revealed by antigen capture with ground ticks and indirect immunofluorescence of haemolymph. The virus was detected in ticks in the first hours post-inoculation (p.i.) and its replication was observed from 36 h p.i. onwards. Virus titre reached a maximum within 3-5 days then decreased slowly to a level of at 2 log LD50/ml for several months until the end of observations. Several specific, non-identified factors seem to favour CCHFV replication in H. truncatum. Long-term virus persistence seems to occur in CCHFV-infected adult ticks.

Phenological and phytochemical changes correlate with differential interactions of Verticillium dahliae with broccoli and cauliflower.

PubMed

Njoroge, S M C; Vallad, G E; Park, S-Y; Kang, S; Koike, S T; Bolda, M; Burman, P; Polonik, W; Subbarao, K V

2011-05-01

Cauliflower (Brassica oleracea var. botrytis subvar. cauliflora) is susceptible to wilt caused by Verticillium dahliae but broccoli (B. oleracea var. italica subvar. cyamosa) is not. Infection of broccoli and cauliflower by a green fluorescent protein-expressing isolate of V. dahliae was examined using epifluorescence and confocal laser-scanning microscopy to follow infection and colonization in relation to plant phenology. Plant glucosinolate, phenolic, and lignin contents were also assayed at 0, 4, 14, and 28 days postinoculation. V. dahliae consistently infected and colonized the vascular tissues of all cauliflower plants regardless of age at inoculation, with the pathogen ultimately appearing in the developing seed; however, colonization decreased with plant age. In broccoli, V. dahliae infected and colonized root and stem xylem tissues of plants inoculated at 1, 2, or 3 weeks postemergence. However, V. dahliae colonized only the root xylem and the epidermal and cortical tissues of broccoli plants inoculated at 4, 5, and 6 weeks postemergence. The frequency of reisolation of V. dahliae from the stems (4 to 22%) and roots (10 to 40%) of mature broccoli plants was lower than for cauliflower stems (25 to 64%) and roots (31 to 71%). The mean level of aliphatic glucosinolates in broccoli roots was 6.18 times higher than in the shoots and did not vary with age, whereas it was 3.65 times higher in cauliflower shoots than in the roots and there was a proportional increase with age. Indole glucosinolate content was identical in both cauliflower and broccoli, and both indole and aromatic glucosinolates did not vary with plant age in either crop. Qualitative differences in characterized glucosinolates were observed between broccoli and cauliflower but no differences were observed between inoculated and noninoculated plants for either broccoli or cauliflower. However, the phenolic and lignin contents were significantly higher in broccoli following inoculation than in noninoculated broccoli or inoculated cauliflower plants. The increased resistance of broccoli to V. dahliae infection was related to the increase in phenolic and lignin contents. Significant differential accumulation of glucosinolates associated with plant phenology may also contribute to the resistant and susceptible reactions of broccoli and cauliflower, respectively, against V. dahliae.

Uropathogenic E.coli (UPEC) Infection Induces Proliferation through Enhancer of Zeste Homologue 2 (EZH2)

PubMed Central

Penna, Frank; Samiei, Alaleh Najdi; Sidler, Martin; Jiang, Jia-Xin; Ibrahim, Fadi; Tolg, Cornelia; Delgado-Olguin, Paul; Rosenblum, Norman; Bägli, Darius J.

2016-01-01

Host-pathogen interactions can induce epigenetic changes in the host directly, as well as indirectly through secreted factors. Previously, uropathogenic Escherichia coli (UPEC) was shown to increase DNA methyltransferase activity and expression, which was associated with methylation-dependent alterations in the urothelial expression of CDKN2A. Here, we showed that paracrine factors from infected cells alter expression of another epigenetic writer, EZH2, coordinate with proliferation. Urothelial cells were inoculated with UPEC, UPEC derivatives, or vehicle (mock infection) at low moi, washed, then maintained in media with Gentamycin. Urothelial conditioned media (CM) and extracellular vesicles (EV) were isolated after the inoculations and used to treat naïve urothelial cells. EZH2 increased with UPEC infection, inoculation-induced CM, and inoculation-induced EV vs. parallel stimulation derived from mock-inoculated urothelial cells. We found that infection also increased proliferation at one day post-infection, which was blocked by the EZH2 inhibitor UNC1999. Inhibition of demethylation at H3K27me3 had the opposite effect and augmented proliferation. CONCLUSION: Uropathogen-induced paracrine factors act epigenetically by altering expression of EZH2, which plays a key role in early host cell proliferative responses to infection. PMID:26964089

Immunization with Salmonella Enteritidis secreting mucosal adjuvant labile toxin confers protection against wild type challenge via augmentation of CD3+CD4+ T-cell proliferation and enhancement of IFN-γ, IL-6 and IL-10 expressions in chicken.

PubMed

Lalsiamthara, Jonathan; Lee, John Hwa

2017-02-01

The protective efficacy and immunological profiles of chickens immunized with an attenuated Salmonella Enteritidis (SE) constitutively secreting double mutant heat labile enterotoxin (dmLT) were investigated. The dmLT is a detoxified variant of Escherichia coli heat labile toxin and is a potent mucosal adjuvant capable of inducing both humoral and cell-mediated immunity. In this study, four-week-old chickens were inoculated with SE-dmLT strain JOL1641, parental SE strain JOL1087 or phosphate buffered saline control. Peripheral blood mononuclear cells of SE-dmLT inoculated birds showed significant proliferation upon stimulation with SE antigens as compared to the control and JOL1087 groups (P⩽0.05). One week post-challenge, the ratio of CD3 + CD4 + to CD3 + CD8 + T-cells showed a significant increase in the immunized groups. Significant increases in IFN-γ levels were observed in JOL1641 birds immunized via oral and intramuscular routes. While immunizations with the JOL1087 strain via the intramuscular route also induced significant increases in IFN-γ, immunization via the oral route did not trigger significant changes. Pro-inflammatory cytokine IL-6 was also elevated significantly in immunized birds; a significant elevation of IL-10 was observed only in oral immunization with JOL1641 (P⩽0.05). JOL1641 immunized birds showed significant reduction of challenge bacterial-organ recovery as compared to JOL1087 and non-immunized birds. Collectively, our results revealed that immunization with the adjuvant-secreting S. Enteritidis confers protection against wild type SE challenge via induction of strong cell proliferative response, augmentation of CD3 + CD4 + : CD3 + CD8 + T-cells ratio and enhancement of IFN-γ, IL-6 and IL-10 cytokine secretion. Copyright © 2016 Elsevier Ltd. All rights reserved.

Neoplastic transformation of SV40-immortalized human urinary tract epithelial cells by in vitro exposure to 3-methylcholanthrene.

PubMed

Reznikoff, C A; Loretz, L J; Christian, B J; Wu, S Q; Meisner, L F

1988-08-01

Normal human urinary tract epithelial cells (HUC) were neoplastically transformed in vitro using a step-wise strategy. First, a partially transformed non-virus-producing cell line was obtained after infection of HUC with simian virus 40 (SV40). This cell line (SV-HUC-1) was demonstrated to be clonal in origin, as 100% of cells contained at least five of seven marker chromosomes. Marker chromosomes were formed by balanced translocations resulting in a 'pseudodiploid' cell line. SV-HUC-1 showed altered growth properties in vitro (e.g. anchorage independent growth) but failed to form tumors in athymic nude mice, even after 3 years in culture (80 passages). In the studies reported here, SV-HUC-1 at early passages (P15-P19) were exposed to 3-methylcholanthrene (MCA) in three separate experiments. After a six-week post-treatment period of cell culture, cells were inoculated s.c. into athymic nude mice. In all experiments, MCA-treated SV-HUC-1 formed carcinomas in mice usually with a latent period of 5-8 weeks. These carcinomas showed heterogeneity with respect to histopathologies and growth properties in the mice and karyotypes. All the tumors retained SV-HUC-1 chromosome markers, but each independent transformant was aneuploid and contained unique new marker chromosomes. Chromosomes usually altered in tumor cells included numbers 3, 5, 6, 9, 11 and 13. Mutations in the ras family of cellular proto-oncogenes resulting in altered mobility of the p21 protein product were not detected in six cell lines established from independently derived tumors. It is not yet known whether other cellular proto-oncogenes are activated in these tumorigenic transformants. Neither control SV-HUC-1 (which were not exposed to MCA), nor early passage HUC exposed to MCA formed tumors when inoculated into mice. Thus, the tumorigenic transformation of HUC resulted from the combined actions of SV40 and MCA.

Species-specificity of equine and porcine Lawsonia intracellularis isolates in laboratory animals

PubMed Central

Sampieri, Francesca; Vannucci, Fabio A.; Allen, Andrew L.; Pusterla, Nicola; Antonopoulos, Aphroditi J.; Ball, Katherine R.; Thompson, Julie; Dowling, Patricia M.; Hamilton, Don L.; Gebhart, Connie J.

2013-01-01

Lawsonia intracellularis infection causes proliferative enteropathy (PE) in many mammalian species, with porcine and equine proliferative enteropathy (PPE and EPE) known worldwide. Hamsters are a well-published animal model for PPE infection studies in pigs. There is no laboratory animal model for EPE infection studies and it is not known whether there is species-specificity for equine or porcine isolates of L. intracellularis in animal models. The objective of this study was to determine whether it is possible to generate typical EPE lesions in hamsters after inoculation with an equine strain of L. intracellularis (EPE strain) and whether it is comparatively possible to generate PPE lesions in rabbits after inoculation with a porcine strain of L. intracellularis (PPE strain). In 2 separate trials, 4-week-old and 3-week-old weanling golden Syrian hamsters were challenged with EPE strains and compared to uninfected (both trials) and PPE-infected controls (Trial 2 only). Concurrently, 6 female New Zealand white juvenile rabbits were infected with PPE strain and observed concomitantly to 8 similar rabbits infected with EPE strain for a different experiment. Hamsters and rabbits were observed for 21 to 24 days post-infection (DPI), depending on the experiment. Neither infected species developed clinical signs. The presence of disease was assessed with diagnostic techniques classically used for pigs and horses: immune-peroxidase monolayer assay on sera; quantitative polymerase chain reaction (qPCR) detection of molecular DNA in feces; and hematoxylin and eosin (H&E) stain and immunohistochemistry (IHC) on intestinal tissues. Our results showed that EPE-challenged hamsters do not develop infection when compared with PPE controls (IHC, P = 0.009; qPCR, P = 0.0003). Conversely, PPE-challenged rabbits do not develop typical intestinal lesions in comparison to EPE-challenged rabbits, with serological response at 14 DPI being significantly lower (P = 0.0023). In conclusion, PPE and EPE strains appear to have different host-specificities for hamsters and rabbits, respectively. PMID:24124268

Toxicity associated with high dosage 9-[(2R,5R-2,5-dihydro-5-phosphonomethoxy)-2-furanyl]adenine therapy off attempts to abort early FIV infection.

PubMed

Hartmann, K; Ferk, G; North, T W; Pedersen, N C

1997-09-01

9-[(2R,5R-2,5-dihydro-5-phosphonomethoxy)-2-furanyl]adenine, or D4API, was tested in the feline immunodeficiency virus (FIV) infection model and found to be significantly more inhibitory in vitro than its parent compound 9-phosphonylmethoxethyl adenine (PMEA). Cytotoxicity was less than for PMEA or azidothymidine (AZT) for culture periods of 7 days, but more toxic after 10 days. D4API was rapidly absorbed by cats following subcutaneous inoculation, with a plasma half-life of less than 1 h after intravenous inoculation and between 2 and 3 h after subcutaneous injection. Peripheral blood mononuclear cells collected from cats given a single dose of D4API were refractory, however, to FIV infection in vitro for up to 24 h. Given its prolonged intracellular phase and high selectivity index, high dose D4API therapy was tested for its ability to abort an acute (i.e. 2 week) FIV infection. A divided daily dose of D4API, which was one-fourth the toxic dose and 125 times the concentration that would totally inhibit virus replication in vitro, completely abrogated the anticipated viremia and antibody responses. Unfortunately, a majority of treated/uninfected and treated/infected test cats died acutely of drug toxicity after 47 days of treatment. Toxicity in vivo mirrored what was observed in vitro, being precipitous and cumulative in nature. Toxic signs included widespread hepatic and lymphoid necrosis. A surviving treated/FIV infected cat remained healthy to day 175 when the study was terminated; antibodies appeared 2 months later than in untreated/infected cats and virus was only detectable at low levels on day 175. In contrast, untreated/infected cats were viremic and antibody positive from 3 to 4 weeks post-infection onwards. Therefore, it was possible to alter, but not abort, an early FIV infection with prolonged, high-dose D4API treatment.

Pre-infection of pigs with Mycoplasma hyopneumoniae induces oxidative stress that influences outcomes of a subsequent infection with a swine influenza virus of H1N1 subtype.

PubMed

Deblanc, C; Robert, F; Pinard, T; Gorin, S; Quéguiner, S; Gautier-Bouchardon, A V; Ferré, S; Garraud, J M; Cariolet, R; Brack, M; Simon, G

2013-03-23

The severity of swine influenza is highly variable and can be exacerbated by many factors, such as a pre-infection of pigs with Mycoplasma hyopneumoniae (Mhp). The aim of this study was to investigate the oxidative stress induced by Mhp and the impact of this stress on the evolution of an infection with the European avian-like swine H1N1 influenza virus. Two experimental trials (E1 and E2), which differed only by the feed delivered to the animals, were conducted on SPF pigs. In each trial, one group of nine 6-week-old pigs was inoculated intra-tracheally with Mhp and H1N1 at 21 days intervals and a mock-infected group (8 pigs) was included. Clinical signs were observed, blood samples were collected throughout the study and pathogens were detected in nasal swabs and lung tissues. Results indicated that Mhp infection induced an oxidative stress in E1 and E2, but its level was more important in E2 than in E1 three weeks post-Mhp inoculation, before H1N1 infection. In both trials, a strong inflammatory response and a response to the oxidative stress previously induced by Mhp appeared after H1N1 infection. However, the severity of influenza disease was significantly more marked in E2 as compared to E1, as revealed by prolonged hyperthermia, stronger reduction in mean daily weight gain and earlier viral shedding. These results suggested that severity of flu syndrome and reduction in animal performance may vary depending on the level of oxidative stress at the moment of the influenza infection, and that host responses could be influenced by the feed. Copyright © 2012 Elsevier B.V. All rights reserved.

Development and activity of Glomus intraradices as affected by co-existence with Glomus claroideum in one root system.

PubMed

Janoušková, Martina; Seddas, Pascale; Mrnka, Libor; van Tuinen, Diederik; Dvořáčková, Anna; Tollot, Marie; Gianinazzi-Pearson, Vivienne; Vosátka, Miroslav; Gollotte, Armelle

2009-08-01

The co-existence of two arbuscular mycorrhizal fungal (AMF) species, Glomus intraradices and Glomus claroideum, in the root systems of plants was investigated in a greenhouse experiment aimed at reconstructing interactions during an early stage of primary succession on a coal-mine spoil bank in Central Europe. Two plant species, Tripleurospermum inodorum and Calamagrostis epigejos, were inoculated either with one or both AMF species. Fungal development, determined by trypan blue and alkaline phosphatase staining as well as by PCR amplification of rRNA genes with species-specific primers, and the expression of five genes with different metabolic functions in the intraradical structures of G. intraradices were followed after 6 and 9 weeks of cultivation. The two AMF closely co-existed in the root systems of both plants possibly through similar colonisation rates and competitivity. Inoculation with the two fungi, however, did not bring any additional benefit to the host plants in comparison with single inoculation; moreover, plant growth depression observed after inoculation with G. claroideum persisted also in mixed inoculation. The expression of all the assayed G. intraradices genes was affected either by host plant or by co-inoculation with G. claroideum. The effects of both factors depended on the time of sampling, which underlines the importance of addressing this topic in time-course studies.

Effectiveness of ivermectin against later 4th-stage Strongylus vulgaris in ponies.

PubMed

Slocombe, J O; McCraw, B M; Pennock, P W; Vasey, J

1982-09-01

Twelve pony foals were reared worm-free and inoculated with Strongylus vulgaris. Approximately 8 weeks after they were inoculated, 6 foals were given ivermectin IM at a dosage rate of 200 micrograms/kg of body weight and 6 were given a placebo. All foals were necropsied 35 days after treatment. Ivermectin was 98.9% effective in eliminating later 4th-stage S vulgaris larvae located near the origin of major intestinal arteries and in reducing clinical signs and permitting resolution of lesions associated with verminous arteritis. One pony foal reared on pasture and with evidence of arteritis of the cranial mesenteric and ileocolic arteries on arteriography was treated with ivermectin at a dosage rate of 200 micrograms/kg of body weight. On arteriographs taken subsequently, there was evidence of regression of the lesion, and at necropsy 9 weeks after treatment, there was no arteritis or larvae in those arteries.

Host plant growth promotion and cadmium detoxification in Solanum nigrum, mediated by endophytic fungi.

PubMed

Khan, Abdur Rahim; Ullah, Ihsan; Waqas, Muhammad; Park, Gun-Seok; Khan, Abdul Latif; Hong, Sung-Jun; Ullah, Rehman; Jung, Byung Kwon; Park, Chang Eon; Ur-Rehman, Shafiq; Lee, In-Jung; Shin, Jae-Ho

2017-02-01

Current investigation conducted to evaluate the associated fungal endophyte interactions of a Cd hyper-accumulator Solanum nigrum Korean ecotype under varying concentrations of Cd. Two indole-3-acetic acid (IAA) producing fungal strains, RSF-4L and RSF-6L, isolated from the leaves of S. nigrum, were initially screened for Cd tolerance and accumulation potential. In terms of dry biomass production, the strain RSF-6L showed higher tolerance and accumulation capacity for Cd toxicity in comparison to RSF-4L. Therefore, RSF-6L was applied in vivo to S. nigrum and grown for six weeks under Cd concentrations of 0, 10, and 30mgKg -1 of dry sand. The effect of fungal inoculation assessed by plant physiological responses, endogenous biochemical regulations, and Cd profile in different tissues. Significant increase were observed in plant growth attributes such as shoot length, root length, dry biomass, leaf area, and chlorophyll contents in inoculated RSF-6L plants in comparison to non-inoculated plants with or without Cd contamination. RSF-6L inoculation decreased uptake of Cd in roots and above ground parts, as evidenced by a low bio-concentration factor (BCF) and improved tolerance index (TI). However, Cd concentration in the leaves remained the same for inoculated and non-inoculated plants under Cd spiking. Fungal inoculation protected the host plants, as evidenced by low peroxidase (POD) and polyphenol peroxidase (PPO) activities and high catalase (CAT) activity. Application of appropriate fungal inoculation that can improve tolerance mechanisms of hyper-accumulators and reduce Cd uptake can be recommended for phyto-stabilisation/immobilisation of heavy metals in crop fields. Copyright © 2016 Elsevier Inc. All rights reserved.

African swine fever convalescent sows: subsequent pregnancy and the effect of colostral antibody on challenge inoculation of their pigs.

PubMed

Schlafer, D H; McVicar, J W; Mebus, C A

1984-07-01

The effect of African swine fever (ASF) virus infection on reproductive performance of recovered sows and their pigs was investigated. Six sows were inoculated with a 1979 ASF isolate from the Dominican Republic. One sow was bred on postinoculation day (PID) 58 and killed on PID 148. Four sows were bred between PID 368 and 419 and were allowed to farrow. One sow did not conceive. Samples collected during pregnancy, at farrowing, and during lactation were tested for virus by tissue culture and animal inoculations to determine whether ASF virus recrudesced during these natural stresses. Virus was recovered only from tissues of the sow killed on PID 148. Virus was not detected in tissue samples from the 4 other sows or from any fetus or neonate. Sow and neonatal pig sera, colostral whey, and milk whey were assayed for antibodies against ASF viral antigens, using an enzyme-linked immunosorbent assay. Antibody values in sows' sera did not change appreciably during pregnancy, farrowing, or lactation. One litter of pigs was raised with their sow. Weekly serum samples were tested for passively acquired antibodies. At 7 weeks of age, the litter was challenge inoculated with the same virus as that used initially to infect their dam. Viremia titers, duration of viremias, and clinical course were reduced. One young pig did not develop fever, viremia, clinical disease, or antibody response to virus challenge exposure. The altered course of infection was attributed to protective effect of passively acquired antibodies.

Effective surveillance for early classical swine fever virus detection will utilize both virus and antibody detection capabilities.

PubMed

Panyasing, Yaowalak; Kedkovid, Roongtham; Thanawongnuwech, Roongroje; Kittawornrat, Apisit; Ji, Ju; Giménez-Lirola, Luis; Zimmerman, Jeffrey

2018-03-01

Early recognition and rapid elimination of infected animals is key to controlling incursions of classical swine fever virus (CSFV). In this study, the diagnostic characteristics of 10 CSFV assays were evaluated using individual serum (n = 601) and/or oral fluid (n = 1417) samples collected from -14 to 28 days post inoculation (DPI). Serum samples were assayed by virus isolation (VI), 2 commercial antigen-capture enzyme-linked immunosorbent assays (ELISA), virus neutralization (VN), and 3 antibody ELISAs. Both serum and oral fluid samples were tested with 3 commercial real-time reverse transcription-polymerase chain reaction (rRT-PCR) assays. One or more serum samples was positive by VI from DPIs 3 to 21 and by antigen-capture ELISAs from DPIs 6 to 17. VN-positive serum samples were observed at DPIs ≥ 7 and by antibody ELISAs at DPIs ≥ 10. CSFV RNA was detected in serum samples from DPIs 2 to 28 and in oral fluid samples from DPIs 4 to 28. Significant differences in assay performance were detected, but most importantly, no single combination of sample and assay was able to dependably identify CSFV-inoculated pigs throughout the 4-week course of the study. The results show that effective surveillance for CSFV, especially low virulence strains, will require the use of PCR-based assays for the detection of early infections (<14 days) and antibody-based assays, thereafter. Copyright © 2018 Elsevier B.V. All rights reserved.

MicroRNA expression analysis of feline and canine parvovirus infection in vivo (felis)

PubMed Central

Zhang, Xin; Zeng, Weijie; Zheng, Qingxu; Hao, Xiangqi; Lin, Xi; Zheng, Yun; Wang, Lifang; Zhang, Guihong; Li, Shoujun

2017-01-01

Feline panleukopenia is a common contagious disease with high morbidity and mortality. At present, feline parvovirus (FPV) and canine parvovirus (CPV) variants are the pathogens of feline panleukopenia. Many studies have shown that miRNAs are involved in virus-host interactions. Nevertheless, miRNA expression profiling of FPV (original virus) or CPV-2b (new virus) in cats has not been reported. To investigate these profiles, three 10-week-old cats were orally inoculated with 106 TCID50 of the viruses (FPV and CPV-2b), and the jejunums of one cat in each group were sectioned for miRNA sequencing at 5 days post-inoculation (dpi). This study is the first attempt to use miRNA analysis to understand the molecular basis of FPV and CPV infection in cats. The miRNA expression profiles of the jejunums of cats infected with FPV and CPV were obtained, and a subset of miRNAs was validated by real-time qPCR. The results show that a variety of metabolism-related pathways, cytokine- and pathogen-host interaction-related pathways, and pathology- and cellar structure-related pathways, as well as others, were affected. Specifically, the JAK-STAT signaling pathway, which is critical for cytokines and growth factors, was enriched. This description of the miRNAs involved in regulating FPV and CPV infection in vivo provides further insight into the mechanisms of viral infection and adaptation and might provide an alternative antiviral strategy for disease control and prevention. PMID:29049413

The duration of immunity in cattle following inoculation of rinderpest cell culture vaccine.

PubMed Central

Plowright, W.

1984-01-01

The duration of immunity following a single administration of rinderpest cell culture vaccine, of 90 or more monolayer passages, was studied in E. African zebu (Boran) and grade (cross-bred European) cattle. All animals were kept for periods of 6-11 years in rinderpest-free environments; groups of them (in all 23 Borans and 10 grades) were then challenged by parenteral or intranasal inoculation of virulent virus or by contact exposure to reacting cattle. Nasal excretion of virus was studied daily over the 10-to 14-day period following challenge, and simultaneous attempts were made to detect viraemia. The neutralizing antibody response was followed at 6-month intervals over the whole post-vaccination period and then daily for 10 days and at longer intervals to 3 weeks after challenge. All 33 animals which were exposed by various routes failed to react clinically and a rinderpest viraemia was never detected. No transmission of virus from the vaccinates to susceptible in-contact controls occurred within 14 or more days, from the 20 animals which could be so tested. Clearcut serological responses to challenge were seen in six cattle (four Borans and two grades) which were challenged after 7 years or more; these reactions were all delayed to the 9th or 10th days, i.e. they were not typically 'anamnestic'. These results are discussed in relation to mass vaccination campaigns for the control of rinderpest and from the comparative viewpoint of measles vaccination in man. PMID:6736639

Effect of ovarian hormones on the phagocytic response of ovariectomized mares.

PubMed

Ganjam, V K; McLeod, C; Klesius, P H; Washburn, S M; Kwapien, R; Brown, B; Fazeli, M H

1982-01-01

The reaction between ovarian hormones and experimental uterine infection (Streptococcus zooepidemicus) was investigated in 3 groups, each containing 6 ovariectomized mares. Group 1 served as controls ('anoestrus'), Group 2 mares were injected with oestrogen ('oestrus') and Group 3 with progesterone ('dioestrus') over a period of 5 weeks. All mares received an intrauterine inoculation of the bacteria 1 week after the start of hormonal treatment, and the results of the challenge were examined by endometrial biopsy and swabs once weekly. At the end of Week 1 no bacteria were recovered from the mares in Group 2. Group 1 mares were free of bacteria at the end of Week 2 but all Group 3 mares remained infected at least for the total period examined. Streptococcal phagocytosis was quantitated by chemiluminescence. Before the challenge-inoculation, phagocytosis was not significantly different in the 3 groups of mares. Bacterial cultures were negative for all three groups. However, within 48 h after infection, there was a significant increase (P less than 0.01) in phagocytosis in Group 2 and a significant suppression (P less than 0.05) in Group 3 mares. Patterns of streptococcal clearance from the uterus closely paralleled the changes in the magnitude of chemiluminescence response. The results suggest that ovarian hormonal status can modulate the phagocytic response in episodes of streptococcal-induced endometritus in mares.

Sodium taurocholate cotransporting polypeptide inhibition efficiently blocks hepatitis B virus spread in mice with a humanized liver

PubMed Central

Nakabori, Tasuku; Hikita, Hayato; Murai, Kazuhiro; Nozaki, Yasutoshi; Kai, Yugo; Makino, Yuki; Saito, Yoshinobu; Tanaka, Satoshi; Wada, Hiroshi; Eguchi, Hidetoshi; Takahashi, Takeshi; Suemizu, Hiroshi; Sakamori, Ryotaro; Hiramatsu, Naoki; Tatsumi, Tomohide; Takehara, Tetsuo

2016-01-01

Sodium taurocholate cotransporting polypeptide (NTCP) is a recently discovered hepatitis B virus (HBV) receptor. In the present study, we used TK-NOG mice with a humanized liver to examine the impact of endogenous NTCP expression on HBV infection. Upon inoculation with HBV, these mice exhibited clear viremia in 2 weeks, and serum HBV DNA levels gradually increased. The frequency of HBsAg-positive hepatocytes in the liver was 5.1 ± 0.6% at 2 weeks and increased with increasing HBV DNA levels, reaching 92.9 ± 2.8% at 10 to 12 weeks. In vivo siRNA-mediated NTCP knockdown before and after HBV inoculation significantly suppressed the levels of HBV replication and the frequency of HBsAg-positive hepatocytes at 2 weeks, whereas NTCP knockdown 13 weeks after infection did not affect these parameters. Similar to the humanized mouse livers in the early phase of HBV infection, human liver samples from chronic hepatitis B patients, especially those treated with nucleos(t)ide analogues, contained a considerable number of hepatocytes that were negative for the anti-HBs antibody. In conclusion, NTCP inhibition prevents the spread of HBV-infected hepatocytes in mice with a humanized liver. NTCP-targeted therapy has potential for regulating HBV infection in patients with chronic hepatitis B. PMID:27278060

Development of a Contemporary Animal Model of Candida albicans-Associated Denture Stomatitis Using a Novel Intraoral Denture System

PubMed Central

Johnson, Clorinda C.; Yu, Alika; Lee, Heeje; Fidel, Paul L.

2012-01-01

Denture stomatitis (DS) is a fungal infection characterized by inflammation of the oral mucosa in direct contact with the denture and affects up to 50% of denture wearers. Despite the prevalence, very little is known about the role of fungal or host factors that contribute to pathogenesis. Recently, we developed a novel intraoral denture system for rodent research. This denture system consists of custom-fitted fixed and removable parts to allow repeated sampling and longitudinal studies. The purpose of this study was to use this denture system to develop a clinically relevant animal model of DS. To establish DS, rats were inoculated with pelleted Candida albicans, which resulted in sustained colonization of the denture and palate for 8 weeks postinoculation. Biofilm formation on the denture was observed by week 4 and on the palate by week 6 postinoculation. Rats were monitored for clinical signs of disease by assigning a clinical score after macroscopic examination of the palate tissue according to Newton's method. By week 4 postinoculation, the majority of inoculated rats with dentures exhibited a clinical score of 1 (pinpoint erythema). By week 6 and week 8 postinoculation, increasing percentages of rats exhibited a clinical score of 2 (diffuse erythema/edema). Histological analysis of palate tissue demonstrated progressively increasing inflammatory cell recruitment throughout the time course of the infection. Palatal biofilm formation was commensurate with development of palatal erythema, which suggests a role for biofilm in the inflammatory response. PMID:22392931

The Phenylpropanoid Pathway and Lignin in Defense against Ganoderma boninense Colonized Root Tissues in Oil Palm (Elaeis guineensis Jacq.)

PubMed Central

Govender, Nisha T.; Mahmood, Maziah; Seman, Idris A.; Wong, Mui-Yun

2017-01-01

Basal stem rot, caused by the basidiomycete fungus, Ganoderma boninense, is an economically devastating disease in Malaysia. Our study investigated the changes in lignin content and composition along with activity and expression of the phenylpropanoid pathway enzymes and genes in oil palm root tissues during G. boninense infection. We sampled control (non-inoculated) and infected (inoculated) seedlings at seven time points [1, 2, 3, 4, 8, and 12 weeks post-inoculation (wpi)] in a randomized design. The expression profiles of phenylalanine ammonia lyase (PAL), cinnamyl alcohol dehydrogenase (CAD), and peroxidase (POD) genes were monitored at 1, 2, and 3 wpi using real-time quantitative polymerase chain reaction. Seedlings at 4, 8, and 12 wpi were screened for lignin content, lignin composition, enzyme activities (PAL, CAD, and POD), growth (weight and height), and disease severity (DS). Gene expression analysis demonstrated up-regulation of PAL, CAD, and POD genes in the infected seedlings, relative to the control seedlings at 1, 2, and 3 wpi. At 2 and 3 wpi, CAD showed highest transcript levels compared to PAL and POD. DS increased progressively throughout sampling, with 5, 34, and 69% at 4, 8, and 12 wpi, respectively. Fresh weight and height of the infected seedlings were significantly lower compared to the control seedlings at 8 and 12 wpi. Lignin content of the infected seedlings at 4 wpi was significantly higher than the control seedlings, remained elicited with no change at 8 wpi, and then collapsed with a significant reduction at 12 wpi. The nitrobenzene oxidation products of oil palm root lignin yielded both syringyl and guaiacyl monomers. Accumulation of lignin in the infected seedlings was in parallel to increased syringyl monomers, at 4 and 8 wpi. The activities of PAL and CAD enzymes in the infected seedlings at DS = 5–34% were significantly higher than the control seedlings and thereafter collapsed at DS = 69%. PMID:28861093

The Phenylpropanoid Pathway and Lignin in Defense against Ganoderma boninense Colonized Root Tissues in Oil Palm (Elaeis guineensis Jacq.).

PubMed

Govender, Nisha T; Mahmood, Maziah; Seman, Idris A; Wong, Mui-Yun

2017-01-01

Basal stem rot, caused by the basidiomycete fungus, Ganoderma boninense , is an economically devastating disease in Malaysia. Our study investigated the changes in lignin content and composition along with activity and expression of the phenylpropanoid pathway enzymes and genes in oil palm root tissues during G. boninense infection. We sampled control (non-inoculated) and infected (inoculated) seedlings at seven time points [1, 2, 3, 4, 8, and 12 weeks post-inoculation (wpi)] in a randomized design. The expression profiles of phenylalanine ammonia lyase (PAL), cinnamyl alcohol dehydrogenase (CAD), and peroxidase (POD) genes were monitored at 1, 2, and 3 wpi using real-time quantitative polymerase chain reaction. Seedlings at 4, 8, and 12 wpi were screened for lignin content, lignin composition, enzyme activities (PAL, CAD, and POD), growth (weight and height), and disease severity (DS). Gene expression analysis demonstrated up-regulation of PAL, CAD, and POD genes in the infected seedlings, relative to the control seedlings at 1, 2, and 3 wpi. At 2 and 3 wpi, CAD showed highest transcript levels compared to PAL and POD. DS increased progressively throughout sampling, with 5, 34, and 69% at 4, 8, and 12 wpi, respectively. Fresh weight and height of the infected seedlings were significantly lower compared to the control seedlings at 8 and 12 wpi. Lignin content of the infected seedlings at 4 wpi was significantly higher than the control seedlings, remained elicited with no change at 8 wpi, and then collapsed with a significant reduction at 12 wpi. The nitrobenzene oxidation products of oil palm root lignin yielded both syringyl and guaiacyl monomers. Accumulation of lignin in the infected seedlings was in parallel to increased syringyl monomers, at 4 and 8 wpi. The activities of PAL and CAD enzymes in the infected seedlings at DS = 5-34% were significantly higher than the control seedlings and thereafter collapsed at DS = 69%.

Eurasian Tree Sparrows, Risk for H5N1 Virus Spread and Human Contamination through Buddhist Ritual: An Experimental Approach

PubMed Central

Gutiérrez, Ramona Alikiiteaga; Sorn, San; Nicholls, John M.; Buchy, Philippe

2011-01-01

Background The Highly Pathogenic Avian Influenza H5N1 virus has dramatically spread throughout Southeast Asia since its first detection in 1997. Merit Release Birds, such as the Eurasian Tree Sparrow, are believed to increase one's positive karma when kissed and released during Buddhist rituals. Since these birds are often in close contact with both poultry and humans, we investigated their potential role in the spread of H5N1 virus. Methodology/Principal Findings Seven series of experiments were conducted in order to investigate the possible interactions between inoculated and exposed birds, including sparrow/sparrow, sparrow/chicken, duck/sparrow. Daily and post-mortem samples collected were tested for H5N1 virus by real-time RT-PCR and egg inoculation. When directly inoculated, Eurasian Tree Sparrows were highly susceptible to the H5N1 virus, with a fatality rate approaching 100% within 5 days post-inoculation. Although transmission of fatal infection between sparrows did not occur, seroconversion of the exposed birds was observed. Up to 100% chickens exposed to inoculated sparrows died of H5N1 infection, depending on the caging conditions of the birds, while a fatality rate of 50% was observed on sparrows exposed to infected ducks. Large quantities of H5N1 virus were detected in the sparrows, particularly in their feathers, from which infectious particles were recovered. Conclusions/Significance Our study indicates that under experimental conditions, Eurasian Tree Sparrows are susceptible to H5N1 infection, either by direct inoculation or by contact with infected poultry. Their ability to transmit H5N1 infection to other birds is also demonstrated, suggesting that the sparrows may play a role in the dissemination of the virus. Finally, the presence of significant quantities of H5N1 virus on sparrows' feathers, including infectious particles, would suggest that Merit Release Birds represent a risk for human contamination in countries where avian influenza virus is circulating and where this religious ritual is practiced. PMID:22164310

[Modeling of mixed infection by tick-borne encephalitis and Powassan viruses in mice].

PubMed

Khozinskaia, G A; Pogodina, V V

1982-01-01

Simultaneous inoculation of mice with tick-borne and Powassan viruses was shown, depending on experimental conditions, to result either in stimulation of infection or its unchanged course as compared with monoinfection and inoculation with the viruses at 2--3-week intervals in cross protection of mice against the superinfecting virus. Simultaneous inoculation of mice with the two viruses was accompanied by their multiplication in the blood and brains of mice and formation of antihemagglutinating antibodies to each of them. In the virus population in the brains of mice there was either formation of a mixture of two viruses or their phenotypic mixing. In cross protection, multiplication of the superinfecting virus in the blood and brain of mice was slightly inhibited, the antihemagglutinating antibody to a second virus either did not form or appeared in low titres.

Rapid CD4+ T-Cell Loss Induced by Human Immunodeficiency Virus Type 1NC in Uninfected and Previously Infected Chimpanzees

PubMed Central

Novembre, Francis J.; de Rosayro, Juliette; Nidtha, Soumya; O'Neil, Shawn P.; Gibson, Terri R.; Evans-Strickfaden, Tammy; Hart, Clyde E.; McClure, Harold M.

2001-01-01

To investigate the pathogenicity of a virus originating in a chimpanzee with AIDS (C499), two chimpanzees were inoculated with a plasma-derived isolate termed human immunodeficiency virus type 1NC (HIV-1NC). A previously uninfected chimpanzee, C534, experienced rapid peripheral CD4+ T-cell loss to fewer than 26 cells/μl by 14 weeks after infection. CD4+ T-cell depletion was associated with high plasma HIV-1 loads but a low virus burden in the peripheral lymph node. The second chimpanzee, C459, infected 13 years previously with HIV-1LAV, experienced a more protracted course of peripheral CD4+ T-cell loss after HIV-1NC inoculation, resulting in fewer than 200 cells/μl by 96 weeks postinoculation. The quantities of viral RNA in the plasma and peripheral lymph node from C459 were below the lower limits of detection prior to inoculation with HIV-1NC but were significantly and persistently increased after superinfection, with HIV-1NC representing the predominant viral genotype. These results show that viruses derived from C499 are more pathogenic for chimpanzees than any other HIV-1 isolates described to date. PMID:11152525

Evaluation of guinea pig model for experimental Salmonella serovar Abortusequi infection in reference to infertility.

PubMed

Singh, B R; Alam, Javed; Hansda, D; Verma, J C; Singh, V P; Yadav, M P

2002-03-01

The present study conclusively revealed the role for Salmonella enterica subspecies enterica serovar Abortusequi in conception failure. None of the 12 guinea pigs conceived when orally exposed to sublethal dose of the pathogen during breeding, while 66.67% of animals in control group were found pregnant during same period of observation under similar conditions. Salmonella carrier animals also had drastic reduction in conception rate (16.67%). During mid pregnancy, S. Abortusequi exposure to guinea pigs through intravaginal, intramuscular and subcutaneous routes induced fetal death followed by resorption. While 2 out of 6 orally inoculated and 3 out of 6 intraperitonially inoculated guinea pigs aborted, in rest of the animals fetal death was followed by meceration and resorption. It was interesting to note that S. Abortusequi could not persist longer than a week in males while in pregnant females it could be detected for >10 weeks after inoculation. In late pregnancy, most of the exposed animals aborted and non aborting animals though had normal parturition, survival rate of their babies was nearly zero in comparison to the control group. The study revealed role for S. Abortusequi in impairing conception, abortion, early fetal deaths, fetal meceration and resorption. Further studies are required to identify factors responsible for increased susceptibility of females particularly during pregnancy.

The oral route in the pathogenesis of paracoccidioidomycosis: an experimental study in BALB/c mice infected with P. brasiliensis conidia.

PubMed

Roldán, J C; Tabares, A M; Gómez, B L; Aristizábal, B E; Cock, A M; Restrepo, A

2001-01-01

Due to the high frequency of oral mucosal lesions observed in paracoccidioidomycosis patients, it was advocated that the infection was acquired by the traumatic implantation of the etiologic agent Paracoccidioides brasiliensis. Although at present this theory is considered invalid, it has not yet been excluded in experimental studies. In order to determine if intra-oral inoculation could explain the pathogenesis of paracoccidioidomycosis, 64 BALB/c mice were inoculated intra-orally with 850.000 viable P. brasiliensis conidia into the mandibular body. Animals were sacrificed at various time intervals up to 20 weeks and cultures were made from gingiva, lungs, spleen, and liver. Additionally, histopathological studies of the mandibular body were also performed. P. brasiliensis was isolated from all gingival tissues during the interval 24-72 h, indicating that the infection was active. During the 5-10 week period, the infection appeared to have been controlled at the inoculation site as cultures showed a significant reduction in colony forming units (CFU); however, at the 15-20 week period such control was lost and the fungus was recovered once more. Dissemination to other body sites was rare; thus, the lungs were involved in just one animal (2%), the liver in two (3%) and the spleen in seven (11%). The infection became established as proven by positive organ cultures, but the dissemination pattern did not correspond to the one observed in humans. Based on these findings, the intra-oral traumatic route does not appear to mimic the natural history of paracoccidioidomycosis.

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

PubMed

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

2008-03-01

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

Testing and Evaluation of a Predeployment Stress Inoculation Training Program (PreSTINT)

DTIC Science & Technology

2016-07-01

PreSTINT training to reduce the risk of PTSD and other psychological distress symptoms. Multivariate analyses assessed differences between experimental...the risk of post-traumatic stress disorder (PTSD) and other post-deployment psychological distress symptoms. Participants reported their stress...Employment or Research Opportunities Jessica Kelley Morgan, MS, a doctoral student in psychology , worked as an intern on this grant. During the last 16

Pre-treatment of soybean plants with calcium stimulates ROS responses and mitigates infection by Sclerotinia sclerotiorum.

PubMed

Arfaoui, Arbia; El Hadrami, Abdelbasset; Daayf, Fouad

2018-01-01

Considering the high incidence of white mold caused by Sclerotinia sclerotiorum in a variety of field crops and vegetables, different control strategies are needed to keep the disease under economical threshold. This study assessed the effect of foliar application of a calcium formulation on disease symptoms, oxalic acid production, and on the oxidative stress metabolism in soybean plants inoculated with each of two isolates of the pathogen that have contrasting aggressiveness (HA, highly-aggressive versus WA, weakly-aggressive). Changes in reactive oxygen species (ROS) levels in soybean plants inoculated with S. sclerotiorum isolates were assessed at 6, 24, 48 and 72 h post inoculation (hpi). Generation of ROS including hydrogen peroxide (H 2 O 2 ), anion superoxide (O 2 - ) and hydroxyl radical (OH) was evaluated. Inoculation with the WA isolate resulted in more ROS accumulation compared to the HA isolate. Pre-treatment with the calcium formulation restored ROS production in plants inoculated with the HA isolate. We also noted a marked decrease in oxalic acid content in the leaves inoculated with the HA isolate in presence of calcium, which coincided with an increase in plant ROS production. The expression patterns of genes involved in ROS detoxification in response to the calcium treatments and/or inoculation with S. Sclerotiorum isolates were monitored by RT-qPCR. All of the tested genes showed a higher expression in response to inoculation with the WA isolate. The expression of most genes tested peaked at 6 hpi, which preceded ROS accumulation in the soybean leaves. Overall, these data suggest that foliar application of calcium contributes to a decrease in oxalic acid production and disease, arguably via modulation of the ROS metabolism. Copyright © 2017 Elsevier Masson SAS. All rights reserved.

Pathological and immunohistochemical studies of subclinical infection of chicken anemia virus in 4-week-old chickens.

PubMed

Haridy, Mohie; Sasaki, Jun; Ikezawa, Mitsutaka; Okada, Kosuke; Goryo, Masanobu

2012-06-01

Subclinical infection of chicken anemia virus (CAV) at 4 to 6 weeks of age, after maternal antibodies have waned, is implicated in several field problems in broiler flocks. In order to understand the pathogenesis of subclinical infection with CAV, an immunopathological study of CAV-inoculated 4-week-old SPF chickens was performed. Sixty 4-week-old SPF chickens were equally divided into CAV and control groups. The CAV group was inoculated intramuscularly with the MSB1-TK5803 strain of CAV. Neither mortality nor anemia was detected in the CAV and control groups. In the CAV group, no signs were observed, except that some chickens were grossly smaller compared with the control group. Sporadic thymus lobes appeared to be reddening and atrophied. Within the first two weeks p.i. of CAV, there was a mild to moderate depletion of lymphocytes in the thymus cortex and spleen in some chickens. Moreover, lymphoid depletion of the bursa of Fabricius, proventriculus and cecal tonsils was observed. Hyperplastic lymphoid foci were observed in the liver, lungs, kidneys and heart at the 4th week p.i. of CAV. Immunohistochemically, a moderate lymphoid depletion of CD4(+)and CD8(+) T cells in the thymus cortex and spleen was observed in some chickens within two weeks p.i. of CAV. CAV inclusions and antigens were detected infrequently in the thymus cortex and spleen. It could be concluded that the immunosuppression in subclinical infection with CAV occurs as a result of reduction of cellular immunity.

Isolation of Toxoplasma gondii from horse meat in Egypt.

PubMed

Shaapan, R M; Ghazy, A A

2007-01-01

Portions of heart, liver, skeletal and diaphragmatic muscles obtained from 150 slaughtered horses at Giza-Zoo abattoir were used for bioassays in mice and cats. T. gondii tachyzoites were isolated successfully from the peritoneal exudates of the inoculated mice 6-8 days post inoculation with pooled horse tissues. Whereas, T. gondii tissue cysts containing bradyzoites were detected in the impression smears of mice brain on the 45th days or more post infection. The oocysts were detected in feces of cats 3-6 days post feeding on horse tissues containing tissue cysts. The oocysts became sporulated within 3-5 days in 2.5% Potassium dichromate. A total of 79 out of 150 horse meat samples were found to be infected with an incidence rate of 52.6 %. This is the first trial for isolation of T. gondii infective stages from horses in Egypt. Moreover, this study pointed out to the high infection rate of T. gondii in horse meat which may be considered as an important source of infection to wild zoo-animals in Egypt and humans in some countries if consumed raw or insufficiently cooked.

Azotobacter chroococcum as a potentially useful bacterial biofertilizer for cotton (Gossypium hirsutum): Effect in reducing N fertilization.

PubMed

Romero-Perdomo, Felipe; Abril, Jorge; Camelo, Mauricio; Moreno-Galván, Andrés; Pastrana, Iván; Rojas-Tapias, Daniel; Bonilla, Ruth

The aim of this research was to evaluate whether the application of two plant growth-promoting (rhizo)bacteria might reduce nitrogen fertilization doses in cotton. We used strains Azotobacter chroococcum AC1 and AC10 for their proven ability to promote seed germination and cotton growth. These microorganisms were characterized by their plant growth-promoting activities. Then, we conducted a glasshouse study to evaluate the plant growth promoting ability of these strains with reduced doses of urea fertilization in cotton. Results revealed that both strains are capable of fixing nitrogen, solubilizing phosphorus, synthesizing indole compounds and producing hydrolytic enzymes. After 12 weeks, the glasshouse experiment showed that cotton growth was positively influenced due to bacterial inoculation with respect to chemical fertilization. Notably, we observed that microbial inoculation further influenced plant biomass (p<0.05) than nitrogen content. Co-inoculation, interestingly, exhibited a greater beneficial effect on plant growth parameters compared to single inoculation. Moreover, similar results without significant statistical differences were observed among bacterial co-inoculation plus 50% urea and 100% fertilization. These findings suggest that co-inoculation of A. chroococcum strains allow to reduce nitrogen fertilization doses up to 50% on cotton growth. Our results showed that inoculation with AC1 and AC10 represents a viable alternative to improve cotton growth while decreasing the N fertilizer dose and allows to alleviate the environmental deterioration related to N pollution. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

Pneumonitis in Syrian golden hamsters (Mesocricetus auratus) infected with Rio Mamoré virus (family Bunyaviridae, genus Hantavirus).

PubMed

Milazzo, Mary Louise; Eyzaguirre, Eduardo J; Fulhorst, Charles F

2014-10-13

Rio Mamoré virus is an etiological agent of hantavirus pulmonary syndrome in South America. The purpose of this study was to determine whether Rio Mamoré virus strain HTN-007 in Syrian golden hamsters is pathogenic. None of 37 adult hamsters infected by intramuscular injection of HTN-007, including 10 animals killed on Day 42 or 43 post-inoculation, exhibited any symptom of disease. Histological abnormalities included severe or moderately severe pneumonitis in 6 (46.2%) of the 13 animals killed on Day 7 or 10 post-inoculation. The primary target of infection in lung was the endothelium of the microvasculature. Collectively, these results indicate that Rio Mamoré virus strain HTN-007 in adult Syrian golden hamsters can cause a nonlethal disease that is pathologically similar to hantavirus pulmonary syndrome. Copyright © 2014 Elsevier B.V. All rights reserved.

Pneumonitis in Syrian golden hamsters (Mesocricetus auratus) infected with Río Mamoré virus (family Bunyaviridae, genus Hantavirus)

PubMed Central

Milazzo, Mary Louise; Eyzaguirre, Eduardo J.; Fulhorst, Charles F.

2014-01-01

Rio Mamoré virus is an etiological agent of hantavirus pulmonary syndrome in South America. The purpose of this study was to determine whether Rio Mamoré virus strain HTN-007 in Syrian golden hamsters is pathogenic. None of 37 adult hamsters infected by intramuscular injection of HTN-007, including 10 animals killed on Day 42 or 43 post-inoculation, exhibited any symptom of disease. Histological abnormalities included severe or moderately severe pneumonitis in 6 (46.2%) of the 13 animals killed on Day 7 or 10 post-inoculation. The primary target of infection in lung was the endothelium of the microvasculature. Collectively, these results indicate that Rio Mamoré virus strain HTN-007 in adult Syrian golden hamsters can cause a nonlethal disease that is pathologically similar to hantavirus pulmonary syndrome. PMID:25064267

Mechanism of demyelination after HSV type I intraocular injection in rabbits.

PubMed

Narang, H K

1980-02-01

Intraocular injection of herpes simplex virus (HSV) type I into the vitreous body of 18-day-old rabbits induced, on the 7th day post-inoculation, neurological signs with marked head jerking and atazia. Examination of semi-serial 1-micrometers thick sections of the whole lengths of right and left optic nerves and chiasma of 4--64 days post-inoculated rabbits revealed a small lesion, restricted to the medial side, which had extended 2--3 mm, during the first 4 days, along the optic nerve. Ahead of the developing lesion marked chromatin changes of neuroglial cells were noticed followed by cuffing of blood vessels, infiltration by macrophages, demyelination and remyelination. The present study indicated that demyelination occurred following the infection of the myelinating cells. It appeared that virus did not become latent and many cells survived the viral attack. Repeated episodies of viral activity caused further damage while repair did not keep pace.

Soil type dependent rhizosphere competence and biocontrol of two bacterial inoculant strains and their effects on the rhizosphere microbial community of field-grown lettuce.

PubMed

Schreiter, Susanne; Sandmann, Martin; Smalla, Kornelia; Grosch, Rita

2014-01-01

Rhizosphere competence of bacterial inoculants is assumed to be important for successful biocontrol. Knowledge of factors influencing rhizosphere competence under field conditions is largely lacking. The present study is aimed to unravel the effects of soil types on the rhizosphere competence and biocontrol activity of the two inoculant strains Pseudomonas jessenii RU47 and Serratia plymuthica 3Re4-18 in field-grown lettuce in soils inoculated with Rhizoctonia solani AG1-IB or not. Two independent experiments were carried out in 2011 on an experimental plot system with three soil types sharing the same cropping history and weather conditions for more than 10 years. Rifampicin resistant mutants of the inoculants were used to evaluate their colonization in the rhizosphere of lettuce. The rhizosphere bacterial community structure was analyzed by denaturing gradient gel electrophoresis of 16S rRNA gene fragments amplified from total community DNA to get insights into the effects of the inoculants and R. solani on the indigenous rhizosphere bacterial communities. Both inoculants showed a good colonization ability of the rhizosphere of lettuce with more than 10(6) colony forming units per g root dry mass two weeks after planting. An effect of the soil type on rhizosphere competence was observed for 3Re4-18 but not for RU47. In both experiments a comparable rhizosphere competence was observed and in the presence of the inoculants disease symptoms were either significantly reduced, or at least a non-significant trend was shown. Disease severity was highest in diluvial sand followed by alluvial loam and loess loam suggesting that the soil types differed in their conduciveness for bottom rot disease. Compared to effect of the soil type of the rhizosphere bacterial communities, the effects of the pathogen and the inoculants were less pronounced. The soil types had a surprisingly low influence on rhizosphere competence and biocontrol activity while they significantly affected the bottom rot disease severity.

Soil Type Dependent Rhizosphere Competence and Biocontrol of Two Bacterial Inoculant Strains and Their Effects on the Rhizosphere Microbial Community of Field-Grown Lettuce

PubMed Central

Schreiter, Susanne; Sandmann, Martin; Smalla, Kornelia; Grosch, Rita

2014-01-01

Rhizosphere competence of bacterial inoculants is assumed to be important for successful biocontrol. Knowledge of factors influencing rhizosphere competence under field conditions is largely lacking. The present study is aimed to unravel the effects of soil types on the rhizosphere competence and biocontrol activity of the two inoculant strains Pseudomonas jessenii RU47 and Serratia plymuthica 3Re4-18 in field-grown lettuce in soils inoculated with Rhizoctonia solani AG1-IB or not. Two independent experiments were carried out in 2011 on an experimental plot system with three soil types sharing the same cropping history and weather conditions for more than 10 years. Rifampicin resistant mutants of the inoculants were used to evaluate their colonization in the rhizosphere of lettuce. The rhizosphere bacterial community structure was analyzed by denaturing gradient gel electrophoresis of 16S rRNA gene fragments amplified from total community DNA to get insights into the effects of the inoculants and R. solani on the indigenous rhizosphere bacterial communities. Both inoculants showed a good colonization ability of the rhizosphere of lettuce with more than 106 colony forming units per g root dry mass two weeks after planting. An effect of the soil type on rhizosphere competence was observed for 3Re4-18 but not for RU47. In both experiments a comparable rhizosphere competence was observed and in the presence of the inoculants disease symptoms were either significantly reduced, or at least a non-significant trend was shown. Disease severity was highest in diluvial sand followed by alluvial loam and loess loam suggesting that the soil types differed in their conduciveness for bottom rot disease. Compared to effect of the soil type of the rhizosphere bacterial communities, the effects of the pathogen and the inoculants were less pronounced. The soil types had a surprisingly low influence on rhizosphere competence and biocontrol activity while they significantly affected the bottom rot disease severity. PMID:25099168

Antibody response to equine coronavirus in horses inoculated with a bovine coronavirus vaccine

PubMed Central

NEMOTO, Manabu; KANNO, Toru; BANNAI, Hiroshi; TSUJIMURA, Koji; YAMANAKA, Takashi; KOKADO, Hiroshi

2017-01-01

A vaccine for equine coronavirus (ECoV) is so far unavailable. Bovine coronavirus (BCoV) is antigenically related to ECoV; it is therefore possible that BCoV vaccine will induce antibodies against ECoV in horses. This study investigated antibody response to ECoV in horses inoculated with BCoV vaccine. Virus neutralization tests showed that antibody titers against ECoV increased in all six horses tested at 14 days post inoculation, although the antibody titers were lower against ECoV than against BCoV. This study showed that BCoV vaccine provides horses with antibodies against ECoV to some extent. It is unclear whether antibodies provided by BCoV vaccine are effective against ECoV, and therefore ECoV challenge studies are needed to evaluate efficacy of the vaccine in the future. PMID:28993568

Antibody response to equine coronavirus in horses inoculated with a bovine coronavirus vaccine.

PubMed

Nemoto, Manabu; Kanno, Toru; Bannai, Hiroshi; Tsujimura, Koji; Yamanaka, Takashi; Kokado, Hiroshi

2017-11-17

A vaccine for equine coronavirus (ECoV) is so far unavailable. Bovine coronavirus (BCoV) is antigenically related to ECoV; it is therefore possible that BCoV vaccine will induce antibodies against ECoV in horses. This study investigated antibody response to ECoV in horses inoculated with BCoV vaccine. Virus neutralization tests showed that antibody titers against ECoV increased in all six horses tested at 14 days post inoculation, although the antibody titers were lower against ECoV than against BCoV. This study showed that BCoV vaccine provides horses with antibodies against ECoV to some extent. It is unclear whether antibodies provided by BCoV vaccine are effective against ECoV, and therefore ECoV challenge studies are needed to evaluate efficacy of the vaccine in the future.

Physiological, structural and molecular traits activated in strawberry plants after inoculation with the plant growth-promoting bacterium Azospirillum brasilense REC3.

PubMed

Guerrero-Molina, M F; Lovaisa, N C; Salazar, S M; Martínez-Zamora, M G; Díaz-Ricci, J C; Pedraza, R O

2015-05-01

The plant growth-promoting strain REC3 of Azospirillum brasilense, isolated from strawberry roots, prompts growth promotion and systemic protection against anthracnose disease in this crop. Hence, we hypothesised that A. brasilense REC3 can induce different physiological, structural and molecular responses in strawberry plants. Therefore, the aim of this work was to study these traits activated in Azospirillum-colonised strawberry plants, which have not been assessed until now. Healthy, in vitro micropropagated plants were root-inoculated with REC3 under hydroponic conditions; root and leaf tissues were sampled at different times, and oxidative burst, phenolic compound content, malondialdehyde (MDA) concentration, callose deposition, cell wall fortification and gene expression were evaluated. Azospirillum inoculation enhanced levels of soluble phenolic compounds after 12 h post-inoculation (hpi), while amounts of cell wall bound phenolics were similar in inoculated and control plants. Other early responses activated by REC3 (at 24 hpi) were a decline of lipid peroxidation and up-regulation of strawberry genes involved in defence (FaPR1), bacterial recognition (FaFLS2) and H₂O₂ depuration (FaCAT and FaAPXc). The last may explain the apparent absence of oxidative burst in leaves after bacterial inoculation. Also, REC3 inoculation induced delayed structural responses such as callose deposition and cell wall fortification (at 72 hpi). Results showed that A. brasilense REC3 is capable of exerting beneficial effects on strawberry plants, reinforcing their physiological and cellular characteristics, which in turns contribute to improve plant performance. © 2014 German Botanical Society and The Royal Botanical Society of the Netherlands.

Growth performance, carcass trait, meat quality and oxidative stability of beef cattle offered alternative silages in a finishing ration.

PubMed

He, L; Yang, J; Chen, W; Zhou, Z; Wu, H; Meng, Q

2018-03-01

As lack of forage resource, alternative roughage sources have been developed for ruminant production and their inclusion would exert a great effect on the dietary nutrition, consequently affecting animal performance. Four silages (corn silage (CS), corn stalk silage (SS), inoculated CS and inoculated SS) were separately offered to 60 Bohai Black cattle (15 cattle/group) during a 24-week finishing period, in which the growth performance, carcass trait, beef quality and oxidative stability of steers were determined. Neither silage material nor silage inoculant exerted a significant effect on the growth performance, carcass trait and oxidative stability of beef cattle (P>0.05). As to beef quality, cattle offered CS had higher (P0.05) on the proximate components and fatty acids profile of beef muscle. There was neither an interaction (P>0.05) between inoculated treatment and silage material. There were no differences (P>0.05) in cholesterol content and meat quality traits in animals fed alternative silages. The collective findings suggest that it is not economical to substitute high-quality forage for relative low-quality forage in a high-concentrate finishing ration of beef cattle and silage inoculant inclusion would not exert a direct effect on animal performance.

Experimental cryptosporidiosis in kids.

PubMed

Koudela, B; Jirí, V

1997-08-01

The clinical, pathological and parasitological features of cryptosporidiosis resulting from experimental inoculation with 6 x 10(6) Cryptosporidium parvum oocysts were studied in kids. Decreased appetite and depression became apparent 72 h post inoculation. Subsequently watery feces with clumps of mucus and color changes from brown to yellow were observed. The mean duration of diarrhea was 4.2 days. Oocyst shedding started 4 days post inoculation (DPI), started to decrease at 7 DPI, and lasted until 12 DPI. The evidence of high infectivity and fast transmission of C. parvum oocysts was observed under standard zoohygienic conditions. The characteristics of intestinal lesions were similar to those found in other neonatal ruminants infected with C. parvum. The most severe lesions were seen in the posterior jejunum and ileum from 3 to 7 DPI, characterized by villus atrophy, villus blunting, fusion of atrophic villi, crypt hyperplasia, inflammatory infiltration in the lamina propria, and metaplasia of mucosal epithelium. Scanning electron microscopy of ileal epithelium revealed ultrastructural changes on the surface of intestinal mucosa. No cryptosporidia or associated pathological lesions were found in the large intestine or other tissues. The distribution of cryptosporidia in the intestine and number of cryptosporidia per ileal villus on different DPI were also estimated for detailed characterization of the infection in kids as a model for experimental cryptosporidiosis.

Intragranulomatous necrosis in pulmonary granulomas is not related to resistance against Mycobacterium tuberculosis infection in experimental murine models induced by aerosol.

PubMed

Guirado, Evelyn; Gordillo, Sergi; Gil, Olga; Díaz, Jorge; Tapia, Gustavo; Vilaplana, Cristina; Ausina, Vicenç; Cardona, Pere-Joan

2006-04-01

Intragranulomatous necrosis is a primary feature in the natural history of human tuberculosis (TB). Unfortunately, this phenomenon is not usually seen in the experimental TB murine model. Artificial induction of this necrosis in pulmonary granulomas (INPG) may be achieved through aerosol inoculation of lipopolysaccharide (LPS) 3 weeks after Mycobacterium tuberculosis infection. At week 9 post-infection, the centre of primary granulomas became larger, showing eosinophilic necrosis. Interestingly, INPG induction was related to mice strains C57BL/6 and 129/Sv, but not to BALB/c and DBA/2. Furthermore, the same pattern was obtained with the induction of infection using a clinical M. tuberculosis strain (UTE 0335R) that naturally induces INPG. In all the mice strains tested, the study of pulmonary mRNA expression revealed a tendency to increase or to maintain the expression of RANTES, interferon-gamma, tumour necrosis factor and iNOS, in both LPS- and UTE 0335R-induced INPG, thus suggesting that this response must be necessary but not sufficient for inducing INPG. Our work supports that INPG induction is a local phenomenon unrelated to the resistant (C57BL/6 and BALB/c) or susceptible (129/Sv and DBA/2) background of mice strains against M. tuberculosis infection.

Comparative Evaluation of Biochemical Changes in Tomato (Lycopersicon esculentum Mill.) Infected by Alternaria alternata and Its Toxic Metabolites (TeA, AOH, and AME).

PubMed

Meena, Mukesh; Zehra, Andleeb; Dubey, Manish K; Aamir, Mohd; Gupta, Vijai K; Upadhyay, Ram S

2016-01-01

In the present study, we have evaluated the comparative biochemical defense response generated against Alternaria alternata and its purified toxins viz. alternariol (AOH), alternariol monomethyl ether (AME), and tenuazonic acid (TeA). The necrotic lesions developed due to treatment with toxins were almost similar as those produced by the pathogen, indicating the crucial role of these toxins in plant pathogenesis. An oxidative burst reaction characterized by the rapid and transient production of a large amount of reactive oxygen species (ROS) occurs following the pathogen infection/toxin exposure. The maximum concentration of hydrogen peroxide (H 2 O 2 ) produced was reported in the pathogen infected samples (22.2-fold) at 24 h post inoculation followed by TeA (18.2-fold), AOH (15.9-fold), and AME (14.1-fold) in treated tissues. 3,3'- Diaminobenzidine staining predicted the possible sites of H 2 O 2 accumulation while the extent of cell death was measured by Evans blue dye. The extent of lipid peroxidation and malondialdehyde (MDA) content was higher (15.8-fold) at 48 h in the sample of inoculated leaves of the pathogen when compared to control. The cellular damages were observed as increased MDA content and reduced chlorophyll. The activities of antioxidative defense enzymes increased in both the pathogen infected as well as toxin treated samples. Superoxide dismutase (SOD) activity was 5.9-fold higher at 24 h post inoculation in leaves followed by TeA (5.0-fold), AOH (4.1-fold) and AME (2.3-fold) treated leaves than control. Catalase (CAT) activity was found to be increased upto 48 h post inoculation and maximum in the pathogen challenged samples followed by other toxins. The native PAGE results showed the variations in the intensities of isozyme (SOD and CAT) bands in the pathogen infected and toxin treated samples. Ascorbate peroxidase (APx) and glutathione reductase (GR) activities followed the similar trend to scavenge the excess H 2 O 2 . The reduction in CAT activities after 48 h post inoculation demonstrate that the biochemical defense programming shown by the host against the pathogen is not well efficient resulting in the compatible host-pathogen interaction. The elicitor (toxins) induced biochemical changes depends on the potential toxic effects (extent of ROS accumulation, amount of H 2 O 2 produced). Thus, a fine tuning occurs for the defense related antioxidative enzymes against detoxification of key ROS molecules and effectively regulated in tomato plant against the pathogen infected/toxin treated oxidative stress. The study well demonstrates the acute pathological effects of A. alternata in tomato over its phytotoxic metabolites.

Interplay between the Gastric Bacterial Microbiota and Candida albicans during Postantibiotic Recolonization and Gastritis

PubMed Central

Mason, Katie L.; Erb Downward, John R.; Falkowski, Nicole R.; Young, Vincent B.; Kao, John Y.

2012-01-01

The indigenous bacterial microbiome of the stomach, including lactobacilli, is vital in promoting colonization resistance against Candida albicans. However, there are gaps in our understanding about C. albicans gastric colonization versus disease, especially during the postantibiotic recovery phase. This study compared the gastric responses to C. albicans strains CHN1 and SC5314 in microbiome-disturbed and germfree mice to elucidate the contribution of the indigenous microbiota in C. albicans colonization versus disease and yeast-bacterium antagonism during the post-cefoperazone recolonization period. C. albicans can prevent the regrowth of Lactobacillus spp. in the stomach after cefoperazone and promote increased colonization by Enterococcus spp. Using a culture-independent analysis, the effects of oral cefoperazone on the gastric bacterial microbiota were observed to last at least 3 weeks after the cessation of the antibiotic. Disturbance of the gastric bacterial community by cefoperazone alone was not sufficient to cause gastritis, C. albicans colonization was also needed. Gastritis was not evident until after day 7 in cefoperazone-treated infected mice. In contrast, in germfree mice which lack a gastric microbiota, C. albicans induced gastric inflammation within 1 week of inoculation. Therefore, the gastric bacterial community in cefoperazone-treated mice during the first week of postantibiotic recolonization was sufficient to prevent the development of gastritis, despite being ineffective at conferring colonization resistance against C. albicans. Altogether, these data implicate a dichotomy between C. albicans colonization and gastric disease that is bacterial microbiome dependent. PMID:21986629

Interplay between the gastric bacterial microbiota and Candida albicans during postantibiotic recolonization and gastritis.

PubMed

Mason, Katie L; Erb Downward, John R; Falkowski, Nicole R; Young, Vincent B; Kao, John Y; Huffnagle, Gary B

2012-01-01

The indigenous bacterial microbiome of the stomach, including lactobacilli, is vital in promoting colonization resistance against Candida albicans. However, there are gaps in our understanding about C. albicans gastric colonization versus disease, especially during the postantibiotic recovery phase. This study compared the gastric responses to C. albicans strains CHN1 and SC5314 in microbiome-disturbed and germfree mice to elucidate the contribution of the indigenous microbiota in C. albicans colonization versus disease and yeast-bacterium antagonism during the post-cefoperazone recolonization period. C. albicans can prevent the regrowth of Lactobacillus spp. in the stomach after cefoperazone and promote increased colonization by Enterococcus spp. Using a culture-independent analysis, the effects of oral cefoperazone on the gastric bacterial microbiota were observed to last at least 3 weeks after the cessation of the antibiotic. Disturbance of the gastric bacterial community by cefoperazone alone was not sufficient to cause gastritis, C. albicans colonization was also needed. Gastritis was not evident until after day 7 in cefoperazone-treated infected mice. In contrast, in germfree mice which lack a gastric microbiota, C. albicans induced gastric inflammation within 1 week of inoculation. Therefore, the gastric bacterial community in cefoperazone-treated mice during the first week of postantibiotic recolonization was sufficient to prevent the development of gastritis, despite being ineffective at conferring colonization resistance against C. albicans. Altogether, these data implicate a dichotomy between C. albicans colonization and gastric disease that is bacterial microbiome dependent.

Evaluation of a commercial ELISA kit for detection of antibodies against Toxoplasma gondii in serum, plasma and meat juice from experimentally and naturally infected sheep

PubMed Central

2013-01-01

Background Toxoplasmosis is one of the most common food borne zoonoses worldwide, and can be a serious life-threatening disease in the congenitally infected fetus and in immunosupressed patients. Among food animals, sheep along with goats and pigs possess the highest incidence of T. gondii cysts in meat, and play a major role as a source of human infection. Methods In this study, a new commercial ELISA kit (PrioCHECK® Toxoplasma Ab SR, Prionics Schlieren-Zurich, Switzerland) for the detection of anti-T. gondii antibodies in serum, plasma and meat juice of sheep, was evaluated by comparing it with the indirect fluorescent antibody test (IFAT), indirect haemagglutination test (IHA) and real-time PCR, on samples from experimentally inoculated and naturally exposed sheep. Results The commercial ELISA detected the infection status in 50% and 100% of sheep orally inoculated with 10,000 T. gondii oocysts (n = 6), from two or three weeks post infection (wpi), respectively, both on serum and plasma samples. Meat juice from all experimentally inoculated sheep collected at slaughter (12 wpi) showed positive ELISA values. In naturally exposed sheep (n = 396), the ELISA showed a very good agreement with IFAT (kappa = 0.91-1.0) and IHA (kappa = 0.96-1.0) performed on serum; and a positive correlation was observed between ELISA values and IFAT titers. By a Receiver Operating Characteristics (ROC) curve analysis, the commercial ELISA had relative sensitivities between 93.33% and 100%, and relative specificities between 96.87% and 100% respect to IFAT and IHA, depending on the considered cut-off value and animal groups tested. Furthermore, the ELISA correctly recognized all animals reacting positive in real-time PCR. The ELISA results on meat juice agreed with those on serum samples in all experimentally inoculated animals, and in 94 out of 96 (97.9%) naturally exposed sheep, when meat juice was tested at a 1:10 dilution. Conclusion The commercial ELISA kit evaluated in this study could represent a valuable tool to improve the surveillance and reporting system for T. gondii in sheep populations at the farm level or for diagnosis at the slaughterhouse, contributing to the control of this widespread zoonosis. PMID:23561035

Using body temperature, food and water consumption as biomarkers of disease progression in mice with Eμ-myc lymphoma.

PubMed

Hunter, J E; Butterworth, J; Perkins, N D; Bateson, M; Richardson, C A

2014-02-18

Non-invasive biomarkers of disease progression in mice with cancer are lacking making it challenging to implement appropriate humane end points. We investigated whether body temperature, food and water consumption could be used to predict tumour burden. Thirty-six male, wild-type C57Bl/6 mice were implanted with subcutaneous RFID temperature sensors and inoculated with Eμ-myc tumours that infiltrate lymphoid tissue. Decrease in body temperature over the course of the study positively predicted post-mortem lymph node tumour burden (R(2)=0.68, F(1,22)=44.8, P<0.001). At experimental and humane end points, all mice that had a mean decrease in body temperature of 0.7 °C or greater had lymph nodes heavier than 0.5 g (100% sensitivity), whereas a mean decrease in body temperature <0.7 °C always predicted lymph nodes lighter than 0.5 g (100% specificity). The mean decrease in food consumption in each cage also predicted mean post-mortem lymph node tumour burden at 3 weeks (R(2)=0.89, F(1,3)=23.2, P=0.017). Temperature, food and water consumption were useful biomarkers of disease progression in mice with lymphoma and could potentially be used more widely to monitor mice with other forms of cancer.

Experimental infection of pregnant goats with swine fever virus.

PubMed

Shimizu, M; Kumagai, T

1989-07-01

Thirteen pregnant goats were inoculated intravenously with the ALD strain of virulent swine fever (SF) virus on Days 64-84 of gestation. Dams showed transient and mild viremia, and produced high serum neutralizing (SN) antibody after inoculation. Six inoculated dams were reared until parturition occurred and bore six apparently normal, one apparently normal but dead, one mummified and three edematous kids. Neutralizing antibody was demonstrated in the pre-colostral sera obtained from all normal kids, but no SF virus was isolated from any of them. The other seven dams were killed on post-inoculation days (PID) 5-61, and fetuses, placenta and amnion were tested for the virus and SN antibody. All fetuses of five dams examined within PID 40 were positive for SF virus, but negative for SN antibody. SF virus was also isolated from one of three fetuses examined on PID 61. Conversely, the other two fetuses examined on PID 61 were negative for SF virus, but positive for SN antibody. Furthermore, SF virus was isolated from the placenta and amnion of all the dams.

The mouse and ferret models for studying the novel avian-origin human influenza A (H7N9) virus.

PubMed

Xu, Lili; Bao, Linlin; Deng, Wei; Zhu, Hua; Chen, Ting; Lv, Qi; Li, Fengdi; Yuan, Jing; Xiang, Zhiguang; Gao, Kai; Xu, Yanfeng; Huang, Lan; Li, Yanhong; Liu, Jiangning; Yao, Yanfeng; Yu, Pin; Yong, Weidong; Wei, Qiang; Zhang, Lianfeng; Qin, Chuan

2013-08-08

The current study was conducted to establish animal models (including mouse and ferret) for the novel avian-origin H7N9 influenza virus. A/Anhui/1/2013 (H7N9) virus was administered by intranasal instillation to groups of mice and ferrets, and animals developed typical clinical signs including body weight loss (mice and ferrets), ruffled fur (mice), sneezing (ferrets), and death (mice). Peak virus shedding from respiratory tract was observed on 2 days post inoculation (d.p.i.) for mice and 3-5 d.p.i. for ferrets. Virus could also be detected in brain, liver, spleen, kidney, and intestine from inoculated mice, and in heart, liver, and olfactory bulb from inoculated ferrets. The inoculation of H7N9 could elicit seroconversion titers up to 1280 in ferrets and 160 in mice. Leukopenia, significantly reduced lymphocytes but increased neutrophils were also observed in mouse and ferret models. The mouse and ferret model enables detailed studies of the pathogenesis of this illness and lay the foundation for drug or vaccine evaluation.

Experimental infection of Bama miniature pigs with a highly virulent classical swine fever virus.

PubMed

Sun, Yuan; Jiang, Qian; Tian, Da-Yong; Lin, Huan; Li, Hong; Han, Qiu-Ying; Han, Wen; Si, Chang-De; Hu, Shou-Ping; Zhang, Zhuo; Qu, Lian-Dong; Qiu, Hua-Ji

2011-09-25

Currently, larger domestic pigs are only animals widely used in vaccine evaluation and pathogenicity study of classical swine fever virus (CSFV). This study was aimed to create an alternative animal experimental infection model of CSFV. Twenty specific-pathogen-free Bama miniature pigs were randomly divided into two groups and rooms, infected and non-infected, and the pigs in the infected group were inoculated intramuscularly with 104, 105 or 106 TCID50 (median tissue culture infective dose) CSFV Shimen strain (n = 5 × 3) or left uninoculated to serve as in-contact pigs (n = 3). The uninfected control pigs (n = 2) were housed in a separate room. Clinical signs, body temperature, viraemia, tissue antigen distribution, pathological changes and seroconversion were monitored. Clinical signs were observed as early as 2 days post-inoculation (dpi) in all infected pigs (though mild in contact pigs), but not non-infected control pigs. All inoculated pigs showed viraemia by 6 dpi. The in-contact pigs showed lower levels of viraemia. At 10 dpi, seroconversion was noted in five of the 15 inoculated pigs. All inoculated or one in-contact pigs died by 15 dpi. These results show that Bama miniature pigs support productive CSFV infection and display clinical signs and pathological changes consistent with CSFV infections observed in larger domestic pigs.

Rapid Start-up and Loading of an Attached Growth, Simultaneous Nitrification/Denitrification Membrane Aerated Bioreactor

NASA Technical Reports Server (NTRS)

Meyer, Caitlin E.; Pensinger, Stuart; Pickering, Karen D.; Barta, Daniel; Shull, Sarah A.; Vega, Letticia M.; Christenson, Dylan; Jackson, W. Andrew

2015-01-01

Membrane aerated bioreactors (MABR) are attached-growth biological systems used for simultaneous nitrification and denitrification to reclaim water from waste. This design is an innovative approach to common terrestrial wastewater treatments for nitrogen and carbon removal and implementing a biologically-based water treatment system for long-duration human exploration is an attractive, low energy alternative to physiochemical processes. Two obstacles to implementing such a system are (1) the "start-up" duration from inoculation to steady-state operations and (2) the amount of surface area needed for the biological activity to occur. The Advanced Water Recovery Systems (AWRS) team at JSC explored these two issues through two tests; a rapid inoculation study and a wastewater loading study. Results from these tests demonstrate that the duration from inoculation to steady state can be reduced to under two weeks, and that despite low ammonium removal rates, the MABRs are oversized.

Experimental infection of conventional dogs with canine parvovirus.

PubMed

McAdaragh, J P; Eustis, S L; Nelson, D T; Stotz, I; Kenefick, K

1982-04-01

Four 6-week-old conventional pups were inoculated with a parvovirus (PV) isolated from the feces of a dog with naturally occurring enteritis. Blood for hematologic studies, virus isolation (VI), and antibody titration and feces for VI and negative-contrast electron microscopy were collected on day 0 and daily until necropsy. Beginning at postinoculation day 2, necropsies were done and specimens were collected for immunofluorescence, VI, and light microscopic examination. The PV infection was confirmed by VI, immunofluorescence, electron microscopy, and seroconversion. Clinical illness was not observed in inoculated pups, although mild intestinal lesions similar to those of naturally occurring PV enteritis were found. The failure to elicit severe disease in conventional pups indicates that one or more factors, such as intercurrent enteric or systemic infections, immune status, age, nutrition, virulence of virus, dose of infectious virus, and route of inoculation influence the clinical and pathologic manifestations of PV infection.

EPIDEMIC TREMOR, AN ENCEPHALOMYELITIS AFFECTING YOUNG CHICKENS

PubMed Central

Jones, E. Elizabeth

1934-01-01

A new disease having a characteristic and well defined symptom complex is described as occurring in young chickens in four New England states. Tremor, principally of the head and neck, and progressive ataxia are the characteristic symptoms, either or both of which may be present in a single bird. Age at onset in field epidemics ranges from 3 days to 6 weeks, with a majority of cases reported at 3 weeks. Morbidity in commercial flocks ranges from 5 to 50 per cent; mortality in affected hatches may be 50 per cent. The disease may or may not recur in successive hatches, and in the same flock in successive years. Although birds may survive an attack of the disease, nervous symptoms persist in a majority of cases. There is no evidence that nutritional factors are involved. Normal chickens have not contracted the disease by contact with affected birds. The disease has been reproduced in normal chickens by intracerebral inoculation of brain and spinal cord from affected birds. Twenty brain-to-brain passages have been made up to the present time. The incubation period in laboratory passages ranges from 6 to 44 days with symptoms appearing usually between 21 and 28 days. The proportion of inoculated birds developing symptoms has increased with successive passages. The infective agent in the brain has survived in 50 per cent glycerine for 69 days. No organism has been cultivated. The disease has been reproduced after inoculation with bacteriologically sterile filtrates obtained with Seitz and Berkefeld N filters. Attempts to demonstrate the presence of the infective agent in the chicken embryo have been inconclusive. Chicks hatched from eggs laid by birds which had survived the disease were not infected, nor were they immune to inoculation at 6 weeks of age. The characteristic lesion of the disease consists of microscopic focal collections of glia cells, perivascular infiltration, degeneration of Purkinje's cells, and degeneration of nerve cells. Foci of infiltration are present throughout the brain and spinal cord. In the viscera of birds from field epidemics, microscopic focal infiltrations of cells of the lymphoid series are often found. Their presence is most notable in the pancreas and heart. No cell inclusions have been demonstrated. PMID:19870279

Efficient transmission of cassava brown streak disease viral pathogens by chip bud grafting.

PubMed

Wagaba, Henry; Beyene, Getu; Trembley, Cynthia; Alicai, Titus; Fauquet, Claude M; Taylor, Nigel J

2013-12-06

Techniques to study plant viral diseases under controlled growth conditions are required to fully understand their biology and investigate host resistance. Cassava brown streak disease (CBSD) presents a major threat to cassava production in East Africa. No infectious clones of the causal viruses, Cassava brown streak virus (CBSV) or Ugandan cassava brown streak virus (UCBSV) are available, and mechanical transmission to cassava is not effective. An improved method for transmission of the viruses, both singly and as co-infections has been developed using bud grafts. Axillary buds from CBSD symptomatic plants infected with virulent isolates of CBSV and UCBSV were excised and grafted onto 6-8 week old greenhouse-grown, disease-free cassava plants of cultivars Ebwanateraka, TME204 and 60444. Plants were assessed visually for development of CBSD symptoms and by RT-PCR for presence of the viruses in leaf and storage root tissues. Across replicated experiments, 70-100% of plants inoculated with CBSV developed CBSD leaf and stem symptoms 2-6 weeks after bud grafting. Infected plants showed typical, severe necrotic lesions in storage roots at harvest 12-14 weeks after graft inoculation. Sequential grafting of buds from plants infected with UCBSV followed 10-14 days later by buds carrying CBSV, onto the same test plant, resulted in 100% of the rootstocks becoming co-infected with both pathogens. This dual transmission rate was greater than that achieved by simultaneous grafting with UCBSV and CBSV (67%), or when grafting first with CBSV followed by UCBSV (17%). The bud grafting method described presents an improved tool for screening cassava germplasm for resistance to CBSD causal viruses, and for studying pathogenicity of this important disease. Bud grafting provides new opportunities compared to previously reported top and side grafting systems. Test plants can be inoculated as young, uniform plants of a size easily handled in a small greenhouse or large growth chamber and can be inoculated in a controlled manner with CBSV and UCBSV, either singly or together. Disease symptoms develop rapidly, allowing better studies of interactions between these viral pathogens, their movement within shoot and root systems, and how they induce their destructive disease symptoms.

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

PubMed

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

2017-02-01

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

Interactive effects of molasses by homofermentative and heterofermentative inoculants on fermentation quality, nitrogen fractionation, nutritive value and aerobic stability of wilted alfalfa (Medicago sativa L) silage.

PubMed

Hashemzadeh-Cigari, F; Khorvash, M; Ghorbani, G R; Ghasemi, E; Taghizadeh, A; Kargar, S; Yang, W Z

2014-04-01

The effect of adding molasses (0, UM or 50 g/kg on DM basis, M) and two types of inoculant including homofermentative (HO) and a combination of homofermentative and propionate-producing bacterial (HOPAB) inoculants on silage fermentation quality, nitrogen fractionation and aerobic stability of pre-bloom, wilted alfalfa (AS) was determined in laboratory silos. The HOPAB inoculant was more effective than HO in reducing the alfalfa silage pH but increased propionate content in the absence of M (p < 0.05). Inoculation of HOPAB reduced (p < 0.01) acid detergent fibre (ADF) and increased (p < 0.01) lactate to acetate ratio compared with uninoculated AS. Acetate concentration was lower (p < 0.01) in HOPAB-inoculated than other AS. This difference was more pronounced in M-added AS (inoculants × M interaction, p = 0.01). Both inoculants reduced (p < 0.01) ammonia-N content in AS added with M, whereas only HOPAB decreased (p < 0.01) ammonia-N concentration in silage without M. Inoculants increased (p < 0.01) B2 fraction in AS with M addition but had no effect on AS without M. Treating silages with HO-UM increased (p < 0.05) C fraction (acid-detergent insoluble-N) but HOPAB decreased C fraction at two levels of M. Treating alfalfa crop with M and HOPAB improved aerobic stability by increasing the concentration of acetate and propionate of AS respectively. Adding M tended (p < 0.10) to increase short-chain fatty acids (SCFA) and cumulative gas production (CGP). HOPAB alone increased DM disappearance at 24 h post-incubation and effective degradability assuming outflow rate of 8%/h relative to untreated AS (p < 0.05). It was concluded that adding M had no pronounced effects on AS fermentation quality, but increased aerobic stability. HOPAB-inoculated AS with no addition of M improved fermentation quality and increased DM degradability compared with HO. Journal of Animal Physiology and Animal Nutrition © 2013 Blackwell Verlag GmbH.

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

PubMed Central

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

1969-01-01

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

Cowpox virus infection of cynomolgus macaques as a model of hemorrhagic smallpox.

PubMed

Johnson, Reed F; Yellayi, Srikanth; Cann, Jennifer A; Johnson, Anthony; Smith, Alvin L; Paragas, Jason; Jahrling, Peter B; Blaney, Joseph E

2011-09-30

Hemorrhagic smallpox was a rare but severe manifestation of variola virus infection that resulted in nearly 100% mortality. Here we describe intravenous (IV) inoculation of cowpox virus Brighton Red strain in cynomolgus macaques (Macaca fascicularis) which resulted in disease similar in presentation to hemorrhagic smallpox in humans. IV inoculation of macaques resulted in a uniformly lethal disease within 12 days post-inoculation in two independent experiments. Clinical observations and hematological and histopathological findings support hemorrhagic disease. Cowpox virus replicated to high levels in blood (8.0-9.0 log(10) gene copies/mL) and tissues including lymph nodes, thymus, spleen, bone marrow, and lungs. This unique model of hemorrhagic orthopoxvirus infection provides an accessible means to further study orthopoxvirus pathogenesis and to identify virus-specific and nonspecific therapies. Such studies will serve to complement the existing nonhuman primate models of more classical poxviral disease. Published by Elsevier Inc.

Two key arginine residues in the coat protein of Bamboo mosaic virus differentially affect the accumulation of viral genomic and subgenomic RNAs.

PubMed

Hung, Chien-Jen; Hu, Chung-Chi; Lin, Na-Sheng; Lee, Ya-Chien; Meng, Menghsiao; Tsai, Ching-Hsiu; Hsu, Yau-Heiu

2014-02-01

The interactions between viral RNAs and coat proteins (CPs) are critical for the efficient completion of infection cycles of RNA viruses. However, the specificity of the interactions between CPs and genomic or subgenomic RNAs remains poorly understood. In this study, Bamboo mosaic virus (BaMV) was used to analyse such interactions. Using reversible formaldehyde cross-linking and mass spectrometry, two regions in CP, each containing a basic amino acid (R99 and R227, respectively), were identified to bind directly to the 5' untranslated region of BaMV genomic RNA. Analyses of the alanine mutations of R99 and R227 revealed that the secondary structures of CP were not affected significantly, whereas the accumulation of BaMV genomic, but not subgenomic, RNA was severely decreased at 24 h post-inoculation in the inoculated protoplasts. In the absence of CP, the accumulation levels of genomic and subgenomic RNAs were decreased to 1.1%-1.5% and 33%-40% of that of the wild-type (wt), respectively, in inoculated leaves at 5 days post-inoculation (dpi). In contrast, in the presence of mutant CPs, the genomic RNAs remained about 1% of that of wt, whereas the subgenomic RNAs accumulated to at least 87%, suggesting that CP might increase the accumulation of subgenomic RNAs. The mutations also restricted viral movement and virion formation in Nicotiana benthamiana leaves at 5 dpi. These results demonstrate that R99 and R227 of CP play crucial roles in the accumulation, movement and virion formation of BaMV RNAs, and indicate that genomic and subgenomic RNAs interact differently with BaMV CP. © 2013 BSPP AND JOHN WILEY & SONS LTD.

Comparison of contamination rates between preserved and preservative-free fluoroquinolone eyedrops.

PubMed

Kim, Mo Sae; Kim, Hong Kyun; Kim, Joon Mo; Choi, Chul Young

2013-03-01

To evaluate the antimicrobial effectiveness of preservative-free fluoroquinolone products compared with benzalkonium chloride containing fluoroquinolones using the challenge test provided by the United States Pharmacopeia (USP) and the in-use test. 1. Challenge test: to compare the growth of microorganisms between different fluoroquinolone preparations, four test organisms, including Staphylococcus aureus, Pseudomonas aeruginosa, Candida albicans, and Aspergillus niger were chosen among five microorganisms listed by USP 2004. The inoculated products were sampled for microbial survivors at days 7, 14, and 28 following initial inoculation at room temperature. The number of surviving organisms were calculated as a Log10 reduction from the original inocula. 2. In-use test: a total of 100 bottles were collected after instillation of preservative-free fluoroquinolone eyedrops in volunteer patients after 1 week of use. The remaining fluid and tips of the bottles were cultured. Colonies on the plates were counted at the end of the incubation period. All microorganisms were identified by Gram staining and biochemical assays. 1. Challenge test: preservative-free gatifloxacin and levofloxacin demonstrated a lower log reduction against A. niger than preserved fluoroquinolones and preservative-free moxifloxacin at all time points. 2. In-use test: There was no contamination identified on plates inoculated by preservative-free quinolone bottles after 1 week of use in this study. Physicians should be aware of the lower antifungal preservative effectiveness of some preservative-free fluoroquinolone preparations than preserved ones.

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

PubMed

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

2015-10-01

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

Comparative performance of three sampling techniques to detect airborne Salmonella species in poultry farms.

PubMed

Adell, Elisa; Moset, Verónica; Zhao, Yang; Jiménez-Belenguer, Ana; Cerisuelo, Alba; Cambra-López, María

2014-01-01

Sampling techniques to detect airborne Salmonella species (spp.) in two pilot scale broiler houses were compared. Broilers were inoculated at seven days of age with a marked strain of Salmonella enteritidis. The rearing cycle lasted 42 days during the summer. Airborne Salmonella spp. were sampled weekly using impaction, gravitational settling, and impingement techniques. Additionally, Salmonella spp. were sampled on feeders, drinkers, walls, and in the litter. Environmental conditions (temperature, relative humidity, and airborne particulate matter (PM) concentration) were monitored during the rearing cycle. The presence of Salmonella spp. was determined by culture-dependent and molecular methods. No cultivable Salmonella spp. were recovered from the poultry houses' surfaces, the litter, or the air before inoculation. After inoculation, cultivable Salmonella spp. were recovered from the surfaces and in the litter. Airborne cultivable Salmonella spp. Were detected using impaction and gravitational settling one or two weeks after the detection of Salmonella spp. in the litter. No cultivable Salmonella spp. were recovered using impingement based on culture-dependent techniques. At low airborne concentrations, the use of impingement for the quantification or detection of cultivable airborne Salmonella spp. is not recommended. In these cases, a combination of culture-dependent and culture-independent methods is recommended. These data are valuable to improve current measures to control the transmission of pathogens in livestock environments and for optimising the sampling and detection of airborne Salmonella spp. in practical conditions.

HSV-mediated gene transfer of vascular endothelial growth factor to dorsal root ganglia prevents diabetic neuropathy

PubMed Central

Chattopadhyay, M; Krisky, D; Wolfe, D; Glorioso, JC; Mata, M; Fink, DJ

2005-01-01

We examined the utility of herpes simplex virus (HSV) vector-mediated gene transfer of vascular endothelial growth factor (VEGF) in a mouse model of diabetic neuropathy. A replication-incompetent HSV vector with VEGF under the control of the HSV ICP0 promoter (vector T0VEGF) was constructed. T0VEGF expressed and released VEGF from primary dorsal root ganglion (DRG) neurons in vitro, and following subcutaneous inoculation in the foot, expressed VEGF in DRG and nerve in vivo. At 2 weeks after induction of diabetes, subcutaneous inoculation of T0VEGF prevented the reduction in sensory nerve amplitude characteristic of diabetic neuropathy measured 4 weeks later, preserved autonomic function measured by pilocarpine-induced sweating, and prevented the loss of nerve fibers in the skin and reduction of neuropeptide calcitonin gene-related peptide and substance P in DRG neurons of the diabetic mice. HSV-mediated transfer of VEGF to DRG may prove useful in treatment of diabetic neuropathy. PMID:15843809

Evaluation of vaccine candidate potential of deltaaroA, deltahtrA and deltaaroAdeltahtrA mutants of Salmonella enterica subspecies enterica serovar Abortusequi in guinea pigs.

PubMed

Singh, Bhoj Raj; Chandra, Mudit; Hansda, Dhananjoy; Alam, Javed; Babu, Narayanan; Siddiqui, Mehtab Z; Agrawal, Ravi K; Sharma, Gautam

2013-04-01

Salmonella enterica subspecies enterica serovar Abortusequi (S. Abortusequi), a host adapted Salmonella causes abortions, still births and foal mortality in equids. Though known since more than 100 years, it is still a problem in many of the developing countries including India. There is dearth of really good vaccine affording immunity lasting at least for one full gestation. In search of a potential vaccine candidate, three defined deletion mutants (deltaaroA, deltahtrA and deltaaroAdeltahtrA) of S. Abortusequi were tested in guinea pig model for attenuation, safety, immunogenicity, humoral immune response, protective efficacy and persistence in host. The deltahtrA and deltaaroAdeltahtrA mutants were found to be safe on oral inoculation in doses as high as 4.2 x 10(9) cfu/animal. Also through subcutaneous inoculation deltaaroAdeltahtrA mutant did not induce any abortion in pregnant guinea pigs. All the three mutants did not induce any illness or death in 1-2 week-old baby guinea pigs except deltahtrA mutant which caused mortality on intraperitoneal inoculation. Inoculation with mutants protected against challenge and increased breeding efficiency of guinea pigs. After >4.5 months of mutant inoculation, guinea pigs were protected against abortifacient dose of wild type S. Abortusequi and mother guinea pigs also conferred resistance to their babies to the similar challenge. Early humoral immune response of S. Abortusequi mutants was characteristic. Faecal excretion of deltaaroA and htrA mutants was detected up to 45 days of inoculation in guinea pigs while deltaaroAdeltahtrA mutant could not be detected after 21 days of inoculation. The results indicated that the double deletion mutant (deltaaroAdeltahtrA) was the most effective and safe candidate for vaccination against S. Abortusequi through mucosal route of inoculation.

Influence of Age and Dose of African Swine Fever Virus Infections on Clinical Outcome and Blood Parameters in Pigs.

PubMed

Post, Jacob; Weesendorp, Eefke; Montoya, Maria; Loeffen, Willie L

African swine fever (ASF) is a fatal disease for domestic pigs, leading to serious economic losses in countries where ASF is endemic. Despite extensive research, efficient vaccines against ASF are lacking. Since peripheral blood cells are important mediators for vaccines, we study the impact of ASF on blood parameters in pigs with different ages and infected with different doses of ASF virus. Four different groups were studied: (1) 12 weeks of age/low virus dose; (2) 12 weeks of age/high virus dose; (3) 18 weeks of age/low virus dose; and (4) 18 weeks of age/high virus dose. By varying in age and/or ASFV inoculation dose, we monitor blood parameters during different degrees of disease. Thirty percent of the pigs survived the infection with a moderately virulent strain of African swine fever virus (ASFV). Animals that did survive infection were generally older, independent from the inoculation dose used. A firm reduction in many different cell types at 3-5 days postinfection (DPI) was accompanied by an increase in body temperature, followed by clinical signs and mortality from day 6 PI. While blood parameters generally normalized in survivors, γδ T cells and IL-10 levels could be related to mortality. These conclusions should be considered in new approaches for protection against ASF.

Antimicrobials for foodborne pathogens inactivation on cantaloupe

USDA-ARS?s Scientific Manuscript database

This study investigated the efficacy of a new generation disinfectant, namely octenidine dihydrochloride (OH) as pre- and post-harvest treatment for reducing Listeria monocytogenes, Salmonella spp., and Escherichia coli O157:H7 on cantaloupe surface. At farm, cantaloupes were dip-inoculated with L. ...

Isolation and purification of Gallid herpesvirus 2 strains currently distributed in Japan.

PubMed

Machida, Yuka; Murata, Shiro; Matsuyama-Kato, Ayumi; Isezaki, Masayoshi; Taneno, Akira; Sakai, Eishi; Konnai, Satoru; Ohashi, Kazuhiko

2017-01-20

Gallid herpesvirus 2 (GaHV-2) causes malignant lymphomas in chickens (Marek's disease, MD). Although MD is controlled through vaccination efforts, field isolates of GaHV-2 have increased in virulence worldwide and even cause MD in vaccinated chickens. GaHV-2 strains are classified into four categories (mild, virulent, very virulent and very virulent +) based on the virulence exhibited in experimental infection in unvaccinated or MD-vaccinated susceptible chickens. Although MD cases are sporadically reported in Japan, the recent field strains of GaHV-2 in Japan have not been characterized. During isolation of recent field strains by using primary chicken kidney cell cultures, a method classically used for GaHV-2 isolation, vaccine strains were simultaneously isolated. Therefore, it is necessary to separate vaccine strains to characterize the virulence and pathogenicity of the GaHV-2 strains currently distributed in Japan. In this study, we prepared cell suspensions from the spleens of MD-symptomatic chickens, inoculated day-old-chicks and isolated GaHV-2 strains by primary chicken kidney cell cultures at 2-3 weeks post inoculation. The isolated strains were passaged several times on chicken embryo fibroblast cells, and PCR analysis revealed that the isolated strains were not contaminated with vaccine strains. Moreover, the contaminant vaccine strains were completely removed by the purification of plaques observed in chicken kidney cells. These procedures are necessary to isolate GaHV-2 field strains from vaccine strains in order to carry out future studies to characterize these strains and glean insights into GaHV-2 virulence and pathogenicity.

Generation of transgenic watermelon resistant to Zucchini yellow mosaic virus and Papaya ringspot virus type W.

PubMed

Yu, Tsong-Ann; Chiang, Chu-Hui; Wu, Hui-Wen; Li, Chin-Mei; Yang, Ching-Fu; Chen, Jun-Han; Chen, Yu-Wen; Yeh, Shyi-Dong

2011-03-01

Zucchini yellow mosaic virus (ZYMV) and Papaya ringspot virus type W (PRSV W) are major limiting factors for production of watermelon worldwide. For the effective control of these two viruses by transgenic resistance, an untranslatable chimeric construct containing truncated ZYMV coat protein (CP) and PRSV W CP genes was transferred to commercial watermelon cultivars by Agrobacterium-mediated transformation. Using our protocol, a total of 27 putative transgenic lines were obtained from three cultivars of 'Feeling' (23 lines), 'China baby' (3 lines), and 'Quality' (1 line). PCR and Southern blot analyses confirmed that the chimeric construct was incorporated into the genomic DNA of the transformants. Greenhouse evaluation of the selected ten transgenic lines of 'Feeling' cultivar revealed that two immune lines conferred complete resistance to ZYMV and PRSV W, from which virus accumulation were not detected by Western blotting 4 weeks after inoculation. The transgenic transcript was not detected, but small interfering RNA (siRNA) was readily detected from the two immune lines and T(1) progeny of line ZW 10 before inoculation, indicating that RNA-mediated post-transcriptional gene silencing (PTGS) is the underlying mechanism for the double-virus resistance. The segregation ratio of T(1) progeny of the immune line ZW10 indicated that the single inserted transgene is nuclearly inherited and associated with the phenotype of double-virus resistance as a dominant trait. The transgenic lines derived from the commercial watermelon cultivars have great potential for control of the two important viruses and can be implemented directly without further breeding.

Rhesus macaque model of chronic opiate dependence and neuro-AIDS: longitudinal assessment of auditory brainstem responses and visual evoked potentials

PubMed Central

Riazi, Mariam; Marcario, Joanne K; Samson, Frank K.; Kenjale, Himanshu; Adany, Istvan; Staggs, Vincent; Ledford, Emily; Marquis, Janet; Narayan, Opendra; Cheney, Paul D.

2013-01-01

Our work characterizes the effects of opiate (morphine) dependence on auditory brainstem and visual evoked responses in a rhesus macaque model of neuro-AIDS utilizing a chronic continuous drug delivery paradigm. The goal of this study was to clarify whether morphine is protective, or if it exacerbates simian immunodeficiency virus (SIV) related systemic and neurological disease. Our model employs a macrophage tropic CD4/CCR5 co-receptor virus, SIVmac239 (R71/E17), which crosses the blood brain barrier shortly after inoculation and closely mimics the natural disease course of human immunodeficiency virus (HIV) infection. The cohort was divided into 3 groups: morphine only, SIV only, and SIV + morphine. Evoked potential (EP) abnormalities in sub-clinically infected macaques were evident as early as eight weeks post-inoculation. Prolongations in EP latencies were observed in SIV-infected macaques across all modalities. Animals with the highest CSF viral loads and clinical disease showed more abnormalities than those with sub-clinical disease, confirming our previous work (Raymond et al, 1998, 1999, 2000). Although some differences were observed in auditory and visual evoked potentials in morphine treated compared to untreated SIV-infected animals, the effects were relatively small and not consistent across evoked potential type. However, morphine treated animals with subclinical disease had a clear tendency toward higher virus loads in peripheral and CNS tissues (Marcario et al., 2008) suggesting that if had been possible to follow all animals to end-stage disease, a clearer pattern of evoked potential abnormality might have emerged. PMID:19283490

The effects of chronic binge alcohol on the genital microenvironment of simian immunodeficiency virus-infected female rhesus macaques.

PubMed

Loganantharaj, Nisha; Nichols, Whitney A; Bagby, Gregory J; Volaufova, Julia; Dufour, Jason; Martin, David H; Nelson, Steve; Amedee, Angela M

2014-08-01

Alcohol abuse is a widespread problem among those at risk for and living with HIV and can impact transmission and disease progression. In this study we sought to use the simian immunodeficiency virus (SIV)-macaque model to evaluate the immunological and virological changes in the genital microenvironment of females exposed to chronic alcohol. Female rhesus macaques were treated with alcohol (n=6) or isocaloric sucrose (n=6) for 3 months and then inoculated with SIVmac251. To assess the effects of chronic alcohol on SIV disease and the genital microenvironment, we quantified plasma and genital SIV levels, measured inflammatory cells in genital fluids, and characterized microbial flora by gram stains over 10 weeks post-SIV infection. Following 3 months of alcohol/sucrose treatment, significant differences were observed in the vaginal microenvironment of alcohol-treated animals as compared to controls. Microbial flora of alcohol-treated animals had decreased levels of lactobacillus morphotypes and increased levels of gram-positive cocci relative to sucrose controls. Alcohol-treated animals were also more likely to have white blood cells in vaginal fluids prior to SIV inoculation, which persisted through viral set point. Similar levels of cell-free SIV were observed in plasma and vaginal fluids of both groups, but alcohol-treated animals had a higher incidence and levels of cell-associated SIV shed in vaginal secretions. Chronic alcohol treatment negatively impacts the genital microenvironment prior to and over the course of SIV infection and may increase the risk of genital virus shedding and transmission.

In vivo imaging and characterization of hypoxia-induced neovascularization and tumor invasion.

PubMed

Lungu, Gina F; Li, Meng-Lin; Xie, Xueyi; Wang, Lihong V; Stoica, George

2007-01-01

Hypoxia is a critical event in tumor progression and angiogenesis. Hypoxia can be detected noninvasively by a novel spectroscopic photoacoustic tomography technology (SPAT) and this finding is supported by our molecular biology investigation aimed to elucidate the etiopathogenesis of SPAT detected hypoxia and angiogenesis. The present study provides an integrated approach to define oxygen status (hypoxia) of intracranial tumor xenografts using spectroscopic photoacoustic tomography. Brain tumors can be identified based on their distorted vascular architecture and oxygen saturation (SO2) images. Noninvasive in vivo tumor oxygenation imaging using SPAT is based on the spectroscopic absorption differences between oxyhemoglobin (O2Hb) and deoxyhemoblobin (HHb). Sprague-Dawley rats inoculated intracranially with ENU1564, a carcinogen-induced rat mammary adenocarcinoma cell line, were imaged with SPAT three weeks post inoculation. Proteins important for tumor angiogenesis and invasion were detected in hypoxic brain foci identified by SPAT and were elevated compared with control brain. Immunohistochemistry, Western blotting, and semi-quantitative RT-PCR showed that HIF-1 alpha, VEGF-A, and VEGFR2 (Flk-1) protein and mRNA expression levels were significantly higher (P < 0.05) in brain tumor tissues compared to normal brain. Gelatin zymography and RT-PCR demonstrated the upregulation of MMP-9 in tumor foci compared with brain control. Together these results suggest the critical role of hypoxia in driving tumor angiogenesis and invasion through upregulation of target genes important for these functions. Moreover this report validates our hypothesis that a novel noninvasive technology (SPAT) developed in our laboratory is suitable for detection of tumors, hypoxia, and angiogenesis.

Experimental SSM-CVB3 infection in macaques.

PubMed

Han, Tiesuo; He, Wenqi; Song, Deguang; Zhao, Kui; Wu, Chenchen; Gao, Feng; Lu, Huijun; Gai, Xianying; Wang, Xinrui; Li, Fei; Ji, Cuicui; Lin, Xijuan

2012-02-01

To evaluate the pathogenicity of SSM-CVB3 in a macaque model. The clinical symptoms of macaques were recorded; hematological, biochemical and histopathological evaluations were completed; viral titers and neutralization titers (NT-titers) in sera were tested; and the mRNA levels of SSM-CVB3, coxsackievirus and adenovirus receptor (CAR) and decay accelerating factor (DAF) were determined. After SSM-CVB3 infection, the macaques showed a lack of activity, a poor appetite, a higher body temperature, and severe diarrhea. The macaques also developed hematuria and albuminuria at 4 to 10 days post-inoculation. Virus titers (5.1-6.5 LogTCID(50)/mL) were higher at 6 to 10 days post-inoculation, and NT-titers (6.5-7.3 Log2) reached plateaus at 8 to 14 days post-inoculation. The infected macaques developed serious anemia with decreased RBC and WBC, but the percentages of LYM were increased. The levels of CK, CK-MB, AST and ALT in the sera were 84-169 U/L, 87.6-271.1 U/L, 43-87 U/L and 43-82 U/L, respectively, and all of those were higher than normal. Histological analysis showed obvious cardiac, hepatic and renal damages in the infected macaques and the mRNA contents of SSM-CVB3, CAR and DAF in the heart, liver and kidneys of infected macaques were higher (P<0.05). This was the first report on experimental SSM-CVB3 infections in macaques with serious hepatic and renal damage, except for myocarditis. The information obtained from this study suggests that the SSM-CVB3 strain and this macaque model could be used for studying CVB3-induced cardiac, hepatic or renal diseases. Copyright © 2011 Elsevier Inc. All rights reserved.

Host Identity Matters in the Amphibian-Batrachochytrium dendrobatidis System: Fine-Scale Patterns of Variation in Responses to a Multi-Host Pathogen

PubMed Central

Gervasi, Stephanie; Gondhalekar, Carmen; Olson, Deanna H.; Blaustein, Andrew R.

2013-01-01

Species composition within ecological assemblages can drive disease dynamics including pathogen invasion, spread, and persistence. In multi-host pathogen systems, interspecific variation in responses to infection creates important context dependency when predicting the outcome of disease. Here, we examine the responses of three sympatric host species to a single fungal pathogen, Batrachochytrium dendrobatidis, which is associated with worldwide amphibian population declines and extinctions. Using an experimental approach, we show that amphibian species from three different genera display significant differences in patterns of pathgen-induced mortality as well as the magnitude and temporal dynamics of infection load. We exposed amphibians to one of four inoculation dose treatments at both larval and post- metamorphic stages and quantified infection load on day 8 and day 15 post-inoculation. Of the three species examined, only one (the Pacific treefrog; Pseudacris regilla) displayed “dose-dependent” responses; survival was reduced and infection load was elevated as inoculation dose was increased. We observed a reduction in survival but no differences in infection load across pathogen treatments in Cascades frogs (Rana cascadae). Western toads (Anaxyrus boreas) displayed differences in infection load but no differences in survival across pathogen treatments. Within species, responses to the pathogen varied with life history stage, and the most heavily infected species at the larval stage was different from the most heavily infected species at the post-metamorphic stage. Temporal changes in infection load were species and life history stage-specific. We show that variation in susceptibility to this multi-host pathogen is complex when viewed at a fine-scale and may be mediated through intrinsic host traits. PMID:23382904

Effects of arbuscular mycorrhizal inoculation and biochar amendment on maize growth, cadmium uptake and soil cadmium speciation in Cd-contaminated soil.

PubMed

Liu, Ling; Li, Jiwei; Yue, Feixue; Yan, Xinwei; Wang, Fayuan; Bloszies, Sean; Wang, Yanfang

2018-03-01

Experiments conducted to understand how arbuscular mycorrhizal (AM) inoculation or biochar application affect plant growth and heavy metal uptake have thus far looked at single applications of either soil amendment. There is little evidence of their synergistic effects, in particular for plants grown in cadmium (Cd) contaminated soil. We conducted a mesocosm experiment to investigate the effect of AM inoculation (Glomus intraradices BEG 141) and/or wheat-straw biochar amendment on maize (Zea mays L. cv. Hongdan No. 897) growth, antioxidant enzymatic activities, and Cd uptake, as well as soil Cd speciation under applications of 0, 3, 6 mg Cd per kg soil. Applying either AM inoculant or biochar alone significantly increased maize growth and reduced Cd uptake. Furthermore, solo AM inoculation alleviating Cd stress more fully than biochar, in turn facilitating maize growth and decreasing soil Cd translocation into plant tissue. Still, solo biochar amendment was more effective at inducing soil alkalinization and contributing to Cd immobilization. Adding biochar together with AM inoculant significantly promoted fungal populations compared to a control. Amending soil with AM inoculant and biochar together produced the largest increase in maize growth and decrease in tissue Cd concentrations. This effect was additive, with 79.1% greater biomass, 51.42%, 82.91%, 43.96% higher activities of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and 50.06%, 67.19%, 58.04% and 76.19% lower Cd concentrations in roots, stems, leaves, and ears, respectively, at a 6 mg kg -1 Cd contamination rate. The combined treatment also had a synergistic effect on inducing soil alkalinization and causing Cd immobilization, and decreasing Cd phytoavailability and post-harvest transfer risks. These results suggest that AM inoculation in combination with biochar application may be applicable not only for maize production but also for phytostabilization of Cd-contaminated soil. Copyright © 2017 Elsevier Ltd. All rights reserved.

In Vivo Volatile Organic Compound Signatures of Mycobacterium avium subsp. paratuberculosis

PubMed Central

Bergmann, Andreas; Trefz, Phillip; Fischer, Sina; Klepik, Klaus; Walter, Gudrun; Steffens, Markus; Ziller, Mario; Schubert, Jochen K.; Reinhold, Petra; Köhler, Heike; Miekisch, Wolfram

2015-01-01

Mycobacterium avium ssp. paratuberculosis (MAP) is the causative agent of a chronic enteric disease of ruminants. Available diagnostic tests are complex and slow. In vitro, volatile organic compound (VOC) patterns emitted from MAP cultures mirrored bacterial growth and enabled distinction of different strains. This study was intended to determine VOCs in vivo in the controlled setting of an animal model. VOCs were pre-concentrated from breath and feces of 42 goats (16 controls and 26 MAP-inoculated animals) by means of needle trap microextraction (breath) and solid phase microextraction (feces) and analyzed by gas chromatography/ mass spectrometry. Analyses were performed 18, 29, 33, 41 and 48 weeks after inoculation. MAP-specific antibodies and MAP-specific interferon-γ-response were determined from blood. Identities of all marker-VOCs were confirmed through analysis of pure reference substances. Based on detection limits in the high pptV and linear ranges of two orders of magnitude more than 100 VOCs could be detected in breath and in headspace over feces. Twenty eight substances differed between inoculated and non-inoculated animals. Although patterns of most prominent substances such as furans, oxygenated substances and hydrocarbons changed in the course of infection, differences between inoculated and non-inoculated animals remained detectable at any time for 16 substances in feces and 3 VOCs in breath. Differences of VOC concentrations over feces reflected presence of MAP bacteria. Differences in VOC profiles from breath were linked to the host response in terms of interferon-γ-response. In a perspective in vivo analysis of VOCs may help to overcome limitations of established tests. PMID:25915653

Study of the potential involvement of pseudorabies virus in swine respiratory disease.

PubMed

Iglesias, G J; Trujano, M; Lokensgard, J; Molitor, T

1992-01-01

In order to investigate the potential involvement of pseudorabies virus (PRV) in swine respiratory disease, nine week old pigs were intranasally inoculated with the PRV strain 4892. Two doses of infection were used: 10(4.5) median tissue culture infectious doses (TCID50)/pig and 10(3.5) TCID50/pig, with ten pigs per group. In the group of pigs inoculated with 10(4.5) TCID50, seven out of ten pigs died within six days after inoculation. The mortality rate in the group of pigs inoculated with the lower dose was only two out of ten and, there were several pigs in this group that showed signs of respiratory distress besides some mild nervous signs. Pseudorabies virus was isolated from various tissues collected postmortem, including alveolar macrophages. Virus localization in tissues was also detected by in situ hybridization. The histopathological examination of the respiratory tract tissues revealed a pathological process that was progressing from mild pneumonia to severe suppurative bronchopneumonia. The isolation of virus from alveolar macrophages provides support to the hypothesis that replication of PRV during the course of infection produces an impairment of the defense mechanisms in the respiratory tract.

Aspergillosis in waterfowl

USGS Publications Warehouse

Herman, Carlton M.; Sladen, William J. L.

1958-01-01

Aspergillosis, a respiratory disease most commonly caused by the fungus Aspergillus fumigatus, although frequently the cause of losses in captive birds, has been little studied in wild waterfowl and other avian species. Evidence indicates this to be of importance in the wild, and studies were conducted to determine factors relating to its epizoology. Field collections from corn and other plants have yielded infective spores of Aspergillus which were inoculated into experimental chickens and ducklings and then re-isolated from characteristic lesions. A technique was developed for inoculating suspensions of known numbers of spores directly into one of the posterior thoracic airsacs. It was demonstrated that less than one million spores of A. fumigatus killed less than one-half of the experimental chickens, 10 million spores killed over 80 per cent and 50 million killed all inoculated chickens as well as ducklings. Older birds were able to survive as many as 500 million spores except when in a weakened condition. Chickens usually started dying within two days after inoculation while those that survived as long as 11 days usually fully recovered by three weeks. Pathological involvement usually was confined to lungs and airsacs. The procedures and techniques involved in these studies were illustrated on a color motion picture.

Impact of experimental human pneumococcal carriage on nasopharyngeal bacterial densities in healthy adults.

PubMed

Shak, Joshua R; Cremers, Amelieke J H; Gritzfeld, Jenna F; de Jonge, Marien I; Hermans, Peter W M; Vidal, Jorge E; Klugman, Keith P; Gordon, Stephen B

2014-01-01

Colonization of the nasopharynx by Streptococcus pneumoniae is a necessary precursor to pneumococcal diseases that result in morbidity and mortality worldwide. The nasopharynx is also host to other bacterial species, including the common pathogens Staphylococcus aureus, Haemophilus influenzae, and Moraxella catarrhalis. To better understand how these bacteria change in relation to pneumococcal colonization, we used species-specific quantitative PCR to examine bacterial densities in 52 subjects 7 days before, and 2, 7, and 14 days after controlled inoculation of healthy human adults with S. pneumoniae serotype 6B. Overall, 33 (63%) of subjects carried S. pneumoniae post-inoculation. The baseline presence and density of S. aureus, H. influenzae, and M. catarrhalis were not statistically associated with likelihood of successful pneumococcal colonization at this study's sample size, although a lower rate of pneumococcal colonization in the presence of S. aureus (7/14) was seen compared to that in the presence of H. influenzae (12/16). Among subjects colonized with pneumococci, the number also carrying either H. influenzae or S. aureus fell during the study and at 14 days post-inoculation, the proportion carrying S. aureus was significantly lower among those who were colonized with S. pneumoniae (p = 0.008) compared to non-colonized subjects. These data on bacterial associations are the first to be reported surrounding experimental human pneumococcal colonization and show that co-colonizing effects are likely subtle rather than absolute.

In vivo induction of neutrophil extracellular traps by Mycobacterium tuberculosis in a guinea pig model.

PubMed

Filio-Rodríguez, Georgina; Estrada-García, Iris; Arce-Paredes, Patricia; Moreno-Altamirano, María M; Islas-Trujillo, Sergio; Ponce-Regalado, M Dolores; Rojas-Espinosa, Oscar

2017-10-01

In 2004, a novel mechanism of cellular death, called 'NETosis', was described in neutrophils. This mechanism, different from necrosis and apoptosis, is characterized by the release of chromatin webs admixed with microbicidal granular proteins and peptides (NETs). NETs trap and kill a variety of microorganisms. Diverse microorganisms, including Mycobacterium tuberculosis, are NET inducers in vitro. The aim of this study was to examine whether M. tuberculosis can also induce NETs in vivo and if the NETs are bactericidal to the microorganism. Guinea pigs were intradermally inoculated with M. tuberculosis H37Rv, and the production of NETs was investigated at several time points thereafter. NETs were detected as early as 30 min post-inoculation and were clearly evident by 4 h post-inoculation. NETs produced in vivo contained DNA, myeloperoxidase, elastase, histones, ROS and acid-fast bacilli. Viable and heat-killed M. tuberculosis, as well as Mycobacterium bovis BCG were efficient NET inducers, as were unilamellar liposomes prepared with lipids from M. tuberculosis. In vitro, guinea pig neutrophils also produced NETs in response to M. tuberculosis. However, neither the in vivo nor the in vitro-produced NETs were able to kill M. tuberculosis. Nevertheless, in vivo, neutrophils might propitiate recruitment and activation of more efficient microbicidal cells.

Examining the safety of respiratory and intravenous inoculation of Bdellovibrio bacteriovorus and Micavibrio aeruginosavorus in a mouse model.

PubMed

Shatzkes, Kenneth; Chae, Richard; Tang, Chi; Ramirez, Gregory C; Mukherjee, Somdatta; Tsenova, Liana; Connell, Nancy D; Kadouri, Daniel E

2015-08-07

Bdellovibrio spp. and Micavibrio spp. are Gram-negative predators that feed on other Gram-negative bacteria, making predatory bacteria potential alternatives to antibiotics for treating multi-drug resistant infections. While the ability of predatory bacteria to control bacterial infections in vitro is well documented, the in vivo effect of predators on a living host has yet to be extensively examined. In this study, respiratory and intravenous inoculations were used to determine the effects of predatory bacteria in mice. We found no reduction in mouse viability after intranasal or intravenous inoculation of B. bacteriovorus 109J, HD100 or M. aeruginosavorus. Introducing predators into the respiratory tract of mice provoked a modest inflammatory response at 1 hour post-exposure, but was not sustained at 24 hours, as measured by RT-qPCR and ELISA. Intravenous injection caused an increase of IL-6 in the kidney and spleen, TNF in the liver and CXCL-1/KC in the blood at 3 hours post-exposure, returning to baseline levels by 18 hours. Histological analysis of tissues showed no pathological changes due to predatory bacteria. Furthermore, qPCR detected predators were cleared from the host quickly and efficiently. This work addresses some of the safety concerns regarding the potential use of predatory bacteria as a live antibiotic.

The development of immune responses in Balb/c mice following inoculation with attenuated or virulent Neospora caninum tachyzoites.

PubMed

Bartley, P M; Wright, S E; Maley, S W; Buxton, D; Nath, M; Innes, E A

2009-07-01

Balb/c mice were inoculated intraperitoneally (i.p.) with either 5 x 10(6) live virulent (group 1) or 5 x 10(6) live attenuated (group 2) tachyzoites, or Vero cells (group 3). Animals were killed at 0, 14, 28 and 42 days post-inoculation (p.i.), with the remaining mice receiving a lethal challenge on day 48 p.i. Serum, spleen and brain samples were collected post-mortem to examine humoral and cell-mediated immune responses as well as pathological lesions and to quantify parasite loads. On day 14 p.i. group 2 (attenuated) demonstrated statistically significant (P < 0.001) lower levels of mean morbidity and weight loss, while also showing significantly (P = 0.01) higher levels of splenocyte proliferation and IFN-gamma production (P = 0.003), compared to group 1 (virulent). Histology of brain samples showed milder lesions and a lower incidence of positive immunohistochemistry, demonstrating tachyzoites and tissue cysts, and statistically significant (P = 0.03) lower mean burdens of parasite DNA in group 2 (attenuated) compared to group 1 (virulent). All mice in group 2 were protected following challenge on day 48 p.i. whereas naïve control mice succumbed to the challenge. No mice from group 1 (virulent) survived beyond day 24 p.i. so they were not included in the challenge.

Novel antimicrobials to control foodborne pathogens on cantaloupe

USDA-ARS?s Scientific Manuscript database

This study investigated the efficacy of a new generation disinfectant, namely octenidine dihydrochloride (OH) as pre- and post-harvest treatment for reducing Listeria monocytogenes, Salmonella spp., and Escherichia coli O157:H7 on cantaloupe surface. At farm, cantaloupes were dip-inoculated with L. ...

Longitudinal study of experimental induction of AA amyloidosis in mice seeded with homologous and heterologous AA fibrils.

PubMed

Muhammad, Naeem; Murakami, Tomoaki; Inoshima, Yasuo; Ishiguro, Naotaka

2016-09-01

To investigate pathogenesis and kinetics of experimentally induced murine AA amyloidosis seeded with homologous (murine) and heterologous (bovine) AA fibrils. Experimental AA amyloidosis was induced by administration of inflammatory stimulus and preformed AA fibrils to a total of 111 female C57/Black mice. In this longitudinal study, heterologous (bovine) as well as homologous (murine) AA fibrils were injected intraperitoneally to mice in various combinations. Re-stimulation was done at 120 or 300 days post first inoculation. To analyze the intensity of amyloid depositions in mice organs, immunohistochemical techniques and image J software were used. Assessment of cytokines level in sera was done using a Mouse Th1/Th2/Th17 Cytokine CBA Kit. Incidence and severity of AA amyloidosis were quite low in mice inoculated with heterologous bovine AA fibrils than homologous murine one. Homologous AA fibrils administration at first and second inoculation caused maximum amount of amyloid depositions and severe systemic form of amyloidosis. Increase in the level of pro-inflammatory cytokine IL-6 was observed after first inoculation, while second inoculation caused a further increase in the level of anti-inflammatory cytokine IL-10. AA amyloidosis can be induced by heterologous as well as homologous AA fibrils. Severity of AA amyloidosis induced with homologous AA fibrils is higher compared to heterologous AA fibrils.

The pathological and molecular but not clinical phenotypes are maintained after second passage of experimental atypical bovine spongiform encephalopathy in cattle.

PubMed

Konold, Timm; Phelan, Laura J; Clifford, Derek; Chaplin, Melanie J; Cawthraw, Saira; Stack, Michael J; Simmons, Marion M

2014-10-02

Atypical bovine spongiform encephalopathies (BSEs), classified as H-type and L-type BSE based on the Western immunoblot profiles, are naturally occurring diseases in cattle, which are phenotypically different to classical BSE. Transmission studies in cattle using the intracerebral route resulted in disease where the phenotypes were maintained irrespective of BSE type but clinically affected cattle with a shorter survival time displayed a nervous form whereas cattle with a longer survival time displayed a dull form. A second transmission study is reported here where four cattle were intracerebrally inoculated with brain tissue from experimentally infected cattle presenting with either the nervous or dull form of H- or L-type BSE to determine whether the phenotype is maintained. The four inoculated cattle were culled at 16.5-19.5 months post inoculation after presenting with difficulty getting up, a positive scratch response (all) and dullness (three cattle), which was not observed in two non-inoculated control cattle, each housed with either group of inoculated cattle. Only the inoculated cattle had detectable prion protein in the brain based on immunohistochemical examination, and the Western immunoblot profile was consistent with the H-type or L-type BSE of the respective donor cattle. Second passage of H-type and L-type BSE in cattle produced a TSE where the majority of cattle displayed the dull form regardless of clinical disease form of the donor cattle. The pathological and molecular phenotypes of H- and L-type BSE were maintained.

Pathogenesis of highly virulent African swine fever virus in domestic pigs exposed via intraoropharyngeal, intranasopharyngeal, and intramuscular inoculation, and by direct contact with infected pigs.

PubMed

Howey, Erin B; O'Donnell, Vivian; de Carvalho Ferreira, Helena C; Borca, Manuel V; Arzt, Jonathan

2013-12-26

To investigate the pathogenesis of African swine fever virus (ASFV), domestic pigs (n=18) were challenged with a range (10(2)-10(6) 50% hemadsorbing doses (HAD50)) of the highly virulent ASFV-Malawi strain by inoculation via the intraoropharyngeal (IOP), intranasopharyngeal (INP), or intramuscular (IM) routes. A subsequent contact challenge experiment was performed in which six IOP-inoculated donor pigs were allowed to have direct contact (DC) with six naïve pigs for exposure times that varied from 24 to 72 h. All challenge routes resulted in clinical progression and postmortem lesions similar to those previously described in experimental and natural infection. The onset of clinical signs occurred between 1 and 7 days post inoculation (dpi) and included pyrexia with variable progression to obtundation, hematochezia, melena, moribundity and death with a duration of 4-11 days. Viremia was first detected between 4 and 5 dpi in all inoculation groups whereas ASFV shedding from the nasal cavity and tonsil was first detected at 3-9 dpi. IM and DC were the most consistent modes of infection, with 12/12 (100%) of pigs challenged by these routes becoming infected. Several clinical and virological parameters were significantly different between IM and DC groups indicating dissimilarity between these modes of infection. Amongst the simulated natural routes, INP inoculation resulted in the most consistent progression of disease across the widest range of doses whilst preserving simulation of natural exposure and therefore may provide a superior system for pathogenesis and vaccine efficacy investigation. Published by Elsevier B.V.

Overcoming nursing barriers to intensive care unit early mobilisation: A quality improvement project.

PubMed

Hunter, Oluwatobi O; George, Elisabeth L; Ren, Dianxu; Morgan, Douglas; Rosenzweig, Margaret; Klinefelter Tuite, Patricia

2017-06-01

To increase adherence with intensive care unit mobility by developing and implementing a mobility training program that addresses nursing barriers to early mobilisation. An intensive care unit mobility training program was developed, implemented and evaluated with a pre-test, immediate post-test and eight-week post-test. Patient mobility was tracked before and after training. A ten bed cardiac intensive care unit. The training program's efficacy was measured by comparing pre-test, immediate post-test and 8-week post-test scores. Patient mobilisation rates before and after training were compared. Protocol compliance was measured in the post training group. Nursing knowledge increased from pre-test to immediate post-test (p<0.0001) and pre-test to 8-week post-test (p<0.0001). Mean test scores decreased by seven points from immediate post-test (80±12) to 8-week post-test (73±14). Fear significantly decreased from pre-test to immediate post-test (p=0.03), but not from pre-test to 8-week post-test (p=0.06) or immediate post-test to 8-week post-test (p=0.46). Post training patient mobility rates increased although not significantly (p=0.07). Post training protocol compliance was 78%. The project successfully increased adherence with intensive care unit mobility and indicates that a training program could improve adoption of early mobility. Copyright © 2016 Elsevier Ltd. All rights reserved.

Biosorption of metal and salt tolerant microbial isolates from a former uranium mining area. Their impact on changes in rare earth element patterns in acid mine drainage.

PubMed

Haferburg, Götz; Merten, Dirk; Büchel, Georg; Kothe, Erika

2007-12-01

The concentration of metals in microbial habitats influenced by mining operations can reach enormous values. Worldwide, much emphasis is placed on the research of resistance and biosorptive capacities of microorganisms suitable for bioremediation purposes. Using a collection of isolates from a former uranium mining area in Eastern Thuringia, Germany, this study presents three Gram-positive bacterial strains with distinct metal tolerances. These strains were identified as members of the genera Bacillus, Micrococcus and Streptomyces. Acid mine drainage (AMD) originating from the same mining area is characterized by high metal concentrations of a broad range of elements and a very low pH. AMD was analyzed and used as incubation solution. The sorption of rare earth elements (REE), aluminum, cobalt, copper, manganese, nickel, strontium, and uranium through selected strains was studied during a time course of four weeks. Biosorption was investigated after one hour, one week and four weeks by analyzing the concentrations of metals in supernatant and biomass. Additionally, dead biomass was investigated after four weeks of incubation. The maximum of metal removal was reached after one week. Up to 80% of both Al and Cu, and more than 60% of U was shown to be removed from the solution. High concentrations of metals could be bound to the biomass, as for example 2.2 mg/g U. The strains could survive four weeks of incubation. Distinct and different patterns of rare earth elements of the inoculated and non-inoculated AMD water were observed. Changes in REE patterns hint at different binding types of heavy metals regarding incubation time and metabolic activity of the cells. (c) 2008 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Effect of manual therapy versus proprioceptive neuromuscular facilitation in dynamic balance, mobility and flexibility in field hockey players. A randomized controlled trial.

PubMed

Espí-López, Gemma V; López-Martínez, Susana; Inglés, Marta; Serra-Añó, Pilar; Aguilar-Rodríguez, Marta

2018-04-22

To compare the effectiveness of a specific Manual Therapy (MT) protocol applied to field hockey players (FHP), versus a Proprioceptive Neuromuscular Facilitation (PNF) protocol, in the improvement of dynamic balance, active range of movement and lumbar flexibility one-week and four-weeks after the treatment. Randomized controlled trial. Participants were assigned to 2 groups: MT and PNF. 30 min' sessions were performed once a week for three weeks. Three evaluations were performed: basal, one-week and four-weeks post-treatment. University of Valencia (Spain). 22 in MT group and 20 in PNF group. Dynamic Balance, measured with Star Excursion Balance Test; Active Range of Motion (ROM), using a manual goniometer and Lumbar Flexibility, assessed with Fingertip-to-floor test. Both groups significantly improved in lateral and medial dynamic balance one-week post-treatment (p < 0.05); but the improvement in the MT group lasted until the fourth-week after treatment in both reaches (lateral and medial) (p < 0.05). MT group also obtained significant improvements in dorsal flexion of the ankle in the fourth-week post-treatment (p < 0.05) and in lumbar flexibility one-week post-treatment (p < 0.05). MT and PNF improve dynamic balance one-week post-treatment; however, the improvement obtained through MT is maintained four-weeks later. Only MT improves dorsal flexion of the ankle four-weeks post-treatment and lumbar flexibility one-week post-treatment. Copyright © 2018 Elsevier Ltd. All rights reserved.

Type 2 diabetes mellitus BALB/c mice are more susceptible to granulomatous amoebic encephalitis: Immunohistochemical study.

PubMed

Omaña-Molina, Maritza; Sanchez-Rocha, Raquel; Hernandez-Martinez, Dolores; Romero Grijalva, Miriam; Salinas-Lara, Citlaltepetl; Rodriguez-Sosa, Miriam; Juarez-Avelar, Imelda; Salazar-Villatoro, Lizbeth; Gonzalez-Robles, Arturo; Mendez-Cruz, Adolfo Rene; Aley-Medina, Patricia; Espinosa-Villanueva, Jesus; Castelan-Ramirez, Ismael; Lorenzo-Morales, Jacob

2017-12-01

Granulomatous amoebic encephalitis (GAE) is a chronic, difficult to resolve infection caused by amphizoic amoebae of the genus Acanthamoeba, which in most cases occurs in immunosuppressed persons or with chronic diseases such as diabetes. In this study, we describe the early events of A. culbertsoni infection of GAE in diabetic mice model. Diabetes was induced in male BALB/c mice, with a dose of streptozotocin (130 mg/kg). Healthy and diabetic mice were inoculated via intranasal with 1 × 10 6 trophozoites of A. culbertsoni. Then were sacrificed and fixed by perfusion at 24, 48, 72 and 96 h post-inoculation, the brains and nasopharyngeal meatus were processed to immunohistochemical analysis. Invasion of trophozoites in diabetic mice was significantly greater with respect to inoculated healthy mice. Trophozoites and scarce cysts were immunolocalized in respiratory epithelial adjacent bone tissue, olfactory nerve packets, Schwann cells and the epineurium base since early 24 h post-inoculation. After 48 h, trophozoites were observed in the respiratory epithelium, white matter of the brain, subcortical central cortex and nasopharyngeal associated lymphoid tissue (NALT). At 72 h, cysts and trophozoites were immunolocalized in the olfactory bulb with the presence of a low inflammatory infiltrate characterized by polymorphonuclear cells. Scarce amoebae were observed in the granular layer of the cerebellum without evidence of inflammation or tissue damage. No amoebas were observed at 96 h after inoculation, suggesting penetration to other tissues at this time. In line with this, no inflammatory infiltrate was observed in the surrounding tissues where the amoebae were immunolocalized, which could contribute to the rapid spread of infection, particularly in diabetic mice. All data suggest that trophozoites invade the tissues by separating the superficial cells, penetrating between the junctions without causing cytolytic effect in the adjacent cells and subsequently reaching the CNS, importantly, diabetes increases the susceptibility to amoebae infection, which could favor the GAE development. Copyright © 2017 Elsevier Inc. All rights reserved.

Early initiation of post-sternotomy cardiac rehabilitation exercise training (SCAR): study protocol for a randomised controlled trial and economic evaluation

PubMed Central

Lobley, Grace; Worrall, Sandra; Powell, Richard; Kimani, Peter K; Banerjee, Prithwish; Barker, Thomas

2018-01-01

Introduction Current guidelines recommend abstinence from supervised cardiac rehabilitation (CR) exercise training for 6 weeks post-sternotomy. This practice is not based on empirical evidence, thus imposing potentially unnecessary activity restrictions. Delayed participation in CR exercise training promotes muscle atrophy, reduces cardiovascular fitness and prolongs recovery. Limited data suggest no detrimental effect of beginning CR exercise training as early as 2 weeks post-surgery, but randomised controlled trials are yet to confirm this. The purpose of this trial is to compare CR exercise training commenced early (2 weeks post-surgery) with current usual care (6 weeks post-surgery) with a view to informing future CR guidelines for patients recovering from sternotomy. Methods and analysis In this assessor-blind randomised controlled trial, 140 cardiac surgery patients, recovering from sternotomy, will be assigned to 8 weeks of twice-weekly supervised CR exercise training commencing at either 2 weeks (early CR) or 6 weeks (usual care CR) post-surgery. Usual care exercise training will adhere to current UK recommendations. Participants in the early CR group will undertake a highly individualised 2–3 week programme of functional mobility, strength and cardiovascular exercise before progressing to a usual care CR programme. Outcomes will be assessed at baseline (inpatient), pre-CR (2 or 6 weeks post-surgery), post-CR (10 or 14 weeks post-surgery) and 12 months. The primary outcome will be change in 6 min walk distance. Secondary outcomes will include measures of functional fitness, quality of life and cost-effectiveness. Ethics and dissemination Recruitment commenced on July 2017 and will complete by December 2019. Results will be disseminated via national governing bodies, scientific meetings and peer-reviewed journals. Trial registration number NCT03223558; Pre-results. PMID:29574443

Inhibition of Salmonella Typhimurium by Cultures of Cecal Bacteria during Aerobic Incubation

USDA-ARS?s Scientific Manuscript database

Two trials were conducted to examine the ability of cecal bacterial cultures from broilers to inhibit growth of Salmonella Typhimurium during aerobic incubation. Cecal broth media was inoculated with 10 µl of cecal contents from 6 week old broilers taken from 2 separate flocks. Cultures were incubat...

Comparative histological and transcriptional analysis of maize kernels infected with Aspergillus flavus and Fusarium verticillioides

USDA-ARS?s Scientific Manuscript database

Aspergillus flavus and Fusarium verticillioides infect maize kernels and contaminate them with the mycotoxins aflatoxin and fumonisin, respectively. Combined histological examination of fungal colonization and transcriptional changes in maize kernels at 4, 12, 24, 48, and 72 hours post inoculation (...

Growth of Aspergillus repens in Flue-Cured Tobacco 1

PubMed Central

Welty, Ronald E.; Nelson, Larry A.

1971-01-01

In laboratory tests, flue-cured tobacco inoculated with Aspergillus repens was stored at 75, 80, 85, 87, and 95% relative humidity at 20 and 30 C. Samples were taken weekly for 4 weeks and evaluated for mold growth (colony count) and moisture content (MC). The weekly rate of fungus increase was slower at 20 C than at 30 C. Tobacco at 20 C with MC between 25 to 30% supported a slight to moderate increase in A. repens after 3 weeks of storage. However, tobacco at the same MC stored at 30 C was subject to rapid invasion by the fungus in as few as 1 to 2 weeks. Tobacco with MC above 30% stored at either 20 or 30 C became moldy in about 1 week. A mold index is proposed for evaluating populations of A. repens in tobacco. PMID:16349905

Growth of Aspergillus repens in Flue-Cured Tobacco.

PubMed

Welty, R E; Nelson, L A

1971-05-01

In laboratory tests, flue-cured tobacco inoculated with Aspergillus repens was stored at 75, 80, 85, 87, and 95% relative humidity at 20 and 30 C. Samples were taken weekly for 4 weeks and evaluated for mold growth (colony count) and moisture content (MC). The weekly rate of fungus increase was slower at 20 C than at 30 C. Tobacco at 20 C with MC between 25 to 30% supported a slight to moderate increase in A. repens after 3 weeks of storage. However, tobacco at the same MC stored at 30 C was subject to rapid invasion by the fungus in as few as 1 to 2 weeks. Tobacco with MC above 30% stored at either 20 or 30 C became moldy in about 1 week. A mold index is proposed for evaluating populations of A. repens in tobacco.

Cocultivation of Legionella pneumophila and free-living amoebae

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

Tyndall, R.L.; Domingue, E.L.

1982-10-01

Studies of the interaction of Legionella pneumophila with free-living amoebae showed that Naegleria lovaniensis and Acanthamoeba royreba could use L. pneumophia as a sole food source. However, growth of the amoebae on nonnutrient agar plates seeded with L. pneumophila was slower than growth on nonnutrient agar plates seeded with Escherichia coli. On inoculation of L. pneumophila into axenic cultures of N. lovaniensis and A. roryba, 99.9% of the L. pneumophila was destroyed within 24 h. After several weeks, however, some amoeba cultures became chronically infected and supported the growth of L. pneumophila. Amoebae exposed to L. pneumophila and containing adheredmore » L. pneumophila, L. pneumophila antigens, or both, showed no increased pathogenic potential on intranasal inoculation of weanling mice. Similarly, L. pneumophila propagated in chronically infected amoeba cultures showed no increase in virulence on intraperitoneal inoculation of guinea pigs relative to L. pneumophila grown in yeast extract broth. 20 references, 1 figure, 4 tables.« less

Experimental Infection of Common Eider Ducklings with Wellfleet Bay Virus, a Newly Characterized Orthomyxovirus

PubMed Central

Ip, Hon Sang; Ballmann, Anne; Hall, Jeffrey S.; Allison, Andrew B.; Ballard, Jennifer; Ellis, Julie C.; Cook, Robert; Gibbs, Samantha E.J.; Dwyer, Chris

2017-01-01

Wellfleet Bay virus (WFBV), a novel orthomyxovirus in the genus Quaranjavirus, was first isolated in 2006 from carcasses of common eider (Somateria mollissima) during a mortality event in Wellfleet Bay (Barnstable County, Massachusetts, USA) and has since been repeatedly isolated during recurrent mortality events in this location. Hepatic, pancreatic, splenic, and intestinal necrosis was observed in dead eiders. We inoculated 6-week-old common eider ducklings with WFBV in an attempt to recreate the naturally occurring disease. Approximately 25% of inoculated eiders had onset of clinical disease and required euthanasia; an additional 18.75% were adversely affected based on net weight loss during the trial. Control ducklings did not become infected and did not have clinical disease. Infected ducklings with clinical disease had pathologic lesions consistent with those observed during natural mortality events. WFBV was reisolated from 37.5% of the inoculated ducklings. Ducklings surviving to 5 days postinoculation developed serum antibody titers to WFBV. PMID:28841405

Experimental Infection of Common Eider Ducklings with Wellfleet Bay Virus, a Newly Characterized Orthomyxovirus.

PubMed

Shearn-Bochsler, Valerie; Ip, Hon Sang; Ballmann, Anne; Hall, Jeffrey S; Allison, Andrew B; Ballard, Jennifer; Ellis, Julie C; Cook, Robert; Gibbs, Samantha E J; Dwyer, Chris

2017-12-01

Wellfleet Bay virus (WFBV), a novel orthomyxovirus in the genus Quaranjavirus, was first isolated in 2006 from carcasses of common eider (Somateria mollissima) during a mortality event in Wellfleet Bay (Barnstable County, Massachusetts, USA) and has since been repeatedly isolated during recurrent mortality events in this location. Hepatic, pancreatic, splenic, and intestinal necrosis was observed in dead eiders. We inoculated 6-week-old common eider ducklings with WFBV in an attempt to recreate the naturally occurring disease. Approximately 25% of inoculated eiders had onset of clinical disease and required euthanasia; an additional 18.75% were adversely affected based on net weight loss during the trial. Control ducklings did not become infected and did not have clinical disease. Infected ducklings with clinical disease had pathologic lesions consistent with those observed during natural mortality events. WFBV was reisolated from 37.5% of the inoculated ducklings. Ducklings surviving to 5 days postinoculation developed serum antibody titers to WFBV.

Molecular biology of Ganoderma pathogenicity and diagnosis in coconut seedlings.

PubMed

Kandan, A; Radjacommare, R; Ramanathan, A; Raguchander, T; Balasubramanian, P; Samiyappan, R

2009-01-01

The pathogenicity of Ganoderma boninense was tested on coconut seedlings under greenhouse conditions and infection confirmed by using immunological and molecular diagnostic tools. Desiccation of older leaves and the emergence of sporophores were observed from pathogen-inoculated seedlings, whereas a control seedling does not show any pathogenic symptoms. Mature sporophores were formed within 10-13 weeks after inoculation. Polyclonal antibodies raised against mycelial proteins of Ganoderma were used for detection of Ganoderma in infected field palm and seedlings through indirect enzyme-linked immunosorbent assay technique. We adopted dot-immunobinding assay for the detection of Ganoderma from greenhouse and field samples. Under nucleic-acid-based diagnosis, G. boninense (167 bp) was detected from artificially inoculated seedlings and infected field palms by polymerase chain reaction. Apart from these, histopathological studies also support the Ganoderma pathogenicity in coconut seedlings. The pathogenicity test and combination of all the three diagnostic methods for Ganoderma could be highly reliable, rapid, sensitive and effective screening of resistance in planting material in the future.

Evidence on Possible Mycoplasma Etiology of Aster Yellows Disease I. Suppression of Symptom Development in Plants by Antibiotics

PubMed Central

Davis, R. E.; Whitcomb, R. F.

1970-01-01

Antibiotics suppressed development of aster yellows (AY) disease symptoms in plants of china aster [Callistephus chinensis (L.) Nees.] and annual chrysanthemum (Chrysanthemum carinatum, Schousb.). When inoculated chrysanthemum plants were treated by any of several techniques with tetracycline antibiotics or chloramphenicol, symptoms failed to appear during treatment but appeared 1 to 4 weeks after treatments were terminated. Under continuous administration of chlortetracycline, aster plants with AY symptoms developed symptomless axillary growth, including flowers. Streptomycin, oleandomycin, kanamycin, tylosin, carbomycin, polymyxin, bacitracin, neomycin, sulfanilamide, penicillin, vancomycin, or cycloserine had no discernible effect on development of AY symptoms. Treatment of plants with tetracycline antibiotics before exposure to inoculative (pathogen-transmitting) vectors delayed the appearance of symptoms or prevented AY infection. Remission of AY symptoms in inoculated plants treated with chlortetracycline was correlated with an inhibition of multiplication of AY agent, as measured by bioassay of extracts. The data give additional support to the hypothesis that aster yellows disease is caused by a mycoplasma-like microorganism. Images PMID:16557820

Experimental infection of common eider ducklings with Wellfleet Bay virus, a newly characterized orthomyxovirus

USGS Publications Warehouse

Shearn-Bochsler, Valerie I.; Ip, Hon S.; Ballmann, Anne; Hall, Jeffrey S.; Allison, Andrew B.; Ballard, Jennifer R.; Ellis, Julie C.; Cook, Robert; Gibbs, Samantha E.J.; Dwyer, Chris P.

2017-01-01

Wellfleet Bay virus (WFBV), a novel orthomyxovirus in the genus Quaranjavirus, was first isolated in 2006 from carcasses of common eider (Somateria mollissima) during a mortality event in Wellfleet Bay (Barnstable County, Massachusetts, USA) and has since been repeatedly isolated during recurrent mortality events in this location. Hepatic, pancreatic, splenic, and intestinal necrosis were observed in dead eiders. We inoculated 6-week-old common eider ducklings with WFBV in an attempt to recreate the naturally occurring disease. Approximately 25% of inoculated eiders had onset of clinical disease and required euthanasia; an additional 18.75% were adversely affected based on net weight loss during the trial. Control ducklings did not become infected and did not have clinical disease. Infected ducklings with clinical disease had pathologic lesions consistent with those observed during natural mortality events. WFBV was re-isolated from 37.5% of the inoculated ducklings. Ducklings surviving to 5 days postinoculation developed serum antibody titers to WFBV.

Rapid Startup and Loading of an Attached Growth, Simultaneous Nitrification/Denitrification Membrane Aerated Bioreactor

NASA Technical Reports Server (NTRS)

Meyer, Caitlin; Vega, Leticia

2014-01-01

The Membrane Aerated Bioreactor (MABR) is an attached-growth biological system for simultaneous nitrification and denitrification. This design is an innovative approach to common terrestrial wastewater treatments for nitrogen and carbon removal. Implementing a biologically-based water treatment system for long-duration human exploration is an attractive, low energy alternative to physiochemical processes. Two obstacles to implementing such a system are (1) the "start-up" duration from inoculation to steady-state operations and (2) the amount of surface area needed for the biological activity to occur. The Advanced Water Recovery Systems (AWRS) team at JSC explored these two issues through two tests; a rapid inoculation study and a wastewater loading study. Results from these tests demonstrate that the duration from inoculation to steady state can be reduced to two weeks and that the surface area to volume ratio baseline used in the Alternative Water Processor (AWP) test was higher than what was needed to remove the organic carbon and ammonium from the system.

Efficacy of vaccination and nisin Z treatments to eliminate intramammary Staphylococcus aureus infection in lactating cows*

PubMed Central

Guan, Ran; Wu, Jun-qiang; Xu, Wei; Su, Xiao-yan; Hu, Song-hua

2017-01-01

This study evaluated the effect of a Staphylococcus aureus bacterin and nisin on bovine subclinical mastitis. A total of 75 Holstein subclinically mastitic cows were randomly allocated to three groups with 25 cows per group. In group I, an intramammary infusion of nisin Z at a dose of 2.50×106 IU was carried out once daily for three days, and an autogenous S. aureus bacterin was inoculated into the supramammary lymph node one week before and one week after nisin treatment. In group II, nisin was administered in the same way as in group I, but no bacterin was inoculated. Group III received no treatment and served as a control. Milk was aseptically sampled from the affected quarters before and 2, 4, and 6 weeks after treatment, for bacteriological examination and analyses of N-acetyl-β-D-glucosaminidase (NAGase) activity, somatic cell count (SCC), and milk protein and fat contents. Results indicated that, compared to the nisin-treated group, nisin-bacterin treatment significantly reduced intramammary S. aureus infections, reduced the number of quarters with milk SCCs of more than 5×105 cells/ml, and increased the protein and fat contents of the milk. Therefore, nisin-bacterin therapy is suggested when subclinical mastitis occurs in lactating cows. PMID:28378574

Pathogenic potential of a Costa Rican strain of 'Candidatus Rickettsia amblyommii' in guinea pigs (Cavia porcellus) and protective immunity against Rickettsia rickettsii.

PubMed

Rivas, Juan J; Moreira-Soto, Andrés; Alvarado, Gilberth; Taylor, Lizeth; Calderón-Arguedas, Olger; Hun, Laya; Corrales-Aguilar, Eugenia; Morales, Juan Alberto; Troyo, Adriana

2015-09-01

'Candidatus Rickettsia amblyommii' is a spotted fever group rickettsia that is not considered pathogenic, although there is serologic evidence of possible infection in animals and humans. The aim of this study was to evaluate the pathogenic potential of a Costa Rican strain of 'Candidatus R. amblyommii' in guinea pigs and determine its capacity to generate protective immunity against a subsequent infection with a local strain of Rickettsia rickettsii isolated from a human case. Six guinea pigs were inoculated with 'Candidatus R. amblyommii' strain 9-CC-3-1 and two controls with cell culture medium. Health status was evaluated, and necropsies were executed at days 2, 4, and 13. Blood and tissues were processed by PCR to detect the gltA gene, and end titers of anti-'Candidatus R. amblyommii' IgG were determined by indirect immunofluorescence. To evaluate protective immunity, another 5 guinea pigs were infected with 'Candidatus R. amblyommii' (IGPs). After 4 weeks, these 5 IGPs and 3 controls (CGPs) were inoculated with pathogenic R. rickettsii. Clinical signs and titers of anti-Rickettsia IgG were determined. IgG titers reached 1:512 at day 13 post-infection with 'Candidatus R. amblyommii'. On day 2 after inoculation, two guinea pigs had enlarged testicles and 'Candidatus R. amblyommii' DNA was detected in testicles. Histopathology confirmed piogranulomatous orchitis with perivascular inflammatory infiltrate in the epididymis. In the protective immunity assay, anti-Rickettsia IgG end titers after R. rickettsii infection were lower in IGPs than in CGPs. IGPs exhibited only transient fever, while CGP showed signs of severe disease and mortality. R. rickettsii was detected in testicles and blood of CGPs. Results show that the strain 9-CC-3-1 of 'Candidatus R. amblyommii' was able to generate pathology and an antibody response in guinea pigs. Moreover, its capacity to generate protective immunity against R. rickettsii may modulate the epidemiology and severity of Rocky Mountain spotted fever in areas where both species circulate. Copyright © 2015 Elsevier GmbH. All rights reserved.

Boron neutron capture therapy (BNCT) for liver metastasis: therapeutic efficacy in an experimental model.

PubMed

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

2012-08-01

Boron neutron capture therapy (BNCT) was proposed for untreatable colorectal liver metastases. The present study evaluates tumor control and potential radiotoxicity of BNCT in an experimental model of liver metastasis. BDIX rats were inoculated with syngeneic colon cancer cells DHD/K12/TRb. Tumor-bearing animals were divided into three groups: BPA-BNCT, boronophenylalanine (BPA) + neutron irradiation; Beam only, neutron irradiation; Sham, matched manipulation. The total absorbed dose administered with BPA-BNCT was 13 ± 3 Gy in tumor and 9 ± 2 Gy in healthy liver. Three weeks post-treatment, the tumor surface area post-treatment/pre-treatment ratio was 0.46 ± 0.20 for BPA-BNCT, 2.7 ± 1.8 for Beam only and 4.5 ± 3.1 for Sham. The pre-treatment tumor nodule mass of 48 ± 19 mg fell significantly to 19 ± 16 mg for BPA-BNCT, but rose significantly to 140 ± 106 mg for Beam only and to 346 ± 302 mg for Sham. For both end points, the differences between the BPA-BNCT group and each of the other groups were statistically significant (ANOVA). No clinical, macroscopic or histological normal liver radiotoxicity was observed. It is concluded that BPA-BNCT induced a significant remission of experimental colorectal tumor nodules in liver with no contributory liver toxicity.

Using body temperature, food and water consumption as biomarkers of disease progression in mice with Eμ-myc lymphoma

PubMed Central

Hunter, J E; Butterworth, J; Perkins, N D; Bateson, M; Richardson, C A

2014-01-01

Background: Non-invasive biomarkers of disease progression in mice with cancer are lacking making it challenging to implement appropriate humane end points. We investigated whether body temperature, food and water consumption could be used to predict tumour burden. Methods: Thirty-six male, wild-type C57Bl/6 mice were implanted with subcutaneous RFID temperature sensors and inoculated with Eμ-myc tumours that infiltrate lymphoid tissue. Results: Decrease in body temperature over the course of the study positively predicted post-mortem lymph node tumour burden (R2=0.68, F(1,22)=44.8, P<0.001). At experimental and humane end points, all mice that had a mean decrease in body temperature of 0.7 °C or greater had lymph nodes heavier than 0.5 g (100% sensitivity), whereas a mean decrease in body temperature <0.7 °C always predicted lymph nodes lighter than 0.5 g (100% specificity). The mean decrease in food consumption in each cage also predicted mean post-mortem lymph node tumour burden at 3 weeks (R2=0.89, F(1,3)=23.2, P=0.017). Conclusion: Temperature, food and water consumption were useful biomarkers of disease progression in mice with lymphoma and could potentially be used more widely to monitor mice with other forms of cancer. PMID:24407190

Effect of two vesicular-arbuscular mycorrhizal fungi on the growth of micropropagated potato plantlets and on the extent of disease caused by Rhizoctonia solani.

PubMed

Yao, M K; Tweddell, R J; Désilets, H

2002-10-01

Two micropropagated potato cultivars, Goldrush and LP89221, were inoculated into sowing trays with either Glomus etunicatum or G. intraradices in a greenhouse. After 2 weeks, plantlets were transplanted into pots and roots were challenged 7 days later with Rhizoctonia solani. At different times after R. solani infection, disease severity, mortality rate, root colonization levels, various growth parameters, and shoot mineral content were evaluated. In Goldrush, only inoculation with G. etunicatum led to a significant reduction in disease severity, ranging between 60.2% and 71.2%, on both shoot and crown. This decrease was not observed in LP89221. Compared with the control plantlets, inoculation of Goldrush with G. etunicatum or G. intraradices reduced significantly the mortality rate by 77% and 26%, respectively, whereas vesicular-arbuscular mycorrhizal (VAM) fungi did not significantly influence the mortality rate in LP89221. In Goldrush, inoculation with G. etunicatum significantly increased shoot fresh weight, root dry weight and the number of tubers produced per plant, whereas G. intraradices only significantly increased the number of tubers. Tuber and root fresh weights of both potato cultivars were significantly reduced by R. solani infection. However, R. solani-infected plantlets of both Goldrush and LP89221, inoculated with G. etunicatum, produced significantly greater tuber fresh weight than non-VAM plantlets. In R. solani-infected plantlets of Goldrush but not LP89221, G. etunicatum and G. intraradices increased root fresh weight by approximately 140.3% and 76.5%, respectively, compared with non-VAM plants. The potato cultivars Goldrush and LP89221 responded differently to VAM fungal inoculation and to R. solani infection in terms of shoot mineral content.

Effects of oral administration of tilmicosin on pulmonary inflammation in piglets experimentally infected with Actinobacillus pleuropneumoniae.

PubMed

Nerland, Erin M; LeBlanc, Justin M; Fedwick, Jason P; Morck, Douglas W; Merrill, John K; Dick, Paul; Paradis, Marie-Anne; Buret, Andre G

2005-01-01

To determine the effects of oral administration of tilmicosin in piglets experimentally infected with Actinobacillus pleuropneumoniae. Forty 3-week-old specific-pathogen free piglets. Piglets were assigned to 1 of 4 groups as follows: 1) uninfected sham-treated control piglets; 2) infected untreated piglets that were intratracheally inoculated with 10(7) CFUs of A pleuropneumoniae; 3) infected treated piglets that were intratracheally inoculated with A pleuropneumoniae and received tilmicosin in feed (400 ppm [microg/g]) for 7 days prior to inoculation; or 4) infected treated piglets that were intratracheally inoculated with A pleuropneumoniae and received chlortetracycline (CTC) in feed (1100 ppm [microg/gl) for 7 days prior to inoculation. Bronchoalveolar lavage (BAL) fluid and lung tissue specimens of piglets for each group were evaluated at 3 or 24 hours after inoculation. For each time point, 4 to 6 piglets/group were studied. Feeding of CTC and tilmicosin decreased bacterial load in lungs of infected piglets. Tilmicosin delivered in feed, but not CTC, enhanced apoptosis in porcine BAL fluid leukocytes. This was associated with a decrease in LTB4 concentrations in BAL fluid of tilmicosin-treated piglets, compared with untreated and CTC-treated piglets, and also with a significant decrease in the number of pulmonary lesions. Tilmicosin inhibited infection-induced increases in rectal temperatures, as measured in untreated and CTC-treated piglets. Pulmonary neutrophil infiltration and prostaglandin E2 concentrations in the BAL fluid were not significantly different among groups at any time. Oral administration of tilmicosin to infected piglets induces apoptosis in BAL fluid leukocytes and decreases BAL fluid LTB4 concentrations and inflammatory lung lesions.

Early physiological responses of Pinus pinea L. seedlings infected by Heterobasidion sp.pl. in an ozone-enriched atmospheric environment.

PubMed

Pollastrini, Martina; Luchi, Nicola; Michelozzi, Marco; Gerosa, Giacomo; Marzuoli, Riccardo; Bussotti, Filippo; Capretti, Paolo

2015-03-01

The presence of the American root-rot disease fungus Heterobasidion irregulare Garbel. & Otrosina was detected in Italian coastal pine forests (Pinus pinea L.) in addition to the common native species Heterobasidion annosum (Fries) Brefeld. High levels of tropospheric ozone (O3) as an atmospheric pollutant are usually experienced in Mediterranean pine forests. To explore the effect of interaction between the two Heterobasidion species and ozone pollution on P. pinea, an open-top chamber (OTC) experiment was carried out. Five-year-old P. pinea seedlings were inoculated with the fungal species considered (H. irregulare, H. annosum and mock-inoculation as control), and then exposed in charcoal-filtered open-top chambers (CF-OTC) and non-filtered ozone-enriched chambers (NF+) from July to the first week of August 2010 at the experimental facilities of Curno (North Italy). Fungal inoculation effects in an ozone-enriched environment were assessed as: (i) the length of the inoculation lesion; (ii) chlorophyll a fluorescence (ChlF) responses; and (iii) analysis of resin terpenes. Results showed no differences on lesion length between fungal and ozone treatments, whereas the short-term effects of the two stress factors on ChlF indicate an increased photosynthetic efficiency, thus suggesting the triggering of compensation/repair processes. The total amount of resin terpenes is enhanced by fungal infection of both species, but depressed by ozone to the levels observed in mock-inoculated plants. Variations in terpene profiles were also induced by stem base inoculations and ozone treatment. Ozone might negatively affect terpene defences making plants more susceptible to pathogens and insects. © The Author 2015. Published by Oxford University Press. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

Body Mass Index, Overweight, and Obesity in Swedish Women Born Post-term.

PubMed

Derraik, José G B; Lundgren, Maria; Cutfield, Wayne S; Ahlsson, Fredrik

2016-07-01

There is increasing evidence that post-term birth (≥42 weeks of gestation) is associated with adverse long-term outcomes. We assessed whether women born post-term displayed increased risk of overweight and obesity in adulthood. Data were collected at first antenatal visit (~10-12 weeks of gestation) on singleton Swedish women aged ≥18 years in 1991-2009 (mean age 26.1 years), who were born post-term (n = 27 153) or at term (37-41 weeks of gestation; n = 184 245). Study outcomes were evaluated for continuous associations with gestational age. Stratified analyses were carried out comparing women born post-term or at term. Analyses were also run with a 2-week buffer between groups to account for possible errors in gestational age estimation, comparing women born very post-term (≥43 weeks of gestation; n = 5761) to those born within a narrower term window (38-40 weeks of gestation; n = 130 110). Increasing gestational age was associated with greater adult weight and body mass index (BMI). Stratified analyses showed that women born post-term were 0.5 kg heavier and had BMI 0.2 kg/m(2) greater than those born at term. Differences were more marked between women born very post-term (≥43 weeks) vs. a narrower term group (38-40 weeks): 1.0 kg and 0.3 kg/m(2) . The adjusted relative risks of overweight/obesity and obesity in women born very post-term were 1.13 and 1.12 times higher, respectively, than in those born at term. Post-term birth is associated with greater BMI and increased risk of overweight and obesity in adulthood, particularly among women born ≥43 weeks of gestation. © 2016 John Wiley & Sons Ltd.

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.

Sanitation of wallboard colonized with Stachybotrys chartarum.

PubMed

Price, D L; Ahearn, D G

1999-07-01

Sections (8 cm2) of unused, nonsterile gypsum wallboard (dry wall) were inoculated with varying densities (10(4) to approximately 10(8)/ml) of conidia from 14- to 21-day cultures of Stachybotrys chartarum grown on cellulose agar. The sections were permitted to air dry and were placed into vessels with 86% or 92% RH and incubated at 22-25 degrees C for up to 12 weeks. The moisture content of the dryboard increased from near 10% to over 35%. Selected sections with confluent surface growth, mainly of S. chartarum, were obtained within 3 weeks. Sections were cleaned with a quaternary or quaternary and chlorine dioxide or a concentrated oxygen-saline solution and treated, in some cases, with a preservative system and returned to humidity vessels. Reemergence of S. chartarum from inoculated and treated surfaces occurred within 5 weeks only with sections treated with the quaternary alone. Other fungi, mostly species of Aspergillus, Chaetomium and Penicillium, slowly colonized (between 9-12 weeks) at least some areas of most treated surfaces and most uninoculated control surfaces. Stachybotrys chartarum was also found on several sections of uninoculated controls. Sections treated with a quaternary/acrylic and placed in a dynamic challenging chamber remained visually free of colonized fungi for over 90 days. These studies indicate that control samples of uninstalled wallboard, available from local distributors, can contain a baseline bioburden, including S. chartarum, that will colonize surfaces under high humidity conditions. Sanitation and preservation treatment of the wallboard can markedly delay regrowth of these fungi, particularly of S. chartarum.

Immunocyte Response to Experimental Mumps Virus Infection in Rhesus Monkeys

PubMed Central

Genco, R. J.; Flanagan, T. D.; Emmings, F. G.

1973-01-01

Nineteen rhesus monkeys were inoculated with mumps virus by retrograde ductal instillation into the parotid gland. Evidence of infection was obtained in all instances. Virus was isolated from buccal swabbings and parotid biopsies for 1 week after inoculation. A vigorous serum-neutralizing antibody response occurred within 3 weeks, and there was marked monocytic infiltration of the parotid stroma. The monocytic infiltrate comprised as much as 60% of the total gland volume 1 week after infection. The predominant inflammatory cells were non-immunoglobulin-containing mononuclear cells resembling lymphocytes. Plasma cells containing immunoglobulin G (IgG), IgA, IgM, and IgE increased in numbers in the gland after infection, the greatest increase occurring in IgG-containing cells. Neutralizing antibodies and interferon were found in extracts prepared from the infected glands. Neutralizing activity was highest in samples taken 3 weeks after infection but was detectable in samples taken as soon as 36 to 48 h after infection. Interferon activity was detected in significant amounts 36 to 48 h after infection. Challenge of previously infected animals resulted in an increase in the monocytic infiltrate as well as an increase in numbers of immunoglobulin-containing plasma cells. However, reinfection did not occur as evidenced by the inability to culture shed virus after challenge. This model should be useful for in vivo study of biochemical mediators which evoke inflammatory cell infiltration and which may be significant both in protection and in tissue damage. PMID:4202659

StressModEx--Physiotherapist-led Stress Inoculation Training integrated with exercise for acute whiplash injury: study protocol for a randomised controlled trial.

PubMed

Ritchie, Carrie; Kenardy, Justin; Smeets, Rob; Sterling, Michele

2015-07-01

Whiplash associated disorders are the most common non-hospitalised injuries following a road traffic crash. Up to 50% of individuals who experience a whiplash injury will not fully recover and report ongoing pain and disability. Most recovery, if it occurs, takes place in the first 2-3 months post injury, indicating that treatment provided in the early stages is critical to long-term outcome. However, early management approaches for people with acute whiplash associated disorders are modestly effective. One reason may be that the treatments have been non-specific and have not targeted the processes shown to be associated with poor recovery, such as post-traumatic stress symptoms. Targeting and modulating these early stress responses in the early management of acute whiplash associated disorders may improve health outcomes. Early aggressive psychological interventions in the form of psychological debriefing may be detrimental to recovery and are now not recommended for management of early post-traumatic stress symptoms. In contrast, Stress Inoculation Training (SIT) is a cognitive behavioural approach that teaches various general problem-solving and coping strategies to manage stress-related anxiety (ie, relaxation training, cognitive restructuring and positive self-statements) and provides important information to injured individuals about the impact of stress on their physical and psychological wellbeing. While referral to a psychologist may be necessary in some cases where acute stress disorder or other more significant psychological reactions to stress are evident, in the case of acute whiplash injuries, it is neither feasible nor necessary for a psychologist to deliver the early stress modulation intervention to all injured individuals. The feasibility of using other specially trained health professionals to deliver psychological interventions has been explored in conditions such as chronic low back pain, chronic whiplash and cancer, but few trials have studied this approach in acute musculoskeletal conditions with the aim of preventing the development of chronic pain. As physiotherapy is the most common intervention received by individuals with a whiplash injury, physiotherapists are ideally placed to provide SIT in conjunction with standard physical rehabilitation. This study (StressModEx) will target individuals in the acute stage of injury and address the stress responses associated with the accident or injury (event-related distress) with the aim of improving both physical and mental health outcomes. Is SIT integrated with standard physiotherapy exercise and delivered by physiotherapists more effective than physiotherapy exercise alone in reducing neck pain and disability in individuals with acute whiplash associated disorders? Parallel randomised controlled trial with blinded outcome assessment. 100 individuals with grade II or III (no fracture/dislocation or neurological loss) acute whiplash associated disorder<4 weeks duration and at least moderate neck pain-related disability and hyper-arousal symptoms will be recruited for the study. Participants will be assessed via online surveys or in-person at a university research laboratory. Interventions will be provided at community physiotherapy practices in Brisbane, Gold Coast, Toowoomba and Mackay, Queensland, Australia. Clinical-guideline-recommended supervised physiotherapy exercise sessions (10 sessions) integrated with six (once per week) SIT sessions. Clinical-guideline-recommended supervised physiotherapy exercise sessions (10) only. Primary (Neck Disability Index) and secondary (Acute Stress Disorder Scale; Post-traumatic Stress Diagnostic Scale; Depression, Anxiety and Stress Scale; Pain Catastrophisingo Scale; Pain Self-Efficacy Questionnaire; Coping Strategies Questionnaire; Global impression of recovery; pain intensity; SF36) outcomes will be measured at baseline, 6 weeks, 6 months and 12 months after randomisation. Data analysis will be blinded and by intention to treat. Outcomes will be analysed using linear mixed and logistic regression models that will include baseline scores as covariates, participants as random effects and treatment conditions as fixed factors. This study will be the first to address early stress responses following acute whiplash injury through a novel intervention that integrates SIT and physiotherapy exercise. Crown Copyright © 2015. Published by Elsevier B.V. All rights reserved.

Construction and immunogenicity of recombinant Mycobacterium bovis BCG expressing GP5 and M protein of porcine reproductive respiratory syndrome virus.

PubMed

Bastos, Reginaldo G; Dellagostin, Odir A; Barletta, Raúl G; Doster, Allan R; Nelson, Eric; Osorio, Fernando A

2002-11-22

Mycobacterium bovis BCG was used to express a truncated form of GP5 (lacking the first 30 NH(2)-terminal residues) and M protein of porcine reproductive and respiratory syndrome virus (PRRSV). The PRRSV proteins were expressed in BCG under control of the mycobacterial hsp60 gene promoter either in the mycobacterial cytoplasm (BCGGP5cyt and BCGMcyt) or as MT19-fusion proteins on the mycobacterial surface (BCGGP5surf and BCGMsurf). Mice inoculated with BCGGP5surf and BCGMsurf developed antibodies against the viral proteins at 30 days post-inoculation (dpi) as detected by ELISA and Western blot. By 60 dpi, the animals developed titer of neutralizing antibodies of 8. A PRRSV-specific gamma interferon response was also detected in splenocytes of recombinant BCG-inoculated mice at 60 and 90 dpi. These results indicate that BCG was able to express antigens of PRRSV and elicit an immune response against the viral proteins in mice.

Whole genome sequencing in the search for genes associated with the control of SIV infection in the Mauritian macaque model.

PubMed

de Manuel, Marc; Shiina, Takashi; Suzuki, Shingo; Dereuddre-Bosquet, Nathalie; Garchon, Henri-Jean; Tanaka, Masayuki; Congy-Jolivet, Nicolas; Aarnink, Alice; Le Grand, Roger; Marques-Bonet, Tomas; Blancher, Antoine

2018-05-08

In the Mauritian macaque experimentally inoculated with SIV, gene polymorphisms potentially associated with the plasma virus load at a set point, approximately 100 days post inoculation, were investigated. Among the 42 animals inoculated with 50 AID 50 of the same strain of SIV, none of which received any preventive or curative treatment, nine individuals were selected: three with a plasma virus load (PVL) among the lowest, three with intermediate PVL values and three among the highest PVL values. The complete genomes of these nine animals were then analyzed. Initially, attention was focused on variants with a potential functional impact on protein encoding genes (non-synonymous SNPs (NS-SNPs) and splicing variants). Thus, 424 NS-SNPs possibly associated with PVL were detected. The 424 candidates SNPs were genotyped in these 42 SIV experimentally infected animals (including the nine animals subjected to whole genome sequencing). The genes containing variants most probably associated with PVL at a set time point are analyzed herein.

The interaction between foliar GA3 application and arbuscular mycorrhizal fungi inoculation improves growth in salinized tomato (Solanum lycopersicum L.) plants by modifying the hormonal balance.

PubMed

Khalloufi, Mouna; Martínez-Andújar, Cristina; Lachaâl, Mokhtar; Karray-Bouraoui, Najoua; Pérez-Alfocea, Francisco; Albacete, Alfonso

2017-07-01

The agriculture industry is frequently affected by various abiotic stresses limiting plant productivity. To decrease the negative effect of salinity and improve growth performance, some strategies have been used, such as exogenous application of plant growth regulators (i.e. gibberellic acid, GA 3 ), or arbuscular mycorrhizal fungi (AMF) inoculation. To gain insights about the cross-talk effect of exogenous GA 3 application and AMF inoculation on growth under salinity conditions, tomato plants (Solanum lycopersicum, cv. TT-115) were inoculated or not with the AMF Rhizophagus irregularis and exposed to different treatments during two weeks: 0M GA 3 +0mM NaCl, 10 -6 M GA 3 +0mM NaCl, 0M GA 3 +100mM NaCl and 10 -6 M GA 3 +100mM NaCl. Results have revealed that AMF inoculation or GA 3 application alone, but especially their interaction, resulted in growth improvement under salinity conditions. The growth improvement observed in AMF-inoculated tomato plants under salinity conditions was mainly associated to ionic factors (higherK concentration and K/Na ratio) while the alleviating effect of GA 3 application and its interaction with AMF appear to be due to changes in the hormonal balance. Foliar GA 3 application was found to increase the active gibberellins (GAs), resulting in a positive correlation between GA 3 and the growth-related parameters. Furthermore, cytokinins, indoleacetic acid and abscisic acid concentrations increased in AMF inoculated or GA 3 treated plants but, notably, in AMF plants treated with GA 3 , which showed improved growth under salinity conditions. This suggests that there is an interactive positive effect between GAs and AMF which alleviates growth impairment under salinity conditions by modifying the hormonal balance of the plant. Copyright © 2017 Elsevier GmbH. All rights reserved.

Impact of the Mass Drug Administration for malaria in response to the Ebola outbreak in Sierra Leone.

PubMed

Aregawi, Maru; Smith, Samuel J; Sillah-Kanu, Musa; Seppeh, John; Kamara, Anitta R Y; Williams, Ryan O; Aponte, John J; Bosman, Andrea; Alonso, Pedro

2016-09-20

As emergency response to the Ebola epidemic, the Government of Sierra Leone and its partners implemented a large-scale Mass Drug Administration (MDA) with artesunate-amodiaquine (ASAQ) covering >2.7 million people in the districts hardest hit by Ebola during December 2014-January 2015. The World Health Organization (WHO) and the National Malaria Control Programme (NMCP) evaluated the impact of the MDA on malaria morbidity at health facilities and the number of Ebola alerts received at District Ebola Command Centres. The coverage of the two rounds of MDA with ASAQ was estimated by relating the number anti-malarial medicines distributed to the estimated resident population. Segmented time-series analysis was applied to weekly data collected from 49 primary health units (PHUs) and 11 hospitals performing malaria parasitological testing during the study period, to evaluate trends of malaria cases and Ebola alerts during the post-MDA weeks compared to the pre-MDA weeks in MDA- and non-MDA-cheifdoms. After two rounds of the MDA, the number of suspected cases tested with rapid diagnostic test (RDT) decreased significantly by 43 % (95 % CI 38-48 %) at week 1 and remained low at week 2 and 3 post-first MDA and at week 1 and 3 post-second MDA; RDT positive cases decreased significantly by 47 % (41-52 %) at week 1 post-first and remained lower throughout all post-MDA weeks; and the RDT test positivity rate (TPR) declined by 35 % (32-38 %) at week 2 and stayed low throughout all post-MDA weeks. The total malaria (clinical + confirmed) cases decreased significantly by 45 % (39-52 %) at week 1 and were lower at week 2 and 3 post-first MDA; and week 1 post-second MDA. The proportion of confirmed malaria cases (out of all-outpatients) fell by 33 % (29-38 %) at week 1 post-first MDA and were lower during all post-MDA weeks. On the contrary, the non-malaria outpatient cases (cases due to other health conditions) either remained unchanged or fluctuated insignificantly. The Ebola alerts decreased by 30 % (13-46 %) at week 1 post-first MDA and much lower during all the weeks post-second MDA. The MDA achieved its goals of reducing malaria morbidity and febrile cases that would have been potentially diagnosed as suspected Ebola cases with increased risk of nosocomial infections. The intervention also helped reduce patient case-load to the severely strained health services at the peak of the Ebola outbreak and malaria transmission. As expected, the effect of the MDA waned in a matter of few weeks and malaria intensity returned to the pre-MDA levels. Nevertheless, the approach was an appropriate public health intervention in the context of the Ebola epidemic even in high malaria transmission areas of Sierra Leone.

Variations in virulence of avian pathogenic Escherichia coli demonstrated by the use of a new in vivo infection model.

PubMed

Pors, Susanne Elisabeth; Olsen, Rikke Heidemann; Christensen, Jens Peter

2014-06-04

Salpingitis and peritonitis are common pathological manifestations observed in egg-laying hens. To improve methods to study these conditions, a surgical model was developed. Initially, eighteen white layers underwent laparotomy with subsequent inoculation of ink, bacteria or sterile broth directly into the oviduct. Eight birds inoculated with 0.1 ml blue ink were euthanized immediately after inoculation and the specific site of inoculation was assessed. In all birds, ink was injected into the oviduct between five and seven cm cranial to the isthmus. To demonstrate the use of this approach to cause infection of the oviduct, five birds were inoculated with 8.6 × 10(6)CFU of a clinical Escherichia coli isolate. Five control birds received broth with no bacteria. Both infected and control birds were euthanized after 48 h followed by a post mortem examination. Infected birds showed diffuse fibrino-purulent peritonitis, E. coli was found in pure culture from one or more positions in the oviduct and the liver. Birds receiving sterile broth did not culture positive and demonstrated no gross lesions. Subsequently, 19 birds were inoculated with an isolate of E. coli ST95 and 20 birds with an isolate of E. coli ST141. Major variation in virulence was observed between the two isolates used in relation to clinical signs, gross lesions and histopathology. In contrast to E. coli ST141, E. coli ST95 caused severe clinical signs, epithelial necrosis of the oviduct and purulent salpingitis. The results of the study show the potential of the model in studies of the pathogenesis of infections and virulence of bacteria of the oviduct. Copyright © 2014 Elsevier B.V. All rights reserved.

Effects of multipurpose solutions on the adhesion of Acanthamoeba to rigid gas permeable contact lenses.

PubMed

Lee, Ga-Hyun; Yu, Hak-Sun; Lee, Ji-Eun

2016-03-01

To evaluate the effect of multipurpose contact lens care solutions (MPSs) on the adhesion of Acanthamoeba to rigid gas permeable (RGP) contact lenses. Acanthamoeba castellanii (AC) trophozoites were inoculated onto untreated RGP contact lenses (FP, Extra, or Menicon Z), and numbers of trophozoites adhering to lenses were counted under a phase contrast microscope at 18 h post-inoculation (controls). Similarly, adhering trophozoites were counted at 6 h post-inoculation on each of the three RGP lens types with one of three MPSs (Boston Simplus, Menicare Plus, and O2 Care). Scanning electron microscopic examinations were performed to compare lens surfaces. Adhesion of AC trophozoites to untreated FP was greater than to untreated Extra or Menicon Z. Surfaces of Extra and Menicon Z lenses were waxier, smoother, and more homogeneous than those of FP lenses. After treatment with Boston Simplus, adhesion of AC trophozoites was significantly reduced for all lens types as compared with controls (p < 0.0001). Treatments with Menicare Plus or O2 Care reduced the number of adherent AC trophozoites significantly on FP lenses only as compared with controls (p < 0.0001). The adhesion rates of AC trophozoites to RGP lenses depended on lens surfaces. Boston Simplus reduced the adhesion rate of AC trophozoites more than Menicare Plus or O2 Care. Appropriate RGP lens and MPS selection could decrease the prevalence of Acanthamoeba keratitis. © 2016 The Authors Ophthalmic & Physiological Optics © 2016 The College of Optometrists.

Global and quantitative proteomic analysis of dogs infected by avian-like H3N2 canine influenza virus

PubMed Central

Su, Shuo; Tian, Jin; Hong, Malin; Zhou, Pei; Lu, Gang; Zhu, Huachen; Zhang, Guihong; Lai, Alexander; Li, Shoujun

2015-01-01

Canine influenza virus A (H3N2) is a newly emerged etiological agent for respiratory infections in dogs. The mechanism of interspecies transmission from avian to canine species and the development of diseases in this new host remain to be explored. To investigate this, we conducted a differential proteomics study in 2-month-old beagles inoculated intranasally with 106 TCID50 of A/canine/Guangdong/01/2006 (H3N2) virus. Lung sections excised at 12 h post-inoculation (hpi), 4 days, and 7 days post-inoculation (dpi) were processed for global and quantitative analysis of differentially expressed proteins. A total of 17,796 proteins were identified at different time points. About 1.6% was differentially expressed between normal and infected samples. Of these, 23, 27, and 136 polypeptides were up-regulated, and 14, 18, and 123 polypeptides were down-regulated, at 12 hpi, 4 dpi, and 7 dpi, respectively. Vann diagram analysis indicated that 17 proteins were up-regulated and one was down-regulated at all three time points. Selected proteins were validated by real-time PCR and by Western blot. Our results show that apoptosis and cytoskeleton-associated proteins expression was suppressed, whereas interferon-induced proteins plus other innate immunity proteins were induced after the infection. Understanding of the interactions between virus and the host will provide insights into the basis of interspecies transmission, adaptation, and virus pathogenicity. PMID:25883591

An experimental study of the pathogenicity of a duck hepatitis A virus genotype C isolate in specific pathogen free ducklings.

PubMed

Zhang, Huanrong; Pi, JinKui; Tang, Cheng; Yue, Hua; Yang, Falong

2012-12-01

Duck hepatitis A virus genotype C (DHAV-C), recognized recently, is one of the pathogens causing fatal duck viral hepatitis in ducklings, especially in Asia. To demonstrate the pathogenesis of the DHAV-C isolate, 3-day-old specific pathogen free ducklings were inoculated subcutaneously with a DHAV-C isolate and the clinical signs were observed. Virus distribution, histological and apoptotic morphological changes of various tissues were examined at different times post inoculation. The serial, characteristic changes included haemorrhage and swelling of the liver. Apoptotic cells and virus antigen staining were found in all of the tissues examined. Where more virus antigen staining was detected, there were more severe histopathological and apoptotic changes. The amount of virus antigen and the histological and apoptotic morphological changes agreed with each other and became increasingly severe with length of time after infection. Apoptotic cells were ubiquitously distributed, especially among lymphocytes, macrophages and monocytes in immune organs such as the bursa of Fabricius, thymus and spleen, and in liver, kidney and cerebral cells. Necrosis was also observed within 72 h post inoculation in all organs examined, except the cerebrum, and was characterized by cell swelling and collapsed plasma membrane. These results suggest that the recent outbreak of disease caused by DHAV-C virus is pantropic, causing apoptosis and necrosis of different organs. The apoptosis and necrosis caused by the DHAV-C field strain in this study is associated with pathogenesis and DHAV-C-induced lesions.

Cytokine gene expression in skin of susceptible guinea-pig infected with Treponema pallidum.

PubMed Central

Wicher, V; Scarozza, A M; Ramsingh, A I; Wicher, K

1998-01-01

Using a semi-quantitative multiplex reverse transcription-polymerase chain reaction assay, we examined cytokine mRNA expression for interleukin-1alpha (IL-1alpha), IL-2, IL-10, IL-12p40, tumour necrosis factor-alpha (TNF-alpha) and transforming growth factor-beta (TGF-beta) in skin samples obtained from C4-deficient (C4D) guinea-pigs inoculated intradermally with virulent Treponema pallidum (VTP). Controls included unmanipulated animals, guinea-pigs injected with T. pallidum-free rabbit inflammatory testicular fluid (ITF) alone, or mixed with heat-killed organisms (HKTP). The expression of IL-1alpha, IL-12p40, and TNF-alpha mRNA [T helper type 1 (Th1)] remained within the normal range in both infected and control animals throughout the experimental period. However, a significant increase (P<0.05) in IL-10 mRNA (Th2) was found exclusively in the VTP-inoculated animals from 3 to 30 days post-infection. Another unique characteristic of the inflammatory response in infected guinea-pigs was the appearance, between 11 and 30 days post-inoculation, of a substantial number of eosinophils in addition to infiltrating mononuclear cells. The results showed a local Th2 response which is consistent with an inadequate immune response. This is reflected by the lengthy and incomplete clearance of the pathogen from the local site of entry and the chronic infection of distant organs. Images Figure 1 Figure 4 PMID:9824482

Impact of Experimental Human Pneumococcal Carriage on Nasopharyngeal Bacterial Densities in Healthy Adults

PubMed Central

Shak, Joshua R.; Cremers, Amelieke J. H.; Gritzfeld, Jenna F.; de Jonge, Marien I.; Hermans, Peter W. M.; Vidal, Jorge E.; Klugman, Keith P.; Gordon, Stephen B.

2014-01-01

Colonization of the nasopharynx by Streptococcus pneumoniae is a necessary precursor to pneumococcal diseases that result in morbidity and mortality worldwide. The nasopharynx is also host to other bacterial species, including the common pathogens Staphylococcus aureus, Haemophilus influenzae, and Moraxella catarrhalis. To better understand how these bacteria change in relation to pneumococcal colonization, we used species-specific quantitative PCR to examine bacterial densities in 52 subjects 7 days before, and 2, 7, and 14 days after controlled inoculation of healthy human adults with S. pneumoniae serotype 6B. Overall, 33 (63%) of subjects carried S. pneumoniae post-inoculation. The baseline presence and density of S. aureus, H. influenzae, and M. catarrhalis were not statistically associated with likelihood of successful pneumococcal colonization at this study’s sample size, although a lower rate of pneumococcal colonization in the presence of S. aureus (7/14) was seen compared to that in the presence of H. influenzae (12/16). Among subjects colonized with pneumococci, the number also carrying either H. influenzae or S. aureus fell during the study and at 14 days post-inoculation, the proportion carrying S. aureus was significantly lower among those who were colonized with S. pneumoniae (p = 0.008) compared to non-colonized subjects. These data on bacterial associations are the first to be reported surrounding experimental human pneumococcal colonization and show that co-colonizing effects are likely subtle rather than absolute. PMID:24915552

Staphylococcus aureus recovery from cotton towels.

PubMed

Oller, Anna R; Mitchell, Ashley

2009-04-30

Staphylococcus aureus is an emerging pathogen afflicting healthy individuals without known risk factors, and methicillin-resistant Staphylococcus aureus has been shown to colonize multiple family members sharing households. Because household items such as towels are often shared by family members, this study investigated whether cotton towel absorbency or washing conditions affect Staphylococcus aureus cell viability or cell retention, and whether the levels may be sufficient for person-to-person transmission. Staphylococcus aureus ATCC 25923 was added to a 48 mm(2) template area on three cotton towel types (terry, pima, and Egyptian), and subjected to hand washing, without manual wringing, in three conditions (water only, bleach addition, or liquid detergent addition). Serial dilutions plated onto mannitol salt plates quantified bacteria for inoculations, pre- and post-wash water samples, towel surfaces, and hand transfer. Hand transfer of bacteria was determined on towels immediately, one, 24, and 48 hours post inoculation. Bleach (p < or = .05) was the most effective at reducing bacterial viability on all towel types compared to detergent and water. Although not statistically significant, more Staphylococcus colonies were recovered from higher absorbency towels and from inside directly inoculated template areas. A paired t-test showed a difference between immediate and one-hour CFUs versus 24- and 48-hour recoveries (0.0002) for hand transfers. Cell viability decreased for over 48 hours on towels, but sufficient quantities may remain for colonization. More absorbent towels may harbor more Staphylococci than less absorbent ones, and may serve as a transmission mechanism for the bacterium.

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

PubMed

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

2014-05-01

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

Cross-protection in nonhuman primates against Argentine hemorrhagic fever.

PubMed Central

Weissenbacher, M C; Coto, C E; Calello, M A; Rondinone, S N; Damonte, E B; Frigerio, M J

1982-01-01

The susceptibility of the marmoset Callithrix jacchus to Tacaribe virus infection was investigated to perform cross-protection studies between Junin and Tacaribe viruses. Five marmosets inoculated with Tacaribe virus failed to show any signs of disease, any alterations in erythrocyte, leukocyte, reticulocyte, and platelet counts or any changes in hematocrit or hemoglobin values. No Tacaribe virus could be recovered from blood at any time postinfection. Anti-Tacaribe neutralizing antibodies appeared 3 weeks postinfection. The five Tacaribe-infected marmosets and four noninfected controls were challenged with the pathogenic strain of Junin virus on day 60 post-Tacaribe infection. The former group showed no signs of disease, no viremia, and no challenge virus replication, whereas the control group exhibited the typical symptoms of Argentine hemorrhagic fever, high viremia, and viral titers in organs. Soon after challenge, the Tacaribe-protected marmosets synthesized neutralizing antibodies against Junin virus. These results indicate that the marmoset C. jacchus can be considered an experimental model for protection studies with arenaviruses and that the Tacaribe virus could be considered as a potential vaccine against Junin virus. PMID:6276301

[Inhibition of Growth of Seed-Borne Fungi and Aflatoxin Production on Stored Peanuts by Allyl Isothiocyanate Vapor].

PubMed

Okano, Kiyoshi; Nishioka, Chikako; Iida, Tetsuya; Ozu, Yuzi; Kaneko, Misao; Watanabe, Yuko; Mizukami, Yuichi; Ichinoe, Masakatsu

2018-01-01

Aspergillus parasiticus contamination of peanuts results in the production of highly toxic metabolites, such as aflatoxin B 1 , B 2 , G 1 and G 2 , and its incidence in imported peanuts is reported to be increasing. Here, we examined whether the antifungal compound allyl isothiocyanate (AIT), which is present in mustard seed, could inhibit the growth of seed-borne fungi and aflatoxin-producing fungi. Peanuts produced in China and Japan were inoculated with A. parasiticus and exposed to AIT vapor released by a commercial mustard seed extract in closed containers under controlled conditions of temperature and humidity. AIT in the inoculated peanut samples reached its highest concentration of 44.8 ng/mL at 3 hr and decreased to 5.6 ng/mL after 9 weeks. Although AIT decreased the growth of the seed-borne fungi during the test period, the inoculated fungi survived. All tested peanuts samples were analyzed for aflatoxin using the HPLC method. There was a correlation between the number of aflatoxin-producing fungi and the total amount of aflatoxin production in the inoculated peanut samples. Our results indicate that AIT was effective in inhibiting the growth of seed-borne fungi and aflatoxin-producing fungi.

Infection and transmission of LPAIV H9N2 viruses in SPF chickens

USDA-ARS?s Scientific Manuscript database

Low pathogenic avian influenza viruses (LPAIV), subtype H9N2 are responsible for economic losses in the poultry industry worldwide. Using multiple strains of H9N2 LPAIV isolates from different years and countries, we inoculated 3-week old SPF laying hens intranasally to evaluate infectivity and tran...

Dietary rice component, Oryzanol, inhibits tumor growth in tumor-bearing Mice

USDA-ARS?s Scientific Manuscript database

Scope: We investigated the effects of rice bran and components on tumor growth in mice. Methods and results: Mice fed standard diets supplemented with rice bran, '-oryzanol, Ricetrienol®, ferulic acid, or phytic acid for 2 weeks were inoculated with CT-26 colon cancer cells and fed the same diet fo...

Effect of sulfur dioxide fumigation on survival of foodborne pathogens on table grapes under standard storage temperature

USDA-ARS?s Scientific Manuscript database

We examined the persistence of Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica Thompson inoculated on freshly-harvested table grapes under standard cold storage with initial and weekly sulfur dioxide (SO2) fumigation. L. monocytogenes and S. enterica Thompson were much more...

Innate and adaptive immune responses to in utero infection with bovine viral diarrhea virus

USDA-ARS?s Scientific Manuscript database

Infection of pregnant cows with noncytopathic (ncp) BVDV induces rapid innate and adaptive immune responses resulting in clearance of the virus in less than 3 weeks. Seven to 14 days after inoculation of the cow, ncpBVDV crosses the placenta and induces a fetal viremia. Establishment of persistent ...

Survival of foodborne pathogens on commercially packed table grapes under simulated refrigerated transit conditions

USDA-ARS?s Scientific Manuscript database

We examined the survival of Listeria monocytogenes, Escherichia coli O157:H7, and Salmonella enterica Thompson inoculated on commercially packed table grapes under simulated in-transit conditions (1.1ºC with 90% RH) for up to three weeks. Grapes were packed in perforated polyethylene cluster bags, w...

Pathogenicity of Nectriaceous Fungi on Avocado in Australia.

PubMed

Parkinson, Louisamarie E; Shivas, Roger G; Dann, Elizabeth K

2017-12-01

Black root rot is a severe disease of young avocado trees in Australia causing black necrotic roots, tree stunting, and leaf drop prior to tree death. Nectriaceous fungi (Nectriaceae, Hypocreales), are commonly isolated from symptomatic roots. This research tested the pathogenicity of 19 isolates from Calonectria, Cylindrocladiella, Dactylonectria, Gliocladiopsis, and Ilyonectria, spp. collected from young avocado trees and other hosts. Glasshouse pathogenicity tests with 'Reed' avocado (Persea americana) seedlings confirmed that Calonectria ilicicola is a severe pathogen of avocado, causing stunting, wilting, and seedling death within 5 weeks of inoculation. Isolates of C. ilicicola from peanut, papaya, and custard apple were also shown to be aggressive pathogens of avocado, demonstrating a broad host range. An isolate of a Calonectria sp. from blueberry and avocado isolates of Dactylonectria macrodidyma, D. novozelandica, D. pauciseptata, and D. anthuriicola caused significant root rot but not stunting within 5 to 9 weeks of inoculation. An isolate of an Ilyonectria sp. from grapevine closely related to Ilyonectria liriodendri, and avocado isolates of Cylindrocladiella pseudoinfestans, Gliocladiopsis peggii, and an Ilyonectria sp. were not pathogenic to avocado.

Infectious bursal disease in New Brunswick.

PubMed

Ide, P R; Stevenson, R G

1973-10-01

A flock of four week old chickens experienced a disease of sudden onset in which the only symptoms were those of depression shortly before death, and in which the predominant histological lesion was necrosis of lymphocytes in the bursa of Fabricius.A virus, designated strain Sk-1, was isolated from pooled bursal tissue of affected birds and was serologically identified as a strain of the infectious bursal agent. This virus was chloroform resistant, did not hemagglutinate guinea pig or chicken erythrocytes and did not produce a cytopathic effect in chick embryo tissue cultures. Equivocal results were obtained in filtration studies but the agent was less than 100nm in diameter. Four week old chicks inoculated with strain Sk-1 developed microscopic lesions in the bursa of Fabricius which were similar to those seen in the original field specimens. Inoculated chick embryos exhibited characteristic macroscopic lesions and necrosis of vascular tissue was a common histological change.A limited serological survey of local poultry flocks indicated that infection by this agent had occurred in four of the ten flocks examined.

Experimental infection of nontarget species of rodents and birds with Brucella abortus strain RB51 vaccine

USGS Publications Warehouse

Januszewski, M.C.; Olsen, S.C.; McLean, R.G.; Clark, L.; Rhyan, Jack C.

2001-01-01

The Brucella abortus vaccine strain RB51 (SRB51) is being considered for use in the management of bnucellosis in wild bison (Bison bison) and elk (Cervus elaphus) populations in the Greater Yellowstone Area (USA). Evaluation of the vaccines safety in non-target species was considered necessary prior to field use. Between June 1998 and December 1999, ground squirrels (Spermophilus richardsonii, n = 21), deer mice (Peromyscus maniculatus, n = 14), prairie voles (Microtus ochrogaster, n = 21), and ravens (Corvus corax, n = 13) were orally inoculated with SRB51 or physiologic saline. Oral and rectal swabs and blood samples were collected for bacteriologic evaluation. Rodents were necropsied at 8 to 10 wk and 12 to 21 wk post inoculation (PI), and ravens at 7 and 11 wk PI. Spleen, liver and reproductive tissues were collected for bacteriologic and histopathologic evaluation. No differences in clinical signs, appetite, weight loss or gain, or activity were observed between saline- and SRB51-inoculated animals in all four species. Oral and rectal swabs from all species were negative throughout the study. In tissues obtained from SRB51-inoculated animals, the organism was isolated from six of seven (86%) ground squirrels, one of six (17%) deer mice, none of seven voles, and one of five (20%) ravens necropsied at 8, 8, 10, and 7 wk PI, respectively. Tissues from four of seven (57%) SRB51-inoculated ground squirrels were culture positive for the organism 12 wk PI; SRB51 was not recovered from deer mice, voles. or ravens necropsied 12, 21, or 11 wk, respectively, PI. SRB51 was not recovered from saline-inoculated ground squirrels, deer mice, or voles at any time but was recovered from one saline-inoculated raven at necropsy, 7 wk PI, likely attributable to contact with SRB51-inoculated ravens in an adjacent aviary room. Spleen was time primary tissue site of colonization in ground squirrels, followed by the liver and reproductive organs. The results indicate oral exposure to SRB51 does not produce morbidity or mortality in ravens, ground squirrels, deer mice, or prairie voles.

Kinetics of lung lesion development and pro-inflammatory cytokine response in pigs with vaccine-associated enhanced respiratory disease induced by challenge with pandemic (2009) A/H1N1 influenza virus.

PubMed

Gauger, P C; Vincent, A L; Loving, C L; Henningson, J N; Lager, K M; Janke, B H; Kehrli, M E; Roth, J A

2012-11-01

The objective of this report was to characterize the enhanced clinical disease and lung lesions observed in pigs vaccinated with inactivated H1N2 swine δ-cluster influenza A virus and challenged with pandemic 2009 A/H1N1 human influenza virus. Eighty-four, 6-week-old, cross-bred pigs were randomly allocated into 3 groups of 28 pigs to represent vaccinated/challenged (V/C), non-vaccinated/challenged (NV/C), and non-vaccinated/non-challenged (NV/NC) control groups. Pigs were intratracheally inoculated with pH1N1 and euthanized at 1, 2, 5, and 21 days post inoculation (dpi). Macroscopically, V/C pigs demonstrated greater percentages of pneumonia compared to NV/C pigs. Histologically, V/C pigs demonstrated severe bronchointerstitial pneumonia with necrotizing bronchiolitis accompanied by interlobular and alveolar edema and hemorrhage at 1 and 2 dpi. The magnitude of peribronchiolar lymphocytic cuffing was greater in V/C pigs by 5 dpi. Microscopic lung lesion scores were significantly higher in the V/C pigs at 2 and 5 dpi compared to NV/C and NV/NC pigs. Elevated TNF-α, IL-1β, IL-6, and IL-8 were detected in bronchoalveolar lavage fluid at all time points in V/C pigs compared to NV/C pigs. These data suggest H1 inactivated vaccines followed by heterologous challenge resulted in potentiated clinical signs and enhanced pulmonary lesions and correlated with an elevated proinflammatory cytokine response in the lung. The lung alterations and host immune response are consistent with the vaccine-associated enhanced respiratory disease (VAERD) clinical outcome observed reproducibly in this swine model.

VEGF induces neuroglial differentiation in bone marrow-derived stem cells and promotes microglia conversion following mobilization with GM-CSF.

PubMed

Avraham-Lubin, Bat-Chen R; Goldenberg-Cohen, Nitza; Sadikov, Tamilla; Askenasy, Nadir

2012-12-01

Evaluation of potential tropic effects of vascular endothelial growth factor (VEGF) on the incorporation and differentiation of bone-marrow-derived stem cells (BMSCs) in a murine model of anterior ischemic optic neuropathy (AION). In the first approach, small-sized subset of BMCs were isolated from GFP donors mice by counterflow centrifugal elutriation and depleted of hematopoietic lineages (Fr25lin(-)). These cells were injected into a peripheral vein (1 × 10(6) in 0.2 ml) or inoculated intravitreally (2 × 10(5)) to syngeneic mice, with or without intravitreal injection of 5 μg/2μL VEGF, simultaneously with AION induction. In a second approach, hematopoietic cells were substituted by myelablative transplant of syngeseic GFP + bone marrow cells. After 3 months, progenitors were mobilized with granulocyte-macrophage colony-stimulating factor (GM-CSF) followed by VEGF inoculation into the vitreous body and AION induction . Engraftment and phenotype were examined by immunohistochemistry and FISH at 4 and 24 weeks post-transplantation, and VEGF receptors were determined by real time PCR. VEGF had no quantitative effect on incorporation of elutriated cells in the injured retina, yet it induced early expression of neuroal markers in cells incorporated in the RGC layer and promoted durable gliosis, most prominent perivascular astrocytes. These effects were mediated by VEGF-R1/Flt-1, which is constitutively expresses in the elutriated fraction of stem cells. Mobilization with GM-CSF limited the differentiation of bone marrow progenitors to microglia, which was also fostered by VEGF. VEGF signaling mediated by Flt-1 induces early neural and sustained astrocytic differentiation of stem cells elutriated from adult bone-marrow, with significant contribution to stabilization retinal architecture following ischemic injury.

Immunogenic and protective efficacy of recombinant protein GtxA-N against Gallibacterium anatis challenge in chickens.

PubMed

Pedersen, Ida J; Pors, Susanne E; Bager Skjerning, Ragnhild J; Nielsen, Søren S; Bojesen, Anders M

2015-10-01

Gallibacterium anatis is a major cause of reproductive tract infections in chickens. Here, we aimed to evaluate the efficacy of the recombinant protein GtxA-N at protecting hens, by addressing three objectives; (i) evaluating the antibody response following immunization (ii) scoring and comparing lesions, following challenge with G. anatis, in immunized and non-immunized hens and (iii) investigating if the anti-GtxA-N antibody titre in individual hens correlated with the observed lesions. Two consecutive experiments were performed in hens. In the first experiment hens were immunized with GtxA-N on day 0 and day 14, infected with G. anatis on day 28 and euthanized on day 56. The GtxA-N antibody response was assessed in pooled serum samples throughout the experiment, using an indirect enzyme-linked immunosorbent assay (ELISA). In the second experiment the GtxA-N antibody titres were assessed in individual hens before and after immunization. Subsequently, the hens were inoculated with G. anatis and finally all hens where euthanized and submitted for post mortem examination 48 h after inoculation. Immunization elicited strong antibody responses that lasted at least 8 weeks (P < .0001). The individual antibody titres observed in response to immunization varied considerably among hens (range: 174,100-281,500). Lesion scores following G. anatis infection were significantly lower in immunized hens compared to non-immunized hens (P = .004). Within the immunized group, no correlation was found between the individual antibody titres and the lesion scores. This study clearly demonstrated GtxA-N as a vaccine antigen able of inducing protective immunity against G. anatis.

Analysis of salicylic acid-dependent pathways in Arabidopsis thaliana following infection with Plasmodiophora brassicae and the influence of salicylic acid on disease.

PubMed

Lovelock, David A; Šola, Ivana; Marschollek, Sabine; Donald, Caroline E; Rusak, Gordana; van Pée, Karl-Heinz; Ludwig-Müller, Jutta; Cahill, David M

2016-10-01

Salicylic acid (SA) biosynthesis, the expression of SA-related genes and the effect of SA on the Arabidopsis-Plasmodiophora brassicae interaction were examined. Biochemical analyses revealed that, in P. brassicae-infected Arabidopsis, the majority of SA is synthesized from chorismate. Real-time monitored expression of a gene for isochorismate synthase was induced on infection. SA can be modified after accumulation, either by methylation, improving its mobility, or by glycosylation, as one possible reaction for inactivation. Quantitative reverse transcription-polymerase chain reaction (qPCR) confirmed the induction of an SA methyltransferase gene, whereas SA glucosyltransferase expression was not changed after infection. Col-0 wild-type (wt) did not provide a visible phenotypic resistance response, whereas the Arabidopsis mutant dnd1, which constitutively activates the immune system, showed reduced gall scores. As dnd1 showed control of the pathogen, exogenous SA was applied to Arabidopsis in order to test whether it could suppress clubroot. In wt, sid2 (SA biosynthesis), NahG (SA-deficient) and npr1 (SA signalling-impaired) mutants, SA treatment did not alter the gall score, but positively affected the shoot weight. This suggests that SA alone is not sufficient for Arabidopsis resistance against P. brassicae. Semi-quantitative PCR revealed that wt, cpr1, dnd1 and sid2 showed elevated PR-1 expression on P. brassicae and SA + P. brassicae inoculation at 2 and 3 weeks post-inoculation (wpi), whereas NahG and npr1 showed no expression. This work contributes to the understanding of SA involvement in the Arabidopsis-P. brassicae interaction. © 2015 BSPP and John Wiley & Sons Ltd.

The effect of environmental and physiological conditions on excystation of Acanthamoeba castellanii belonging to the T4 genotype.

PubMed

Lakhundi, Sahreena; Khan, Naveed Ahmed; Siddiqui, Ruqaiyyah

2014-08-01

Excystation in Acanthamoeba is an important property for the onset of infection as well as infection recurrence, post-treatment. The overall aim of this study was to determine the effects of several environmental and physiological parameters on excystation in Acanthamoeba castellanii belonging to the T4 genotype. Cysts were prepared by inoculating A. castellanii trophozoites on non-nutrient agar plates for up to 2 weeks. To determine the effects of various conditions on excystation, A. castellanii cysts were inoculated in growth medium i.e. PYG and incubated at varying temperatures (4-40 °C), various pHs (4-9), artificial light/dark cycles and 5% of CO2. Optimum excystation was observed when cysts were incubated at 30 °C in growth medium at neutral pH. Extremes of temperature and pH reduced excystation, while light/dark cycles had no effect on excystation of A. castellanii. On the other hand, 5% of CO2 enhanced excystation and growth of excysting amoebae. To determine the effect of serum on A. castellanii excystation, assays were performed in the presence of varying concentrations of heat-inactivated foetal bovine serum (FBS) (5-100%). The results revealed that FBS promoted excystation. The involvement of G proteins in excystation was also determined. Using propranolol hydrochloride, a G protein inhibitor, the results revealed that G proteins play a role in A. castellanii differentiation. Furthermore, organic solvents (methanol/ethanol) completely blocked excystation. None of the aforementioned conditions had any effect on the viability of A. castellanii. A complete understanding of excystation in A. castellanii will be of value to counter infection recurrence.

Apoptosis induction and release of inflammatory cytokines in the oviduct of egg-laying hens experimentally infected with H9N2 avian influenza virus.

PubMed

Wang, Jingyu; Tang, Chao; Wang, Qiuzhen; Li, Ruiqiao; Chen, Zhanli; Han, Xueying; Wang, Jing; Xu, Xingang

2015-06-12

The H9N2 subtype avian influenza virus (AIV) can cause serious damage to the reproductive tract of egg-laying hens, leading to severe egg-drop and poor egg shell quality. However, previous studies in relation to the oviductal-dysfunction resulted from this agent have not clearly been elucidated. In this study, apoptosis and pathologic changes in the oviducts of egg-laying hens caused by H9N2 AIV were evaluated. To understand the immune response in the pathogenic processes, 30-week old specific pathogen free (SPF) egg-laying hens inoculated with H9N2 subtype of AIV through combined intra-ocular and intra-nasal routes. H9N2 AIV infection resulted in oviductal lesions, triggered apoptosis and expression of immune related genes accompanied with infiltration of CD3(+)CD4(+) and CD3(+)CD8α(+) cells. Significant tissue damage and apoptosis were observed in the five oviductal parts (infundibulum, magnum, isthmus, uterus and vagina) at 5 days post-inoculation (dpi). Furthermore, immune-related genes, including chicken TLR3 (7, 21), MDA5, IL-2, IFN-β, CXCLi1, CXCLi2, XCL1, XCR1 and CCR5 showed variation in the egg-laying hens infected with H9N2 AIV. Notably, mRNA expression of IFN-α was suppressed during the infection. These results show distinct expression patterns of inflammatory cytokines and chemokines amongst segments of the oviduct. Differential gene expression of inflammatory cytokines and lymphocytes aggregation occurring in oviducts may initiate the infected tissue in response to virus replication which may eventually lead to excessive cellular apoptosis and tissue damage. Copyright © 2015 Elsevier B.V. All rights reserved.

Intracerebral infection of Cebus apella with the XJ-Clone 3 strain of Junín virus.

PubMed

Carballal, G; Oubiña, J R; Molinas, F C; Nagle, C; de la Vega, M T; Videla, C; Elsner, B

1987-03-01

To assess the usefulness of the South American primate Cebus apella as a model for neurovirulence of Junín virus, eight monkeys were inoculated with 10(5) LD50 of the attenuated XJ-Clone 3 Junín virus strain by the intrathalamic route. After the second week, weight loss and polyadenopathies were observed in most animals, one-half of which had a transient leukothrombocytopenia. Moderate clinical central nervous system (CNS) involvement was present in four of eight monkeys, while the rest had only mild neurologic signs. All recovered except one, which developed a deep coma and was killed in a pre-mortem stage at 18 days post-infection (pi). Junín virus was isolated from the throat from five, from the blood from three, and from the brain from two monkeys. In the most severely ill animal, virus titers higher than viremia were detected in both inoculated and contralateral brain hemispheres, as well as in lung, lymph node, and small intestine. Junín antigens and "in vivo" bound immunoglobulins were detected by immunofluorescence (IF) in the brain of four animals at 18, 21, 40, and 155 days pi. Moderate lymphocytic parenchymal and meningeal infiltration were observed in the brain of four animals, and gliosis was also present in the most affected monkey. Although the clinical response to infection was not uniform, all infected monkeys developed high IF antibodies. Cebus apella cannot be used as a highly sensitive model for Argentine hemorrhagic fever (AHF). However, the results obtained show that the XJ-Clone 3 strain can replicate in the primate CNS and to induce lesions and immunoglobulin deposition. In addition, viral persistence is suggested by the late detection of viral antigens in brain at 40 and 155 days pi.

Tomato type and post-treatment water rinse affect efficacy of acid washes against Salmonella enterica inoculated on stem scars of tomatoes and product quality.

PubMed

Fan, Xuetong; Gurtler, Joshua B; Sokorai, Kimberly J B

2018-09-02

A study was conducted to evaluate the effects of post-treatment rinsing with water on the inactivation efficacy of acid treatments against Salmonella inoculated onto stem scar areas of two types of tomatoes. In addition, impact on fruit quality was investigated during 21 days post-treatment storage at 10 °C. A four-strain cocktail of Salmonella enterica (S. Montevideo, S. Newport, S. Saintpaul, and S. Typhimurium) was inoculated onto stem scar areas of grape and large round tomatoes. The inoculated fruits were then treated for 2 min with the following solutions: water, 2% lactic acid +2% acetic acid +2% levulinic acid, 1.7% lactic acid +1.7% acetic acid +1.7% levulinic acid, and 3% lactic acid +3% acetic acid. After treatments, half of the fruits were rinsed with water while another half were not rinsed. Non-inoculated grape tomatoes for quality analysis were treated with the same solutions with and without subsequent water rinse. Results demonstrated that the acid combinations reduced populations of Salmonella enterica on the stem scar area of grape tomatoes by 1.52-1.90 log CFU/fruit, compared with the non-treated control while water wash and rinse removed the bacterium by only 0.23-0.30 log CFU/fruit. On the stem scar of large round tomatoes, the same acid treatments achieved 3.54 log CFU/fruit reduction of the pathogen. The varying response to the acid washes between grape and large round tomatoes seems to be related to the differences in surface characteristics of stem scar areas observed with SEM. Rinsing with water after acid combination treatments did not significantly affect the efficacy of the treatments in either grape or large round tomatoes. Acidic off-odor was detected on fruits treated with acid combination without water rinse 1 day after treatment while water rinse eliminated the off-odor. The acid treatments with and without water rinse did not consistently affect appearance, color, firmness, or lycopene or ascorbic acid contents of tomatoes during 21-days storage at 10 °C. Considering the similarity in antimicrobial efficacy between the fruits with and without water rinse following acid treatments, and the elimination of acidic odor by water rinse, fruits should be rinsed with water after acid treatments. Overall, our results demonstrated that the acids were more effective in inactivating Salmonella on large round tomatoes than on grape tomatoes, and water rinses following acid treatments eliminated the acidic odor without affecting the efficacy of the acids against Salmonella. Published by Elsevier B.V.

Biological behavior of tumors and associated retroviremia in cats inoculated with Snyder-Theilen fibrosarcoma virus and the phenomenon of tumor recurrence after primary regression.

PubMed Central

Pedersen, N C; Johnson, L; Theilen, G H

1984-01-01

The fate of tumors and associated retroviremia was studied in 111 cats infected with the Snyder-Theilen strain of feline sarcoma virus (FeSV). Tumors appeared at the site of inoculation within 7 to 10 days. A retroviremia, due mainly to the associated feline leukemia virus helper virus (FeLV-helper), developed at the same time as tumors. Of the cats, 44 developed progressively growing tumors and therefore had to be killed, and 67 developed tumors that regressed. There was a strong correlation between the persistence of the accompanying retroviremia and the growth of the tumors. The 44 cats with progressively growing fibrosarcomas remained retroviremic until death. Conversely, 53 of the 67 cats with solitary, regressing tumors were only transiently retroviremic. Tumor regression in these cats paralleled the disappearance of retrovirus from the blood. The fate of tumors and retroviremia was not always the same, however. Twelve cats remained persistently retroviremic after all signs of gross tumors disappeared. Two other kittens became nonviremic within 20 days after inoculation, yet tumors continued to grow and even metastasize for another 3 to 5 weeks before regressing. Fibrosarcomas recurred 3 weeks to 8 months later in 8 of 12 persistently retroviremic cats with regressed tumors. Although the blood and bone marrow from these cats contained predominantly FeLV-helper, tumor cells yielded both FeSV and FeLV-helper. Of 53 animals, 3 developed recurrent fibrosarcomas 5 weeks to 8 months after all signs of tumors and retroviremia had disappeared. Cells cultured from these tumors appeared initially like normal fibroblasts and were virus nonproducers. After one to three passages in culture, however, cells became malignantly transformed and replicated both FeSV and FeLV-helper. Cultures of the bone marrow from these and other nonviremic cats with regressed tumors yielded only FeLV-helper. PMID:6319286

Diverse pathogenicity of equine herpesvirus 1 (EHV-1) isolates in CBA mouse model.

PubMed

Yu, Mi Htay Htay; Kasem, Samy Gomaa Ahmed; Tsujimura, Koji; Matsumura, Tomio; Yanai, Tokuma; Yamaguchi, Tsuyoshi; Ohya, Kenji; Fukushi, Hideto

2010-03-01

The pathogenicity of equine herpesvirus 1 (EHV-1) isolates of Japan were evaluated by using the CBA mouse model. CBA mice were inoculated with eight Japanese EHV-1 strains (89c1, 90c16, 90c18, 97c11, 98c12, 00c19, 01c1 and HH-1) and one British strain (Ab4p). 89c1 caused slight body weight loss and nervous signs in mice at 8 days post infection (dpi). Severe weight loss and nervous signs were observed in mice inoculated with Ab4p at 6 dpi. The other strains did not cause apparent clinical signs. Infectious viruses were recovered from the lungs of all groups at 2 dpi. Histopathological analysis revealed interstitial pneumonia in the lungs of all mice inoculated with EHV-1. Encephalitis or meningoencephalitis was observed in the brains of mice inoculated with 89c1, 90c18, 97c11, 98c12, 01c1 and Ab4p. Japanese EHV-1 strains showed low pathogenicity in CBA mice, whereas the sequential affects of infection are similar to those of the highly pathogenic strain Ab4p. These results suggest that field isolates of EHV-1 have varying degrees of pathogenicity in CBA mice.

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

PubMed

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

1997-01-01

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

A mouse model for Chlamydia suis genital infection.

PubMed

Donati, Manuela; Di Paolo, Maria; Favaroni, Alison; Aldini, Rita; Di Francesco, Antonietta; Ostanello, Fabio; Biondi, Roberta; Cremonini, Eleonora; Ginocchietti, Laura; Cevenini, Roberto

2015-02-01

A mouse model for Chlamydia suis genital infection was developed. Ninety-nine mice were randomly divided into three groups and intravaginally inoculated with chlamydia: 45 mice (group 1) received C. suis purified elementary bodies (EBs), 27 (group 2) were inoculated with C. trachomatis genotype E EBs and 27 mice (group 3) with C. trachomatis genotype F EBs. Additionally, 10 mice were used as a negative control. At seven days post-infection (dpi) secretory anti-C. suis IgA were recovered from vaginal swabs of all C. suis inoculated mice. Chlamydia suis was isolated from 93, 84, 71 and 33% vaginal swabs at 3, 5, 7 and 12 dpi. Chlamydia trachomatis genotype E and F were isolated from 100% vaginal swabs up to 7 dpi and from 61 and 72%, respectively, at 12 dpi. Viable C. suis and C. trachomatis organisms were isolated from uterus and tubes up to 16 and 28 dpi, respectively. The results of the present study show the susceptibility of mice to intravaginal inoculation with C. suis. A more rapid course and resolution of C. suis infection, in comparison to C. trachomatis, was highlighted. The mouse model could be useful for comparative investigations involving C. suis and C. trachomatis species. © FEMS 2014. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

Predicting key malaria transmission factors, biting and entomological inoculation rates, using modelled soil moisture in Kenya.

PubMed

Patz, J A; Strzepek, K; Lele, S; Hedden, M; Greene, S; Noden, B; Hay, S I; Kalkstein, L; Beier, J C

1998-10-01

While malaria transmission varies seasonally, large inter-annual heterogeneity of malaria incidence occurs. Variability in entomological parameters, biting rates and entomological inoculation rates (EIR) have been strongly associated with attack rates in children. The goal of this study was to assess the weather's impact on weekly biting and EIR in the endemic area of Kisian, Kenya. Entomological data collected by the U.S. Army from March 1986 through June 1988 at Kisian, Kenya was analysed with concurrent weather data from nearby Kisumu airport. A soil moisture model of surface-water availability was used to combine multiple weather parameters with landcover and soil features to improve disease prediction. Modelling soil moisture substantially improved prediction of biting rates compared to rainfall; soil moisture lagged two weeks explained up to 45% of An. gambiae biting variability, compared to 8% for raw precipitation. For An. funestus, soil moisture explained 32% variability, peaking after a 4-week lag. The interspecies difference in response to soil moisture was significant (P < 0.00001). A satellite normalized differential vegetation index (NDVI) of the study site yielded a similar correlation (r = 0.42 An. gambiae). Modelled soil moisture accounted for up to 56% variability of An. gambiae EIR, peaking at a lag of six weeks. The relationship between temperature and An. gambiae biting rates was less robust; maximum temperature r2 = -0.20, and minimum temperature r2 = 0.12 after lagging one week. Benefits of hydrological modelling are compared to raw weather parameters and to satellite NDVI. These findings can improve both current malaria risk assessments and those based on El Niño forecasts or global climate change model projections.

Effect of a Pheromone on Stress-Associated Reactivation of Feline Herpesvirus-1 in Experimentally Inoculated Kittens.

PubMed

Contreras, Elena T; Hodgkins, E; Tynes, V; Beck, A; Olea-Popelka, F; Lappin, M R

2018-01-01

Stress contributes to reactivation of feline herpesvirus-1 (FHV-1). The usage of pheromones to decrease stress in FHV-1 experimentally inoculated kittens has not previously been investigated. To determine whether a feline pheromone would lessen stress, resulting in decreased recurrence of FHV-1-associated illness in kittens. Twelve 5-month-old, purpose-bred kittens. Randomized, double-blind, placebo-controlled clinical trial. Kittens previously infected with the same dose of FHV-1 were randomized into 2 separate but identical group rooms. After a 2-week equilibration period, a diffuser containing either the pheromone or placebo was placed in each of the rooms, and the kittens acclimated for an additional 2 weeks. Every 2 weeks thereafter, for the 8-week study period, housing was alternated between kennel- and group housing. Blinded observers applied a standardized clinical and behavioral scoring rubric daily. After each 2-week period, serum cortisol concentrations and quantitative PCR for FHV-1 and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) ratios were evaluated. Clinical, behavioral, and laboratory test results were compared between groups within individual and combined study periods. Sneezing occurred more frequently in the placebo group during individual (P = 0.006) and combined study periods (P = 0.001). Sleep at the end of observation periods occurred more frequently in the pheromone group during individual (P = 0.006) and combined study periods (P < 0.001). The findings suggest that the pheromone decreased stress, and the decrease in stress response may have resulted in decreased sneezing associated with FHV-1. Copyright © 2017 The Authors. Journal of Veterinary Internal Medicine published by Wiley Periodicals, Inc. on behalf of the American College of Veterinary Internal Medicine.

Differential expression proteomics to investigate responses and resistance to Orobanche crenata in Medicago truncatula

PubMed Central

Castillejo, Ma Ángeles; Maldonado, Ana M; Dumas-Gaudot, Eliane; Fernández-Aparicio, Mónica; Susín, Rafael; Diego, Rubiales; Jorrín, Jesús V

2009-01-01

Background Parasitic angiosperm Orobanche crenata infection represents a major constraint for the cultivation of legumes worldwide. The level of protection achieved to date is either incomplete or ephemeral. Hence, an efficient control of the parasite requires a better understanding of its interaction and associated resistance mechanisms at molecular levels. Results In order to study the plant response to this parasitic plant and the molecular basis of the resistance we have used a proteomic approach. The root proteome of two accessions of the model legume Medicago truncatula displaying differences in their resistance phenotype, in control as well as in inoculated plants, over two time points (21 and 25 days post infection), has been compared. We report quantitative as well as qualitative differences in the 2-DE maps between early- (SA 27774) and late-resistant (SA 4087) genotypes after Coomassie and silver-staining: 69 differential spots were observed between non-inoculated genotypes, and 42 and 25 spots for SA 4087 and SA 27774 non-inoculated and inoculated plants, respectively. In all, 49 differential spots were identified by peptide mass fingerprinting (PMF) following MALDI-TOF/TOF mass spectrometry. Many of the proteins showing significant differences between genotypes and after parasitic infection belong to the functional category of defense and stress-related proteins. A number of spots correspond to proteins with the same function, and might represent members of a multigenic family or post-transcriptional forms of the same protein. Conclusion The results obtained suggest the existence of a generic defense mechanism operating during the early stages of infection and differing in both genotypes. The faster response to the infection observed in the SA 27774 genotype might be due to the action of proteins targeted against key elements needed for the parasite's successful infection, such as protease inhibitors. Our data are discussed and compared with those previously obtained with pea [1] and transcriptomic analysis of other plant-pathogen and plant-parasitic plant systems. PMID:19575787

Incorporation of a recombinant Eimeria maxima IMP1 antigen into nanoparticles confers protective immunity against E. Maxima challenge infection.

PubMed

Jenkins, Mark C; Stevens, Laura; O'Brien, Celia; Parker, Carolyn; Miska, Katrzyna; Konjufca, Vjollca

2018-02-14

The purpose of this study was to determine if conjugating a recombinant Eimeria maxima protein, namely EmaxIMP1, into 20 nm polystyrene nanoparticles (NP) could improve the level of protective immunity against E. maxima challenge infection. Recombinant EmaxIMP1 was expressed in Escherichia coli as a poly-His fusion protein, purified by NiNTA chromatography, and conjugated to 20 nm polystyrene NP (NP-EmaxIMP1). NP-EMaxIMP1 or control non-recombinant (NP-NR) protein were delivered per os to newly-hatched broiler chicks with subsequent booster immunizations at 3 and 21 days of age. In battery cage studies (n = 4), chickens immunized with NP-EMaxIMP1 displayed complete protection as measured by weight gain (WG) against E. maxima challenge compared to chickens immunized with NP-NR. WG in the NP-EMaxIMP1-immunized groups was identical to WG in chickens that were not infected with E. maxima infected chickens. In floor pen studies (n = 2), chickens immunized with NP-EMaxIMP1 displayed partial protection as measured by WG against E. maxima challenge compared to chickens immunized with NP-NR. In order to understand the basis for immune stimulation, newly-hatched chicks were inoculated per os with NP-EMaxIMP1 or NP-NR protein, and the small intestine, bursa, and spleen, were examined for NP localization at 1 h and 6 h post-inoculation. Within 1 h, both NP-EMaxIMP1 and NP-NR were observed in all 3 tissues. An increase was observed in the level of NP-EmaxIMP1 and NP-NR in all tissues at 6 h post-inoculation. These data indicate that 20 nm NP-EmaxIMP1 or NP-NR reached deeper tissues within hours of oral inoculation and elicited complete to partial immunity against E. maxima challenge infection. Published by Elsevier Ltd.

Identifying ingredients that delay outgrowth of Listeria monocytogenes in natural, organic, and clean-label ready-to-eat meat and poultry products.

PubMed

McDonnell, Lindsey M; Glass, Kathleen A; Sindelar, Jeffrey J

2013-08-01

The objective of this study was to identify ingredients that inhibit Listeria monocytogenes in natural, organic, or clean-label ready-to-eat meat and poultry products. Fourteen ingredients were screened in uncured (no-nitrate-or-nitrite-added), traditional-cured (156 ppm of purified sodium nitrite), cultured (alternative cured, natural nitrate source, and Staphylococcus carnosus), or preconverted (alternative cured, natural nitrite source) turkey slurries. Slurries were cooked, cooled, inoculated to yield 3 log CFU/ml L. monocytogenes, stored at 4°C, and tested weekly for 4 weeks. Three antimicrobial ingredients, 1.5 % vinegar-lemon-cherry powder blend, 2.5 % buffered vinegar, and 3.0 % cultured sugar-vinegar blend, were incorporated into alternative-cured ham and uncured roast beef and deli-style turkey breast. Controls included all three meat products without antimicrobial ingredients and a traditional-cured ham with 2.8 % sodium lactate-diacetate. Cooked, sliced products were inoculated with 3 log CFU/g L. monocytogenes, vacuum packed, and stored at 4 or 7°C, for up to 12 weeks. For control products without antimicrobial agents stored at 4°C, a 2-log L. monocytogenes increase was observed at 2 weeks for ham and turkey and at 4 weeks for roast beef. Growth (>1-log increase) in the sodium lactate-diacetate was delayed until week 6. Compared with the control, the addition of either vinegar-lemon-cherry powder blend or buffered vinegar delayed L. monocytogenes growth for an additional 2 weeks, while the addition of cultured sugar-vinegar blend delayed growth for an additional 4 weeks for both ham and turkey. The greatest L. monocytogenes delay was observed in roast beef containing any of the three antimicrobial ingredients, with no growth detected through 12 weeks at 4°C for all the treatments. As expected, L. monocytogenes grew substantially faster in products stored at 7°C than at 4°C. These data suggest that antimicrobial ingredients from a natural source can enhance the safety of ready-to-eat meat and poultry products, but their efficacy is improved in products containing nitrite and with lower moisture and pH.

Xylella fastidiosa infection of grapevines affects xylem levels of phenolic compounds and pathogenesis-related proteins

USDA-ARS?s Scientific Manuscript database

Pierce’s disease (PD), caused by the xylem-dwelling pathogen Xylella fastidiosa (X.f.), is a serious threat to grape production. The effects of X.f. infection six months post-inoculation on defense-associated proteins and phenolic compounds found in xylem sap and tissue were evaluated. Defense-assoc...

Intranasal inoculation of white-tailed deer (Odocoileus virginianus) with lyophilized chronic wasting disease prion particulate complexed to montmorillonite clay

USDA-ARS?s Scientific Manuscript database

Chronic wasting disease (CWD) is the transmissible spongiform encephalopathy or TSE of deer and elk, occurring primarily in North America. The TSEs are fatal neurodegenerative disorders associated with conversion of a normal cell protein to a pathogenic and potentially infectious agent by post trans...

9 CFR 113.328 - Fowl Laryngotracheitis Vaccine.

Code of Federal Regulations, 2010 CFR

2010-01-01

.... Disregard all deaths during the first 24 hours post-injection. To be a valid test, at least four embryos in... pathogens by the chicken embryo inoculation test prescribed in § 113.37, except that, if the test is inconclusive because of vaccine virus override, the test may be repeated and if the repeat test is inconclusive...

9 CFR 113.208 - Avian Encephalomyelitis Vaccine, Killed Virus.

Code of Federal Regulations, 2011 CFR

2011-01-01

... vaccine. For a valid test, at least 80 percent of the embryos shall survive for 48 hours post-inoculation... requirements prescribed in this section. Any serial found unsatisfactory by a prescribed test shall not be released. (a) Safety tests. (1) The prechallenge part of the potency test prescribed in paragraph (b) of...

9 CFR 113.208 - Avian Encephalomyelitis Vaccine, Killed Virus.

Code of Federal Regulations, 2010 CFR

2010-01-01

... vaccine. For a valid test, at least 80 percent of the embryos shall survive for 48 hours post-inoculation... requirements prescribed in this section. Any serial found unsatisfactory by a prescribed test shall not be released. (a) Safety tests. (1) The prechallenge part of the potency test prescribed in paragraph (b) of...

9 CFR 113.208 - Avian Encephalomyelitis Vaccine, Killed Virus.

Code of Federal Regulations, 2012 CFR

2012-01-01

... vaccine. For a valid test, at least 80 percent of the embryos shall survive for 48 hours post-inoculation... requirements prescribed in this section. Any serial found unsatisfactory by a prescribed test shall not be released. (a) Safety tests. (1) The prechallenge part of the potency test prescribed in paragraph (b) of...

Effect of storage at 4 and 10 C on growth of listeria monocytogenes in Queso Fresco

USDA-ARS?s Scientific Manuscript database

A five-strain rifampicin - resistant Listeria monocytogenes cocktail (ca. 3.0 loglOCFU/g) was introduced as a post-pasteurization contaminant in Queso Fresco (QF) that was manufactured using a commercial make procedure. L. monocytogenes was either inoculated into (IN) the curds before slicing or on...

Horizontal transmission of Salmonella and Campylobacter among caged and cage-free laying hens

USDA-ARS?s Scientific Manuscript database

In each of five trials, laying hens (56-72 wk-of-age) were challenged orally, intracolonally, and intravaginally with Salmonella and Campylobacter. One wk post inoculation, challenged hens (n=3) were commingled with non-challenged hens (n=12) in conventional wire cages, on all wire slats, or on all...

Detailed analysis of BALB/c mice challenged with wild type rotavirus EDIM provide an alternative for infection model of rotavirus.

PubMed

Du, Jialiang; Lan, Zhiling; Liu, Yueyue; Liu, Yan; Li, Yanchao; Li, Xiangming; Guo, Tai

2017-01-15

Mouse is one of the infection animal models for rotavirus. Since the optimal age of mouse sensitive to rotavirus infection thus far has not been unified, we elucidated clinical symptoms, immune responses and pathological changes of mice in different ages after challenged by murine rotavirus wild strain EDIM (Epidemic Diarrhea of Infant Mice) to provide data for the estimation. One-week-old, two-week-old, and three-week-old BALB/c mice were inoculated with EDIM in the challenge dose of 235 ID50, 470 ID50 and 705 ID50 respectively and were compared to mock-infected controls. Diarrhea illness, mobility, bodyweight were recorded, viral shedding and immune responses including serum IgA, fecal sIgA were detected, and small intestine tissue was evaluated for virus distribution and pathological changes. All the mice in one-week-old and two-week-old groups were completely unavoidable to be infected by EDIM and have been found to be malaise, activity reduced and even diarrhea, while three-week-old mice partly resist the challenge with 40% mice free from diarrhea. Meanwhile, EDIM infection has greater impact to the bodyweight of two-week-old group than those of one-week-old, three-week-old (0.9860 vs 1.2340, 1.2375g/day). One peak of virus shedding in three groups was observed in day 1-2 post infection, but the duration shortened with age increase. Feces sIgA in both two-week-old and three-week-old groups began to increase in day 4, 2-3days earlier than that in one-week-old group, and grow to the peak in day 8, which is about 2 fold of that in one-week-old group. Stronger serum IgA response was found in two-week-old group, it increased to the peak in day 15 and the level was 2 fold of three-week-old group and 4 fold of one-week-old group. The pathological changes included vacuolar degeneration, edema and congestion of intestinal wall, integrity destruction of enteric epithelium, and the changes relieved with the increase of age. Besides, rotavirus particles were found in small intestine tissues, especially in the surface and crypt of villi. In conclusion, the two-week-old mice were more sensitive to EDIM infection and initiated more effective immune response. In combination with that 14days old mice equals to 2 months infant when the first dose of rotavirus vaccine should be administrated, two-week-old mice is preferred to be used as infection model for the study of pathogenicity and immunogenicity of rotavirus. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

Rapid Dissemination of SIV Follows Multisite Entry after Rectal Inoculation

PubMed Central

Prétet, Jean-Luc; Michel-Salzat, Alice; Messent, Valérie; Bogdanova, Anna; Couëdel-Courteille, Anne; Souil, Evelyne; Cheynier, Rémi; Butor, Cécile

2011-01-01

Receptive ano-rectal intercourse is a major cause of HIV infection in men having sex with men and in heterosexuals. Current knowledge of the mechanisms of entry and dissemination during HIV rectal transmission is scarce and does not allow the development of preventive strategies. We investigated the early steps of rectal infection in rhesus macaques inoculated with the pathogenic isolate SIVmac251 and necropsied four hours to nine days later. All macaques were positive for SIV. Control macaques inoculated with heat-inactivated virus were consistently negative for SIV. SIV DNA was detected in the rectum as early as four hours post infection by nested PCR for gag in many laser-microdissected samples of lymphoid aggregates and lamina propria but never in follicle-associated epithelium. Scarce SIV antigen positive cells were observed by immunohistofluorescence in the rectum, among intraepithelial and lamina propria cells as well as in clusters in lymphoid aggregates, four hours post infection and onwards. These cells were T cells and non-T cells that were not epithelial cells, CD68+ macrophages, DC-SIGN+ cells or fascin+ dendritic cells. DC-SIGN+ cells carried infectious virus. Detection of Env singly spliced mRNA in the mucosa by nested RT-PCR indicated ongoing viral replication. Strikingly, four hours post infection colic lymph nodes were also infected in all macaques as either SIV DNA or infectious virus was recovered. Rapid SIV entry and dissemination is consistent with trans-epithelial transport. Virions appear to cross the follicle-associated epithelium, and also the digestive epithelium. Viral replication could however be more efficient in lymphoid aggregates. The initial sequence of events differs from both vaginal and oral infections, which implies that prevention strategies for rectal transmission will have to be specific. Microbicides will need to protect both digestive and follicle-associated epithelia. Vaccines will need to induce immunity in lymph nodes as well as in the rectum. PMID:21573012

Orally administered live attenuated Salmonella Typhimurium protects mice against lethal infection with H1N1 influenza virus.

PubMed

Kamble, Nitin Machindra; Hajam, Irshad Ahmed; Lee, John Hwa

2017-03-01

Pre-stimulation of toll-like receptors (TLRs) by agonists has been shown to increase protection against influenza virus infection. In this study, we evaluated the protective response generated against influenza A/Puerto Rico/8/1934 (PR8; H1N1) virus by oral and nasal administration of live attenuated Salmonella enterica serovar Typhimurium, JOL911 strain, in mice. Oral and nasal inoculation of JOL911 significantly increased the mRNA copy number of TLR-2, TLR4 and TLR5, and downstream type I interferon (IFN) molecules, IFN-α and IFN-β, both in peripheral blood mononuclear cells (PBMCs) and in lung tissue. Similarly, the mRNA copy number of interferon-inducible genes (ISGs), Mx and ISG15, were significantly increased in both the orally and the nasally inoculated mice. Post PR8 virus lethal challenge, the nasal JOL911 and the PBS control group mice showed significant loss of body weight with 70% and 100% mortality, respectively, compared to only 30% mortality in the oral JOL911 group mice. Post sub-lethal challenge, the significant reduction in PR8 virus copy number in lung tissue was observed in oral [on day 4 and 6 post-challenge (dpc)] and nasal (on 4dpc) than the PBS control group mice. The lethal and sub-lethal challenge showed that the generated stimulated innate resistance (StIR) in JOL911 inoculated mice conferred resistance to acute and initial influenza infection but might not be sufficient to prevent the PR8 virus invasion and replication in the lung. Overall, the present study indicates that oral administration of attenuated S. Typhimurium can pre-stimulate multiple TLR pathways in mice to provide immediate early StIR against a lethal H1N1 virus challenge. Copyright © 2017 Elsevier B.V. All rights reserved.

Alcohol consumption suppresses metastasis of B16-BL6 melanoma in mice.

PubMed

Meadows, G G; Elstad, C A; Blank, S E; Gallucci, R M; Pfister, L J

1993-03-01

Female C57BL/6 mice were fed a defined, pelleted diet and given 10% w/v or 20% w/v ethanol in their drinking water. Natural killer (NK) cell cytolytic activity was compared between water-drinking and ethanol-consuming mice and in mice that were also treated with polyinosinic-polycytidylic acid (poly I:C) to augment NK cell activity or with anti-NK1.1 antibody to decrease activity. NK cell cytolytic activity was not altered in mice given 10% ethanol, but was decreased in mice given 20% ethanol compared to water-drinking mice. Poly I:C treatment increased and anti-NK1.1 antibody treatment decreased NK cell activity in both water-drinking and 20% ethanol-consuming mice. Experimental and spontaneous metastases of B16-BL6 melanoma were evaluated as a function of the duration of ethanol consumption before tumor inoculation and as a function of altered NK cell activity. Experimental metastasis was inhibited after 4 and also after 6.5 weeks of ethanol exposure. Poly I:C treatment inhibited tumor lung colonization irrespective of ethanol consumption. Anti-NK1.1 antibody treatment increased metastasis, although to a lesser degree in mice consuming 10% ethanol. Spontaneous metastasis was inhibited in mice consuming 10% ethanol for 4 weeks, and in mice consuming 20% ethanol for 1 and 4 weeks before melanoma inoculation.

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

PubMed Central

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

2002-01-01

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

Effects of pre- or post-processing storage conditions on high-hydrostatic pressure inactivation of Vibrio parahaemolyticus and V. vulnificus in oysters.

PubMed

Ye, Mu; Huang, Yaoxin; Gurtler, Joshua B; Niemira, Brendan A; Sites, Joseph E; Chen, Haiqiang

2013-05-15

The effects of storage conditions on subsequent high-hydrostatic pressure (HHP) inactivation of Vibrio parahaemolyticus and Vibrio vulnificus in oysters were investigated. Live oysters were inoculated with V. parahaemolyticus or V. vulnificus to ca. 7-8 log MPN/g by feeding and stored at varying conditions (i.e., 21 or 35 °C for 5h, 4 or 10 °C for 1 and 2 days and -18 °C for 2 weeks). Oyster meats were then treated at 225-300 MPa for 2 min at 4, 21 or 35 °C. HHP at 300 MPa for 2 min achieved a >5-log MPN/g reduction of V. parahaemolyticus, completely inactivating V. vulnificus (negative by enrichment) in oysters. Treatment temperatures of 4, 21 and 35 °C did not significantly affect pressure inactivation of V. parahaemolyticus or V. vulnificus (P>0.05). Cold storage at -18, 4 and 10 °C, prior to HHP, decreased V. parahaemolyticus or V. vulnificus populations by 1.5-3.0 log MPN/g, but did not increase their sensitivity to subsequent HHP treatments. The effects of cold storage after HHP on inactivation of V. parahaemolyticus in oysters were also determined. Oysters were inoculated with V. parahaemolyticus and stored at 21 °C for 5h or 4 °C for 1 day. Oyster meats were then treated at 250-300 MPa for 2 min at 21 or 35 °C and stored for 15 days in ice or in a freezer. V. parahaemolyticus populations in HHP-treated oysters gradually decreased during post-HHP ice or frozen storage. A validation study using whole-shell oysters was conducted to determine whether the presence of oyster shells influenced HHP inactivation of V. parahaemolyticus. No appreciable differences in inactivation between shucked oyster meat and whole-shell oysters were observed. HPP at 300 MPa for 2 min at 21 °C, followed by 5-day ice storage or 7-day frozen storage, and HPP at 250 MPa for 2 min at 21 °C, followed by 10-day ice or 7-day frozen storage, completely inactivated V. parahaemolyticus in whole-shell oysters (>7 log reductions). The combination of HHP at a relatively low pressure (e.g., 250 MPa) followed by short-term frozen storage (7 days) could potentially be applied by the shellfish industry as a post-harvest process to eliminate V. parahaemolyticus in oysters. Copyright © 2013 Elsevier B.V. All rights reserved.

Microbial community development in a dynamic gut model is reproducible, colon region specific, and selective for Bacteroidetes and Clostridium cluster IX.

PubMed

Van den Abbeele, Pieter; Grootaert, Charlotte; Marzorati, Massimo; Possemiers, Sam; Verstraete, Willy; Gérard, Philippe; Rabot, Sylvie; Bruneau, Aurélia; El Aidy, Sahar; Derrien, Muriel; Zoetendal, Erwin; Kleerebezem, Michiel; Smidt, Hauke; Van de Wiele, Tom

2010-08-01

Dynamic, multicompartment in vitro gastrointestinal simulators are often used to monitor gut microbial dynamics and activity. These reactors need to harbor a microbial community that is stable upon inoculation, colon region specific, and relevant to in vivo conditions. Together with the reproducibility of the colonization process, these criteria are often overlooked when the modulatory properties from different treatments are compared. We therefore investigated the microbial colonization process in two identical simulators of the human intestinal microbial ecosystem (SHIME), simultaneously inoculated with the same human fecal microbiota with a high-resolution phylogenetic microarray: the human intestinal tract chip (HITChip). Following inoculation of the in vitro colon compartments, microbial community composition reached steady state after 2 weeks, whereas 3 weeks were required to reach functional stability. This dynamic colonization process was reproducible in both SHIME units and resulted in highly diverse microbial communities which were colon region specific, with the proximal regions harboring saccharolytic microbes (e.g., Bacteroides spp. and Eubacterium spp.) and the distal regions harboring mucin-degrading microbes (e.g., Akkermansia spp.). Importantly, the shift from an in vivo to an in vitro environment resulted in an increased Bacteroidetes/Firmicutes ratio, whereas Clostridium cluster IX (propionate producers) was enriched compared to clusters IV and XIVa (butyrate producers). This was supported by proportionally higher in vitro propionate concentrations. In conclusion, high-resolution analysis of in vitro-cultured gut microbiota offers new insight on the microbial colonization process and indicates the importance of digestive parameters that may be crucial in the development of new in vitro models.

Boric acid-phenolic relationships within the Pinus echinata-Pisolithus tinctorius ectomycorrhizal association

Treesearch

Mary Anne Sword; Harold E. Garrett

1994-01-01

At germination, container-grown shortleaf pine seedlings were inoculated with Pisolithus tinctorius (Pers.) Coker & Couch or left uninoculated, and both groups were fertilized semiweekly with a modified Hoagland’s solution supplemented with 0 or 0.4 mM boric acid. After 12, 16 and 24 weeks, seedling root tissue was analyzed for...

Benomyl Stimulates Ectomycorrhizal Development by Pisolithus Tinctorius on Shortleaf Pine Grown in Containers

Treesearch

William H. Pawuk; James P. Barnett

1981-01-01

Container-grown shortleaf pine (Pinus echinata Mill.) seedlings inoculated with Pisolithus tinctorius and drenched with benomyl formed more mycorrhizal roots than undrenched seedlings. Seedlings were drenched (2.5, 5, and 10 mg ai in 15 ml of water per individual) prior to sowing and at either 2-, 4-, or 8-week intervals....

Impaired anti-fibrotic effect of bone marrow-derived mesenchymal stem cell in a mouse model of pulmonary paracoccidioidomycosis

PubMed Central

Arango, Julián Camilo; Puerta-Arias, Juan David; Pino-Tamayo, Paula Andrea; Salazar-Peláez, Lina María; Rojas, Mauricio

2017-01-01

Bone marrow-derived mesenchymal stem cells (BMMSCs) have been consider as a promising therapy in fibrotic diseases. Experimental models suggest that BMMSCs may be used as an alternative therapy to treat chemical- or physical-induced pulmonary fibrosis. We investigated the anti-fibrotic potential of BMMSCs in an experimental model of lung fibrosis by infection with Paracoccidioides brasiliensis. BMMSCs were isolated and purified from BALB/c mice using standardized methods. BALB/c male mice were inoculated by intranasal infection of 1.5x106 P. brasiliensis yeasts. Then, 1x106 BMMSCs were administered intra venous at 8th week post-infection (p.i.). An additional group of mice was treated with itraconazole (ITC) two weeks before BMMSCs administration. Animals were sacrificed at 12th week p.i. Histopathological examination, fibrocytes counts, soluble collagen and fibrosis-related genes expression in lungs were evaluated. Additionally, human fibroblasts were treated with homogenized lung supernatants (HLS) to determine induction of collagen expression. Histological analysis showed an increase of granulomatous inflammatory areas in BMMSCs-treated mice. A significant increase of fibrocytes count, soluble collagen and collagen-3α1, TGF-β3, MMP-8 and MMP-15 genes expression were also observed in those mice. Interestingly, when combined therapy BMMSCs/ITC was used there is a decrease of TIMP-1 and MMP-13 gene expression in infected mice. Finally, human fibroblasts stimulated with HLS from infected and BMMSCs-transplanted mice showed a higher expression of collagen I. In conclusion, our findings indicate that late infusion of BMMSCs into mice infected with P. brasiliensis does not have any anti-fibrotic effect; possibly because their interaction with the fungus promotes collagen expression and tissue remodeling. PMID:29040281

Enhanced protective immunity of the chimeric vector-based vaccine rAdV-SFV-E2 against classical swine fever in pigs by a Salmonella bacterial ghost adjuvant.

PubMed

Xia, Shui-Li; Lei, Jian-Lin; Du, Mingliang; Wang, Yimin; Cong, Xin; Xiang, Guang-Tao; Li, Lian-Feng; Yu, Shenye; Du, Enqi; Liu, Siguo; Sun, Yuan; Qiu, Hua-Ji

2016-06-14

Classical swine fever (CSF) is a highly contagious swine disease caused by classical swine fever virus (CSFV). Previously, we demonstrated that rAdV-SFV-E2, an adenovirus-delivered, Semliki Forest virus replicon-vectored marker vaccine against CSF, is able to protect pigs against lethal CSFV challenge. From an economical point of view, it will be beneficial to reduce the minimum effective dose of the vaccine. This study was designed to test the adjuvant effects of Salmonella enteritidis-derived bacterial ghosts (BG) to enhance the protective immunity of rAdV-SFV-E2 in pigs. Groups of 5-week-old pigs (n = 4) were immunized intramuscularly twice with 10(5) median tissue culture infective doses (TCID50) rAdV-SFV-E2 combined with 10(10) colony forming units (CFU) BG, 10(6) or 10(5) TCID50 rAdV-SFV-E2 alone or 10(10) CFU BG alone at an interval of 3 weeks, and challenged with the highly virulent CSFV Shimen strain at 1 week post-booster immunization. The results show that the pigs inoculated with 10(5) TCID50 rAdV-SFV-E2 plus BG or 10(6) TCID50 rAdV-SFV-E2 alone were completely protected from lethal CSFV challenge, in contrast with the pigs vaccinated with 10(5) TCID50 rAdV-SFV-E2 or BG alone, which displayed partial or no protection following virulent challenge. The data indicate that BG are a promising adjuvant to enhance the efficacy of rAdV-SFV-E2 and possibly other vaccines.

Potential of the TCE-degrading endophyte Pseudomonas putida W619-TCE to improve plant growth and reduce TCE phytotoxicity and evapotranspiration in poplar cuttings.

PubMed

Weyens, Nele; Truyens, Sascha; Dupae, Joke; Newman, Lee; Taghavi, Safiyh; van der Lelie, Daniel; Carleer, Robert; Vangronsveld, Jaco

2010-09-01

The TCE-degrading poplar endophyte Pseudomonas putida W619-TCE was inoculated in poplar cuttings, exposed to 0, 200 and 400 mg l(-1) TCE, that were grown in two different experimental setups. During a short-term experiment, plants were grown hydroponically in half strength Hoagland nutrient solution and exposed to TCE for 3 days. Inoculation with P. putida W619-TCE promoted plant growth, reduced TCE phytotoxicity and reduced the amount of TCE present in the leaves. During a mid-term experiment, plants were grown in potting soil and exposed to TCE for 3 weeks. Here, inoculation with P. putida W619-TCE had a less pronounced positive effect on plant growth and TCE phytotoxicity, but resulted in strongly reduced amounts of TCE in leaves and roots of plants exposed to 400 mg l(-1) TCE, accompanied by a lowered evapotranspiration of TCE. Dichloroacetic acid (DCAA) and trichloroacetic acid (TCAA), which are known intermediates of TCE degradation, were not detected. Copyright (c) 2010 Elsevier Ltd. All rights reserved.

Biodegradation of chlorobenzene by indigenous bacteria

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

Nishino, S.F.; Spain, J.C.; Pettigrew, C.A.

Soil and ground water from four sites chronically contaminated with chlorobenzenes were examined to determine whether indigenous bacteria could degrade the contaminants and whether the addition of specific chlorobenzene-degrading bacteria enhanced the degradation rate. At each site, chlorobenzene-degrading bacteria were readily isolated from chlorobenzene-contaminated wells, whereas similar samples from noncontaminated wells yielded no chlorobenzene-degrading bacteria. Isolates were tested for growth on a variety of substrates. At a site contaminated with several solvents, a bioreactor was inoculated with the chlorobenzene-degrading Pseudomonas sp. strain JS150. Contaminated water was pumped through this bioreactor and a control bioreactor that had been colonized by inmore » indigenous microorganisms. The contaminants were removed from both bioreactors; however, JS150 could not be recovered from the inoculated bioreactor after three weeks of operation. A follow-up lab study using ground water from the contaminated site confirmed the field results. The authors conclude that chlorobenzene contamination of soil causes the development of indigenous degradative populations that have a competitive advantage over inoculated strains. The mechanism and time course of this acclimation are poorly understood and require additional study.« less

Coast Guard All Hands

Science.gov Websites

. In this post, Rachel gets some answers from the experts on military moves! U.S. Coast Guard photo by messages to publish in a weekly post to help raise awareness about specific messages and useful information . General Messages Weekly Roundup All Hands selects several messages to publish in a weekly post to help

Vertical Transmission of H9N2 Avian Influenza Virus in Goose.

PubMed

Yu, Guanliu; Wang, Aihua; Tang, Yi; Diao, Youxiang

2017-01-01

During a study on high mortality cases of goose embryo in Shandong Province, China (2014-2015), we isolated an H9N2 avian influenza virus (AIV) strain (A/goose/Shandong/DP01/2014, DP01), which was supposedly the causative agent for goose embryo death. Sequence analysis revealed that DP01 shared 99.9% homology in the HA gene with a classic immune suppression strain SD06. To study the potential vertical transmission ability of the DP01 strain in breeder goose, a total of 105 Taizhou breeder geese, which were 360 days old, were equally divided into five groups (A, B, C, D, and E) for experimental infection. H9N2 AIV (DP01) was used for inoculating through intravenous (group A), intranasal instillation (group B), and throat inoculation (group C) routes, respectively. The geese in group D were inoculated with phosphate buffer solution (PBS) and those in group E were the non-treated group. At 24 h post inoculation, H9N2 viral RNA could be detected at vitelline membrane, embryos, and allantoic fluid of goose embryos from H9N2 inoculated groups. Furthermore, the HA gene of H9N2 virus from vitelline membrane, embryo, allantoic fluid, and gosling shared almost 100% homology with an H9N2 virus isolated from the ovary of breeder goose, which laid these eggs, indicating that H9N2 AIV can be vertically transmitted in goose. The present research study provides evidence that vertical transmission of H9N2 AIV from breeding goose to goslings is possible.

Binaural Beat Technology: A Complementary Path to Post Deployment Wellness

DTIC Science & Technology

2017-03-16

for at least three consecutive nights per week, for four weeks. A 20-minute pre and post -intervention heartrate variability (HRV) stress test and...four weeks. A 20-minute pre and post -intervention heartrate variability (HRV) stress test and daily perceived stress assessed intervention efficacy...DATES COVERED (Jul 7 2012 – Dec 31 2016) Binaural Beat Technology: A Complementary Path to Post Deployment Wellness 5a. CONTRACT NUMBER N/A 5b

Evaluation of chimeric yellow fever 17D/dengue viral replication in ticks.

PubMed

Kazimírová, Mária; Mantel, Nathalie; Raynaud, Sandrine; Slovák, Mirko; Ustaniková, Katarína; Lang, Jean; Guy, Bruno; Barban, Veronique; Labuda, Milan

2012-11-01

Chimeric yellow fever 17D/DENV-1-4 viruses (CYD-1-4) have been developed as a tetravalent dengue vaccine candidate which is currently being evaluated in efficacy trials in Asia and America. While YF 17D and DENV are mosquito-borne flaviviruses, it has been shown that CYD-1-4 do not replicate after oral infection in mosquitoes and are not transmitted to new hosts. To further document the risk of environmental dissemination of these viruses, we evaluated the replication of CYD-1-4 in ticks, the vector of tick-borne encephalitis virus (TBEV), another member of the flavivirus family. Females of two hard tick species, Ixodes ricinus and Rhipicephalus appendiculatus, were inoculated intracoelomically with CYD-1-4 viruses and parent viruses (DENV-1-4 and YF 17D). Virus persistence and replication was assessed 2, 16, and 44 days post-inoculation by plaque titration and qRT-PCR. CYD-1-4 viruses were detected in I. ricinus ticks at early time points post-inoculation, but with infectious titers at least 100-fold lower than those observed in TBEV-infected ticks. Unlike TBEV, complete viral clearance occurred by day 44 in most ticks except for CYD-2, which had a tendency to decline. In addition, while about 70% of TBEV-infected I. ricinus nymphs acquired infection by co-feeding with infected tick females on non-viremic hosts, no co-feeding transmission of CYD-2 virus was detected. Based on these results, we conclude that the risk of dissemination of the candidate vaccine viruses by tick bite is highly unlikely.

Do Holocaust survivors show increased vulnerability or resilience to post-Holocaust cumulative adversity?

PubMed

Shrira, Amit; Palgi, Yuval; Ben-Ezra, Menachem; Shmotkin, Dov

2010-06-01

Prior trauma can hinder coping with additional adversity or inoculate against the effect of recurrent adversity. The present study further addressed this issue by examining whether a subsample of Holocaust survivors and comparison groups, drawn from the Israeli component of the Survey of Health, Ageing, and Retirement in Europe, were differentially affected by post-Holocaust cumulative adversity. Post-Holocaust cumulative adversity had a stronger effect on the lifetime depression of Holocaust survivors than on that of comparisons. However, comparisons were more negatively affected by post-Holocaust cumulative adversity when examining markers of physical and cognitive functioning. Our findings suggest that previous trauma can both sensitize and immunize, as Holocaust survivors show general resilience intertwined with specific vulnerability when confronted with additional cumulative adversity.

Bacterial adaptation to the gut environment favors successful colonization: microbial and metabonomic characterization of a simplified microbiota mouse model.

PubMed

Rezzonico, Enea; Mestdagh, Renaud; Delley, Michèle; Combremont, Séverine; Dumas, Marc-Emmanuel; Holmes, Elaine; Nicholson, Jeremy; Bibiloni, Rodrigo

2011-01-01

Rodent models harboring a simple yet functional human intestinal microbiota provide a valuable tool to study the relationships between mammals and their bacterial inhabitants. In this study, we aimed to develop a simplified gnotobiotic mouse model containing 10 easy-to-grow bacteria, readily available from culture repositories, and of known genome sequence, that overall reflect the dominant commensal bacterial makeup found in adult human feces. We observed that merely inoculating a mix of fresh bacterial cultures into ex-germ free mice did not guarantee a successful intestinal colonization of the entire bacterial set, as mice inoculated simultaneously with all strains only harbored 3 after 21 d. Therefore, several inoculation procedures were tested and levels of individual strains were quantified using molecular tools. Best results were obtained by inoculating single bacterial strains into individual animals followed by an interval of two weeks before allowing the animals to socialize to exchange their commensal microbes. Through this procedure, animals were colonized with almost the complete bacterial set (9/10). Differences in the intestinal composition were also reflected in the urine and plasma metabolic profiles, where changes in lipids, SCFA, and amino acids were observed. We conclude that adaptation of bacterial strains to the host's gut environment (mono-colonization) may predict a successful establishment of a more complex microbiota in rodents.

Utilizing fluorescence hyperspectral imaging to differentiate corn inoculated with toxigenic and atoxigenic fungal strains

NASA Astrophysics Data System (ADS)

Yao, Haibo; Hruska, Zuzana; Kincaid, Russell; Brown, Robert L.; Bhatnagar, Deepak; Cleveland, Thomas E.

2012-05-01

Naturally occurring Aspergillus flavus strains can be either toxigenic or atoxigenic, indicating their ability to produce aflatoxin or not, under specific conditions. Corn contaminated with toxigenic strains of A. flavus can result in great losses to the agricultural industry and pose threats to public health. Past research showed that fluorescence hyperspectral imaging could be a potential tool for rapid and non-invasive detection of aflatoxin contaminated corn. The objective of the current study was to assess, with the use of a hyperspectral sensor, the difference in fluorescence emission between corn kernels inoculated with toxigenic and atoxigenic inoculums of A. flavus. Corn ears were inoculated with AF13, a toxigenic strain of A. flavus, and AF38, an atoxigenic strain of A. flavus, at dough stage of development and harvested 8 weeks after inoculation. After harvest, single corn kernels were divided into three groups prior to imaging: control, adjacent, and glowing. Both sides of the kernel, germplasm and endosperm, were imaged separately using a fluorescence hyperspectral imaging system. It was found that the classification accuracies of the three manually designated groups were not promising. However, the separation of corn kernels based on different fungal inoculums yielded better results. The best result was achieved with the germplasm side of the corn kernels. Results are expected to enhance the potential of fluorescence hyperspectral imaging for the detection of aflatoxin contaminated corn.

Effects of self-compassion workbook training on trauma-related guilt in a sample of homeless veterans: a pilot study.

PubMed

Held, Philip; Owens, Gina P

2015-06-01

The present pilot study examined the effects of a 4-week-long self-administered self-compassion training on trauma-related guilt and compared it to a stress inoculation control group. A total of 47 homeless male veterans who were living in transitional housing facilities volunteered to participate in this study. Participants were randomly assigned to either a self-compassion (N = 13) or a stress inoculation (N = 14) group and were asked to complete pre-, mid-, and postintervention assessments measuring changes in self-compassion, trauma-related guilt, and posttraumatic stress disorder severity. Participants in both interventions reported increased levels of self-compassion and equal reductions in trauma-related guilt. No other significant changes were noted. The results from this pilot study provide preliminary evidence for the use of self-compassion and stress inoculation trainings as effective interventions for trauma-related guilt. The findings also suggest that self-administered trainings in the form of workbooks may be a viable, cost-effective form of intervention for disadvantaged populations, such as homeless veterans in transitional housing, who may lack resources or access to professionals or paraprofessionals. The effects of both self-compassion training and stress inoculation training on the study variables and directions for future research on self-compassion and trauma-related guilt are discussed. © 2015 Wiley Periodicals, Inc.

The effect of moisture on infection of Rhododendron 'Cunningham's White' and Viburnum tinus by zoospores of Phytophthora ramorum

Treesearch

Paul W. Tooley; Marsha Browning

2017-01-01

We performed studies to determine the effect of leaf wetness on infection of whole plants of Rhododendron 'Cunningham's White' and Viburnum tinus by zoospores of Phytophthora ramorum. We also evaluated the effect of a post-inoculation drying period on infectivity of the two host species with...

Effect of storage at 4 and 10 C on growth of Listeria monocytogenes in Queso Fresco

USDA-ARS?s Scientific Manuscript database

A five-strain rifampicin – resistant Listeria monocytogenes cocktail (ca. 3.0 log10CFU/g) was introduced as a post-pasteurization contaminant in Queso Fresco (QF) that was manufactured using a commercial make procedure. L. monocytogenes was either inoculated into (IN) the curds before slicing or on...

A simplified technique to detect variations of leaf chlorogenic acid levels between and within plants caused by maturation or biological stress

USDA-ARS?s Scientific Manuscript database

The composition of phenolics in the plant apoplast vary during the first 5 hr post inoculation with different bacteria, depending on the plant-bacterial interaction. In general these responsive phenolics are not found at high concentrations in the symplast, within the cell. Recently we found that ...

Antibody response to chicken parvovirus following inoculation with inactivated virus and recombinant viruses expressing chicken parvovirus viral protein 2(VP2).

USDA-ARS?s Scientific Manuscript database

We reported earlier that day-old broiler chickens showed typical runting-stunting syndrome (RSS) post infection with chicken parvovirus (ChPV). There was also evidence that ChPV-specific maternal antibodies could provide significant protection against parvovirus induced enteric disease. Here, we st...

Increased lymphocyte subpopulations and macrophages in the ovaries and oviducts of laying hens infected with Salmonella enterica serovar Enteritidis.

PubMed

Withanage, G S K; Sasai, K; Fukata, T; Miyamoto, T; Lillehoj, H S; Baba, E

2003-12-01

Salmonella enterica serovar Enteritidis (SE) is a causative agent for human food poisoning cases throughout the world. The ovaries and the oviducts of the laying hens are the major sites of SE colonization from which vertical transmission to eggs occurs. In this study, Salmonella-induced changes in T lymphocytes, B lymphocytes and macrophages in the ovaries and oviducts were assessed after primary and secondary experimental inoculations of laying hen with SE. Statistically significant increases in the numbers of T cells (both CD4+ and CD8+) and macrophages were observed 7 to 14 days after primary inoculation, followed by a peak in B-cell numbers from the 14th day post-primary inoculation onwards in the secretory areas of the oviducts. The peak in lymphocyte numbers immediately preceded a decline in the rate of SE recovery from the reproductive tract beginning at day 14. The correlation of decreased Salmonella recovery with elevated lymphocyte and macrophage numbers strongly suggests that local cell-mediated immunity is involved in controlling SE injection in the ovaries and oviducts.

Biochar and Glomus caledonium Influence Cd Accumulation of Upland Kangkong (Ipomoea aquatica Forsk.) Intercropped with Alfred Stonecrop (Sedum alfredii Hance)

NASA Astrophysics Data System (ADS)

Hu, Junli; Wu, Fuyong; Wu, Shengchun; Lam, Cheung Lung; Lin, Xiangui; Wong, Ming Hung

2014-04-01

Both biochar application and mycorrhizal inoculation have been proposed to improve plant growth and alter bioaccumulation of toxic metals. A greenhouse pot trial was conducted to investigate growth and Cd accumulation of upland kangkong (Ipomoea aquatica Forsk.) intercropped with Alfred stonecrop (Sedum alfredii Hance) in a Cd-contaminated soil inoculated with Glomus caledonium and/or applied with biochar. Compared with the monocultural control, intercropping with stonecrop (IS) decreased kangkong Cd acquisition via rhizosphere competition, and also decreased kangkong yield. Gc inoculation (+M) accelerated growth and Cd acquisition of stonecrop, and hence resulted in further decreases in kangkong Cd acquisition. Regardless of IS and +M, biochar addition (+B) increased kangkong yield via elevating soil available P, and decreased soil Cd phytoavailability and kangkong Cd concentration via increasing soil pH. Compared with the control, the treatment of IS + M + B had a substantially higher kangkong yield (+25.5%) with a lower Cd concentration (-62.7%). Gc generated additive effects on soil alkalinization and Cd stabilization to biochar, causing lower DTPA-extractable (phytoavailable) Cd concentrations and post-harvest transfer risks.

Susceptibility of openbill storks (Anastomius oscitans) to highly pathogenic avian influenza virus subtype H5N1.

PubMed

Chaichoun, Kridsada; Wiriyarat, Withawat; Phonaknguen, Rassmeepen; Sariya, Ladawan; Taowan, Nam-aoy; Chakritbudsabong, Warunya; Chaisilp, Natnapat; Eiam-ampai, Krirat; Phuttavatana, Pilaipan; Ratanakorn, Parntep

2013-09-01

This investigation detailed the clinical disease, gross and histologic lesions in juvenile openbill storks (Anastomus oscitans) intranasally inoculated with an avian influenza virus, A/chicken/Thailand/vsmu-3 (H5N1), which is highly pathogenic for chickens. High morbidity and mortality were observed in openbill storks inoculated with HPAI H5N1 virus. Gross lesions from infected birds were congestion and brain hemorrhage (10/20), pericardial effusions, pericarditis and focal necrosis of the cardiac muscle (2/20), pulmonary edema and pulmonary necrosis, serosanguineous fluid in the bronchis (16/20), liver congestion (6/20), bursitis (5/20), subcutaneous hemorrhages (2/20) and pinpoint proventiculus hemorrhage (2/20). Real time RT-PCR demonstrated the presence of viral RNA in organs associated with the lesions: brain, trachea, lungs, liver, spleen and intestines. Similar to viral genome detection, virus was also isolated from these vital organs. Antibodies to influenza virus detected with a hemagglutination inhibition test, were found only in the openbill storks who died 8 days post-inoculation.

Fetal Cortical Transplants in Adult Rats Subjected to Experimental Brain Injury

PubMed Central

Soares, Holly; McIntosh, Tracy K.

1991-01-01

Fetal cortical tissue was injected into injured adult rat brains following concussive fluid percussion (FP) brain injury. Rats subjected to moderate FP injury received E16 cortex transplant injections into lesioned motor cortex 2 days, 1 week, 2 weeks, and 4 weeks post injury. Histological assessment of transplant survival and integration was based upon Nissl staining, glial fibrillary acidic protein (GFAP) immunocytochemistry, and staining for acetylcholinesterase. In addition to histological analysis, the ability of the transplants to attenuate neurological motor deficits associated with concussive FP brain injury was also tested. Three subgroups of rats receiving transplant 1 week, 2 weeks, and 4 weeks post injury Were chosen for evaluation of neurological motor function. Fetal cortical tissue injected into the injury site 4 weeks post injury failed to incorporate with injured host brain, did not affect glial scar formation, and exhibited extensive GFAP immunoreactivity. No improvement in neurological motor function was observed in animals receiving transplants 4 weeks post injury. Conversely, transplants injected 2 days, 1 week, or 2 weeks post injury survived, incorporated with host brain, exhibited little GFAP immunoreactivity, and successfully attenuated glial scarring. However, no significant improvement in motor function was observed at the one week or two week time points. The inability of the transplants to attenuate motor function may indicate inappropriate host/transplant interaction. Our results demonstrate that there exists a temporal window in which fetal cortical transplants can attenuate glial scarring as well as be successfully incorporated into host brains following FP injury. PMID:1782253

Pulmonary Abnormalities in Mice with Paracoccidioidomycosis: A Sequential Study Comparing High Resolution Computed Tomography and Pathologic Findings

PubMed Central

Hidalgo, José Miguel; de Oliveira Pascarelli, Bernardo Miguel; Patiño, Jairo Hernando; Lenzi, Henrique Leonel; Restrepo, Angela; Cano, Luz Elena

2010-01-01

Background Human paracoccidioidomycosis (PCM) is an endemic fungal disease of pulmonary origin. Follow-up of pulmonary lesions by image studies in an experimental model of PCM has not been previously attempted. This study focuses on defining patterns, topography and intensity of lung lesions in experimentally infected PCM mice by means of a comparative analysis between High Resolution Computed Tomography (HRCT) and histopathologic parameters. Methodology Male BALB/c mice were intranasally inoculated with 3×106 Paracoccidioides brasiliensis (Pb) conidia (n = 50) or PBS (n = 50). HRCT was done every four weeks to determine pulmonary lesions, quantify lung density, reconstruct and quantify lung air structure. Lungs were also analyzed by histopathology and histomorphometry. Results Three different patterns of lesions were evidenced by HRCT and histopathology, as follows: nodular-diffuse, confluent and pseudo-tumoral. The lesions were mainly located around the hilus and affected more frequently the left lung. At the 4th week post-challenge HRCT showed that 80% of the Pb-infected mice had peri-bronchial consolidations associated with a significant increase in upper lung density when compared with controls, (−263±25 vs. −422±10 HU, p<0.001). After the 8th and 12th weeks, consolidation had progressed involving also the middle regions. Histopathology revealed that consolidation as assessed by HRCT was equivalent histologically to a confluent granulomatous reaction, while nodules corresponded to individual compact granulomas. At the 16th week of infection, confluent granulomas formed pseudotumoral masses that obstructed large bronchi. Discrete focal fibrosis was visible gradually around granulomas, but this finding was only evident by histopathology. Conclusions/Significance This study demonstrated that conventional HRCT is a useful tool for evaluation and quantification of pulmonary damage occurring in experimental mouse PCM. The experimental design used decreases the need to sacrifice a large number of animals, and serves to monitor treatment efficacy by means of a more rational approach to the study of human lung disease. PMID:20614019

Assessment of the vaginal residence time of biomarkers of semen exposure.

PubMed

Thurman, Andrea; Jacot, Terry; Melendez, Johan; Kimble, Thomas; Snead, Margaret; Jamshidi, Roxanne; Wheeless, Angie; Archer, David F; Doncel, Gustavo F; Mauck, Christine

2016-11-01

The primary objective of this pilot study is to determine and compare the residence time in the vagina of biomarkers of semen exposure for up to 15 days post exposure. The biomarkers are prostate-specific antigen (PSA), Y chromosome DNA, the sex determining region of the Y chromosome (SRY) and testis-specific protein Y-encoded 4 (TSPY4). The secondary objectives are to determine if biomarker concentrations differed between intercourse and inoculation groups, to establish whether the sampling frequency post exposure affected biomarker concentrations and decay profile and to determine if biomarker concentrations in vaginal swabs obtained by the participant at home were similar to swabs obtained by the nurse in the clinic. We randomized healthy women to unprotected intercourse (n=17) versus vaginal inoculation with the male partner's semen in the clinic (n=16). Women were then further randomized to have vaginal swabs obtained at either 7 or 4 time points after semen exposure, up to 15 days post exposure, either obtained at home by the participant or in the clinic by the research nurse. PSA and SRY were markers of recent semen exposure. TSPY4 was detectable in approximately 50% of participants at 15 days post exposure. Unprotected intercourse resulted in significantly higher concentrations of select biomarkers. Sampling frequency and home versus clinic sampling had no significant effect on biomarker concentrations. Objective biomarkers of recent or distant semen exposure may have great utility for verifying protocol compliance in a variety of clinical trials. Copyright © 2016 Elsevier Inc. All rights reserved.

Assessment of the vaginal residence time of biomarkers of semen exposure☆,☆☆,★

PubMed Central

Thurman, Andrea; Jacot, Terry; Melendez, Johan; Kimble, Thomas; Snead, Margaret; Jamshidi, Roxanne; Wheeless, Angie; Archer, David F.; Doncel, Gustavo F.; Mauck, Christine

2016-01-01

Objective The primary objective of this pilot study is to determine and compare the residence time in the vagina of biomarkers of semen exposure for up to 15 days post exposure. The biomarkers are prostate-specific antigen (PSA), Y chromosome DNA, the sex determining region of the Y chromosome (SRY) and testis-specific protein Y-encoded 4 (TSPY4). The secondary objectives are to determine if biomarker concentrations differed between intercourse and inoculation groups, to establish whether the sampling frequency post exposure affected biomarker concentrations and decay profile and to determine if biomarker concentrations in vaginal swabs obtained by the participant at home were similar to swabs obtained by the nurse in the clinic. Study design We randomized healthy women to unprotected intercourse (n=17) versus vaginal inoculation with the male partner’s semen in the clinic (n=16). Women were then further randomized to have vaginal swabs obtained at either 7 or 4 time points after semen exposure, up to 15 days post exposure, either obtained at home by the participant or in the clinic by the research nurse. Results PSA and SRY were markers of recent semen exposure. TSPY4 was detectable in approximately 50% of participants at 15 days post exposure. Unprotected intercourse resulted in significantly higher concentrations of select biomarkers. Sampling frequency and home versus clinic sampling had no significant effect on biomarker concentrations. Conclusions Objective biomarkers of recent or distant semen exposure may have great utility for verifying protocol compliance in a variety of clinical trials. PMID:27259675

Escherichia coli O78 isolated from septicemic lambs shows high pathogenicity in a zebrafish model.

PubMed

Kjelstrup, Cecilie K; Barber, Amelia E; Norton, J Paul; Mulvey, Matthew A; L'Abée-Lund, Trine M

2017-01-25

The pathogenicity of Escherichia coli O78 strain K46, originally isolated from an outbreak of septicemia in neonatal lambs, was investigated in zebrafish embryo and murine models of infection. Its biofilm potential, cellulose production, and the expression of type 1 pili and curli fimbriae were measured by in vitro assays. The strain was highly pathogenic in the zebrafish embryo model of infection, where it killed all embryos within 24 h post inoculation (hpi) at doses as low as 1000 colony forming units. Zebrafish embryos inoculated with similar doses of commensal E. coli strains showed no signs of disease, and cleared the bacteria within 24 h. E. coli K46 colonized the murine gut at the same level as the uropathogenic E. coli (UPEC) reference strain CFT073 in CBA/J mice after oral inoculation, but infected the murine bladder significantly less than CFT073 after transurethral inoculation. Type 1 pili were clearly expressed by E. coli K46, while curli fimbriae and cellulose production were weakly expressed. The ability to produce biofilm varied in different growth media, but overall E. coli K46 was a poorer biofilm producer compared to the reference strain E. coli UTI89. In conclusion, the zebrafish lethality model provides further evidence that E. coli K46 is highly pathogenic and might be useful in future studies to identify bacterial virulence factors.

Tissue localization, shedding, virus carriage, antibody response, and aerosol transmission of porcine epidemic diarrhea virus (PEDV) following inoculation of 4 week-old feeder pigs

USDA-ARS?s Scientific Manuscript database

Porcine epidemic diarrhea virus (PEDV) emerged in the U.S. in April 2013 and caused significant losses to the swine industry. The purpose of this investigation was to determine tissue localization, shedding patterns, virus carriage, antibody response, and aerosol transmission of PEDV following inocu...

Test method for assessing resistance of pine lumber and waferboard to mold

Treesearch

Carol A. Clausen; Michael West

2005-01-01

Methods are needed to evaluate the ability of framing lumber and composite construction materials to withstand mold growth when they are exposed to rain between manufacture and installation. A laboratory-controlled rain chamber was developed to expose biocide-treated specimens of pine lumber and waferboard to bi-weekly wetting followed by re-inoculation with test fungi...

No evidence for differential dose effects of hydrocortisone on intrusive memories in female patients with complex post-traumatic stress disorder--a randomized, double-blind, placebo-controlled, crossover study.

PubMed

Ludäscher, Petra; Schmahl, Christian; Feldmann, Robert E; Kleindienst, Nikolaus; Schneider, Miriam; Bohus, Martin

2015-10-01

Post-traumatic stress disorder is characterized by intrusive traumatic memories. Presently, a controversial debate is ongoing regarding whether reduced cortisol secretion in post-traumatic stress disorder promotes an automatic retrieval of trauma-associated memories. Hence, a pharmacological elevation of cortisol was proposed to decrease post-traumatic stress disorder symptoms, particularly intrusions. The present study investigated the impact of two different doses of hydrocortisone on automatic memory retrieval using a randomized, double-blind, placebo-controlled, crossover study in 30 inpatients with post-traumatic stress disorder. All participants were female and received various psychotropic medications. They were randomly assigned to one of two groups within a crossover design: they received either 1 week placebo followed by 1 week hydrocortisone 10/d, followed by 1 week placebo, followed by hydrocortisone 30 mg/d (15 participants) or 1 week hydrocortisone 30 mg/d, followed by 1 week placebo, followed by 1 week hydrocortisone 10 mg/d, followed by 1 week placebo (15 participants). The outcome measures were the frequency and the intensity of intrusions, the overall symptomatology of post-traumatic stress disorder and the general psychopathology. We did not find any differences in the frequency and the intensity of post-traumatic stress disorder-related intrusions between the 10 mg hydrocortisone, the 30 mg hydrocortisone and the placebo condition. All effect sizes for the hydrocortisone condition vs. placebo were very small. Additionally, the overall symptomatology of post-traumatic stress disorder and the general psychopathology did not differ between the hydrocortisone therapies and placebo. Our results do not show any effect of the hydrocortisone administration on intrusions in complex post-traumatic stress disorder. © The Author(s) 2015.

Arbuscular mycorrhizal fungi affect both penetration and further life stage development of root-knot nematodes in tomato.

PubMed

Vos, Christine; Geerinckx, Katleen; Mkandawire, Rachel; Panis, Bart; De Waele, Dirk; Elsen, Annemie

2012-02-01

The root-knot nematode Meloidogyne incognita poses a worldwide threat to agriculture, with an increasing demand for alternative control options since most common nematicides are being withdrawn due to environmental concerns. The biocontrol potential of arbuscular mycorrhizal fungi (AMF) against plant-parasitic nematodes has been demonstrated, but the modes of action remain to be unraveled. In this study, M. incognita penetration of second-stage juveniles at 4, 8 and 12 days after inoculation was compared in tomato roots (Solanum lycopersicum cv. Marmande) pre-colonized or not by the AMF Glomus mosseae. Further life stage development of the juveniles was also observed in both control and mycorrhizal roots at 12 days, 3 weeks and 4 weeks after inoculation by means of acid fuchsin staining. Penetration was significantly lower in mycorrhizal roots, with a reduction up to 32%. Significantly lower numbers of third- and fourth-stage juveniles and females accumulated in mycorrhizal roots, at a slower rate than in control roots. The results show for the first time that G. mosseae continuously suppresses root-knot nematodes throughout their entire early infection phase of root penetration and subsequent life stage development.

Experimental infection of a US spike-insertion deletion porcine epidemic diarrhea virus in conventional nursing piglets and cross-protection to the original US PEDV infection.

PubMed

Lin, Chun-Ming; Annamalai, Thavamathi; Liu, Xinsheng; Gao, Xiang; Lu, Zhongyan; El-Tholoth, Mohamed; Hu, Hui; Saif, Linda J; Wang, Qiuhong

2015-11-20

Although the original US porcine epidemic diarrhea virus (PEDV) was confirmed as highly virulent by multiple studies, the virulence of spike-insertion deletion (S-INDEL) PEDV strains is undefined. In this study, 3-4 day-old conventional suckling piglets were inoculated with S-INDEL PEDV Iowa106 (4 pig litters) to study its virulence. Two litters of age-matched piglets were inoculated with either the original US PEDV PC21A or mock as positive and negative controls, respectively. Subsequently, all pigs were challenged with the original US PEDV PC21A on 21-29 days post-inoculation (dpi) to assess cross-protection. All S-INDEL Iowa106- and the original US PC21A-inoculated piglets developed diarrhea. However, the severity of clinical signs, mortality (0-75%) and fecal PEDV RNA shedding titers varied among the four S-INDEL Iowa106-inoculated litters. Compared with the original PC21A, piglets euthanized/died acutely from S-INDEL Iowa106 infection had relatively milder villous atrophy, lower antigen scores and more limited intestinal infection. Two of four S-INDEL Iowa106-infected sows and the original PC21A-infected sow showed anorexia and watery diarrhea for 1-4 days. After the original PC21A challenge, a subset (13/16) of S-INDEL Iowa106-inoculated piglets developed diarrhea, whereas all (5/5) and no (0/4) pigs in the mock and original PC21A-inoculated pigs had diarrhea, respectively. Our results suggest that the virulence of S-INDEL PEDV Iowa106 was less than the original US PEDV PC21A in suckling pigs, with 100% morbidity and 18% (6/33) overall (0-75%) mortality in suckling pigs depending on factors such as the sow's health and lactation and the piglets' birth weight. Prior infection by S-INDEL Iowa106 provided partial cross-protection to piglets against the original PC21A challenge at 21-29 dpi.

78 FR 34103 - Proposed Data Collections Submitted for Public Comment and Recommendations

Federal Register 2010, 2011, 2012, 2013, 2014

2013-06-06

... mine personnel about their behaviors the previous week, and (4) a post-test concerning miners' attitude... pre- and post-test and be interviewed upon completion of the intervention period. The operators at... Worksheet 109 1 15/60 27 Week 4 Worksheet 109 1 15/60 27 Week 5 Worksheet 109 1 15/60 27 Post-test Survey...

Combined Inoculation with Multiple Arbuscular Mycorrhizal Fungi Improves Growth, Nutrient Uptake and Photosynthesis in Cucumber Seedlings.

PubMed

Chen, Shuangchen; Zhao, Hongjiao; Zou, Chenchen; Li, Yongsheng; Chen, Yifei; Wang, Zhonghong; Jiang, Yan; Liu, Airong; Zhao, Puyan; Wang, Mengmeng; Ahammed, Golam J

2017-01-01

Mycorrhizal inoculation stimulates growth, photosynthesis and nutrient uptake in a wide range of host plants. However, the ultimate effects of arbuscular mycorrhyzal (AM) symbiosis vary with the plants and fungal species involved in the association. Therefore, identification of the appropriate combinations of AM fungi (AMF) that interact synergistically to improve their benefits is of high significance. Here, three AM fungal compositions namely VT ( Claroideoglomus sp., Funneliformis sp., Diversispora sp., Glomus sp., and Rhizophagus sp.) and BF ( Glomus intraradices , G. microageregatum BEG and G. Claroideum BEG 210), and Funneliformis mosseae (Fm) were investigated with respect to the growth, gas exchange parameters, enzymes activities in Calvin cycles and related gene expression in cucumber seedlings. The results showed that VT, BF and Fm could successfully colonize cucumber root to a different degree with the colonization rates 82.38, 74.65, and 70.32% at 46 days post inoculation, respectively. The plant height, stem diameter, dry weight, root to shoot ratio of cucumber seedlings inoculated with AMF increased significantly compared with the non-inoculated control. Moreover, AMF colonization greatly increased the root activity, chlorophyll content, net photosynthetic rate, light saturated rate of the CO 2 assimilation ( A sat), maximum carboxylation rate ( V cmax ) and maximum ribulose-1,5-bis-phosphate (RuBP) regeneration rate ( J max), which were increased by 52.81, 30.75, 58.76, 47.00, 69.15, and 65.53% when inoculated with VT, respectively. The activities of some key enzymes such RuBP carboxylase/oxygenase (RuBisCO), D-fructose-1,6-bisphosphatase (FBPase), D-fructose-6-phosphatase (F6P) and ribulose-5-phosphate kinase (Ru5PK), and related gene expression involved in the Calvin cycle including RCA , FBPase , FBPA , SBPase , rbcS and rbcL were upregulated by AMF colonization. AMF inoculation also improved macro- and micro nutrient contents such as N, P, K, S, Ca, Cu, Fe, Mn, Mg, and Zn in roots. Further analysis revealed that inoculation with VT had relatively better effect on growth of cucumber seedling followed by BF and Fm, indicating that AMF composition consisting of distant AMF species may have a better effect than a single or closely related AMF spp. This study advances the understanding of plant responses to different AM fungi toward development of strategies on AMF-promoted vegetable production.

Prevalence of neuro-musculoskeletal pain and dysfunction in open-heart surgical patients preoperatively and at 6 and 12 weeks postoperatively: a prospective longitudinal observation study.

PubMed

Bellet, R Nicole; Lamb, Rhonda L; Gould, Tonya D; Bartlett, Harold J

2017-01-01

Chronic neuro-musculoskeletal pain is an important complication of open-heart surgery (OHS). To better understand the development and natural course of neuro-musculoskeletal pain in the immediate post-OHS period, this prospective longitudinal study assessed the prevalence and degree of pain and shoulder disability, and areas of pain pre- and post-OHS. Usual medical, nursing, and physiotherapy care was provided including early extubation, education, walking, sitting out of bed, and upper, lower limb, and trunk exercises from day 1 post-operation. Of 114 elective patients who provided consent, 98 subjects were surveyed preoperatively, and at week 6 and week 12 post-OHS. Open and closed questions encompassed numerical rating of pain scales for various body areas summed as a total pain score (TPS), the shoulder disability score (SDS), exercise compliance, and sternal clicking. Usual care comprised mobility exercises, walking program, and cardiac rehabilitation referral. Survey return rates were 100%, 88%, and 82%, respectively. Of the 76 (78%) subjects with complete data sets, 68% subjects reported a history of previous neuro-musculoskeletal injuries/conditions preoperatively while prevalence for neuro-musculoskeletal pain was 64%, 88%, and 67% and 38%, 63%, and 42% for shoulder disability, at the three assessments. In all, 11% subjects reported sternal clicking at week 6 and 7% at week 12. Pain commonly occurred in the lower back and neck preoperatively, and in front of the chest, neck, rib cage, upper back, and left shoulder at week 6. Rib cage pain alone remained significantly greater than preoperative levels by week 12 post-OHS. Preoperative SDS was positively correlated with post-OHS length of stay; women had higher SDSs than men at week 6 and week 12 and week 12 SDS was negatively correlated with height. Surgical risk score was negatively correlated with change in SDS and TPS from pre-operation to week 12. In conclusion, neuro-musculoskeletal pain and shoulder disability were common preoperatively and while prevalence increased at week 6 post-OHS, overall preoperative levels were restored by week 12.

Prevalence of neuro-musculoskeletal pain and dysfunction in open-heart surgical patients preoperatively and at 6 and 12 weeks postoperatively: a prospective longitudinal observation study

PubMed Central

Bellet, R Nicole; Lamb, Rhonda L; Gould, Tonya D; Bartlett, Harold J

2017-01-01

Chronic neuro-musculoskeletal pain is an important complication of open-heart surgery (OHS). To better understand the development and natural course of neuro-musculoskeletal pain in the immediate post-OHS period, this prospective longitudinal study assessed the prevalence and degree of pain and shoulder disability, and areas of pain pre- and post-OHS. Usual medical, nursing, and physiotherapy care was provided including early extubation, education, walking, sitting out of bed, and upper, lower limb, and trunk exercises from day 1 post-operation. Of 114 elective patients who provided consent, 98 subjects were surveyed preoperatively, and at week 6 and week 12 post-OHS. Open and closed questions encompassed numerical rating of pain scales for various body areas summed as a total pain score (TPS), the shoulder disability score (SDS), exercise compliance, and sternal clicking. Usual care comprised mobility exercises, walking program, and cardiac rehabilitation referral. Survey return rates were 100%, 88%, and 82%, respectively. Of the 76 (78%) subjects with complete data sets, 68% subjects reported a history of previous neuro-musculoskeletal injuries/conditions preoperatively while prevalence for neuro-musculoskeletal pain was 64%, 88%, and 67% and 38%, 63%, and 42% for shoulder disability, at the three assessments. In all, 11% subjects reported sternal clicking at week 6 and 7% at week 12. Pain commonly occurred in the lower back and neck preoperatively, and in front of the chest, neck, rib cage, upper back, and left shoulder at week 6. Rib cage pain alone remained significantly greater than preoperative levels by week 12 post-OHS. Preoperative SDS was positively correlated with post-OHS length of stay; women had higher SDSs than men at week 6 and week 12 and week 12 SDS was negatively correlated with height. Surgical risk score was negatively correlated with change in SDS and TPS from pre-operation to week 12. In conclusion, neuro-musculoskeletal pain and shoulder disability were common preoperatively and while prevalence increased at week 6 post-OHS, overall preoperative levels were restored by week 12. PMID:29066939

Screening and identification of resistance related proteins from apple leaves inoculated with Marssonina coronaria (EII. & J. J. Davis)

PubMed Central

2014-01-01

Background Apple, an invaluable fruit crop worldwide, is often prone to infection by pathogenic fungi. Identification of potentially resistance-conferring apple proteins is one of the most important aims for studying apple resistance mechanisms and promoting the development of disease-resistant apple strains. In order to find proteins which promote resistance to Marssonina coronaria, a deadly pathogen which has been related to premature apple maturation, proteomes from apple leaves inoculated with M. coronaria were characterized at 3 and 6 days post-inoculation by two dimensional electrophoresis (2-DE). Results Overall, 59 differentially accumulated protein spots between inoculation and non-inoculation were successfully identified and aligned as 35 different proteins or protein families which involved in photosynthesis, amino acid metabolism, transport, energy metabolism, carbohydrate metabolism, binding, antioxidant, defense and stress. Quantitative real-time PCR (qRT-PCR) was also used to examine the change of some defense and stress related genes abundance under inoculated conditions. Conclusions In a conclusion, different proteins in response to Marssonina coronaria were identified by proteomic analysis. Among of these proteins, there are some PR proteins, for example class III endo-chitinase, beta-1,3-glucanase and thaumatine-like protein, and some antioxidant related proteins including aldo/keto reductase AKR, ascorbate peroxidase and phi class glutathione S-transferase protein that were associated with disease resistance. The transcription levels of class III endo-chitinase (L13) and beta-1, 3-glucanase (L17) have a good relation with the abundance of the encoded protein’s accumulation, however, the mRNA abundance of thaumatine-like protein (L22) and ascorbate peroxidase (L28) are not correlated with their protein abundance of encoded protein. To elucidate the resistant mechanism, the data in the present study will promote us to investigate further the expression regulation of these target proteins. PMID:24507458

Post discharge problems in women recovering from coronary artery bypass graft surgery.

PubMed

Gallagher, Robyn; McKinley, Sharon; Dracup, Kathleen

2004-11-01

This study was conducted to describe the types and frequency of problems Australian women experience when recovering at home in the first 6 weeks following coronary artery bypass graft (CABG) surgery and the relationship between symptom experience and psychological distress. A convenience sample of 52 women (mean age 66.31 years, range 53-79 years) who had uncomplicated CABG surgery was selected from two tertiary hospitals in Sydney. A descriptive design was used with information related to post-operative problems collected by telephone interview at 1, 3 and 6 weeks post discharge using a semistructured questionnaire. Psychological distress was assessed at 12 weeks post discharge using the Hospital Anxiety and Depression Scale (HADS). Responses were categorised, collapsed and described using frequencies and percentages. Relationships were assessed by Spearman's r. The most common problems in the first and third weeks post discharge were sleeplessness and nausea or poor appetite and chest incision pain. Although problems improved over the first 6 weeks post-operatively, approximately one-quarter of the women still reported chest incision pain and almost 40% reported problems with leg wounds and oedema. The number of problems experienced at 6 weeks was significantly correlated with depression at 12 weeks. These findings support the importance of a preoperative education programme that includes anticipation of physical problems in the immediate post-operative period and a follow-up of female patients in the early transition period following hospital discharge.

The quest for a non-vector psyllid: Natural variation in acquisition and transmission of the huanglongbing pathogen ‘Candidatus Liberibacter asiaticus’ by Asian citrus psyllid isofemale lines

PubMed Central

Ammar, El-Desouky; Hall, David G.; Hosseinzadeh, Saeed

2018-01-01

Genetic variability in insect vectors is valuable to study vector competence determinants and to select non-vector populations that may help reduce the spread of vector-borne pathogens. We collected and tested vector competency of 15 isofemale lines of Asian citrus psyllid, Diaphorina citri, vector of ‘Candidatus Liberibacter asiaticus’ (CLas). CLas is associated with huanglongbing (citrus greening), the most serious citrus disease worldwide. D. citri adults were collected from orange jasmine (Murraya paniculata) hedges in Florida, and individual pairs (females and males) were caged on healthy Murraya plants for egg laying. The progeny from each pair that tested CLas-negative by qPCR were maintained on Murraya plants and considered an isofemale line. Six acquisition tests on D. citri adults that were reared as nymphs on CLas-infected citrus, from various generations of each line, were conducted to assess their acquisition rates (percentage of qPCR-positive adults). Three lines with mean acquisition rates of 28 to 32%, were classified as ‘good’ acquirers and three other lines were classified as ‘poor’ acquirers, with only 5 to 8% acquisition rates. All lines were further tested for their ability to inoculate CLas by confining CLas-exposed psyllids for one week onto healthy citrus leaves (6–10 adults/leaf/week), and testing the leaves for CLas by qPCR. Mean inoculation rates were 19 to 28% for the three good acquirer lines and 0 to 3% for the three poor acquirer lines. Statistical analyses indicated positive correlations between CLas acquisition and inoculation rates, as well as between CLas titer in the psyllids and CLas acquisition or inoculation rates. Phenotypic and molecular characterization of one of the good and one of the poor acquirer lines revealed differences between them in color morphs and hemocyanin expression, but not the composition of bacterial endosymbionts. Understanding the genetic architecture of CLas transmission will enable the development of new tools for combating this devastating citrus disease. PMID:29652934

Fall risk and function in older women after gynecologic surgery.

PubMed

Miller, Karen L; Richter, Holly E; Graybill, Charles S; Neumayer, Leigh A

2017-11-01

To examine change in balance-related fall risk and daily functional abilities in the first 2 post-operative weeks and up to 6 weeks after gynecologic surgery. Prospective cohort study in gynecologic surgery patients age 65 and older. Balance confidence (Activities-specific Balance Confidence Scale) and functional status (basic and instrumental activities of daily living) were recorded pre- and post-operatively daily for 1 week and twice the second week. Physical performance balance and functional mobility were measured pre- and 1 week post-operatively using the Tinetti Fall Risk Scale, Timed Up and Go, and 6-Minute Walk test. Measures were repeated 6 weeks after surgery. Non-parametric tests for paired data were used comparing scores baseline to post-operative (POD) 7 and to POD 42. Median age was 72 years (range 65-88). Fall risk was elevated during the first 2 post-operative weeks, greatest on the median discharge day, POD 2 (p<0.01). Balance performance and functional mobility at 1 week were significantly lower than baseline (p<0.01). Functional abilities declined, including new dependence in medication management at home in 22% of these independent and cognitively intact women. After gynecologic surgery, older women's fall risk is highest on POD 2 and remains elevated from baseline for 2 weeks. Functional limitations in the early home recovery period include the anticipated (bathing, cooking, etc.) and some unanticipated (medication management) ones. This information may help with post-operative discharge planning. Copyright © 2017 The Author(s). Published by Elsevier B.V. All rights reserved.

Experimental Traumatic Brain Injury Induces Bone Loss in Rats.

PubMed

Brady, Rhys D; Shultz, Sandy R; Sun, Mujun; Romano, Tania; van der Poel, Chris; Wright, David K; Wark, John D; O'Brien, Terence J; Grills, Brian L; McDonald, Stuart J

2016-12-01

Few studies have investigated the influence of traumatic brain injury (TBI) on bone homeostasis; however, pathophysiological mechanisms involved in TBI have potential to be detrimental to bone. The current study assessed the effect of experimental TBI in rats on the quantity and quality of two different weight-bearing bones, the femur and humerus. Rats were randomly assigned into either sham or lateral fluid percussion injury (FPI) groups. Open-field testing to assess locomotion was conducted at 1, 4, and 12 weeks post-injury, with the rats killed at 1 and 12 weeks post-injury. Bones were analyzed using peripheral quantitative computed tomography (pQCT), histomorphometric analysis, and three-point bending. pQCT analysis revealed that at 1 and 12 weeks post-injury, the distal metaphyseal region of femora from FPI rats had reduced cortical content (10% decrease at 1 week, 8% decrease at 12 weeks; p < 0.01) and cortical thickness (10% decrease at 1 week, 11% decrease at 12 weeks p < 0.001). There was also a 23% reduction in trabecular bone volume ratio at 1 week post-injury and a 27% reduction at 12 weeks post-injury in FPI rats compared to sham (p < 0.001). There were no differences in bone quantity and mechanical properties of the femoral midshaft between sham and TBI animals. There were no differences in locomotor outcomes, which suggested that post-TBI changes in bone were not attributed to immobility. Taken together, these findings indicate that this rat model of TBI was detrimental to bone and suggests a link between TBI and altered bone remodeling.

Storage Time and Temperature Effects on Histamine Production in Tuna Salad Preparations.

PubMed

McCarthy, Susan; Bjornsdottir-Butler, Kristin; Benner, Ronald

2015-07-01

Scombrotoxin fish poisoning (SFP), also known as histamine (Hst) poisoning, has been associated with consumption of scombroid-type fish, including tuna and tuna fish products. Preparation of commercial tuna salad contaminated with Hstproducing bacteria (HPB), combined with time-temperature abuse, can present a food safety hazard. A potential source of HPB is raw ingredients, such as celery and onions. The objectives of this study were to determine whether raw ingredients can be a source of HPB and to ascertain the effects of storage time (up to 4 days or 4 weeks) and temperature (4, 10, 18, 25, 30°C) on growth and Hst production by high-HPB (>1,000 ppm of Hst) in tuna salad preparations. Pantoea-Erwinia, Erwinia persicinus, Erwinia spp., and Enterobacter pyrinus isolated from celery in this study were used to inoculate tuna salad and tuna salad with celery or onion. HPB numbers were 0.7 to 4.3 log most probable number per g higher in the presence of celery or onion versus plain tuna salad (3:1 tuna:mayonnaise). E. pyrinus-inoculated plain tuna salad and tuna salad with celery and onion had >500 ppm of Hst after 2 days at 30°C and 4 days at 25°C. E. pyrinus-inoculated salad with celery and onion had >500 ppm of Hst after 4 days at 18°C and 2 weeks at 10°C. Raw celery can introduce HPB into tuna salad, which can cause SFP if the product is time-temperature abused. Tuna salad products must be refrigerated at ≤4°C to prevent growth and Hst production by the HPB used in this study, to protect consumers from potential SFP.

Agrobacterium tumefaciens-mediated transformation of blueberry (Vaccinium corymbosum L.).

PubMed

Song, Guo-Qing; Sink, K C

2004-12-01

Transient expression studies using blueberry leaf explants and monitored by beta-glucuronidase (GUS) assays indicated Agrobacterium tumefaciens strain EHA105 was more effective than LBA4404 or GV3101; and the use of acetosyringone (AS) at 100 microM for inoculation and 6 days co-cultivation was optimum compared to 2, 4, 8, 10 or 12 days. Subsequently, explants of the cultivars Aurora, Bluecrop, Brigitta, and Legacy were inoculated with strain EHA105 containing the binary vector pBISN1 with the neomycin phosphotransferase gene (nptII) and an intron-interrupted GUS gene directed by the chimeric super promoter (Aocs)3AmasPmas. Co-cultivation was for 6 days on modified woody plant medium (WPM) plus 100 microM AS. Explants were then placed on modified WPM supplemented with 1.0 mg l(-1) thidiazuron, 0.5 mg l(-1) alpha-naphthaleneacetic, 10 mg l(-1) kanamycin (Km), and 250 mg l(-1) cefotaxime. Selection for Km-resistant shoots was carried out in the dark for 2 weeks followed by culture in the light at 30 microE m(-2) s(-1) at 25 degrees C. After 12 weeks, selected shoots that were both Km resistant and GUS positive were obtained from 15.3% of the inoculated leaf explants of cultivar Aurora. Sixty-eight independent clones derived from such shoots all tested positive by the polymerase chain reaction using a nptII primer. Eight of eight among these 68 clones tested positive by Southern hybridization using a gusA gene derived probe. The transformation protocol also yielded Km-resistant, GUS-positive shoots that were also PCR positive at frequencies of 5.0% for Bluecrop, 10.0% for Brigitta and 5.6% for Legacy.

Enrofloxacin and Macrolides Alone or in Combination with Rifampicin as Antimicrobial Treatment in a Bovine Model of Acute Chlamydia psittaci Infection

PubMed Central

Prohl, Annette; Lohr, Markus; Ostermann, Carola; Liebler-Tenorio, Elisabeth; Berndt, Angela; Schroedl, Wieland; Rothe, Michael; Schubert, Evelyn; Sachse, Konrad; Reinhold, Petra

2015-01-01

Chlamydia psittaci is a zoonotic bacterium with a wide host range that can cause respiratory disease in humans and cattle. In the present study, effects of treatment with macrolides and quinolones applied alone or in combination with rifampicin were tested in a previously established bovine model of respiratory C. psittaci infection. Fifty animals were inoculated intrabronchially at the age of 6–8 weeks. Seven served as untreated controls, the others were assigned to seven treatment groups: (i) rifampicin, (ii) enrofloxacin, (iii) enrofloxacin + rifampicin, (iv) azithromycin, (v) azithromycin + rifampicin, (vi) erythromycin, and (vii) erythromycin + rifampicin. Treatment started 30 hours after inoculation and continued until 14 days after inoculation (dpi), when all animals were necropsied. The infection was successful in all animals and sufficient antibiotic levels were detected in blood plasma and tissue of the treated animals. Reisolation of the pathogen was achieved more often from untreated animals than from other groups. Nevertheless, pathogen detection by PCR was possible to the same extent in all animals and there were no significant differences between treated and untreated animals in terms of local (i.e. cell count and differentiation of BALF-cells) and systemic inflammation (i.e. white blood cells and concentration of acute phase protein LBP), clinical signs, and pathological findings at necropsy. Regardless of the reduced reisolation rate in treated animals, the treatment of experimentally induced respiratory C. psittaci infection with enrofloxacin, azithromycin or erythromycin alone or in combination with rifampicin was without obvious benefit for the host, since no significant differences in clinical and pathological findings or inflammatory parameters were detected and all animals recovered clinically within two weeks. PMID:25768665

Poliomyelitis in transgenic mice expressing CD155 under the control of the Tage4 promoter after oral and parenteral poliovirus inoculation

PubMed Central

Khan, Shaukat; Toyoda, Hidemi; Linehan, Melissa; Iwasaki, Akiko; Nomoto, Akio; Bernhardt, Günter; Wimmer, Eckard

2014-01-01

An important step in poliovirus (PV) infection by the oral route in humans is replication of the virus in lymphatic tissues of the gastrointestinal (GI) tract, thought to be mainly in the Peyer’s patches of the small intestine. No immunocompetent transgenic (tg) mice that express human PV receptor (CD155) under the control of different promoters can be infected orally. The mouse orthologue of human CD155 is Tage4, a protein expressed at the surface of enterocytes and in the Peyer’s patches. We describe here the generation of a tg mouse model in which the Tage4 promoter was used to drive expression of the human PV receptor-coding region (Tage4-CD155tg mice). In this model, CD155 expression was observed by immunostaining in different regions in the Peyer’s patches but not in their germinal centres. Although a similar pattern of staining was observed between 3- and 6-week-old Tage4-CD155tg mice, poliomyelitis was only seen in the younger mice after PV infection by the oral route. When compared with TgPVR21 mice that expressed CD155 driven by its human promoter, 3-week-old Tage4-CD155tg mice were more susceptible to gut infection and paralysis following feeding with PV. Also, Tage4-CD155tg mice exhibited higher susceptibility to poliomyelitis after parenteral inoculation of PV. Remarkably, the LD50 after intracerebral inoculation of PV was similar in both CD155 tg mouse strains. The CD155 tg mouse model reported here, although moderately susceptible to oral infection, may be suitable to study mechanisms of PV replication in the gastrointestinal tract and to dissect important aspects of PV neuroinvasiveness. PMID:24784416

Poliomyelitis in transgenic mice expressing CD155 under the control of the Tage4 promoter after oral and parenteral poliovirus inoculation.

PubMed

Khan, Shaukat; Toyoda, Hidemi; Linehan, Melissa; Iwasaki, Akiko; Nomoto, Akio; Bernhardt, Günter; Cello, Jeronimo; Wimmer, Eckard

2014-08-01

An important step in poliovirus (PV) infection by the oral route in humans is replication of the virus in lymphatic tissues of the gastrointestinal (GI) tract, thought to be mainly in the Peyer's patches of the small intestine. No immunocompetent transgenic (tg) mice that express human PV receptor (CD155) under the control of different promoters can be infected orally. The mouse orthologue of human CD155 is Tage4, a protein expressed at the surface of enterocytes and in the Peyer's patches. We describe here the generation of a tg mouse model in which the Tage4 promoter was used to drive expression of the human PV receptor-coding region (Tage4-CD155tg mice). In this model, CD155 expression was observed by immunostaining in different regions in the Peyer's patches but not in their germinal centres. Although a similar pattern of staining was observed between 3- and 6-week-old Tage4-CD155tg mice, poliomyelitis was only seen in the younger mice after PV infection by the oral route. When compared with TgPVR21 mice that expressed CD155 driven by its human promoter, 3-week-old Tage4-CD155tg mice were more susceptible to gut infection and paralysis following feeding with PV. Also, Tage4-CD155tg mice exhibited higher susceptibility to poliomyelitis after parenteral inoculation of PV. Remarkably, the LD50 after intracerebral inoculation of PV was similar in both CD155 tg mouse strains. The CD155 tg mouse model reported here, although moderately susceptible to oral infection, may be suitable to study mechanisms of PV replication in the gastrointestinal tract and to dissect important aspects of PV neuroinvasiveness. © 2014 The Authors.

Astrovirus, reovirus and rotavirus concomitant infection causes decreased weight gain in broad-breasted white poults

USDA-ARS?s Scientific Manuscript database

Turkey astrovirus type-2 (TAstV-2), turkey rotavirus (TRotV) and turkey reovirus (TReoV) were evaluated for pathogenesis in 3 day-old turkey poults in all possible combinations of one, two or three viruses. Body-weights were recorded at 2, 4, 7, 10 and 14 days post inoculation (PI) and were decreas...

Experimental transmission and tissue tropism of white spot syndrome virus (WSSV) in two species of lobsters, Panulirus homarus and Panulirus ornatus.

PubMed

Syed Musthaq, S; Sudhakaran, R; Balasubramanian, G; Sahul Hameed, A S

2006-10-01

The susceptibility of two species of lobsters, Panulirus homarus and Panulirus ornatus to white spot syndrome virus (WSSV) was tested by oral route and intramuscular injection. The results revealed that these lobsters were as highly susceptible as marine shrimp when the WSSV was administered intramuscularly. The WSSV caused 100% mortality in both Panulirus homarus and Panulirus ornatus, at 168 and 120 h, respectively, after intramuscular injection and failed to cause mortality when given orally. The presence of WSSV in moribund lobsters was confirmed by single-step and nested PCR, Western blot, histology, and bioassay test. It was found in eyestalk, gill, head muscle, tail muscle, hemolymph, appendages, and stomach. In lobsters with oral route infection, all tested organs except stomach and head muscle was negative for WSSV by nested PCR at 120 h post-inoculation. The stomach and head muscle was positive by nested PCR at 120 h p.i., but negative at 168 h p.i. Western blot analysis was negative in all the tested organs of both species of lobster at 120 h post-inoculation by oral route.

Short interfering RNAs targeting a vampire-bat related rabies virus phosphoprotein mRNA.

PubMed

Ono, Ekaterina Alexandrovna Durymanova; Taniwaki, Sueli Akemi; Brandão, Paulo

The aim of this study was to assess the in vitro and in vivo effects of short-interfering RNAs (siRNAs) against rabies virus phosphoprotein (P) mRNA in a post-infection treatment for rabies as an extension of a previous report (Braz J Microbiol. 2013 Nov 15;44(3):879-82). To this end, rabies virus strain RABV-4005 (related to the Desmodus rotundus vampire bat) were used to inoculate BHK-21 cells and mice, and the transfection with each of the siRNAs was made with Lipofectamine-2000™. In vitro results showed that siRNA 360 was able to inhibit the replication of strain RABV-4005 with a 1log decrease in virus titter and 5.16-fold reduction in P mRNA, 24h post-inoculation when compared to non-treated cells. In vivo, siRNA 360 was able to induce partial protection, but with no significant difference when compared to non-treated mice. These results indicate that, despite the need for improvement for in vivo applications, P mRNA might be a target for an RNAi-based treatment for rabies. Copyright © 2017 Sociedade Brasileira de Microbiologia. Published by Elsevier Editora Ltda. All rights reserved.

The inoculating role of previous exposure to potentially traumatic life events on coping with prolonged exposure to rocket attacks: A lifespan perspective.

PubMed

Palgi, Yuval; Gelkopf, Marc; Berger, Rony

2015-06-30

Relatively little research have addressed the effect of prolonged exposure to rocket attacks with a lifespan perspective and only a handful of these studies focused on the effect of this exposure as a function of aging. The present study examined the effects of seven years of rocket attacks fired toward the south of Israel on adult participants of different ages. We examined whether potentially traumatic life events (PTLEs) unrelated to rocket attacks moderated the association between post-traumatic stress (PTS) symptoms and age. Data were obtained from a 2007 telephone survey using the Random Digit Dialing method and including 343 individuals (76.7% participation rate). Exposure to rockets, PTLEs, global distress, and post-traumatic symptomatology were assessed. Older age was associated with a higher level of PTS symptoms. Higher PTLE levels attenuated the association between age and PTS symptoms. Our results suggest that age is a risk factor for developing PTS symptoms under prolonged exposure to rocket attacks. However, previous levels of exposure to other negative events, as well as gender, appear to inoculate a person to stress, thus modulating the age-PTS association. Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Efficacy of gaseous ozone to counteract postharvest table grape sour rot.

PubMed

Pinto, L; Caputo, L; Quintieri, L; de Candia, S; Baruzzi, F

2017-09-01

This work aims at studying the efficacy of low doses of gaseous ozone in postharvest control of the table grape sour rot, a disease generally attributed to a consortium of non-Saccharomyces yeasts (NSY) and acetic acid bacteria (AAB). Sour rot incidence of wounded berries, inoculated with 8 NSYstrains, or 7 AAB, or 56 yeast-bacterium associations, was monitored at 25 °C up to six days. Sour rot incidence in wounded berries inoculated with yeast-bacterium associations resulted higher than in berries inoculated with one single NSY or AAB strain. Among all NSY-AAB associations, the yeast-bacterium association composed of Candida zemplinina CBS 9494 (Cz) and Acetobacter syzygii LMG 21419 (As) showed the highest prevalence of sour rot; thus, after preliminary in vitro assays, this simplified As-Cz microbial consortium was inoculated in wounded berries that were stored at 4 °C for ten days under ozone (2.14 mg m -3 ) or in air. At the end of cold storage, no berries showed sour-rot symptoms although ozonation mainly affected As viable cell count. After additional 12 days at 25 °C, the sour rot index of inoculated As-Cz berries previously cold-stored under ozone or in air accounted for 22.6 ± 3.7% and 66.7 ± 4.5%, respectively. Molecular analyses of dominant AAB and NSY populations of both sound and rotten berries during post-refrigeration period revealed the appearance of new strains mainly belonging to Gluconobacter albidus and Hanseniaspora uvarum species, respectively. Cold ozonation resulted an effective approach to extend the shelf-life of table grapes also after cold storage. Copyright © 2017 Elsevier Ltd. All rights reserved.

Cap analog and Potato virus A HC-Pro silencing suppressor improve GFP transient expression using an infectious virus vector in Nicotiana benthamiana.

PubMed

Tahmasebi, Amin-Alah; Afsharifar, Alireza

2017-06-01

Transient expression of proteins in plants has become a choice to facilitate recombinant protein production with its fast and easy application. On the other hand, host defensive mechanisms have been reported to reduce the efficiency of transient expression in plants. Hence, this study was designed to evaluate the effect of cap analog and Potato virus A helper component proteinase (PVA HC-Pro) on green fluorescent protein (GFP) expression efficiency. N . benthamiana leaves were inoculated with capped or un-capped RNA transcripts of a Turnip crinkle virus (TCV) construct containing a green fluorescent protein reporter gene (TCV-sGFP) in place of its coat protein (CP) ORF. PVA HC-Pro as a viral suppressor of RNA silencing was infiltrated in trans by Agrobacterium tumefaciens , increased the GFP foci diameter to six and even more cells in both capped and un capped treatments. The expression level of GFP in inoculated plants with TCV-sGFP transcript pre-infiltrated with PVA HC-Pro was 12.97-fold higher than the GFP accumulation level in pre-infiltrated leaves with empty plasmid (EP) control. Also, the yield of GFP in inoculated N. benthamiana plants with capped TCV-sGFP transcript pre-infiltrated with EP and PVA HC-Pro was 1.54 and 1.2-fold respectively, greater than the level of GFP expressed without cap analog application at 5 days post inoculation (dpi). In addition, the movement of TCV-sGFP was increased in some cells of inoculated leaves with capped transcripts. Results of this study indicated that PVA HC-Pro and mRNA capping can increase GFP expression and its cell to cell movement in N. benthamiana .

From Rare to Dominant: a Fine-Tuned Soil Bacterial Bloom during Petroleum Hydrocarbon Bioremediation.

PubMed

Fuentes, Sebastián; Barra, Bárbara; Caporaso, J Gregory; Seeger, Michael

2016-02-01

Hydrocarbons are worldwide-distributed pollutants that disturb various ecosystems. The aim of this study was to characterize the short-lapse dynamics of soil microbial communities in response to hydrocarbon pollution and different bioremediation treatments. Replicate diesel-spiked soil microcosms were inoculated with either a defined bacterial consortium or a hydrocarbonoclastic bacterial enrichment and incubated for 12 weeks. The microbial community dynamics was followed weekly in microcosms using Illumina 16S rRNA gene sequencing. Both the bacterial consortium and enrichment enhanced hydrocarbon degradation in diesel-polluted soils. A pronounced and rapid bloom of a native gammaproteobacterium was observed in all diesel-polluted soils. A unique operational taxonomic unit (OTU) related to the Alkanindiges genus represented ∼ 0.1% of the sequences in the original community but surprisingly reached >60% after 6 weeks. Despite this Alkanindiges-related bloom, inoculated strains were maintained in the community and may explain the differences in hydrocarbon degradation. This study shows the detailed dynamics of a soil bacterial bloom in response to hydrocarbon pollution, resembling microbial blooms observed in marine environments. Rare community members presumably act as a reservoir of ecological functions in high-diversity environments, such as soils. This rare-to-dominant bacterial shift illustrates the potential role of a rare biosphere facing drastic environmental disturbances. Additionally, it supports the concept of "conditionally rare taxa," in which rareness is a temporary state conditioned by environmental constraints. Copyright © 2016, American Society for Microbiology. All Rights Reserved.

From Rare to Dominant: a Fine-Tuned Soil Bacterial Bloom during Petroleum Hydrocarbon Bioremediation

PubMed Central

Fuentes, Sebastián; Barra, Bárbara; Caporaso, J. Gregory

2015-01-01

Hydrocarbons are worldwide-distributed pollutants that disturb various ecosystems. The aim of this study was to characterize the short-lapse dynamics of soil microbial communities in response to hydrocarbon pollution and different bioremediation treatments. Replicate diesel-spiked soil microcosms were inoculated with either a defined bacterial consortium or a hydrocarbonoclastic bacterial enrichment and incubated for 12 weeks. The microbial community dynamics was followed weekly in microcosms using Illumina 16S rRNA gene sequencing. Both the bacterial consortium and enrichment enhanced hydrocarbon degradation in diesel-polluted soils. A pronounced and rapid bloom of a native gammaproteobacterium was observed in all diesel-polluted soils. A unique operational taxonomic unit (OTU) related to the Alkanindiges genus represented ∼0.1% of the sequences in the original community but surprisingly reached >60% after 6 weeks. Despite this Alkanindiges-related bloom, inoculated strains were maintained in the community and may explain the differences in hydrocarbon degradation. This study shows the detailed dynamics of a soil bacterial bloom in response to hydrocarbon pollution, resembling microbial blooms observed in marine environments. Rare community members presumably act as a reservoir of ecological functions in high-diversity environments, such as soils. This rare-to-dominant bacterial shift illustrates the potential role of a rare biosphere facing drastic environmental disturbances. Additionally, it supports the concept of “conditionally rare taxa,” in which rareness is a temporary state conditioned by environmental constraints. PMID:26590285

Remyelination of central nervous system lesions in experimental genital herpes simplex virus infection.

PubMed

Soffer, D; Martin, J R

1988-08-01

To study spinal cord remyelination in a model of genital herpes simplex virus type 2 (HSV-2) infection, adult female mice were inoculated by a vaginal route. At intervals up to 6 months after infection, cord tissues were removed and examined by light and electron microscopy and by immunohistochemical methods. As a consequence of acute infection, 60% of mice developed multifocal central nervous system (CNS) demyelinative lesions in the lower thoracic, lumbar, or upper sacral cord. These lesions, already present 10 days after infection, contained naked axons and mononuclear cells, including macrophages. At 2 weeks, while active myelin breakdown was still ongoing, numerous Schwann cells were present in lesions and surrounded denuded axons. At 3 weeks, the earliest remyelination was seen, and was carried out by Schwann cells and to a lesser extent by oligodendrocytes. Remyelination was extensive by 6-10 weeks and was apparently completed after 3 months. Immunocytochemical studies using antisera to myelin proteins showed relatively distinct zones of central and peripheral remyelination in some lesions, whereas remyelination was of mixed type in others. Thus the remyelinative response following experimental HSV-2-induced CNS demyelination begins promptly, proceeds briskly and goes to completion. With a natural route of inoculation and a relatively avirulent strain of this human pathogen, we have produced a model of CNS white matter injury and repair in a high proportion of infected mice that may be useful in understanding mechanisms of human demyelinative disease.

Single dose parenteral hyposensitization to poison ivy urushiol in guinea pigs.

PubMed

Walker, L A; Watson, E S; elSohly, M A

1995-08-01

Studies were carried out in guinea pigs to evaluate the potential for single dose hyposensitization to poison ivy urushiol dermatitis. Sensitization was induced by topical application of 1 mg of poison ivy urushiol to the back of the neck. In the first series of studies, three different analogs of poison ivy urushiol were studied: 1) a mixture of pentadecyl and heptadecyl catechols (PDC/HDC), the saturated side chain analog of the natural urushiol mixture; 2) a mixture of the diacetate esters of PDC and HDC (PDC/HDC Ac), the esterified form of the saturated sidechain analogs; 3) 2-n-pentadecyl hydroquinone diacetate (HQ Ac). Each of these compounds was administered as 5 mg of the free catechol i.m. each week for three weeks. A vehicle group received only corn oil injections. Reactivity to poison ivy urushiol (PIU) challenge was evaluated in skin tests at 1 and 5 weeks post-treatment. PDC/HDC Ac induced a marked reduction in both the incidence and the severity of lesions induced by PIU at both 1 and at 5 weeks post-treatment. Other analogs were ineffective at 5 weeks post-treatment, and were less effective than PDC/HDC Ac at 1 week post-treatment. In a second series of experiments, the efficacy of PDC/HDC Ac was evaluated in both single and multiple dose regiments. One treatment group received 5 mg of PDC/HDC Ac intramuscularly each week for 4 weeks, while another treatment group received a single dose of 20 mg PDC/HDC Ac i.m. Corresponding vehicle control groups were also included. At 1 week post-treatment in the single dose group, the PDC/HDC Ac was only modestly effective, with some reduction of severity of lesions at the higher challenge doses of PIU. However, at 4 and 7 weeks post-treatment, both the incidence and the severity of the lesions at all challenge doses were reduced. In the multiple dose group, the incidence and severity of lesions are reduced at 1 week and 4 weeks post-treatment (4 weeks and 7 weeks after the initial dose) but were not significantly different from the single dose group. These findings indicate that the diacetate ester of PDC/HDC is an effective hyposensitizer to poison ivy urushiol, and that this hyposensitization can be reasonably accomplished in a single dose treatment regimen.

A recombinant pseudorabies virus expressing rabies virus glycoprotein: safety and immunogenicity in dogs.

PubMed

Yuan, Ziguo; Zhang, Shoufeng; Liu, Ye; Zhang, Fei; Fooks, Anthony R; Li, Qianxue; Hu, Rongliang

2008-03-04

Several recombinant vaccines expressing the rabies virus glycoprotein have been developed, particularly for the oral vaccination of wildlife. While these vaccines induce protective immunity in some animal species such as foxes, they are less effective in others. Pseudorabies virus (PRV) has been licensed for use as a live vaccine in pigs and possesses an excellent safety and efficacy record. We have used it to construct a recombinant virus, rPRV/eGFP/rgp, expressing the rabies virus glycoprotein. This recombinant virus has been shown to be safe for dogs by oral and intramuscular routes of inoculation and was demonstrated to induce immune responses against both pseudorabies and rabies in dogs after a single oral dose of 2 x 10(7.0) plaque forming units (PFU). Neutralizing antibody titers against rabies reached > 0.5 IU/ml and 1:64-1:128 against pseudorabies by 5 weeks post-vaccination in all dogs, indicating that the pseudorabies virus vector infected dogs and replicated in vivo, and that the rabies virus glycoprotein had been expressed and an effective immune response elicited. Antibody titers were maintained for over 6 months. This suggests that pseudorabies virus could be an effective live vector for recombinant rabies oral vaccination.

Establishment of Mimosa biuncifera (Fabaceae) inoculated with arbuscular mycorrhizal fungi in greenhouse and field drought conditions.

PubMed

Peña-Becerril, Juan C; Monroy-Ata, Arcadio; Orozco-Almanza, María del Socorro; García-Amador, Esther Matiana

2016-06-01

Mexico is dominated by arid or semi-arid ecosystems, predominantly characterized as xeric shrublands. These areas are frequently deteriorated due to agriculture or over-grazing by livestock (sheep and goats). The vegetation type mainly consists of thorny plant species, and among these, the dominant one in overgrazed areas is catclaw (Mimosa biuncifera). This is a nurse plant that facilitates establishment of other vegetation and promotes plant succession. Catclaw plants form a mutualistic association with arbuscular mycorrhizal fungi (AMF), which improves uptake of nutrients and water. The objective of this study was to determine the effect of inoculating catclaw plants with native AMF and starting their growth under a low water availability treatment in a greenhouse, and later transplanting them to field conditions of drought and deterioration. Field plants were evaluated according to their survivorship and growth. The seeds of catclaw plants and soil with AMF spores were collected in the Mezquital Valley of Hidalgo State, in Central Mexico. Seedlings were grown in individual pots in a greenhouse. The experimental design consisted of two levels of pot irrigation, wet (W) and dry (D), as well as the presence (M+) or absence (M-) of AMF inoculum, with 20 replicates for each treatment. The following plant parameters were recorded every week: height, number of leaves and pinnae, and mean diameter of coverage. After 20 weeks in the greenhouse, determination was made of fresh and dry biomass, relative growth rate (RGR), root/shoot ratio, real evapotranspiration (RET), water-use efficiency (WUE), and percentage of mycorrhizal colonization. The remaining plants growing under the dry treatment (M+ and M-) were then transplanted to a semi-arid locality in the Mezquital Valley. During one year, monthly records were kept of their height, number of leaves, mean diameter of coverage and survival. Results showed that compared to greenhouse plants under other treatments, those under the wet mycorrhizal (WM+) treatment were taller, had more pinnae, and were characterized by greater coverage, faster RGR, and greater fresh and dry biomass. Moreover, inoculated plants (WM+ and DM+) showed higher WUE than those uninoculated (WM- and DM-, respectively). After one year in field conditions, there was a higher survival rate for previously inoculated versus uninoculated plants. Hence, mycorrhization of M. biuncifera with native AMF inoculum increased plant efficiency in biomass production, thus favoring establishment and survival in field conditions. We concluded that inoculation of catclaw plants is recommendable for revegetation programs in deteriorated semi-arid zones.

Fate of selected microorganisms when introduced as cross-contamination inocula into simulated food trash compartment waste

NASA Astrophysics Data System (ADS)

Strayer, Richard; Hummerick, Mary; Richards, Jeffrey; Birmele, Michele; Roberts, Michael

AdHocReviewCycleID-309796538 NewReviewCycle EmailSubjectPlease review this (?today?) AuthorEm Richard F. (KSC)[DYNAMAC CORP] ReviewingToolsShownOnceurn:schemas-microsoft-com:office:smart One goal of Exploration Life Support solid waste processing is to stabilize wastes for storage, mitigate crew risks, and enable resource recovery. Food and crew fecal wastes contain easily biodegraded organic components that support microbial growth. Our objective is to determine a baseline for the fate of selected microbes in wastes prior to processing treatments. Challenge microbes, including human-associated pathogens, were added to unsterilized, simulated food trash solid waste containing a mixed microbial community. The fate of the microbial community and challenge microbes was determined over a 6 week time course of waste storage. Challenge microbes were selected from a list of microorganisms common to residual food or fecal wastes and included: Escherichia coli, Salmonella enterica serovar typhimurium, Staphylococcus aureus, Pseudomonas aeruginosa, Aspergillus niger (a common mold), and Bacillus pumilus SAFR-032, a spore-forming bacterium previously isolated from spacecraft assembly facilities selected for its resistance to heat, uv, and desiccation. The trash model simulant contained 80% food trash (food waste and containers) and 20% hygiene wipes. Cultures of challenge microbes were grown overnight on Nutrient Agar (Difco), harvested, re-suspended in physiological saline, and diluted to achieve the desired optical density for inoculation. The six organisms were pooled and inoculated into the simulated food wastes and packaging before manual mixing. Inoculated simulated waste was stored in custom FlexfoilTM gas sampling bags (SKC, Inc.) which were then connected to a gas analysis system designed to supply fresh air to each bag to maintain O2 above 1%. Bag headspace was monitored for CO2 (PP Systems) and O2 (Maxtec). Total microbes were quantified by microscopic direct counts and general cultivation-based methods. Detection and enumeration of challenge microbes was accomplished by cultivation-based microbiological methods with specific selective media and by molecular methods using quantitative stocktickerPCR (qPCR) with stocktickerDNA primers specific for each challenge organism. stocktickerDNA was extracted and purified from residual wastes with a stocktickerDNA isolation kit (Mo Bio), and quantified (NanoDrop) from standard curves prepared from pure culture isolates of each challenge organism. QPCR was conducted on a Roche LightCycler 480 using the Roche stocktickerSYBR Green Master Mix Kit. The identity of all challenge microbes in recovered isolates was verified by stocktickerDNA sequencing (stocktickerABI 3130 Genetic Analyzer - Applied Biosystems). To date, concentrations of challenge microbial populations at concentrations ranging from ˜107 - 108 have been added to simulated food waste and extracted either immediately after mixing or after 1 week of storage. Cultivation-based counts indicated that 5 of 6 challenge microbes could be recovered from simulated food wastes after inoculation for both concentrations. Only S. enterica serovar typhimurium could not be detected at week 0 for the 107 inoculum. Between week 0 and 1, challenge microbes increased in density: S. aureus, E. coli, and P. aeruginosa increasing up to 4 orders of magnitude from the 107 inoculum. Molecular results for the week 0 and week 1 stored samples indicated that the relative concentrations of target stocktickerDNA for the challenge microbes had increased between 1 and 3 orders of magnitude. These preliminary studies demonstrate that potential problems regarding pathogens as cross-contaminants from other waste streams could develop during storage of space mission solid wastes. Ongoing studies are examining longer storage times up to 6 weeks. The results can be used to determine requirements and criteria for waste treatment prior to storage and provides a means of testing the ability of treatment technologies to limit contaminant survival and proliferation.

Highly pathogenic avian influenza virus H5N1 infection in a long-distance migrant shorebird under migratory and non-migratory states.

PubMed

Reperant, Leslie A; van de Bildt, Marco W G; van Amerongen, Geert; Buehler, Debbie M; Osterhaus, Albert D M E; Jenni-Eiermann, Susi; Piersma, Theunis; Kuiken, Thijs

2011-01-01

Corticosterone regulates physiological changes preparing wild birds for migration. It also modulates the immune system and may lead to increased susceptibility to infection, with implications for the spread of pathogens, including highly pathogenic avian influenza virus (HPAIV) H5N1. The red knot (Calidris canutus islandica) displays migratory changes in captivity and was used as a model to assess the effect of high plasma concentration of corticosterone on HPAIV H5N1 infection. We inoculated knots during pre-migration (N = 6), fueling (N = 5), migration (N = 9) and post-migration periods (N = 6). Knots from all groups shed similar viral titers for up to 5 days post-inoculation (dpi), peaking at 1 to 3 dpi. Lesions of acute encephalitis, associated with virus replication in neurons, were seen in 1 to 2 knots per group, leading to neurological disease and death at 5 to 11 dpi. Therefore, the risk of HPAIV H5N1 infection in wild birds and of potential transmission between wild birds and poultry may be similar at different times of the year, irrespective of wild birds' migratory status. However, in knots inoculated during the migration period, viral shedding levels positively correlated with pre-inoculation plasma concentration of corticosterone. Of these, knots that did not become productively infected had lower plasma concentration of corticosterone. Conversely, elevated plasma concentration of corticosterone did not result in an increased probability to develop clinical disease. These results suggest that birds with elevated plasma concentration of corticosterone at the time of migration (ready to migrate) may be more susceptible to acquisition of infection and shed higher viral titers--before the onset of clinical disease--than birds with low concentration of corticosterone (not ready for take-off). Yet, they may not be more prone to the development of clinical disease. Therefore, assuming no effect of sub-clinical infection on the likelihood of migratory take-off, this may favor the spread of HPAIV H5N1 by migratory birds over long distances.

Higher frequency of PEDV shedding and lesions in suckling pigs compared to nursery pigs and protective immunity after homologous re-challenge

USDA-ARS?s Scientific Manuscript database

Porcine epidemic diarrhea virus (PEDV) causes enteric disease in pigs and is known to spread rapidly after entering naïve pig populations. The objectives were to 1) compare the disease course following inoculation with PEDV isolate US/Colorado/2013 in naïve 10-day and 8-week-old pigs, and 2) contras...

Neospora caninum infection during early pregnancy in cattle: how the isolate influences infection dynamics, clinical outcome and peripheral and local immune responses

PubMed Central

2014-01-01

This work studies the influence of Neospora caninum intra-species diversity on abortion outcome, infection dynamics in terms of parasite dissemination and peripheral-local immune responses in pregnant cattle. Animals were intravenously inoculated at day 70 of pregnancy with 107 tachyzoites of two isolates showing marked differences in virulence in vitro and in pregnant mouse models: Nc-Spain7, a high virulence isolate, and Nc-Spain8, a low-to-moderate virulence isolate. After inoculation, pregnancy was monitored, and dams were culled when foetal death was detected. Foetal mortality occurred in all infected heifers between days 24 and 49 post-infection (pi), however, it was detected sooner in Nc-Spain7-infected animals (median day = 34) than those inoculated with Nc-Spain8 (median day = 41) with a trend towards significance (P < 0.11). Similar histological lesions were observed in placentomes and in most of the foetuses from the two infected groups. However, parasites were more frequently detected in the placenta and foetuses by PCR and in the foetal brain by immunohistochemistry in Nc-Spain7-infected animals. Specific antibodies were detected starting at day 13 post-infection in all infected cattle, with higher IgG levels in Nc-Spain7-infected group. IFN-γ and IL-4 profiles also varied between infected groups in PBMC stimulation assays. Infected animals showed significant increases in their cytokine mRNA levels (IFN-γ, IL-4, IL-10, IL-12p40 and TNF-α) in the caruncle at time of foetal death. Differences between the infected groups were also observed for cytokine profiles. These results demonstrate the influence of the N. caninum isolate on foetal death outcome, infection dynamics and immune responses in cattle. PMID:24479988

Independent Preharvest Applications of Methyl Jasmonate and Chitosan Elicit Differential Upregulation of Defense-Related Genes with Reduced Incidence of Gray Mold Decay during Postharvest Storage of Fragaria chiloensis Fruit

PubMed Central

Saavedra, Gabriela M.; Sanfuentes, Eugenio; Figueroa, Carlos R.

2017-01-01

The Chilean strawberry (Fragaria chiloensis) fruit has interesting organoleptic properties, but its postharvest life is affected by gray mold decay caused by Botrytis cinerea. The effect of preharvest applications of methyl jasmonate (MeJA) or chitosan on the molecular defense-related responses and protection against gray mold decay were investigated in Chilean strawberry fruit during postharvest storage. Specifically, we inoculated harvested fruit with B. cinerea spores and studied the expression of genes encoding for the pathogenesis-related (PR) proteins β-1,3-glucanases (FcBG2-1, FcBG2-2 and FcBG2-3) and chitinases (FcCHI2-2 and FcCHI3-1), and for polygalacturonase inhibiting proteins (FcPGIP1 and FcPGIP2) at 0, 2, 24, 48, and 72 h post inoculation (hpi). Remarkably, MeJA- and chitosan-treated fruit exhibited a lower incidence of B. cinerea infection than the control-treated at 48 and 72 hpi. At the molecular level, both are efficient elicitors for priming in F. chiloensis fruit since we observed an upregulation of the FcBG2-1, FcBG2-3, FcPGIP1, and FcPGIP2 at 0 hpi. Moreover, a chitosan-mediated upregulation of FcPGIPs at early times post inoculation (2–24 hpi) and MeJA upregulated FcBGs (24–72 hpi) and FcPGIP1 at later times could contribute to reduce B. cinerea incidence by differential upregulation of defense genes. We concluded that preharvest applications of MeJA or chitosan had a long-lasting effect on the reduction of B. cinerea incidence during postharvest as well as an enhancer effect on the induction of PR and PGIP gene expression. PMID:28671619

Independent Preharvest Applications of Methyl Jasmonate and Chitosan Elicit Differential Upregulation of Defense-Related Genes with Reduced Incidence of Gray Mold Decay during Postharvest Storage of Fragaria chiloensis Fruit.

PubMed

Saavedra, Gabriela M; Sanfuentes, Eugenio; Figueroa, Pablo M; Figueroa, Carlos R

2017-07-03

The Chilean strawberry ( Fragaria chiloensis ) fruit has interesting organoleptic properties, but its postharvest life is affected by gray mold decay caused by Botrytis cinerea . The effect of preharvest applications of methyl jasmonate (MeJA) or chitosan on the molecular defense-related responses and protection against gray mold decay were investigated in Chilean strawberry fruit during postharvest storage. Specifically, we inoculated harvested fruit with B. cinerea spores and studied the expression of genes encoding for the pathogenesis-related (PR) proteins β-1,3-glucanases ( FcBG2-1 , FcBG2-2 and FcBG2-3 ) and chitinases ( FcCHI2-2 and FcCHI3-1 ), and for polygalacturonase inhibiting proteins ( FcPGIP1 and FcPGIP2 ) at 0, 2, 24, 48, and 72 h post inoculation (hpi). Remarkably, MeJA- and chitosan-treated fruit exhibited a lower incidence of B. cinerea infection than the control-treated at 48 and 72 hpi. At the molecular level, both are efficient elicitors for priming in F. chiloensis fruit since we observed an upregulation of the FcBG2-1 , FcBG2-3 , FcPGIP1, and FcPGIP2 at 0 hpi. Moreover, a chitosan-mediated upregulation of FcPGIP s at early times post inoculation (2-24 hpi) and MeJA upregulated FcBG s (24-72 hpi) and FcPGIP1 at later times could contribute to reduce B. cinerea incidence by differential upregulation of defense genes. We concluded that preharvest applications of MeJA or chitosan had a long-lasting effect on the reduction of B. cinerea incidence during postharvest as well as an enhancer effect on the induction of PR and PGIP gene expression.

Chlamydia suis and Chlamydia trachomatis induce multifunctional CD4 T cells in pigs.

PubMed

Käser, T; Pasternak, J A; Delgado-Ortega, M; Hamonic, G; Lai, K; Erickson, J; Walker, S; Dillon, J R; Gerdts, V; Meurens, F

2017-01-03

Chlamydia trachomatis infections are the most prominent bacterial sexually-transmitted disease world-wide and a lot of effort is put into the development of an effective vaccine. Pigs have been shown to be a valuable animal model for C. trachomatis vaccine development. The aim of this study was to decipher the T-cell-mediated immune response to chlamydial infections including C. trachomatis and C. suis, the chlamydia species naturally infecting pigs with a demonstrated zoonotic potential. Vaginal infection of pigs with C. suis and C. trachomatis lasted from 3 to 21days and intra-uterine infection was still present after 21days in 3 out of 5 C. suis- and 4 out of 5 C. trachomatis-inoculated animals and caused severe pathological changes. Humoral immune responses including neutralizing antibodies were found predominantly in response to C. suis starting at 14days post inoculation. The T-cell-mediated immune responses to C. trachomatis and C. suis-infections started at 7days post inoculation and consisted mainly of CD4 + T cells which were either IFN-γ single cytokine-producing or IFN-γ/TNF-α double cytokine-producing T-helper 1 cells. IL-17-producing CD4 + T cells were rare or completely absent. The T-cell-mediated immune responses were triggered by both homologous or heterologous re-stimulation indicating that cross-protection between the two chlamydia species is possible. Thus, having access to a working genital C. suis and C. trachomatis infection model, efficient monitoring of the host-pathogen interactions, and being able to accurately assess the responses to infection makes the pig an excellent animal model for vaccine development which also could bridge the gap to the clinical phase for C. trachomatis vaccine research. Copyright © 2016 Elsevier Ltd. All rights reserved.

Sugarcane genes differentially expressed in response to Puccinia melanocephala infection: identification and transcript profiling.

PubMed

Oloriz, María I; Gil, Víctor; Rojas, Luis; Portal, Orelvis; Izquierdo, Yovanny; Jiménez, Elio; Höfte, Monica

2012-05-01

Brown rust caused by the fungus Puccinia melanocephala is a major disease of sugarcane (Saccharum spp.). A sugarcane mutant, obtained by chemical mutagenesis of the susceptible variety B4362, showed a post-haustorial hypersensitive response (HR)-mediated resistance to the pathogen and was used to identify genes differentially expressed in response to P. melanocephala via suppression subtractive hybridization (SSH). Tester cDNA was derived from the brown rust-resistant mutant after inoculation with P. melanocephala, while driver cDNAs were obtained from the non-inoculated resistant mutant and the inoculated susceptible donor variety B4362. Database comparisons of the sequences of the SSH recombinant clones revealed that, of a subset of 89 non-redundant sequences, 88% had similarity to known functional genes, while 12% were of unknown function. Thirteen genes were selected for transcript profiling in the resistant mutant and the susceptible donor variety. Genes involved in glycolysis and C4 carbon fixation were up-regulated in both interactions probably due to disturbance of sugarcane carbon metabolism by the pathogen. Genes related with the nascent polypeptide associated complex, post-translational proteome modulation and autophagy were transcribed at higher levels in the compatible interaction. Up-regulation of a putative L-isoaspartyl O-methyltransferase S-adenosylmethionine gene in the compatible interaction may point to fungal manipulation of the cytoplasmatic methionine cycle. Genes coding for a putative no apical meristem protein, S-adenosylmethionine decarboxylase, non-specific lipid transfer protein, and GDP-L-galactose phosphorylase involved in ascorbic acid biosynthesis were up-regulated in the incompatible interaction at the onset of haustorium formation, and may contribute to the HR-mediated defense response in the rust-resistant mutant.

Identifying Early Target Cells of Nipah Virus Infection in Syrian Hamsters.

PubMed

Baseler, Laura; Scott, Dana P; Saturday, Greg; Horne, Eva; Rosenke, Rebecca; Thomas, Tina; Meade-White, Kimberly; Haddock, Elaine; Feldmann, Heinz; de Wit, Emmie

2016-11-01

Nipah virus causes respiratory and neurologic disease with case fatality rates up to 100% in individual outbreaks. End stage lesions have been described in the respiratory and nervous systems, vasculature and often lymphoid organs in fatal human cases; however, the initial target organs of Nipah virus infection have not been identified. Here, we detected the initial target tissues and cells of Nipah virus and tracked virus dissemination during the early phase of infection in Syrian hamsters inoculated with a Nipah virus isolate from Malaysia (NiV-M) or Bangladesh (NiV-B). Syrian hamsters were euthanized between 4 and 48 hours post intranasal inoculation and tissues were collected and analyzed for the presence of viral RNA, viral antigen and infectious virus. Virus replication was first detected at 8 hours post inoculation (hpi). Nipah virus initially targeted type I pneumocytes, bronchiolar respiratory epithelium and alveolar macrophages in the lung and respiratory and olfactory epithelium lining the nasal turbinates. By 16 hpi, virus disseminated to epithelial cells lining the larynx and trachea. Although the pattern of viral dissemination was similar for both virus isolates, the rate of spread was slower for NiV-B. Infectious virus was not detected in the nervous system or blood and widespread vascular infection and lesions within lymphoid organs were not observed, even at 48 hpi. Nipah virus initially targets the respiratory system. Virus replication in the brain and infection of blood vessels in non-respiratory tissues does not occur during the early phase of infection. However, virus replicates early in olfactory epithelium and may serve as the first step towards nervous system dissemination, suggesting that development of vaccines that block virus dissemination or treatments that can access the brain and spinal cord and directly inhibit virus replication may be necessary for preventing central nervous system pathology.

220D-F2 from Rubus ulmifolius kills Streptococcus pneumoniae planktonic cells and pneumococcal biofilms.

PubMed

Talekar, Sharmila J; Chochua, Sopio; Nelson, Katie; Klugman, Keith P; Quave, Cassandra L; Vidal, Jorge E

2014-01-01

Streptococcus pneumoniae (pneumococcus) forms organized biofilms to persist in the human nasopharynx. This persistence allows the pneumococcus to produce severe diseases such as pneumonia, otitis media, bacteremia and meningitis that kill nearly a million children every year. While bacteremia and meningitis are mediated by planktonic pneumococci, biofilm structures are present during pneumonia and otitis media. The global emergence of S. pneumoniae strains resistant to most commonly prescribed antibiotics warrants further discovery of alternative therapeutics. The present study assessed the antimicrobial potential of a plant extract, 220D-F2, rich in ellagic acid, and ellagic acid derivatives, against S. pneumoniae planktonic cells and biofilm structures. Our studies first demonstrate that, when inoculated together with planktonic cultures, 220D-F2 inhibited the formation of pneumococcal biofilms in a dose-dependent manner. As measured by bacterial counts and a LIVE/DEAD bacterial viability assay, 100 and 200 µg/ml of 220D-F2 had significant bactericidal activity against pneumococcal planktonic cultures as early as 3 h post-inoculation. Quantitative MIC's, whether quantified by qPCR or dilution and plating, showed that 80 µg/ml of 220D-F2 completely eradicated overnight cultures of planktonic pneumococci, including antibiotic resistant strains. When preformed pneumococcal biofilms were challenged with 220D-F2, it significantly reduced the population of biofilms 3 h post-inoculation. Minimum biofilm inhibitory concentration (MBIC)50 was obtained incubating biofilms with 100 µg/ml of 220D-F2 for 3 h and 6 h of incubation. 220D-F2 also significantly reduced the population of pneumococcal biofilms formed on human pharyngeal cells. Our results demonstrate potential therapeutic applications of 220D-F2 to both kill planktonic pneumococcal cells and disrupt pneumococcal biofilms.

The bovine lung in biomedical research: visually guided bronchoscopy, intrabronchial inoculation and in vivo sampling techniques.

PubMed

Prohl, Annette; Ostermann, Carola; Lohr, Markus; Reinhold, Petra

2014-07-03

There is an ongoing search for alternative animal models in research of respiratory medicine. Depending on the goal of the research, large animals as models of pulmonary disease often resemble the situation of the human lung much better than mice do. Working with large animals also offers the opportunity to sample the same animal repeatedly over a certain course of time, which allows long-term studies without sacrificing the animals. The aim was to establish in vivo sampling methods for the use in a bovine model of a respiratory Chlamydia psittaci infection. Sampling should be performed at various time points in each animal during the study, and the samples should be suitable to study the host response, as well as the pathogen under experimental conditions. Bronchoscopy is a valuable diagnostic tool in human and veterinary medicine. It is a safe and minimally invasive procedure. This article describes the intrabronchial inoculation of calves as well as sampling methods for the lower respiratory tract. Videoendoscopic, intrabronchial inoculation leads to very consistent clinical and pathological findings in all inoculated animals and is, therefore, well-suited for use in models of infectious lung disease. The sampling methods described are bronchoalveolar lavage, bronchial brushing and transbronchial lung biopsy. All of these are valuable diagnostic tools in human medicine and could be adapted for experimental purposes to calves aged 6-8 weeks. The samples obtained were suitable for both pathogen detection and characterization of the severity of lung inflammation in the host.

A polyvalent Clade B virus-like particle HIV vaccine combined with partially protective oral preexposure prophylaxis prevents simian-human immunodeficiency virus Infection in macaques and primes for virus-amplified immunity.

PubMed

Ross, Ted M; Pereira, Lara E; Luckay, Amara; McNicholl, Janet M; García-Lerma, J Gerardo; Heneine, Walid; Eugene, Hermancia S; Pierce-Paul, Brooke R; Zhang, Jining; Hendry, R Michael; Smith, James M

2014-11-01

Vaccination and preexposure prophylaxis (PrEP) with antiretrovirals have shown only partial protection from HIV-1 infection in human trials. Oral Truvada (emtricitabine/tenofovir disoproxil fumarate) is FDA approved as PrEP but partial adherence reduces efficacy. If combined as biomedical preventions (CBP), an HIV vaccine could protect when PrEP adherence is low and PrEP could prevent vaccine breakthroughs. The efficacy of combining oral PrEP with an HIV vaccine has not been evaluated in humans. We determined the efficacy of combining a DNA/virus-like particle (VLP) vaccine with partially effective intermittent PrEP in Indian rhesus macaques (RM). Eight RM received intramuscular inoculations of five DNA plasmids encoding four HIV-1 Clade B primary isolate Envs and SIVmac239 Gag (at weeks 0 and 4), followed by intramuscular and intranasal inoculations of homologous Gag VLPs and four Env VLPs (at weeks 12, 16, and 53). At week 61, we initiated weekly rectal exposures with heterologous SHIV162p3 (10 TCID50) along with oral Truvada (TDF, 22 mg/kg; FTC 20 mg/kg) dosing 2 h before and 22 h after each exposure. This PrEP regimen previously demonstrated 50% efficacy. Five controls (no vaccine, no PrEP) received weekly SHIV162p3. All controls were infected after a median of four exposures; the mean peak plasma viral load (VL) was 3.9×10(7) vRNA copies/ml. CBP protected seven of eight (87.5%) RM. The one infected CBP RM had a reduced peak VL of 8.8×10(5) copies/ml. SHIV exposures during PrEP amplified Gag and Env antibody titers in protected RM. These results suggest that combining oral PrEP with HIV vaccines could enhance protection against HIV-1 infection.

Six Weeks of Massage Therapy Produces Changes in Balance, Neurological and Cardiovascular Measures in Older Persons

PubMed Central

Sefton, JoEllen M.; Yarar, Ceren; Berry, Jack W.

2012-01-01

Objectives: Falls in older adults represent a primary cause of decreased mobility and independence, increased morbidity, and accidental death. Research and clinical reports indicate that therapeutic massage (TM) may positively influence suggested causative factors. The second in a two-part study, this project assessed the effects of six weeks of TM treatment on balance, nervous system, and cardiovascular measures in older adults. Design: A randomized controlled trial assessed the effects of six weekly 60-minute sessions of TM on balance, cardiovascular, and nervous system measures. Thirty-five volunteers (19 male and 16 female; ages 62.9 ± 4.6) were randomly assigned to relaxation control or TM groups. A 2 × 4 [treatment condition X time (week 1 and 6)] mixed factorial experimental design was utilized for cardiovascular/balance variables assessed at pretreatment baseline, immediate post-treatment, and 20- and 60-minutes post-treatment; nervous system measures were assessed only at pretreatment and at 60-minute follow-up (2 × 2 mixed design). Long-term benefits were assessed by comparing the TM and control groups on pretreatment baseline measures at week six and a follow-up assessment at week seven (2 × 3 mixed design). Setting: Laboratory Intervention: Six weekly 60-minute, full-body TM. Outcome Measures: Postural control/cardiovascular measures were assessed weeks one, six, and seven; pretreatment and immediate, 20- and 60-minutes post-treatment. Motoneuron pool excitability was assessed pretreatment and 60 minutes post-treatment. Results: The TM group showed significant differences relative to controls in cardiovascular and displacement area/velocity after the week six session, with decreasing blood pressure and increasing stability over time from immediate post-TM to 60 minutes post-TM. The TM group revealed lower H-max/M-max ratios 60-minutes post-treatment. Long-term differences between the groups were detected at week seven in displacement area/velocity and systolic blood pressure. Conclusions: Results suggest six weeks of TM resulted in immediate and long-term improvements in postural stability and blood pressure, compared to a controlled condition. PMID:23087776

Short-term effects of splenectomy on serum fibrosis indexes in liver cirrhosis patients.

PubMed

Kong, Degang; Chen, Xiuli; Lu, Shichun; Guo, Qingliang; Lai, Wei; Wu, Jushan; Lin, Dongdong; Zeng, Daobing; Duan, Binwei; Jiang, Tao; Cao, Jilei

2015-01-01

To determine the changing patterns of 4 liver fibrosis markers pre and post splenectomy (combined with pericardial devascularization [PCDV]) and to examine the short-term effects of splenectomy on liver fibrosis. Four liver fibrosis markers of 39 liver cirrhosis patients were examined pre, immediately post, 2 days post, and 1 week post (15 cases) splenectomy (combined with PCDV). The laminin (LN) level decreased immediately post surgery compared with the preoperative LN level (P < 0.05). The type IV collagen level decreased immediately post surgery compared with that pre surgery (P < 0.05), it significantly increased (P < 0.05) 2 days post surgery and significantly decreased 1 week post surgery (P < 0.05). Hyaluronic acid and the procollagen III N-terminal peptide levels increased significantly 2 days post surgery compared with that pre and immediately post surgery, they significantly decreased 1 week post surgery compared to 2 days post surgery (P < 0.05). In the short-term, the 4 liver fibrosis markers and the FibroScans post splenectomy showed characteristic changes, splenectomy may transiently initiate the degradation process of liver fibrosis.

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

PubMed

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

2015-01-01

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

Arbuscular mycorrhizal fungi influence water relations, gas exchange, abscisic acid and growth of micropropagated chile ancho pepper (Capsicum annuum) plantlets during acclimatization and post-acclimatization.

PubMed

Estrada-Luna, Andrés A; Davies, Fred T

2003-09-01

Little is known about the role of arbuscular mycorrhiza fungi (AMF) on physiological changes of micropropagated plantlets during acclimatization and post-acclimatization. Using chile ancho pepper (Capsicum annuum L. cv. San Luis), measurements were made of water relations, gas exchange, abscisic acid (ABA), plantlet growth and AMF development. Plantlets had low photosynthetic rates (A) and poor initial growth during acclimatization. Relative water content (RWC) decreased during the first days after transfer from tissue culture containers to ex vitro conditions. Consequently, transpiration rates (E) and stomatal conductance (gs) declined, confirming that in vitro formed stomata were functional and able to respond ex vitro to partial desiccation--thus avoiding excessive leaf dehydration and plant death. Colonization by AMF occurred within 3 days after inoculation. Colonized plantlets had lower leaf ABA and higher RWC than noncolonized (NonAMF) plantlets during peak plant dehydration (6 days after plant transfer)--and a higher A and gs as early as days 5 and 7. During post-acclimatization [after day 8, when RWC increased and stabilized], A increased in all plantlets; however, more dramatic changes occurred with AMF plantlets. Within 48 days, 45% of the roots sampled of inoculated plantlets were colonized and had extensive arbuscule development. At this time, AMF plantlets also had greater E, A, leaf chlorophyll, leaf elemental N, P and K, leaf dry biomass and leaf area, fruit production and differences in carbon partitioning [lower root/shoot ratio and higher leaf area ratio] compared with NonAMF plantlets. Rapid AMF colonization enhanced physiological adjustments, which helped plantlets recover rapidly during acclimatization and obtain greater growth during post-acclimatization.

Low-dose naltrexone augmentation of nicotine replacement for smoking cessation with reduced weight gain: a randomized trial.

PubMed

Toll, Benjamin A; White, Marney; Wu, Ran; Meandzija, Boris; Jatlow, Peter; Makuch, Robert; O'Malley, Stephanie S

2010-10-01

Fear of weight gain is a significant obstacle to smoking cessation, preventing some smokers from attempting to quit. Several previous studies of naltrexone yielded promising results for minimization of post-quit weight gain. Given these encouraging findings, we endeavored to test whether minimization of weight gain might translate to better quit outcomes for a population that is particularly concerned about gaining weight upon quitting. Smokers (N=172) in this investigation were prospectively randomized to receive either 25 mg naltrexone or placebo for 27 weeks (1 week pre-, 26 weeks post-quit) for minimization of post-quit weight gain and smoking cessation. All participants received open label therapy with the nicotine patch for the first 8 weeks post-quit and behavioral counseling over the 27-week treatment. The 2 pre-specified primary outcomes were change in weight for continuously abstinent participants and biologically verified end-of-treatment 7-day point-prevalence abstinence at 26 weeks after the quit date. The difference in weight at 26 weeks post-quit between the naltrexone and placebo groups (naltrexone: 6.8 lbs ± 8.94 vs placebo: 9.7 lbs ± 9.19, p = 0.45) was not statistically different. Seven-day point-prevalence smoking abstinence rates at 26 weeks post-quit was not significantly different between the 2 groups (naltrexone: 22% vs placebo: 27%, p = 0.43). For smokers high in weight concern, the relatively small reduction in weight gain with low-dose naltrexone is not worth the potential for somewhat lower rates of smoking abstinence. Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.

Characterizing the recovery trajectories of knee range of motion for one year after total knee replacement.

PubMed

Mehta, Saurabh; Rigney, Andrew; Webb, Kyle; Wesney, Jacob; Stratford, Paul W; Shuler, Franklin D; Oliashirazi, Ali

2018-06-13

Retrospective analysis of routinely collected clinical data. This study modeled the recovery in knee flexion and extension range of motion (ROM) over 1 year after total knee replacement (TKR). Recovery after TKR has been characterized for self-reported pain and functional status. Literature describing target knee ROM at different follow-up periods after TKR is scarce. Data were extracted for patients who had undergone TKR at a tertiary care hospital at 2, 8, 12, 26, and 52 weeks after TKR. A linear mixed-effects growth model was constructed that investigated the following covariates age, sex, pre-TKR range, body mass index, duration of symptoms, and their interaction with weeks post TKR. Of the 559 patients included (age 64.8 ± 8.5 years), 370 were women and 189 were men. Knee ROM showed the greatest change during the first 12 weeks after TKR, plateauing by 26 weeks. For an average patient, knee flexion increased from approximately 100º 2 weeks post TKR to 117º 52 weeks post TKR. Knee extension increased from approximately 3º knee flexion 2 weeks post TKR to 1º flexion 52 weeks post TKR. The results showed that the maximum gains in knee ROM should be expected within the first 12 weeks with small changes occurring up to 26 weeks after TKR. In addition, age and presurgery knee ROM are associated with the gains in knee ROM and should be factored into the estimation of expected knee ROM at a given follow-up interval after TKR.

Role of chicken astrovirus as a causative agent of gout in commercial broilers in India.

PubMed

Bulbule, N R; Mandakhalikar, K D; Kapgate, S S; Deshmukh, V V; Schat, K A; Chawak, M M

2013-01-01

Several outbreaks of gout were reported in commercial broilers in India during 2011 and 2012, causing up to 40% mortality in the birds. Gross and histopathological observations confirmed gout. Quantitative reverse transcriptase polymerase chain reaction (qRT-PCR) analysis from kidney samples of gout-affected birds indicated the presence of chicken astrovirus (CAstV) in 41.7% of cases and a mixed infection of CAstV and avian nephritis virus (ANV) in 36.4% of cases. CAstV isolated from gout-affected kidneys by inoculating embryonated specific pathogen free (SPF) eggs showed dwarfing in embryos and a cytopathic effect in chicken embryo kidney cells. Inoculation of 1-day-old SPF and broiler chicks with CAstVs caused gout and mortality between 4 and 10 days post inoculation. Virus isolation and qRT-PCR analysis showed the presence of only CAstV in inoculated chicks. Sequence analysis of capsid genes indicated a major group of Indian CAstVs that displayed 92.0 to 99.2% intergroup amino acid identity and 83.9 to 90.4% identity with subgroup Bi CAstVs of UK origin. We designated this group Indian Bi. Analysis of the partial polymerase amino acid sequences of our isolates indicated two groups of CAstVs (Indian 1 and 2) that displayed 90.2 to 95.5% amino acid identity between them. We thus report for the first time that, in addition to infectious bronchitis virus and ANV, CAstVs are a causative agent of gout.

Profiling of Disease-Related Metabolites in Grapevine Internode Tissues Infected with Agrobacterium vitis

PubMed Central

Jung, Sung-Min; Hur, Youn-Young; Preece, John E.; Fiehn, Oliver; Kim, Young-Ho

2016-01-01

Green shoot cuttings of 10 different grapevine species were inoculated with Agrobacterium vitis to find disease-related metabolites in the grapevine. Crown galls formed 60 days after inoculation varied in gall severity (GS) evaluated by gall incidence (GI) and gall diameter (GD), which were classified into three response types as RR (low GI and small GD), SR (high GI and small GD), and SS (high GI and large GD), corresponding to resistant, moderately resistant, and susceptible responses, respectively. In this, 4, 4, and 2 Vitis species were classified into RR, SR, and SS, respectively. Gas chromatography mass spectrometry (GC-MS) analysis of the grapevine stem metabolites with A. vitis infection showed 134 metabolites in various compound classes critically occurred, which were differentially clustered with the response types by the principal component analysis. Multivariate analysis of the metabolite profile revealed that 11 metabolites increased significantly in relation to the response types, mostly at post-inoculation stages, more prevalently (8 metabolites) at two days after inoculation than other stages, and more related to SS (7 metabolites) than RR (3 metabolites) or SR (one metabolite). This suggests most of the disease-related metabolites may be rarely pre-existing but mostly induced by pathogen infection largely for facilitating gall development except stilbene compound resveratrol, a phytoalexin that may be involved in the resistance response. All of these aspects may be used for the selection of resistant grapevine cultivars and their rootstocks for the control of the crown gall disease of the grapevine. PMID:27904455

Experimental infection with the Toxoplasma gondii ME-49 strain in the Brazilian BR-1 mini pig is a suitable animal model for human toxoplasmosis.

PubMed

Miranda, Farlen José Bebber; Souza, Diogo Benchimol de; Frazão-Teixeira, Edwards; Oliveira, Fábio Conceição de; Melo, João Cardoso de; Mariano, Carlos Magno Anselmo; Albernaz, Antonio Peixoto; Carvalho, Eulógio Carlos Queiróz de; Oliveira, Francisco Carlos Rodrigues de; Souza, Wanderley de; DaMatta, Renato Augusto

2015-02-01

Toxoplasma gondii causes toxoplasmosis, a worldwide disease. Experimentation with pigs is necessary for the development of new therapeutic approaches to human diseases. BR-1 mini pigs were intramuscularly infected with T. gondii with tachyzoites (RH strain) or orally infected with cysts (ME-49 strain). Haematology and serum biochemistry were analysed and buffy coat cells were inoculated in mice to determine tachyzoite circulation. No alterations were observed in erythrocyte and platelet values; however, band neutrophils increased seven days after infection with ME-49. Serology of the mice inoculated with pig blood leucocytes revealed circulating ME-49 or RH strain tachyzoites in the pigs' peripheral blood at two and seven or nine days post-infection. The tachyzoites were also directly observed in blood smears from the infected pigs outside and inside leucocytes for longer periods. Alanine-aminotransferase was high at days 21 and 32 in the RH infected pigs. After 90 days, the pigs were euthanised and their tissue samples were processed and inoculated into mice. The mice serology revealed the presence of parasites in the hearts, ileums and mesenteric lymph nodes of the pigs. Additionally, cysts in the mice were only observed after pig heart tissue inoculation. The infected pigs presented similar human outcomes with relatively low pathogenicity and the BR-1 mini pig model infected with ME-49 is suitable to monitor experimental toxoplasmosis.

Experimental infection with the Toxoplasma gondii ME-49 strain in the Brazilian BR-1 mini pig is a suitable animal model for human toxoplasmosis

PubMed Central

Miranda, Farlen José Bebber; de Souza, Diogo Benchimol; Frazão-Teixeira, Edwards; de Oliveira, Fábio Conceição; de Melo, João Cardoso; Mariano, Carlos Magno Anselmo; Albernaz, Antonio Peixoto; de Carvalho, Eulógio Carlos Queiróz; de Oliveira, Francisco Carlos Rodrigues; de Souza, Wanderley; DaMatta, Renato Augusto

2015-01-01

Toxoplasma gondii causes toxoplasmosis, a worldwide disease. Experimentation with pigs is necessary for the development of new therapeutic approaches to human diseases. BR-1 mini pigs were intramuscularly infected with T. gondii with tachyzoites (RH strain) or orally infected with cysts (ME-49 strain). Haematology and serum biochemistry were analysed and buffy coat cells were inoculated in mice to determine tachyzoite circulation. No alterations were observed in erythrocyte and platelet values; however, band neutrophils increased seven days after infection with ME-49. Serology of the mice inoculated with pig blood leucocytes revealed circulating ME-49 or RH strain tachyzoites in the pigs' peripheral blood at two and seven or nine days post-infection. The tachyzoites were also directly observed in blood smears from the infected pigs outside and inside leucocytes for longer periods. Alanine-aminotransferase was high at days 21 and 32 in the RH infected pigs. After 90 days, the pigs were euthanised and their tissue samples were processed and inoculated into mice. The mice serology revealed the presence of parasites in the hearts, ileums and mesenteric lymph nodes of the pigs. Additionally, cysts in the mice were only observed after pig heart tissue inoculation. The infected pigs presented similar human outcomes with relatively low pathogenicity and the BR-1 mini pig model infected with ME-49 is suitable to monitor experimental toxoplasmosis. PMID:25742268

Growth-promoting Sphingomonas paucimobilis ZJSH1 associated with Dendrobium officinale through phytohormone production and nitrogen fixation

PubMed Central

Yang, Suijuan; Zhang, Xinghai; Cao, Zhaoyun; Zhao, Kaipeng; Wang, Sai; Chen, Mingxue; Hu, Xiufang

2014-01-01

Growth-promoting Sphingomonas paucimobilis ZJSH1, associated with Dendrobium officinale, a traditional Chinese medicinal plant, was characterized. At 90 days post-inoculation, strain ZJSH1 significantly promoted the growth of D. officinale seedlings, with increases of stems by 8.6% and fresh weight by 7.5%. Interestingly, the polysaccharide content extracted from the inoculated seedlings was 0.6% higher than that of the control. Similar growth promotion was observed with the transplants inoculated with strain ZJSH1. The mechanism of growth promotion was attributed to a combination of phytohormones and nitrogen fixation. Strain ZJSH1 was found using the Kjeldahl method to have a nitrogen fixation activity of 1.15 mg l−1, which was confirmed by sequencing of the nifH gene. Using high-performance liquid chromatography-mass spectrometry, strain ZJSH1 was found to produce various phytohormones, including salicylic acid (SA), indole-3-acetic acid (IAA), Zeatin and abscisic acid (ABA). The growth curve showed that strain ZJSH1 grew well in the seedlings, especially in the roots. Accordingly, much higher contents of SA, ABA, IAA and c-ZR were detected in the inoculated seedlings, which may play roles as both phytohormones and ‘Systemic Acquired Resistance’ drivers. Nitrogen fixation and secretion of plant growth regulators (SA, IAA, Zeatin and ABA) endow S. paucimobilis ZJSH1 with growth-promoting properties, which provides a potential for application in the commercial growth of D. officinale. PMID:25142808

A morel improved growth and suppressed Fusarium infection in sweet corn.

PubMed

Yu, Dan; Bu, Fangfang; Hou, Jiaojiao; Kang, Yongxiang; Yu, Zhongdong

2016-12-01

A post-fire morel collected from Populus simonii stands in Mt. Qingling was identified as Morchella crassipes Mes-20 by using nuclear ribosomal DNA internal transcribed spacer phylogeny. It was inoculated into sweet corn to observe colonized roots in purified culture and in greenhouse experiments. The elongation and maturation zones of sweet corn were remarkably colonized at the cortex intercellular and intracellular cells, vessel cells, and around the Casparian strip, forming ectendomycorrhiza-like structures. Colonization was also observed in the zone of cell division proximal to the root cap. Greenhouse assays with sweet corn showed that this morel stimulated the development of the root system and significantly increased the dry root biomass. M. crassipes also significantly reduced the incidence of Fusarium verticillioides in the kernels of mature ears when inoculated into young ears before Fusarium inoculation and prevented Fusarium infection in corn ears compared with that of the control in the greenhouse. When grown under axenic conditions, M. crassipes produced the phytohormones abscisic acid, indole-3-acetic acid, and salicylic acid. The benefits to plants elicited by M. crassipes may result from these phytohormones which may improve the drought resistance, biomass growth and resistance to Fusarium.

Biochar and Glomus caledonium Influence Cd Accumulation of Upland Kangkong (Ipomoea aquatica Forsk.) Intercropped with Alfred Stonecrop (Sedum alfredii Hance)

PubMed Central

Hu, Junli; Wu, Fuyong; Wu, Shengchun; Lam, Cheung Lung; Lin, Xiangui; Wong, Ming Hung

2014-01-01

Both biochar application and mycorrhizal inoculation have been proposed to improve plant growth and alter bioaccumulation of toxic metals. A greenhouse pot trial was conducted to investigate growth and Cd accumulation of upland kangkong (Ipomoea aquatica Forsk.) intercropped with Alfred stonecrop (Sedum alfredii Hance) in a Cd-contaminated soil inoculated with Glomus caledonium and/or applied with biochar. Compared with the monocultural control, intercropping with stonecrop (IS) decreased kangkong Cd acquisition via rhizosphere competition, and also decreased kangkong yield. Gc inoculation (+M) accelerated growth and Cd acquisition of stonecrop, and hence resulted in further decreases in kangkong Cd acquisition. Regardless of IS and +M, biochar addition (+B) increased kangkong yield via elevating soil available P, and decreased soil Cd phytoavailability and kangkong Cd concentration via increasing soil pH. Compared with the control, the treatment of IS + M + B had a substantially higher kangkong yield (+25.5%) with a lower Cd concentration (−62.7%). Gc generated additive effects on soil alkalinization and Cd stabilization to biochar, causing lower DTPA-extractable (phytoavailable) Cd concentrations and post-harvest transfer risks. PMID:24728157

Defense Health: Coordinating Authority Needed for Psychological Health and Traumatic Brain Injury Activities

DTIC Science & Technology

2012-01-01

Placebo-Controlled Trial of the Dopamine Beta Hydroxylase (DBH) Inhibitor, Nepicastat, for the Treatment of PTSD in Operation Iraqi Freedom (OIF...Operation Enduring Freedom (OEF) Veterans 1 A Randomized, Placebo-Controlled Trial of the Dopamine -?-Hydroxylase (DBH) Inhibitor, Nepicastat for the...Reduction: Predeployment Stress Inoculation Training 1 Combat, Sexual Assault, and Post-Traumatic Stress in OIF/OEF Military Women 1 Comparing

The primary immune response to Vaccinia virus vaccination includes cells with a distinct cytotoxic effector CD4 T-cell phenotype.

PubMed

Munier, C Mee Ling; van Bockel, David; Bailey, Michelle; Ip, Susanna; Xu, Yin; Alcantara, Sheilajen; Liu, Sue Min; Denyer, Gareth; Kaplan, Warren; Suzuki, Kazuo; Croft, Nathan; Purcell, Anthony; Tscharke, David; Cooper, David A; Kent, Stephen J; Zaunders, John J; Kelleher, Anthony D

2016-10-17

Smallpox was eradicated by a global program of inoculation with Vaccinia virus (VV). Robust VV-specific CD4 T-cell responses during primary infection are likely essential to controlling VV replication. Although there is increasing interest in cytolytic CD4 T-cells across many viral infections, the importance of these cells during acute VV infection is unclear. We undertook a detailed functional and genetic characterization of CD4 T-cells during acute VV-infection of humans. VV-specific T-cells were identified by up-regulation of activation markers directly ex vivo and through cytokine and co-stimulatory molecule expression. At day-13-post primary inoculation with VV, CD38highCD45RO+ CD4 T-cells were purified by cell sorting, RNA isolated and analysed by microarray. Differential expression of up-regulated genes in activated CD4 T-cells was confirmed at the mRNA and protein levels. We compared analyses of VV-specific CD4 T-cells to studies on 12 subjects with primary HIV infection (PHI). VV-specific T-cells lines were established from PBMCs collected post vaccination and checked for cytotoxicity potential. A median 11.9% CD4 T-cells were CD38highCD45RO+ at day-13 post-VV inoculation, compared to 3.0% prior and 10.4% during PHI. Activated CD4 T-cells had an up-regulation of genes related to cytolytic function, including granzymes K and A, perforin, granulysin, TIA-1, and Rab27a. No difference was seen between CD4 T-cell expression of perforin or TIA-1 to VV and PHI, however granzyme k was more dominant in the VV response. At 25:1 effector to target ratio, two VV-specific T-cell lines exhibited 62% and 30% cytotoxicity respectively and CD107a degranulation. We show for the first time that CD4 CTL are prominent in the early response to VV. Understanding the role of CD4 CTL in the generation of an effective anti-viral memory may help develop more effective vaccines for diseases such as HIV. Crown Copyright © 2016. Published by Elsevier Ltd. All rights reserved.

Zika virus transmission to mouse ear by mosquito bite: a laboratory model that replicates the natural transmission process.

PubMed

Secundino, Nagila Francinete Costa; Chaves, Barbara Aparecida; Orfano, Alessandra Silva; Silveira, Karine Renata Dias; Rodrigues, Nilton Barnabe; Campolina, Thais Bonifácio; Nacif-Pimenta, Rafael; Villegas, Luiz Eduardo Martinez; Silva, Breno Melo; Lacerda, Marcus Vinícius Guimarães; Norris, Douglas Eric; Pimenta, Paulo Filemon Paolucci

2017-07-20

Zika disease has transformed into a serious global health problem due to the rapid spread of the arbovirus and alarming severity including congenital complications, microcephaly and Guillain-Barré syndrome. Zika virus (ZIKV) is primarily transmitted to humans through the bite of an infective mosquito, with Aedes aegypti being the main vector. We successfully developed a ZIKV experimental transmission model by single infectious Ae. aegypti bite to a laboratory mouse using circulating Brazilian strains of both arbovirus and vector. Mosquitoes were orally infected and single Ae. aegypti were allowed to feed on mouse ears 14 days post-infection. Additionally, salivary gland (SG) homogenates from infected mosquitoes were intrathoracically inoculated into naïve Ae. aegypti. Mosquito and mouse tissue samples were cultured in C6/36 cells and processed by quantitative real-time PCR. A total of 26 Ae. aegypti were allowed to feed individually on mouse ears. Of these, 17 mosquitoes fed, all to full engorgement. The transmission rate of ZIKV by bite from these engorged mosquitoes to mouse ears was 100%. The amount of virus inoculated into the ears by bites ranged from 2 × 10 2 -2.1 × 10 10 ZIKV cDNA copies and was positively correlated with ZIKV cDNA quantified from SGs dissected from mosquitoes post-feeding. Replicating ZIKV was confirmed in macerated SGs (2.45 × 10 7 cDNA copies), mouse ear tissue (1.15 × 10 3 cDNA copies, and mosquitoes 14 days post-intrathoracic inoculation (1.49 × 10 7 cDNA copies) by cytopathic effect in C6/36 cell culture and qPCR. Our model illustrates successful transmission of ZIKV by an infectious mosquito bite to a live vertebrate host. This approach offers a comprehensive tool for evaluating the development of infection in and transmission from mosquitoes, and the vertebrate-ZIKV interaction and progression of infection following a natural transmission process.

NON-PARALYTIC POLIOMYELITIS IN THE CHIMPANZEE

PubMed Central

Bodian, David; Howe, Howard A.

1945-01-01

1. Thirteen cases of non-paralytic poliomyelitis infection in chimpanzees are described. Nine of these animals were excreting virus in. their stools at periods of from 3 days to 8 weeks following inoculation. 2. All animals killed during the acute stage showed lesions in the brain distributed in centers usually involved in, and compatible with the presence of, poliomyelitic infection. In 2 chimpanzees typical cord lesions were also present. No lesions were found in the brains of 4 control chimpanzees which had had no virus contact as far as known. The occurrence of a purely systemic or peripheral form of poliomyelitis, without lesions in the central nervous system, has thus not been established. 3. Four instances of arrest of the pathological process near the portal of entry into the brain, indicating partial resistance, are included in this series. One was a chimpanzee inoculated intranasally (A1-75) who had severe tuberculosis at the time of inoculation. The second was an animal convalescent after intracerebral inoculation (A1-74), who sustained a second infection limited to the olfactory bulbs when inoculated intranasally 2 months later with homologous virus. The third (A5-01) was inoculated orally with human stool, but contammation of the olfactory area resulted with infection of the olfactory bulbs and of the forebrain; virus was present in the stools of this animal. The fourth chimpanzee (A48) had suffered an initial non-paralytic attack after stomach tube inoculation, followed by a second attack about 9 months later after oral inoculation with part of the same virus-containing pool (human stools). The second attack consisted of a facial paralysis, with arrest of the pathological process near the facial nucleus. 4. Although cerebral lesions were light in some of the non-paralytic and inapparent infections, their presence in all indicates the action of virus on the central nervous system with the possibihty of production of at least partial local resistance. It is not unreasonable to assume that this may occur in inapparent human cases, although the point is, of course, not susceptible to critical proof in man. 5. The degree of severity of pathological involvement in non-paralytic cases varies from a fully developed distribution of lesions in brain and spinal cord in some chimpanzees, to mild and scattered lesions in the brains of others. This suggests that if the extent of pathological reaction is an indicator of subsequent local resistance to reinfection, the degree of protection afforded by a non-paralytic attack of poliomyelitis to even homologous virus must be variable. PMID:19871456

Comparison of the efficacy of aroma-acupressure and aromatherapy for the treatment of dementia-associated agitation.

PubMed

Yang, Man-Hua; Lin, Li-Chan; Wu, Shiao-Chi; Chiu, Jen-Hwey; Wang, Pei-Ning; Lin, Jaung-Geng

2015-03-29

One of the most common symptoms observed in patients with dementia is agitation, and several non-pharmacological treatments have been used to control this symptom. However, because of limitations in research design, the benefit of non-pharmacological treatments has only been demonstrated in certain cases. The purpose of this study was to compare aroma-acupressure and aromatherapy with respect to their effects on agitation in patients with dementia. In this experimental study, the participants were randomly assigned to three groups: 56 patients were included in the aroma-acupressure group, 73 patients in the aromatherapy group, and 57 patients in the control group who received daily routine as usual without intervention. The Cohen-Mansfield Agitation Inventory (CMAI) scale and the heart rate variability (HRV) index were used to assess differences in agitation. The CMAI was used in the pre-test, post-test and post-three-week test, and the HRV was used in the pre-test, the post-test and the post-three-week test as well as every week during the four-week interventions. The CMAI scores were significantly lower in the aroma-acupressure and aromatherapy groups compared with the control group in the post-test and post-three-week assessments. Sympathetic nervous activity was significantly lower in the fourth week in the aroma-acupressure group and in the second week in the aromatherapy group, whereas parasympathetic nervous activity increased from the second week to the fourth week in the aroma-acupressure group and in the fourth week in the aromatherapy group. Aroma-acupressure had a greater effect than aromatherapy on agitation in patients with dementia. However, agitation was improved in both of the groups, which allowed the patients with dementia to become more relaxed. Future studies should continue to assess the benefits of aroma-acupressure and aromatherapy for the treatment of agitation in dementia patients. ChiCTR-TRC-14004810; Date of registration: 2014/6/12.

The influence of physical therapy and anti-botulinum toxin antibody on the efficacy of botulinum toxin-A injections in children with spastic cerebral palsy.

PubMed

Jang, Dae-Hyun; Sung, In Young

2014-12-01

To identify factors associated with the efficacy of botulinum toxin-A (BoNT-A) injections. Thirty-eight children with spastic cerebral palsy (CP) received BoNT-A injections into the gastrocnemius. The baseline anti-botulinum antibodies were checked. The Static dorsiflexion range of motion (ROM), Modified Tardieu Scale (MTS) and Physician Rating Scale (PRS) were assessed at pre-injection as well as 4- and 12-week post-injection. No samples contained anti-botulinum antibodies. Greater baseline MTS dynamic range was associated with greater changes in MTS dynamic ranges at 4-week post-injection. More frequent physical therapy was associated with greater changes in static dorsiflexion ROM at 4-week post-injection and greater changes in PRS at 4- and 12-week post-injection. The improvement in PRS at 12-week post-injection was associated with the frequency of physical therapy. Therefore, intensive physical therapy programs may be necessary to maintain the beneficial effects of BoNT-A injections in children with CP.

A DNA vaccine against dolphin morbillivirus is immunogenic in bottlenose dolphins.

PubMed

Vaughan, Kerrie; Del Crew, Jason; Hermanson, Gary; Wloch, Mary K; Riffenburgh, Robert H; Smith, Cynthia R; Van Bonn, William G

2007-12-15

The immunization of exotic species presents considerable challenges. Nevertheless, for facilities like zoos, animal parks, government facilities and non-profit conservation groups, the protection of valuable and endangered species from infectious disease is a growing concern. The rationale for immunization in these species parallels that for human and companion animals; to decrease the incidence of disease. The U.S. Navy Marine Mammal Program, in collaboration with industry and academic partners, has developed and evaluated a DNA vaccine targeting a marine viral pathogen - dolphin morbillivirus (DMV). The DMV vaccine consists of the fusion (F) and hemagglutinin (H) genes of DMV. Vaccine constructs (pVR-DMV-F and pVR-DMV-H) were evaluated for expression in vitro and then for immunogenicity in mice. Injection protocols were designed for application in Atlantic bottlenose dolphins (Tursiops truncatus) to balance vaccine effectiveness with clinical utility. Six dolphins were inoculated, four animals received both pDMV-F and pDMV-H and two animals received a mock vaccine (vector alone). All animals received an inoculation week 0, followed by two booster injections weeks 8 and 14. Vaccine-specific immune responses were documented in all four vaccinated animals. To our knowledge, this is the first report of pathogen-specific immunogenicity to a DNA vaccine in an aquatic mammal species.

Effects of combined therapy with silymarin and glucantime on leishmaniasis induced by Leishmania major in BALB/c mice.

PubMed

Jabini, R; Jaafari, M R; Vahdati Hasani, F; Ghazizadeh, F; Khamesipour, A; Karimi, G

2015-03-01

Leishmania major is resistant to the traditional treatments in many parts of the world. PgpA, a member of (ABC) transporter superfamily, has been identified in Leishmania involved in antimony resistance. Silymarin can inhibit PgpA. The aim of this study was to determine the effect of combined therapy with glucantime and silymarin on Cutaneous Leishmaniasis. The effects of silymarin on response of L. major to glucantime were evaluated with amastigote macrophage and mice model of leishmaniasis. Immediately after injection in mice inoculated into footpads with L. major amastigote, systemic treatment was performed and the size of footpad swelling was measured twice a week. 4 and 8 weeks after the beginning of the treatment, splenic parasite burden was done. Silymarin showed no significant effect on the response of L. major promastigotes to glucantime. 2 formulations (glucantime 25 µm with silymarin 25 µm or 12.5 µm) reduced cell death in amastigote assays. The effect of silymarin on footpad swelling was detected when the combination of low-dose glucantime (20 mg/kg) with 25-50 mg/kg silymarin (especially 50 mg/kg) were used at day 22 of post infection (P<0.05). According to the parasite burden data, use of silymarin in the presence of different doses of glucantime, did not show significant effect compared to glucantime alone. The results of this study suggest that silymarin in conjunction with glucantime may have benefit effects in murine model of cutaneous leishmaniasis. © Georg Thieme Verlag KG Stuttgart · New York.

Tumour xenograft detection through quantitative analysis of the metabolic profile of urine in mice

NASA Astrophysics Data System (ADS)

Moroz, Jennifer; Turner, Joan; Slupsky, Carolyn; Fallone, Gino; Syme, Alasdair

2011-02-01

The metabolic content of urine from NIH III nude mice (n = 22) was analysed before and after inoculation with human glioblastoma multiforme (GBM) cancer cells. An age- and gender-matched control population (n = 14) was also studied to identify non-tumour-related changes. Urine samples were collected daily for 6 weeks, beginning 1 week before cell injection. Metabolite concentrations were obtained via targeted profiling with Chenomx Suite 5.1, based on nuclear magnetic resonance (NMR) spectra acquired on an Oxford 800 MHz cold probe NMR spectrometer. The Wilcoxon rank sum test was used to evaluate the significance of the change in metabolite concentration between the two time points. Both the metabolite concentrations and the ratios of pairs of metabolites were studied. The complicated inter-relationships between metabolites were assessed through partial least-squares discriminant analysis (PLS-DA). Receiver operating characteristic (ROC) curves were generated for all variables and the area under the curve (AUC) calculated. The data indicate that the number of statistically significant changes in metabolite concentrations was more pronounced in the tumour-bearing population than in the control animals. This was also true of the ratios of pairs of metabolites. ROC analysis suggests that the ratios were better able to differentiate between the pre- and post-injection samples compared to the metabolite concentrations. PLS-DA models produced good separation between the populations and had the best AUC results (all models exceeded 0.937). These results demonstrate that metabolomics may be used as a screening tool for GBM cells grown in xenograft models in mice.

Involvement of rainbow trout leucocytes in the pathogenesis of infectious hematopoietic necrosis

USGS Publications Warehouse

Chilmonczyk, S.; Winton, J.R.

1994-01-01

Rainbow trout Oncorhynchus myluss leucocytes were tested for their ability to support replication of infectious hematopoietic necrosis virus (IHNV). Viral replication occurred in vitro uslng leucocytes cultured from peripheral blood, kidney, and thymus where viral titers peaked at 2 to 4 d post-inoculation. Leucocytes collected from trout following waterborne challenge with IHNV were cocultured on EPC cell monolayers. These assays detected IHNV in leucocytes infected in vivo as early as 6 h post-exposure before the challenge virus had undergone replication. These data showed that leucocyte populations could serve as target cells in the initial phase of IHNV infection.

The effect of different grain diets on fecal shedding of Escherichia coli O157:H7 by steers.

PubMed

Buchko, S J; Holley, R A; Olson, W O; Gannon, V P; Veira, D M

2000-11-01

Three groups of six yearling steers (three rumen fistulated plus three nonfistulated) fed one of three different grain diets (85% cracked corn, 15% whole cottonseed and 70% barley, or 85% barley) were inoculated with 10(10) CFU of Escherichia coli O157:H7 strain 3081, and the presence of the inoculated strain was followed in the rumen fluid and feces for a 10-week period. E. coli O157:H7 was rapidly eliminated from the rumen of the animals on all three diets but persisted in the feces of some animals up to 67 days after inoculation, suggesting that the bovine hindgut is the site of E. coli O157:H7 persistence. A significant difference existed in the levels of E. coli O157:H7 shed by the animals among diets on days 5, 7, 49, and 63 after inoculation (P < 0.05). No significant difference was found between the levels shed among diets on days 9 through 42 and on day 67 (P > 0.05). The number of animals that were culture positive for E. coli O157:H7 strain 3081 during the 10-week period was significantly higher for the barley fed group (72 of 114 samplings) as opposed to the corn fed group (44 of 114 samplings) (P < 0.005) and the cottonseed and barley fed group (57 of 114 samplings) (P < 0.05). The fecal pH of the animals fed the corn diet was significantly lower (P < 0.05) than the fecal pH of the animals fed the cottonseed and barley and barley diets, likely resulting in a less suitable environment for E. coli O157:H7 in the hindgut of the corn fed animals. E. coli O157:H7 strain 3081 was present in 3 of 30 (corn, 1 of 10; cottonseed, 1 of 10; barley, 1 of 10) animal drinking water samples, 3 of 30 (corn, 1 of 10; cottonseed, 0 of 10; barley, 2 of 10) water trough biofilm swabs, 5 of 30 (corn, 0 of 10; cottonseed, 2 of 10; barley, 3 of 10) feed samples, and 30 of 30 manure samples taken from the pens during the entire experimental period. Mouth swabs of the steers were also culture positive for E. coli O157:H7 strain 3081 in 30 of 180 samples (corn, 7 of 60; cottonseed, 4 of 60; barley, 19 of 60) taken during the 10-week period. Minimizing environmental dissemination of E. coli O157:H7 in conjunction with diet modification may reduce numbers of E. coli O157:H7-positive cattle.

Elevated Atmospheric CO2 and Strain of Rhizobium Alter Freezing Tolerance and Cold-induced Molecular Changes in Alfalfa (Medicago sativa)

PubMed Central

Bertrand, Annick; Prévost, Danielle; Bigras, Francine J.; Castonguay, Yves

2007-01-01

Background and Aims The objective of the study was to assess the impact of elevated CO2 in interaction with rhizobial strains on freezing tolerance and cold-induced molecular changes in alfalfa. Methods Alfalfa inoculated with two different strains of rhizobium (A2 and NRG34) was grown and cold acclimated (2 weeks at 2 °C) under either 400 (ambient) or 800 µmol mol−1 (elevated) CO2. Key Results Plants acclimated under 400 µmol mol−1 CO2 were more freezing tolerant than those maintained under 800 µmol mol−1. Cryoprotective sugars typically linked with the acquisition of freezing tolerance such as sucrose, stachyose and raffinose increased in roots in response to low temperature but did not differ between CO2 treatments. Similarly high CO2 did not alter the expression of many cold-regulated (COR) genes although it significantly increased the level of transcripts encoding a COR gene homologous to glyceraldehyde-3-phosphate-dehydrogenase (GAPDH). A significant effect of rhizobial strain was observed on both freezing tolerance and gene expression. Plants of alfalfa inoculated with strain A2 were more freezing tolerant than those inoculated with strain NRG34. Transcripts of COR genes homologous to a pathogenesis-related protein (PR-10) and to a nuclear-targeted protein were markedly enhanced in roots of alfalfa inoculated with strain A2 as compared with strain NRG34. Transcripts encoding the vegetative storage proteins (VSPs) β-amylase and chitinase were more abundant in roots of non-acclimated plants inoculated with strain NRG34 than with strain A2. Conclusions Taken together, the results suggest that elevated CO2 stimulates plant growth and reduces freezing tolerance. The acquisition of cold tolerance is also influenced by the rhizobial strain, as indicated by lower levels of expression of COR genes and sustained accumulation of VSP-encoding transcripts in alfalfa inoculated with strain NRG34 as compared with strain A2. PMID:17218341

Biocontrol of the Brown-Banded Cockroach, Supella longipalpa F. (Blattaria: Blattellidae), with Entomopathogenic Fungus, Metharhizium anisopliae

PubMed Central

Sharififard, Mona; Mossadegh, Mohammad Saeed; Vazirianzadeh, Babak; Latifi, Seyed Mahmood

2016-01-01

Background: Considering to the high distribution of cockroaches as urban pests, the efficacy of different formulations of Metarhizium anisopliae strain Iran 437C were assessed against the brown-banded cockroach, Supella longipalpa F. under laboratory and field conditions. Methods: Metarhizium anisopliae isolates were screened with immersing adults of the brown-banded cockroachs in aqueous suspension of 108 conidia ml−1 followed by surface or bait treated with different doses of the most virulent isolate against the nymphs. Then formulations of conidia oil-in-water were examined versus cockroach nymphs using different plant oils and paraffin. Then they were evaluated and compared with aqueous suspension and control group. On a large-scale, the sunflower oil-in-water formulation of conidia was sprayed at houses using a hand sprayer. Results: Metarhizium anisopliae IRAN 437C was the most virulent isolate against the brown-banded cockroach, causing 100% mortality in adults at seven days post-exposure. Inoculated bait with this isolate was not enough pathogenic against the cockroach even at two weeks after treatment. Treated surface with conidia as aqueous suspension or oil-in-water formulation was more effective than the bait formulation against the cockroach caused 39.4–97.2% mortality compared with 2.5% mortality in control group after two days. Spraying the conidia formulated with sunflower oil was an effective formulation causing 76.1% reduction in the cockroach density on the third day post treatment in the houses. Conclusion: The oil-in-water formulation of M. anisopliae IRAN 437C could be recommended as a promising alternative for cockroach control. PMID:27308292

Tendon Mineralization Is Progressive and Associated with Deterioration of Tendon Biomechanical Properties, and Requires BMP-Smad Signaling in the Mouse Achilles Tendon Injury Model

PubMed Central

Zhang, Kairui; Asai, Shuji; Hast, Michael W.; Liu, Min; Usami, Yu; Iwamoto, Masahiro; Soslowsky, Louis J.; Enomoto-Iwamoto, Motomi

2016-01-01

Ectopic tendon mineralization can develop following tendon rupture or trauma surgery. The pathogenesis of ectopic tendon mineralization and its clinical impact have not been fully elucidated yet. In this study, we utilized a mouse Achilles tendon injury model to determine whether ectopic tendon mineralization alters the biomechanical properties of the tendon and whether BMP signaling is involved in this condition. A complete transverse incision was made at the midpoint of the right Achilles tendon in 8-week-old CD1 mice and the gap was left open. Ectopic cartilaginous mass formation was found in the injured tendon by 4 weeks post-surgery and ectopic mineralization was detected at 8–10 weeks post-surgery. Ectopic mineralization grew over time and volume of the mineralized materials of 25-weeks samples was about 2.5 fold bigger than that of 10-weeks samples, indicating that injury-induced ectopic tendon mineralization is progressive. In vitro mechanical testing showed that max force, max stress and mid-substance modulus in the 25-weeks samples were significantly lower than the 10-weeks samples. We observed substantial increases in expression of bone morphogenetic protein family genes in injured tendons 1 week post-surgery. Immunohistochemical analysis showed that phosphorylation of both Smad1 and Smad3 were highly increased in injured tendons as early as 1 week post-injury and remained high in ectopic chondrogenic lesions 4 weeks post-injury. Treatment with the BMP receptor kinase inhibitor (LDN193189) significantly inhibited injury-induced tendon mineralization. These findings indicate that injury-induced ectopic tendon mineralization is progressive, involves BMP signaling and associated with deterioration of tendon biomechanical properties. PMID:26825318

Effect of cutaneous and digestive colonization in the induction of anti-Candida albicans antibodies: experimental study.

PubMed

Nogueira, J M; Garcia-de-Lomas, J; Buesa, F J; Prat, J; Mir, A; Camarena, J J

1985-10-01

Candida albicans colonization induces antibodies, which must be taken into account in the serological diagnosis of candidiasis. In order to determine the degree of this effect, an experimental study in rabbits free of specific anti-Candida antibodies by cutaneous and digestive inoculation has been carried out. The evolution of humoral response was studied over 8 weeks by indirect immunofluorescence (IIF), direct agglutination (DA), counterimmunoelectrophoresis (CIE) and double diffusion (DD). The cutaneous colonization detectable by culture was maintained until the second week in 70% of the animals and the presence of antibodies detectable by IIF and DA was observed after the 2nd week. The highest antibody titre by IFF and DA was 1/64, and was reached in the 5th week, with a tendency to drop in the following weeks. Precipitins were only detected by CIE in 15% of the animals in the 7th week. Elimination of yeast in stools continued only in 20% of the animals in the 2nd week of the experiment. Antibodies were detected by IIF and DA after the 2nd week, with the highest titres detectable by IFF in the 5th week. Precipitant antibodies detectable by CIE appeared in 15% of the animals in the 8th week.

Selenium Status Alters the Immune Response and Expulsion of Adult Heligmosomoides bakeri Worms in Mice

PubMed Central

Cheung, Lumei; Beshah, Ethiopia; Shea-Donohue, Terez; Urban, Joseph F.

2013-01-01

Heligmosomoides bakeri is a nematode with parasitic development exclusively in the small intestine of infected mice that induces a potent STAT6-dependent Th2 immune response. We previously demonstrated that host protective expulsion of adult H. bakeri worms from a challenge infection was delayed in selenium (Se)-deficient mice. In order to explore mechanisms associated with the delayed expulsion, 3-week-old female BALB/c mice were placed on a torula yeast-based diet with or without 0.2 ppm Se, and after 5 weeks, they were inoculated with H. bakeri infective third-stage larvae (L3s). Two weeks after inoculation, the mice were treated with an anthelmintic and then rested, reinoculated with L3s, and evaluated at various times after reinoculation. Analysis of gene expression in parasite-induced cysts and surrounding tissue isolated from the intestine of infected mice showed that the local-tissue Th2 response was decreased in Se-deficient mice compared to that in Se-adequate mice. In addition, adult worms recovered from Se-deficient mice had higher ATP levels than worms from Se-adequate mice, indicating greater metabolic activity in the face of a suboptimal Se-dependent local immune response. Notably, the process of worm expulsion was restored within 2 to 4 days after feeding a Se-adequate diet to Se-deficient mice. Expulsion was associated with an increased local expression of Th2-associated genes in the small intestine, intestinal glutathione peroxidase activity, secreted Relm-β protein, anti-H. bakeri IgG1 production, and reduced worm fecundity and ATP-dependent metabolic activity. PMID:23649095

An evaluation of aerobic and anaerobic composting of banana peels treated with different inoculums for soil nutrient replenishment.

PubMed

Kalemelawa, Frank; Nishihara, Eiji; Endo, Tsuneyoshi; Ahmad, Zahoor; Yeasmin, Rumana; Tenywa, Moses M; Yamamoto, Sadahiro

2012-12-01

This study sought to evaluate the efficacy of aerobic and anaerobic composting of inoculated banana peels, and assess the agronomic value of banana peel-based compost. Changes in the chemical composition under aerobic and anaerobic conditions were examined for four formulations of banana peel-based wastes over a period of 12 weeks. The formulations i.e. plain banana peel (B), and a mixture with either cow dung (BC), poultry litter (BP) or earthworm (BE) were separately composted under aerobic and anaerobic conditions under laboratory conditions. Inoculation with either cow dung or poultry litter significantly facilitated mineralization in the order: BP>BC>B. The rate of decomposition was significantly faster under aerobic than in anaerobic composting conditions. The final composts contained high K (>100 g kg(-1)) and TN (>2%), indicating high potential as a source of K and N fertilizer. Copyright © 2012 Elsevier Ltd. All rights reserved.

Agrobacterium-mediated transformation of protocorm-like bodies in Cymbidium.

PubMed

Chin, Dong Poh; Mishiba, Kei-ichiro; Mii, Masahiro

2007-06-01

Genetically transformed plants of Cymbidium were regenerated after cocultivating protocorm-like bodies (PLB) with Agrobacterium tumefaciens strain EHA101 (pIG121Hm) that harbored genes for beta-glucuronidase (gus), hygromycin phosphotransferase (hpt) and neomycin phosphotransferase II (nptII). PLB of three genotypes maintained in liquid new Dogashima medium (NDM), were subjected to transformation experiments. The PLB inoculated with Agrobacterium produced secondary PLB, 4 weeks after transfer onto 2.5 g L(-1) gellan gum-solidified NDM containing 10 g L(-1) sucrose, 20 mg L(-1) hygromycin and 40 mg L(-1) meropenem. Transformation efficiency was affected by genotype and the presence of acetosyringone during cocultivation. The highest transformation efficiency was obtained when PLB from the genotype L4 were infected and cocultivated with Agrobacterium on medium containing 100 muM acetosyringone. Transformation of the hygromycin-resistant plantlets regenerated from different sites of inoculated PLB was confirmed by histochemical GUS assay, PCR analysis and Southern blot hybridization.

The Cold Hardiness of Phrynocephalus erythrurus, the Lizard Living at Highest Altitude in the World.

PubMed

Li, X T; Wang, Y; Lu, S S; Li, M; Men, S K; Bai, Y C; Tang, X L; Chen, Q

Phrynocephalus erythrurus living at Qinghai-Tibet Plateau, is believed to be the highest lizard in the world, but we know little about how these lizards cope with very low temperatures in winter. The aim of this study was to find the difference of the lizards before and after cold acclimatization. In this study the limit of supercooling and inoculative freezing, the concentration of four organic osmolytes, and the activity of lactate dehydrogenase in the plasma were measured in samples shortly after capture and in other samples after 7~8 weeks of acclimatization at 2~4 degree C. Animals acquired an ability to undergo deeper supercooling and inoculative freezing through the course of acclimatization. We find no regular changes of the four organic osmolytes after the acclimatization. We think that this species of lizard is partly freeze-tolerant and conclude that it uses supercooling to survive in winter.

Acute HBV infection in humanized chimeric mice has multiphasic viral kinetics

DOE PAGES

Ishida, Yuji; Chung, Tje Lin; Imamura, Michio; ...

2018-03-23

Background: Chimeric uPA/SCID mice reconstituted with humanized livers are useful for studying HBV infection in the absence of an adaptive immune response. However, the detailed characterization of HBV infection kinetics necessary to enable in-depth mechanistic studies in this novel in vivo HBV infection model is lacking. Methods: To characterize HBV kinetics post-inoculation (p.i.) to steady state, 42 mice were inoculated with HBV. Serum HBV DNA was frequently measured from 1 minute to 63 days p.i. Total intrahepatic HBV DNA, HBV cccDNA, and HBV RNA was measured in a subset of mice at 2, 4, 6, 10, and 13 weeks p.i.more » HBV half-life (t 1/2) was estimated using a linear mixed-effects model. Results: During the first 6 h p.i. serum HBV declined in repopulated uPA/SCID mice with a t 1/2=62 min [95%CI=59-67min]. Thereafter, viral decline slowed followed by a 2 day lower plateau. Subsequent viral amplification was multiphasic with an initial mean doubling time of t 2= 8±3 h followed by an interim plateau before prolonged amplification (t 2=2±0.5 days) to a final HBV steady state of 9.3 ± 0.3 log copies/ml. Serum HBV and intrahepatic HBV DNA were positively correlated (R2=0.98). Conclusions: HBV infection in uPA/SCID chimeric mice is highly dynamic despite the absence of an adaptive immune response. The serum HBV t 1/2 in humanized uPA/SCID mice was estimated to be ~1 h regardless of inoculum size. Finally, the HBV acute infection kinetics presented here is an important step in characterizing this experimental model system so that it can be effectively used to elucidate the dynamics of the HBV lifecycle and thus possibly reveal effective antiviral drug targets.« less

Acute HBV infection in humanized chimeric mice has multiphasic viral kinetics

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

Ishida, Yuji; Chung, Tje Lin; Imamura, Michio

Background: Chimeric uPA/SCID mice reconstituted with humanized livers are useful for studying HBV infection in the absence of an adaptive immune response. However, the detailed characterization of HBV infection kinetics necessary to enable in-depth mechanistic studies in this novel in vivo HBV infection model is lacking. Methods: To characterize HBV kinetics post-inoculation (p.i.) to steady state, 42 mice were inoculated with HBV. Serum HBV DNA was frequently measured from 1 minute to 63 days p.i. Total intrahepatic HBV DNA, HBV cccDNA, and HBV RNA was measured in a subset of mice at 2, 4, 6, 10, and 13 weeks p.i.more » HBV half-life (t 1/2) was estimated using a linear mixed-effects model. Results: During the first 6 h p.i. serum HBV declined in repopulated uPA/SCID mice with a t 1/2=62 min [95%CI=59-67min]. Thereafter, viral decline slowed followed by a 2 day lower plateau. Subsequent viral amplification was multiphasic with an initial mean doubling time of t 2= 8±3 h followed by an interim plateau before prolonged amplification (t 2=2±0.5 days) to a final HBV steady state of 9.3 ± 0.3 log copies/ml. Serum HBV and intrahepatic HBV DNA were positively correlated (R2=0.98). Conclusions: HBV infection in uPA/SCID chimeric mice is highly dynamic despite the absence of an adaptive immune response. The serum HBV t 1/2 in humanized uPA/SCID mice was estimated to be ~1 h regardless of inoculum size. Finally, the HBV acute infection kinetics presented here is an important step in characterizing this experimental model system so that it can be effectively used to elucidate the dynamics of the HBV lifecycle and thus possibly reveal effective antiviral drug targets.« less

Matrix Metalloproteinases as a Therapeutic Target to Improve Neurologic Recovery after Spinal Cord Injury

DTIC Science & Technology

2012-10-01

pressure (maximal vesical pressure in an animal without voluntary voiding, prior to urine overflow), voiding duration, and voiding interval (the frequency...until euthanasia . Neurological status was evaluated at 1 and 3 days post injury and weekly thereafter for 3 weeks. At 4 weeks post-injury, awake

Characterization of the rhesus macaque (Macaca mulatta) scrub typhus model: Susceptibility to intradermal challenge with the human pathogen Orientia tsutsugamushi Karp

PubMed Central

Sunyakumthorn, Piyanate; Somponpun, Suwit J.; Im-erbsin, Rawiwan; Anantatat, Tippawan; Jenjaroen, Kemajittra; Dunachie, Susanna J.; Lombardini, Eric D.; Burke, Robin L.; Blacksell, Stuart D.; Jones, James W.; Mason, Carl J.; Richards, Allen L.; Day, Nicholas P. J.

2018-01-01

Background Scrub typhus is an important endemic disease in tropical Asia caused by Orientia tsutsugamushi for which no effective broadly protective vaccine is available. The successful evaluation of vaccine candidates requires well-characterized animal models and a better understanding of the immune response against O. tsutsugamushi. While many animal species have been used to study host immunity and vaccine responses in scrub typhus, only limited data exists in non-human primate (NHP) models. Methodology/Principle findings In this study we evaluated a NHP scrub typhus disease model based on intradermal inoculation of O. tsutsugamushi Karp strain in rhesus macaques (n = 7). After an intradermal inoculation with 106 murine LD50 of O. tsutsugamushi at the anterior thigh (n = 4) or mock inoculum (n = 3), a series of time course investigations involving hematological, biochemical, molecular and immunological assays were performed, until day 28, when tissues were collected for pathology and immunohistochemistry. In all NHPs with O. tsutsugamushi inoculation, but not with mock inoculation, the development of a classic eschar with central necrosis, regional lymphadenopathy, and elevation of body temperature was observed on days 7–21 post inoculation (pi); bacteremia was detected by qPCR on days 6–18 pi; and alteration of liver enzyme function and increase of white blood cells on day 14 pi. Immune assays demonstrated raised serum levels of soluble cell adhesion molecules, anti-O. tsutsugamushi-specific antibody responses (IgM and IgG) and pathogen-specific cell-mediated immune responses in inoculated macaques. The qPCR assays detected O. tsutsugamushi in eschar, spleen, draining and non-draining lymph nodes, and immuno-double staining demonstrated intracellular O. tsutsugamushi in antigen presenting cells of eschars and lymph nodes. Conclusions/Significance These data show the potential of using rhesus macaques as a scrub typhus model, for evaluation of correlates of protection in both natural and vaccine induced immunity, and support the evaluation of future vaccine candidates against scrub typhus. PMID:29522521

Early activation of wheat polyamine biosynthesis during Fusarium head blight implicates putrescine as an inducer of trichothecene mycotoxin production

PubMed Central

2010-01-01

Background The fungal pathogen Fusarium graminearum causes Fusarium Head Blight (FHB) disease on wheat which can lead to trichothecene mycotoxin (e.g. deoxynivalenol, DON) contamination of grain, harmful to mammalian health. DON is produced at low levels under standard culture conditions when compared to plant infection but specific polyamines (e.g. putrescine and agmatine) and amino acids (e.g. arginine and ornithine) are potent inducers of DON by F. graminearum in axenic culture. Currently, host factors that promote mycotoxin synthesis during FHB are unknown, but plant derived polyamines could contribute to DON induction in infected heads. However, the temporal and spatial accumulation of polyamines and amino acids in relation to that of DON has not been studied. Results Following inoculation of susceptible wheat heads by F. graminearum, DON accumulation was detected at two days after inoculation. The accumulation of putrescine was detected as early as one day following inoculation while arginine and cadaverine were also produced at three and four days post-inoculation. Transcripts of ornithine decarboxylase (ODC) and arginine decarboxylase (ADC), two key biosynthetic enzymes for putrescine biosynthesis, were also strongly induced in heads at two days after inoculation. These results indicated that elicitation of the polyamine biosynthetic pathway is an early response to FHB. Transcripts for genes encoding enzymes acting upstream in the polyamine biosynthetic pathway as well as those of ODC and ADC, and putrescine levels were also induced in the rachis, a flower organ supporting DON production and an important route for pathogen colonisation during FHB. A survey of 24 wheat genotypes with varying responses to FHB showed putrescine induction is a general response to inoculation and no correlation was observed between the accumulation of putrescine and infection or DON accumulation. Conclusions The activation of the polyamine biosynthetic pathway and putrescine in infected heads prior to detectable DON accumulation is consistent with a model where the pathogen exploits the generic host stress response of polyamine synthesis as a cue for production of trichothecene mycotoxins during FHB disease. However, it is likely that this mechanism is complicated by other factors contributing to resistance and susceptibility in diverse wheat genetic backgrounds. PMID:21192794

Early Changes of Articular Cartilage and Subchondral Bone in The DMM Mouse Model of Osteoarthritis.

PubMed

Fang, Hang; Huang, Lisi; Welch, Ian; Norley, Chris; Holdsworth, David W; Beier, Frank; Cai, Daozhang

2018-02-12

To examine the early changes of articular cartilage and subchondral bone in the DMM mouse model of osteoarthritis, mice were subjected to DMM or SHAM surgery and sacrificed at 2-, 5- and 10-week post-surgery. Catwalk gait analyses, Micro-Computed Tomography, Toluidine Blue, Picrosirius Red and Tartrate-Resistant Acid Phosphatase (TRAP) staining were used to investigate gait patterns, joint morphology, subchondral bone, cartilage, collagen organization and osteoclasts activity, respectively. Results showed OA progressed over 10-week time-course. Gait disparity occurred only at 10-week post-surgery. Osteophyte formed at 2-week post-surgery. BMDs of DMM showed no statistical differences comparing to SHAM at 2 weeks, but BV/TV is much higher in DMM mice. Increased BMD was clearly found at 5- and 10-week post-surgery in DMM mice. TRAP staining showed increased osteoclast activity at the site of osteophyte formation of DMM joints at 5- and 10-week time points. These results showed that subchondral bone turnover might occurred earlier than 2 weeks in this mouse DMM model. Gait disparity only occurred at later stage of OA in DMM mice. Notably, patella dislocation could occur in some of the DMM mice and cause a different pattern of OA in affected knee.

The Effect of Clinical Pilates on Functional Movement in Recreational Runners.

PubMed

Laws, Anna; Williams, Sean; Wilson, Cassie

2017-09-01

Biomechanical imbalances and inefficient functional movements are considered contributing factors to running-related injuries. Clinical Pilates uses a series of exercises focused on retraining normal movement patterns. This study investigated whether a 6-week course of Clinical Pilates improves functional movement and thereby, potentially, reduces the risk of running-related injuries associated with movement dysfunction. A modified functional movement screen was used to analyze the functional movement ability of forty runners. Forty participants completed a 6-week course of Clinical Pilates delivered by a Clinical Pilates instructor. The movement screen was carried out 3 times for each runner: 6 weeks pre-intervention (baseline), within one week pre-intervention (pre) and within one week post-intervention (post). Repeated-measures analysis of variance and post-hoc tests found significant increases in scores between baseline and post (mean±SD; 13.4±2.4 vs. 17.0±1.7, p<0.01) and pre and post (mean±SD; 13.5±2.5 vs. 17.0±1.7, p<0.01), but no significant difference between baseline and pre (p=0.3). A 6-week course of Clinical Pilates significantly improves functional movement in recreational runners, and this may lead to a reduction in the risk of running-related injuries. © Georg Thieme Verlag KG Stuttgart · New York.