TY - JOUR
T1 - An improved culture method for selective isolation of Campylobacter jejuni from wastewater
AU - Kim, Jinyong
AU - Oh, Euna
AU - Banting, Graham S.
AU - Braithwaite, Shannon
AU - Chui, Linda
AU - Ashbolt, Nicholas J.
AU - Neumann, Norman F.
AU - Jeon, Byeonghwa
N1 - Publisher Copyright:
© 2016 Kim, Oh, Banting, Braithwaite, Chui, Ashbolt, Neumann and Jeon.
PY - 2016/8/26
Y1 - 2016/8/26
N2 - 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 Ct 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.
AB - 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 Ct 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.
KW - Antibiotics
KW - Campylobacter jejuni
KW - Isolation
KW - QRT-PCR
KW - Wastewater
UR - http://www.scopus.com/inward/record.url?scp=84989314259&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84989314259&partnerID=8YFLogxK
U2 - 10.3389/fmicb.2016.01345
DO - 10.3389/fmicb.2016.01345
M3 - Article
AN - SCOPUS:84989314259
SN - 1664-302X
VL - 7
JO - Frontiers in Microbiology
JF - Frontiers in Microbiology
IS - AUG
M1 - 1345
ER -