A Comparative Evaluation of Four Different Immunoassays in the Diagnosis of Cystic Echinococcosis Using a Crude and Purified Hydatid Cyst Fluid Antigen.

PURPOSE: Cystic echinococcosis (CE) is one of the most neglected tropical diseases as per WHO which has an immense public health significance. Diagnosis of CE is difficult as specific clinical signs are manifested only after the hydatid cyst attains a considerable size. Immunodiagnosis is a reliable method of diagnosing CE. METHODS: SDS-PAGE was performed for the hydatid cyst fluid antigens. The antigen purity was tested by Western blotting and four different immunoassays were evaluated using these two antigens in sheep and buffalo in diagnosis of CE. RESULTS: SDS-PAGE revealed four bands of 72, 64, 48 and 24 kDa for crude antigen and a single 72 kDa band for purified antigen. Among sheep sera, ELISA was most sensitive (70%) using crude antigen and also while using the purified antigen (80%). In case of buffaloes, ELISA, DID and CIEP were more sensitive (83.3%) using crude antigen, whereas DID and CIEP were more sensitive (83.3%) using purified antigen. CONCLUSION: In sheep, while using the crude antigen ELISA was the most sensitive assay and IHA was the least sensitive assay. While using the purified antigen also, ELISA was the most sensitive and others were absolutely specific except for IHA being less sensitive. In buffaloes, using crude antigen, all the immunoassays CIEP, DID and ELISA were highly sensitive in diagnosing CE infection except IHA, whereas using the purified antigen, both CIEP and DID were more sensitive than ELISA and IHA which were comparatively less sensitive in detecting CE in buffalo sera.

Genetic diversity, virulence and distribution of antimicrobial resistance among Listeria monocytogenes isolated from milk, beef, and bovine farm environment.

BACKGROUND: Listeria monocytogenes is an opportunistic intracellular foodborne pathogen and is ubiquitous in nature. The occurrence of L. monocytogenes in animal production units coupled with their presence in milk, faeces, feed, water, sewage, and soil is a contributory factor for foodborne listeriosis in humans and animals. AIMS: The study was aimed to characterize genotype and serogroup of L. monocytogenes recovered from different types of samples and also to study antimicrobial patterns by phenotypic and genotypic methods. METHODS: Multiplex polymerase chain reaction (PCR) was used for the confirmation of L. monocytogenes, the identification of its serogroup and lineage, and the detection of virulence markers. Enterobacterial repetitive intergenic consensus (ERIC), and randomly amplified polymorphic DNA (RAPD)-PCR were used to characterize those isolates, and antimicrobial patterns were studied phenotypically by Kirby-Bauer method and genotypically by PCR. RESULTS: Out of the screened 474 samples (274 milk and 50 each of soil, feed, sewage, and beef), ten L. monocytogenes isolates (milk=8, soil=1, and beef=1) were confirmed by PCR targeting the hlyA gene and found to belong to the 1/2a, 3a serogroup and fall under type II lineage. Virulence potential assessment revealed that all the ten isolates harbored the iap gene while the presence of plcA and plcB genes were noticed in seven and eight isolates respectively. Six isolates from milk were found to group in the same cluster by ERIC and RAPD fingerprinting, suggesting both methods to be efficient molecular typing tools for L. monocytogenes. Genotypic characterization of antimicrobial resistance (AMR) genes revealed that seven isolates were positive for tetM, five for mefA, four for msrA, and one for lnuA genes while none of the isolates showed tetK, ermA, ermB, and lnuB genes. CONCLUSION: The presence of L. monocytogenes in bovine farm environments coupled with virulence markers, and multidrug resistance from the study area suggest a possible transmission from the environment to humans and animals which needs to be monitored regularly to ensure food safety and the well-being of animals and humans.