Burden assessment and characterization of thermophilic Campylobacter in broiler chickens
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Date
2019-07
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G.B. Pant University of Agriculture and Technology, Pantnagar - 263145 (Uttarakhand)
Abstract
Campylobacter is considered to be a major pathogen in terms of food safety concern since it causes gastrointestinal problems worldwide. As these organisms have an intricate epidemiological cycle and the sources of contamination are varied in nature, necessary steps should be taken for controlling the organism below the level of risk for humans. The present research was organized in such a manner to screen and detect thermophilic Campylobacters from a total of seven farms of poultry (Haldwani, Pantnagar, Lalkuan, Kiccha, Jawaharnagar, Shanthipuri farm 1, and Shanthipuri farm 2) and certain retail meat shops (pantnagar) located in the state of Uttarakhand. Totally 645 samples which includes 15 poultry meat samples, 446 poultry fecal samples and 184 environmental samples were processed according to their standard protocol for accomplishing the objectives of the study undertaken. Bacteria showing characteristic morphological and staining characteristics were confirmed biochemically. 16S rRNA gene (816 bp) was targeted for genus identification while lpxA gene was targeted for species differentiation of C. jejuni (331 bp) and C. coli (391 bp). Through simplex PCR 73 isolates were confirmed to be belonging to Campylobacter genus with an overall prevalence of 11.32%. Maximum recovery was found at lalkuan farm (33.77%) followed by Haldwani farm (16.13%), Pantnagar farm (11.93%), Shanthipuri farm1 (8.79%), Shanthipuri farm2 (6.76%), Pantnagar meat shops (6.67%) while least prevalence in Jawaharnagar farm (5.1%). No Campylobacter isolates were recovered from Kiccha. Prevalence of C. jejuni (52.05%) species was observed to be more than C. Coli (47.95%). All the 73 isolates were screened for the presence of ten virulence genes among which cadF, flaA and dnaJ showed 100% prevalence followed by cdtB(87.67%), cgtB(45.68%), pldA(31.51%) and wlaN(16.44%). None of the isoates were harbouring ciaB, iam, virB. Only 40 isolates sustained out of 73 isolates which were further processed for Kirby Bauer disc diffusion assay in which antibiotic sensitivity testing of Cefoxitin had highest resistance as 95% followed by Ciprofloxacin 80%, Nalidixicacid25%, Cefatoxime-22.5%, Ampicillin-20%, Tetracycline-12.5%, Clindamycin-7.5%, Erythromycin-5%. The antibiotics Gatifloxacin, Streptomycin were susceptible to all the isolates screened but some intermediate resistance was found in Sulfisaxazole and Levofloxacin. The isolates which developed resistance phenotypically were checked for their respective Antimicrobial Resistance Genes (ARGs). β-lactam resistance gene blaOXA-61 was prevalent in 32 isolates (86.49%), cmeB was found in 24 isolates (72.73%) , tet (O) was detected in 3 isolates (60%), ermB was detected in 1 isolate (50%). The resistance towards cephalosporin and fluoroquinolone group of antibiotics is increasing representing the over usage of antibiotics in poultry farms which should be taken care off and necessary measures to be undertaken to reduce its usage to avoid AMR among humans.
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