Clostridioides difficile in food and food products of animal origin in Assam, India.

OBJECTIVE: Considering the paucity of information about food-associated Clostridioides difficile from India, a study was undertaken to establish the prevalence of C. difficile in a variety of foods of animal origin, together with molecular strain characterization and antimicrobial resistance. METHODS: A total of 235 samples comprising raw meat and meat products, fish products, and milk and milk products were screened for C. difficile. Toxin genes and other parts of PaLoc were amplified in isolated strains. The resistance pattern towards commonly used antimicrobial agents was studied by the Epsilometric test. RESULTS: C. difficile was isolated from 17(7.23%) different food samples of animal origin, including toxigenic (6) and non-toxigenic (11) isolates. In four toxigenic strains, the tcdA gene could not be detected under used conditions (tcdA-tcdB+). However, all strains had binary toxin-associated genes (cdtA and cdtB). The antimicrobial resistance was highest in non-toxigenic C. difficile isolates in food of animal origin. CONCLUSION: Meat, meat products and dry fish, but not milk and milk products were contaminated with C. difficile. Contamination rates were low with diverse toxin profiles and antibiotic resistance patterns among the C. difficile strains.

Evaluation of different antigenic preparations against necrotic enteritis in broiler birds using a novel Clostridium perfringens type G strain.

OBJECTIVE: Keeping in view, the constraints faced by the Indian broiler industry with lack of a suitable vaccine against Necrotic Enteritis (NE), a study has been proposed to explore the prevalence and detail characterization of C. perfringens type G in NE suspected broiler chicken in the process of suitable vaccine development. METHODS: Intestinal scrapings/faecal contents of NE suspected broiler chickens were screened to establish the prevalence of C.perfringens type G in broiler birds. A most pathogenic, highly resistant type G isolate of C. perfringens, bearing both tpeL and gapC gene was selected for preparation of three different vaccine formulations, and to evaluate their immunogenic potential in broiler birds. RESULTS: Screening of clinical samples of NE suspected broiler birds revealed C. perfringens type G, bearing gapC gene in 51.22% samples, of which 47.62% revealed tpeL gene. Seven of the tpeLpos type G isolates were comparatively more pathogenic for mice, of which, one exhibited multidrug resistance towards ciprofloxacin, norfloxacin, tetracycline and levofloxacin. The sonicated supernatant (SS) prepared from the selected tpeL and gapC positive isolate could maintain a significantly higher protective IgG response than toxoid and bacterin preparation from the 21st to 28thday of age in immunized birds. CONCLUSION: The additional TpeL toxin in C. perfringens type G has been proved to be an additional key biological factor in the pathogenesis of NE in broiler chickens. Considering the release of more immunogenic proteins, the SS proved to be a better immunogenic preparation against NE with a multiple immunization dose.

Pathodynamics of Circulating Strains of Duck Enteritis Virus: A Step Forward to Understand Its Pathogenesis.

Duck enteritis virus (DEV) causes an acute and contagious infection in duck. The present study was carried out to evaluate the pathogenicity and pathodynamics of DEV isolates from different natural outbreaks in the Assam Province of India. A total of six wild-type isolates of DEV were revived in ducklings to determine its biologic characterization. Postmortem examination of infected ducklings revealed DEV-specific gross lesions in different organs. The presence of DEV was confirmed by its genome amplification and the presence of viral antigens from collected tissue samples by indirect fluorescent antibody test. All the isolates revived in ducklings were further propagated in duck embryo fibroblast cells. Highly virulent and low virulent isolates of DEV were selected for further study based on median duck infectivity dose (DID50) and median tissue culture infectivity dose (TCID50). The highly virulent isolate of DEV had values of 102 DID50/ml and 106.33 TCID50/ml, whereas the low virulent strain had titers of 10 DID50/ml and 104.83 TCID50/ml in the cell culture. Our results showed replication of DEV in ducks with the highest and lowest viral titers in the thymus and bursa of Fabricius, respectively. In addition, microscopic analysis revealed necrosis and degeneration of submucosal esophageal glands and glandular epithelium. The study will be useful to understand the organ tropism and pathologic alteration among the virulent DEV isolates.

Patodinámica de las cepas circulantes del virus de la enteritis del pato: Un paso adelante para comprender su patogenia. El virus de la enteritis del pato (DEV) causa una infección aguda y contagiosa en el pato. El presente estudio se llevó a cabo para evaluar la patogenicidad y la patodinámica de los aislamientos del virus de la enteritis del pato de diferentes brotes naturales en la provincia de Assam en la India. Se replicaron un total de seis aislamientos del virus de la enteritis del pato de tipo silvestre en patitos para su caracterización biológica. El examen post mortem de los patitos infectados reveló lesiones macroscópicas específicas de la enteritis viral en diferentes órganos. La presencia del virus de la enteritis viral de pato fue confirmada por su amplificación del genoma y por la presencia de antígenos virales mediante la prueba indirecta de anticuerpos fluorescentes con muestras de tejido recolectadas. Todos los aislamientos replicados en patitos se propagaron adicionalmente en células de fibroblastos de embriones de pato. Se seleccionaron aislamientos del virus de la enteritis del pato altamente virulentos y poco virulentos para un estudio adicional basado en la dosis de infectividad en el pato (DID50) y la dosis de infectividad de cultivo de tejidos (TCID50). El aislado altamente virulento del virus de la enteritis del pato mostró valores de 102 DID50/ml y 106.33 TCID50/ml, mientras que la cepa virulenta baja tenía títulos de 10 DID50/ml y 104.83 TCID50/ml en cultivo celular. Nuestros resultados mostraron la replicación del virus de la enteritis viral en patos con los títulos virales más altos y más bajos en el timo y en la bolsa de Fabricio, respectivamente. Además, el análisis microscópico reveló necrosis y degeneración de las glándulas esofágicas submucosas y del epitelio glandular. El estudio será útil para comprender el tropismo de los órganos y la alteración patológica entre los aislados virulentos del virus de la enteritis viral del pato.