Upadhyay, A.K.Bisht, Piyush2019-08-032019-08-032019-07http://krishikosh.egranth.ac.in/handle/1/5810118892Campylobacters are Gram negative helical bacilli belonging to the family Campylobacteraceae and are of major public health significance as they are one of the leading cause of food borne gastroenteritis worldwide. The present study was carried out to find a novel technique for isolation of Campylobacter spp. and and determine the prevalence of thermophilic Campylobacter spp. in poultry and their living environment at different poultry farms and meat vendors located in Kumaon region of Uttarakhand state. Most of the Campylobacter culture media described in the literature are supplemented with sheep or horse blood, therefore, this study examined the use of goat blood as potential substitutes for sheep blood in blood-supplemented bacteriologic media commonly used in clinical microbiology laboratories. A total of 381 samples comprising of 156 poultry caeca, 86 poultry faeces, 25 goat faeces, 10 sheep faeces, 24 meat swabs and 80 environmental samples viz.; water (n=32) and litter (n=48) were collected from three (n=3) poultry farms and meat shops (n=4) were analyzed, of which 49 samples showed characteristic colonies, either having a spreading or watery nature. They were typical Gram negative spiral rods and had characteristic cork screw motility. All the 49 Campylobacter isolates were confirmed using biochemical and molecular assays. In latex agglutination test, all the isolates produced characteristic agglutination. Genus-specific PCR amplification of 16SrRNA gene yielded expected product of 816 bp in all the isolates. In multiplex PCR assay conducted targeting lpxA gene for the identification of C. jejuni (331 bp) and C. coli (391bp) was used. According to research, the mean of the viable count of bacteria obtained from media supplemented with Goat blood was 0.921x108 c.f.u./ml and sheep blood was 0.936x108 c.f.u./ml. The Chi square analysis (ϗ2) was performed which proves the result to be non-significant showing similarities between the results obtained by both the media. Hence, the data indicates that goat blood can also be used as alternative and all the studies were carried on goat blood. The prevalence rate of thermophilic campylobacters sample was found to be 12.79% (11/86) in poultry faeces, 21.15% (33/156) in poultry caeca, 12.5% (3/24) in meat swabs and 4.16% (2/48) in litter from poultry farms. No thermophilic Campylobacter was isolated from goat faeces, sheep faeces and water samples from the poultry farms. Farmwise, the highest prevalence in poultry farms was detected in Anandpur (11.7%) followed by Haldwani (7.5%), and Pantnagar (5.3%). Among the chicken meat shops, highest prevalence was detected in Pantnagar (24%) followed by Haldwani (23.3%), Lalkuan (15%) and Nainital (6.6%). Out of the total of 49 thermophilic Campylobacter isolated, comprising of 36 C. jejuni (67.34%) and 13 C. coli (32.65%). As poultry serve as important source of Campylobacter, awareness among the poultry farmers and chicken meat retailers should be created to prevent the further transmission of this zoonotic agent.ennullThesis