Assuntos
Endometrite/veterinária , Infecções por Bactérias Gram-Negativas/veterinária , Doenças dos Cavalos/microbiologia , Taylorella equigenitalis/isolamento & purificação , Animais , Cruzamento , Endometrite/diagnóstico , Endometrite/epidemiologia , Feminino , França/epidemiologia , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/epidemiologia , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/epidemiologia , Cavalos , Masculino , Estudos RetrospectivosRESUMO
The objective of this study was to examine the degree of phenotypic and genotypic diversity between 43 French Taylorella asinigenitalis strains isolated from 22 jacks, two stallions and one mare between 1995 and 2008 by culturing genital swabs obtained during routine diagnosis for contagious equine metritis. This retrospective analysis revealed the existence of T. asinigenitalis species since 1995 and the natural colonization of a mare's genital tract in 2001. Despite the presence of 27 different patterns revealed by the combination of API ZYM, antibiogram and 16S rDNA profiles, we show that T. asinigenitalis is a highly homogeneous species. API ZYM diversity only concerns acid phosphatase and naphthol-AS-BI-phosphohydrolase activity. The majority of strains are susceptible to a wide range of antimicrobial agents but most are streptomycin-resistant (95.5%), ampicillin-resistant (88.4%), and four strains are atypical due to a high degree of resistance to at least eight antimicrobial agents. 16S rDNA sequence analysis showed only two clusters and revealed similarity of 99.3-100% between T. asinigenitalis strains. The geographic origin of the 43 isolates correlates to the two 16S rDNA clusters.
Assuntos
Variação Genética , Infecções por Bactérias Gram-Negativas/veterinária , Cavalos/microbiologia , Taylorella/genética , Animais , DNA Bacteriano/genética , DNA Ribossômico/genética , Farmacorresistência Bacteriana , Feminino , Infecções por Bactérias Gram-Negativas/diagnóstico , Masculino , Testes de Sensibilidade Microbiana , Fenótipo , RNA Ribossômico 16S/genética , Estudos Retrospectivos , Análise de Sequência de DNA , Taylorella/classificação , Taylorella/isolamento & purificaçãoRESUMO
Contagious equine metritis is a horse disease that causes endometrial inflammation due to Taylorella equigenitalis. Since Taylorella asinigenitalis was characterized, genital swab culture has proved to be an insufficient method for distinguishing between the two Taylorella species. Here, we developed an indirect immunofluorescence (IIF) test using polyclonal antibodies. Specificity, sensitivity, and detection limit were assessed using isolated bacteria (55 T. equigenitalis strains, 46 T. asinigenitalis strains and 18 other bacterial species), experimental and genital swabs in comparison to bacterial culture and polymerase chain reaction (PCR) testing. Our results indicated that IIF using polyclonal antibodies allows T. equigenitalis to be discriminated from T. asinigenitalis. This test constitutes a rapid, sensitive and specific tool for confirming presumptive colonies of T. equigenitalis.
Assuntos
Infecções por Bactérias Gram-Negativas/veterinária , Doenças dos Cavalos/microbiologia , Taylorella equigenitalis/isolamento & purificação , Animais , Anticorpos Antibacterianos/análise , Antígenos de Bactérias/análise , Técnica Indireta de Fluorescência para Anticorpo , Infecções por Bactérias Gram-Negativas/diagnóstico , Doenças dos Cavalos/diagnóstico , Cavalos , Coelhos/imunologia , Taylorella equigenitalis/imunologiaRESUMO
Equine herpesviruses type 1 and 4 (EHV-1 and EHV-4) are ubiquitous in the equine population. One of their main properties is their ability to establish life-long latent infections in their hosts even in those with natural or vaccine-induced immunity. However, effect of vaccination status on prevalence and tissue tropism was not established. In this study, EHV-1 and EHV-4 were detected by polymerase chain reaction and by classical virus isolation from neural, epithelial and lymphoid tissues collected from unvaccinated (33) or vaccinated (23) horses. The percentage of EHV-1- and EHV-4-positive horses between vaccinates and unvaccinates was similar. Both viruses were detected in all tissues of both groups; in particular, lymph nodes draining the respiratory tract, nasal epithelium and nervous ganglia [i.e. trigeminal ganglia (TG)], which represent the main positive sites for EHV-1 and EHV-4. In vaccinated animals, the nervous ganglia (i.e. TG) were less frequently positive than in unvaccinated animals. Detection of positive TG was strongly correlated to the presence of EHV-1 in nasal epithelium.
Assuntos
Infecções por Herpesviridae/veterinária , Herpesvirus Equídeo 1/imunologia , Herpesvirus Equídeo 4/imunologia , Doenças dos Cavalos/epidemiologia , Doenças dos Cavalos/prevenção & controle , Vacinas Virais , Animais , Autopsia , Primers do DNA , DNA Viral/genética , França/epidemiologia , Infecções por Herpesviridae/epidemiologia , Infecções por Herpesviridae/prevenção & controle , Herpesvirus Equídeo 1/genética , Herpesvirus Equídeo 4/genética , Doenças dos Cavalos/virologia , Cavalos , Reação em Cadeia da Polimerase/veterináriaRESUMO
The specific detection and enumeration of Lactobacillus brevis LB62, Carnobacterium divergens V14 and Carnobacterium piscicola VI were studied by in situ hybridization-flow cytometry. The method was performed on the exponential growth phase with three probes targeting 16S rRNA labelled with fluorescein isothicyanate (FITC): EUB338 probe universal for Eubacteria, Lb probe specific for Lact. brevis and Cb probe specific for the genus Carnobacterium. EUB338 was used to determine the permeabilization and hybridization conditions for the cells. The Lb probe gave no hybridization signal whereas the Cb probe allowed the detection and quantification by flow cytometry at 520 nm of the two Carnobacterium strains in pure culture or in mixtures with Listeria innocua F.