Disseminated mycosis in layer cockerels and pullets

Eight-wk-old layer cockerels and pullets were presented to the diagnostic lab with a history of increased mortality, ruffled feathers, lameness, and recent vaccination. At necropsy, the birds had large multifocal granulomas in multiple tissues. Only light bacterial growth was seen on culture. On histopathology, a mixed population of fungi was seen within the granulomas including zygomycetes and Aspergillus, with the zygomycetes being the predominant organism. Because of the coinfection with Aspergillus and Penicillium, obtaining the zygomycetes in pure culture was unsuccessful. The source of the zygomycete fungi remains unknown; however, zygomycetes are known to be ubiquitous. Serology was performed to evaluate the flock's immune status. There was no evidence of immunosuppression caused by chicken anemia virus or bursal disease infections. No flock treatment was initiated.     

Effects of dietary n-6 and n-3 fatty acids and vitamin E on the immune response of healthy geriatric dogs

OBJECTIVE: To determine the effect of dietary n-6 to n-3 fatty acid ratios and alpha-tocopheryl acetate concentration on immune functions andT cell subpopulations in healthy dogs. ANIMALS: Thirty-two 7- to 10-year old female Beagles. PROCEDURE: For 17 weeks, dogs were fed food that contained low (1.4:1) or high (40:1) ratios of n-6 to n-3 fatty acids in combination with 3 concentrations of all rac-alpha-tocopheryl acetate (low, 17 mg/kg of food; medium, 101 mg/kg; high, 447 mg/kg). Dogs were inoculated twice with a keyhole limpet hemocyanin suspension at 13 and 15 weeks. RESULTS: After 12 weeks, dogs consuming low concentrations of alpha-tocopheryl acetate had lower percentages of CD8+ T cells, compared with dogs consuming medium or high alpha-tocopheryl acetate concentrations. Also, dogs consuming low alpha-tocopheryl acetate concentrations had higher CD4+ to CD8+ T cell ratios. On day 4 of week 15, the percentage of CD8+ T cells was highest in dogs fed medium concentrations of alpha-tocopheryl acetate, compared with other dogs; however, the CD4+ to CD8+ T cell ratio was higher only in dogs fed low concentrations of alpha-tocopheryl acetate with high concentrations of n-3 fatty acids. Dogs consuming low concentrations of n-3 fatty acids with medium concentrations of alpha-tocopheryl acetate had the largest delayed-type hypersensitivity (DTH) skin test response. CONCLUSIONS AND CLINICAL RELEVANCE: An optimum amount of dietary alpha-tocopheryl acetate concentration, regardless of the dietary n-6 to n-3 fatty acid ratio, stimulates the CD8+ T cell population. Effects of an optimum amount of dietary alpha-tocopheryl acetate concentration on the DTH response are blunted by dietary n-3 fatty acids.     

Expressed sequence tags from the phytopathogenic fungus Botrytis cinerea

A large set of genes was identified in the phytopathogenic fungus Botrytis cinerea by using an expressed sequence tag approach. The fungus was grown in axenic culture and a cDNA library was produced. From this library, 6559 ESTs were obtained. The combined sequences represent 3026 unisequences that corresponds to approximately one-quarter of the estimated total number of genes in B. cinerea. Approximately 18% of the ESTs showed significant similarities with genes coding for proteins with known functions,~56% were similar to genes coding for proteins with unknown functions and ~26% were orphans. A substantial B. cinerea gene inventory including putative virulence factors was therefore obtained and is now available at the Génoplante-Info Database interface (http://urgi.infobiogen.fr///Projects/GPiDB/Interface/).     

Long-term monitoring of classical swine fever in wild boar (Sus scrofa sp.) using serological data

In the European Community, epizootics of classical swine fever (CSF) in the wild boar (Sus scrofa) are compulsorily monitored because transmission may occur between wild boars and domestic pigs, causing heavy economic losses to the pork industry. The estimation of incidence in populations of wild boars is generally based on viroprevalence. However, viral isolation becomes rare when the incidence is low because the virus cannot be detected for more than a few weeks following infection. On the contrary, seroprevalence is detectable at low incidence levels, because antibodies can be detected for the lifetime of the infected animal. We thus attempted to analyse the long-term evolution of CSF incidence using serological data. The data came from France, where CSF had been monitored from 1992 to 2002, and where the virus has not been detected since 1997. We assumed that the overall seroprevalence would estimate the proportion of immune wild boars, that seroprevalence in juveniles would approximate incidence and that seroprevalence in different age classes would show the evolution of incidence in a given cohort. Spatial and temporal trends of incidence and seroprevalence were explored using logistic modelling and the spatial trend was analysed using polynomial regression. In 1992, incidence peaked in the northern area. After 1993, incidence decreased but remained the highest in the northern area. After 2000, no seropositive juvenile was observed, suggesting the extinction of the epizootic. Our results support the reliability of serological monitoring since it allowed a longer detection of viral transmission and provided more information on the spatio-temporal evolution of incidence than did viral isolation. We advocate that the highest persistence of infection in northeastern France is not independent from infection persistence in Reinland-Pfalz (Germany). Such persistence may be due to favourable local conditions and/or the social organisation of wild boars.     

Avirulence of viable but non-culturable Listeria monocytogenes cells demonstrated by in vitro and in vivo models

The virulence of Viable But Non-Culturable (VBNC) cells of 4 strains of Listeria monocytogenes was investigated in both a human adenocarcinoma cell line (HT-29) and a mouse model. LO 28, ATCC 19115 and CNL 895807 strains of Listeria monocytogenes became VBNC when incubated in microcosm water at 20 degrees C and Scott A strain at 4 degrees C. No culturable bacteria were detected in the VBNC state, although 104 active cells/mL were found by the Direct Viable Count (DVC) and CTC-DAPI double staining methods. A comparison of virulence in both human adenocarcinoma cell line HT-29 and the mouse model showed that culturable controls were more virulent than VBNC cells, which appeared to be avirulent regardless of the virulence methods applied. Pathogenicity was tested in each model and was lost concomitantly with culturability, whereas some cells were still metabolically active (determined by CTC and DVC). Moreover, amplification of a 388 bp fragment with Immunocapture-PCR revealed the presence of Listeria monocytogenes DNA in all mixed spleen samples after intravenous injection of VBNC cells. These results demonstrate that VBNC cells were present in the mouse spleens. The results of the study suggest that Listeria monocytogenes strains might remain in the aquatic environment for prolonged periods in the VBNC state but these cells were not pathogenic in the conditions tested. These findings demonstrate the value of VBNC studies and show the need to investigate the role of VBNC cells in environmental transmission of Listeria monocytogenes. Further studies are needed in order to investigate the virulence of VBNC cells of Listeria monocytogenes after recovery of a culturable state.     

Characteristics of Salmonella enteritidis contamination in eggs after oral,aerosol, and intravenous inoculation of laying hens

Experimental infection models are useful tools for understanding how Salmonella enteritidis is deposited in eggs and for testing potential strategies to control eggborne transmission of disease to humans. Oral inoculation of laying hens is presumed to provide the closest simulation of naturally occurring infections, but alternatives such as intravenous or aerosol inoculation have sometimes been recommended as options to induce higher incidences of egg contamination. The present study compared the frequency, level, and location of S. enteritidis deposition in egg contents after experimental inoculation by three different routes. In two replicate trials, specific-pathogen-free laying hens were infected with an S. enteritidis culture mixture prepared to optimize invasive behavior. Groups of hens received either an oral dose of 10(9) S. enteritidis, an aerosol dose of 10(9) S. enteritidis, or an intravenous dose of 10(5)-10(7) S. enteritidis. Oral inoculation led to the highest incidence of fecal shedding of S. enteritidis, whereas intravenous inoculation produced the highest specific antibody titers. Eggs laid during the first 21 days postinoculation were cultured to detect and enumerate S. enteritidis in the yolk and albumen. No significant differences were observed among the three inoculation routes in the frequencies of isolation of S. enteritidis from either yolk or albumen. For all three routes of administration, S. enteritidis was recovered more often from yolk (at frequencies ranging from 4% to 7%) than from albumen (0 to 2%). Over 73% of contaminated eggs harbored fewer than 1 colony-forming unit (CFU) of S. enteritidis per milliliter, and only 3% of such eggs contained more than 100 CFUs/ml. Significantly higher levels of S. enteritidis contaminants were associated with intravenous inoculation than with the other routes. No advantage of using aerosol or intravenous administration of S. enteritidis as an alternative to oral inoculation for inducing the production of contaminated eggs was evident in this study.     

Comparison of two formaldehyde administration methods of in ovo-injected eggs

Formaldehyde administration in the hatchery can be very useful in decreasing microbial numbers. However, its use is controversial because of the adverse effects that can occur to chicks and people. This study was designed to look at alternative methods of application of formaldehyde in the hatchery. In addition, the study compared the effects of these methods of application on in ovo-and non-in ovo-injected eggs. All in ovo-injected eggs were given diluent only with no vaccine or antibiotic added. In hatchers containing both in ovo-injected eggs and non-in ovo-injected eggs, formaldehyde was administered two ways, dose (DOSE) and constant rate infusion (CRI). In the DOSE hatcher, 12 ml of formaldehyde was administered at one time every 12 hr, whereas in the CRI hatcher, the same volume was administered at a rate of 1 ml/hr over a 12-hr period. A control (CONT) hatcher received 12 ml of distilled water at the same time that the DOSE hatcher was given formaldehyde. In the DOSE hatcher, a peak concentration of formaldehyde of 102 ppm was reached. The CRI was maintained at approximately 20 ppm of formaldehyde. At pipping, the aerosol bacterial load in the hatchers receiving formaldehyde (DOSE, 130 colony-forming units [CFU]/m3; CRI, 82.5 CFU/m3) was significantly less than in the CONT hatcher (235 CFU/m3). At hatch, the CRI (337.5 CFU/m3) was not able to control bacterial levels and only the DOSE hatcher (150 CFU/m3) had a significantly lower aerosol bacterial count. The CRI non-in ovo-injected eggs (93.39%) had a significantly higher percentage of hatch of fertile compared with non-in ovo-injected eggs exposed to water (84.27%). In ovo-injected eggs in CONT and DOSE treatment groups contained significantly higher percentages of visual contamination than non-in on-injected eggs in the same hatchers. This difference had numerical significance only in the treatment groups within the CRI hatcher. The chicks were then placed into replicate treatment groups and grown for 14 days. Chicks from the CRI in ovo-injected eggs had a statistically significant improvement in feed conversion ratio (1.24) at 14 days when compared with chicks from CONT non-in ovo-injected eggs (1.29). All formaldehyde-exposed chicks had numerically lower feed conversion ratios compared with the CONT exposed chicks.