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Campylobacter jejuni  is one of the leading foodborne pathogens worldwide.  C. jejuni  is isolated from a wide range of foods, domestic animals, wildlife, and environmental sources. The currently available culture-based isolation methods are not highly effective for wastewater samples due to the low number of  C. jejuni  in the midst of competing bacteria. To detect and isolate  C. jejuni  from wastewater samples, in this study, we evaluated a few different enrichment conditions using five different antibiotics (i.e., cefoperazone, vancomycin, trimethoprim, polymyxin B, and rifampicin), to which  C. jejuni  is intrinsically resistant. The selectivity of each enrichment condition was measured with  C  t  value using quantitative real-time PCR, and multiplex PCR to determine  Campylobacter  species. In addition, the efficacy of  Campylobacter  isolation on different culture media after selective enrichment was examined by growing on Bolton and Preston agar plates. The addition of polymyxin B, rifampicin, or both to the Bolton selective supplements enhanced the selective isolation of  C. jejuni . The results of 16S rDNA sequencing also revealed that  Enterococcus  spp. and  Pseudomonas aeruginosa  are major competing bacteria in the enrichment conditions. Although it is known to be difficult to isolate  Campylobacter  from samples with heavy contamination, this study well exhibited that the manipulation of antibiotic selective pressure improves the isolation efficiency of fastidious  Campylobacter  from wastewater.

An Improved Culture Method for Selective Isolation of Campylobacter jejuni from Wastewater