ISOLATION AND MOLECULAR CHARACTERIZATION OF Campylobacter spp. FROM CHICKEN
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Date
2021
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Abstract
Campylobacter spp. are the zoonotic bacteria which are the most common cause of food borne gastroenteritis around the world. The link between human campylobacteriosis and poultry has been well established. This study was aimed to isolate Campylobacter spp. from chicken and characterize them with molecular methods like multiplex PCR, Loop Mediated Isothermal Amplification assay (LAMP) and Multilocus Sequence Typing (MLST). A total of 191 chicken caecal mucosal scraping were collected from Chennai, Namakkal, Krishnagiri, Erode and Coimbatore districts of Tamil Nadu. Bacterial isolation was done by plating on blood free Campylobacter broth added with 3% agar agar, CCDA supplement and Campylobacter growth supplement IV. Identification of Campylobacter species was done by multiplex PCR and they were also subjected to LAMP assay. The prevalence of Campylobacter spp., C. coli and C. jejuni was found to be 28% (54/191), 7% (14/191), 0.5% (1/191), respectively. The minimum inhibitory concentration (MIC) values for eight antimicrobials – azithromycin, chloramphenicol, ciprofloxacin, clindamycin, erythromycin, gentamicin, nalidixic acid and tetracycline were obtained by microdilution resazurin assay. Fourteen C. coli isolates showed 100% resistance to nalidixic acid and higher resistance to tetracycline (92.8%), erythromycin (71.4%), clindamycin (71.4%) and azithromycin (64.2%). All C. coli isolates were sensitive to chloramphenicol and higher sensitivity to ciprofloxacin (78.5%) and gentamicin (71.4%) was observed. One C. jejuni isolate was resistant to azithromycin, tetracycline, nalidixic acid and sensitive to all other antimicrobials used. Multilocus sequence typing was carried out for five representative C. coli isolates. Three C. coli isolates had the existing sequence types ST-899, ST- 872 and ST-9108. Two novel sequence types were found in this study and they were ST-10872 and ST-11031. However, all the STs belonged to the same clonal complex of ST-828. This is the first report with characterization of C. coli isolates by MLST in India