Method Comparison for Enhanced Recovery, Isolation and Qualitative Detection of C. jejuni and C. coli from Wastewater Effluent Samples

Abstract

Seeking a sensitive protocol, culture-dependent methods were compared to detect thermophilic Campylobacter species in untreated urban effluents. We evaluated various combinations of selective media, with and without an enrichment steps, as well as an extra filtration step. Culture-independent real-time quantitative PCR was also included and all detected isolates underwent antimicrobial susceptibility testing. All tested water samples contained Campylobacter DNA, but only 64% were positive after culture. Although enrichment using Preston broth resulted in better recovery of potentially stressed Campylobacter than Bolton or Campyfood broth (CFB), there was no significant increase in efficiency compared to direct plating. The type of selective agar media used, on the other hand, had a significant effect, with CASA plates performing better than mCCDA or CFA ones. Inclusion of an enrichment step increased the ratio of C. coli vs. C. jejuni being isolated. Resistances against all antimicrobials tested were observed in C. coli, but fewer instances of resistance were found in C. jejuni isolates. Whether this difference was the result of selection during the enrichment step could not be determined. The presence of Campylobacter in urban effluents can be considered as a valuable proxy for Campylobacter populations present in urban environments.

Characterization and Phylogenetic Analysis of Campylobacter Species Isolated from Paediatric Stool and Water Samples in the Northwest Province, South Africa

“…Increase in tetracycline resistance has been reported to emerge from the extensive use of these antibiotics as prophylaxis and therapy of human and animal infections and in promoting animal growth [77]. Thus, Campylobacter resistance to tetracycline has been frequently reported in humans, animals and aquatic environments [78,79,80]. The current study observed a resistance rate of 32% of clinical Campylobacter isolates to tetracycline and 55% from water isolates.…”

Contamination Status of Salmonella spp., Shigella spp. and Campylobacter spp. in Surface and Groundwater of the Kelani River Basin, Sri Lanka

“…However, Premarathne et al [80] recorded that the favored environmental place for Campylobacter spp. is to be the intestinal tract of most of birds and mammals; Ugarte-Ruiz et al [81] recorded that survival of Campylobacter spp. in surface water for a higher recovery of the organism in wide range of water quality changes.…”

Gene pool transmission of multidrug resistance among Campylobacter from livestock, sewage and human disease

“…Human samples (n = 152; 118 C. jejuni and 34 C. coli) were associated with campylobacteriosis cases in hospitals in the regions of Castilla y Leon, Extremadura and Andalucía between 2013 and 2016. Campylobacter isolates of urban effluent origin (n = 53; 6 C. jejuni and 47 C. coli) were collected from the wastewater treatment plants in the city of Madrid (Spain) between 2011 and 2013 (Ugarte-Ruiz et al, 2015). All isolates were obtained using culture based methods (Moreno et al, 2000;Ugarte-Ruiz et al, 2015;Hormeño et al, 2016) and speciated as C. jejuni or C. coli using a conventional multiplex PCR as previously described (Ugarte-Ruiz et al, 2012).…”