Moleculer typing of Campylobacter jejuni and Campylobacter coli isolated from cattle and sheep

The objectives of this study were to investigate the presence ofCampylobacter jejuni and Campylobacter coli, which are recognized as the majoragents of bacterial gastroenteritis in humans, from internal organs and faecalsamples of clinically healthy cattle and sheep by means of a multiplex PolymeraseChain Reaction (multiplex PCR) and to determine the degree of geneticheterogeneity among these isolates by using Flagellin A (flaA) typing andRandom Amplified Polymorphic DNA (RAPD) assays.Intestinal content, gall bladder, liver and faecal samples collected from atotal number of 1154 clinically healthy cattle were inoculated onto PrestonEnrichment broth and Preston Campylobacter Selective agar. Of the cattlesamples, 301 (26.1 %) were determined to be positive for Campylobacter spp. Inthe analysis of the isolates by multiplex PCR assay, C. jejuni and C. coli wereidentified from 59.5 % (179/301) and 10 % (30/301), respectively. None of theliver samples examined was found to be positive for either C. jejuni or C. coli.The isolates identified as C. jejuni or C. coli were subtyped by RestrictionFragment Length Polymorphism (RFLP) analysis (flaA typing) of 1.7 kbamplicon portion of the flaA gene and by RAPD based on a single random 10-merprimer. Although all the isolates were successfully typed by flaA typing, noproductive results could be obtained from C. jejuni isolates of intestinal contentsand faecal samples by RAPD analysis. The total number of band patterns definedby flaA typing and RAPD were 28 and 42, respectively in cattle.In addition, intestinal content, gall bladder and faecal samples collectedfrom a total number of 610 clinically healthy sheep were examined for thepresence of C. jejuni and C. coli strains. Of the sheep samples, 302 (49.5 %) weredetermined to be positive for Campylobacter spp. by conventional culturemethods. In the analysis of the isolates by multiplex PCR assay, C. jejuni and C.coli were identified from 34.1 % (103/302) and 33.1 % (100/302), respectively.All the isolates identified as C. jejuni or C. coli were successfully subtyped byflaA typing and RAPD assay. The total number of band patterns defined by flaAtyping and RAPD were 48 and 45, respectively in sheep.In conclusion, it was observed that a high degree of heterogeneity existedamong C. jejuni and C. coli isolates of clinically healthy cattle and sheep originand some of the flaA and RAPD profiles were represented by remarkably highpercentages of the isolates. The results of the present study also suggest thathealthy cattle and sheep can play significant role in the contamination ofenvironment and human food chain by Campylobacter spp. and these animalsshould therefore be considered seriously in order to have a better understanding ofthe epidemiology of Campylobacter infections.Key Words: Cattle, sheep, C. jejuni, C. coli, multiplex PCR, RFLP,RAPD.