Development of novel isothermal-based DNA amplification assay for detection of pig tissues in adulterated meat.

For the first time, we describe an innovative polymerase spiral reaction (PSR) assay for the rapid, simple, and accurate detection of pig tissues or pork in adulterated meat including heat-treated and processed ones. The PSR assay specifically targeting the mitochondrial cytochrome b (cyt-b) gene of the pig was successfully optimized permitting assay results in 65 min time. The developed detection method was 100% specific amplifying only the cyt-b gene and displaying negative results with all the tested non-pork meats. The sensitivity of the developed PSR (760 fg porcine DNA) was tenfold better than the end-point PCR and able to detect heat-treated (121 °C) and adulterated (0.5% pork in beef) meat and processed pork products such as sausages, salami, meatball, soup, curry, etc. The developed PSR-based method can be used for point-of-care detection with minimum instrumentation and technical expertise to guarantee instant clearance of exported and imported meat products. This is the first time that PSR has been adapted for food authenticity purposes.

Exploring the potential hazard of Mycobacterium avium subspecies paratuberculosis as a cause for Crohn's disease.

BACKGROUND: Mycobacterium avium subspecies paratuberculosis (MAP) is the causative agent of Johne's disease (JD), or paratuberculosis in ruminants has been suspected to be implicated in the pathogenesis of Crohn's disease (CD) in humans with chronic inflammatory intestinal changes. As the hypothesis is now fast being recognized that MAP could possibly be the etiological agent of CD which is found to be excreted in milk of dairy animals subclinically or terminally ill with JD. AIM: The present study was aimed to detect MAP in milk by polymerase chain reaction (PCR) targeting IS900 and to describe the excretion pattern of MAP in milk from asymptomatic lactating cows and does with relevance to the public health significance. MATERIALS AND METHODS: A total of 77 milk samples were collected randomly from lactating animals which include cows (45) and does (32). All the 77 milk samples were processed to identify the presence of MAP by employing the direct IS900 PCR as per the standard protocol. RESULTS: Out of 77 milk samples from asymptomatic lactating animals, 12 (15.58%) were showed positivity for IS900 PCR in which 5 (11.11%) were from lactating cows and 7 (21.87%) were from lactating does. CONCLUSION: In our study, 15.58% of milk samples showed IS900 positivity which indicates the presence of subclinical MAP infection in lactating animals. Hence, there is a possibility for excretion of MAP through milk which can be a potential threat for CD in humans by raw milk consumption. Therefore, the prevention of MAP in the food chain need to be assured by sourcing raw products from animal herds free of MAP infection.

Evaluation of recombinant outer membrane protein based vaccine against Salmonella Typhimurium in birds.

Food-borne diseases caused by Salmonella enterica from poultry sources represent an important public health problem and no reliable control by vaccination has proved effective despite research. The aim of the present study was to evaluate the use of recombinant OmpC protein for immunization of birds to elucidate its protection against virulent Salmonella Typhimurium. The recombinant OmpC protein was prepared after cloning and expressing ompC gene and was characterized by SDS-PAGE and Western blot analyses. The protein preparations were tested as vaccine candidate in layer birds by comparing the immune response, protection and organ clearance against crude lysate and control. The biologically functional recombinant 43 kDa truncated OmpC protein proved to be a good immunogen which induced a significantly high humoral immune response than control. At the same time, it primed a stable cell-mediated immune response. A protective index (based on faecal shedding of organism) of rOmpC based preparations ranged between 50 and 75% as observed for 3 weeks after challenge. Therefore, the protein preparations conferred satisfactory protection against challenge infections with virulent strains of S. Typhimurium as evidenced by limited faecal shedding and minimal detection of Salmonella from edible tissues and eggs. These findings suggest the possibility to explore the use of S. enterica OMP protein for the production of novel vaccine.