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Int. J. Environ. Res. Public Health 2015, 12(3), 2749-2764; doi:10.3390/ijerph120302749

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

1
VISAVET Health Surveillance Centre, Complutense University of Madrid, Avda. Puerta de Hierro, s/n. 28040 Madrid, Spain
2
Animal Health Department, Veterinary Faculty, Complutense University of Madrid, Avda. Puerta de Hierro, s/n. 28040 Madrid, Spain
3
Molecular Microbiology and Genomics Consultants, Zotzenheim 55576, Germany
*
Author to whom correspondence should be addressed.
Academic Editor: Dietmar Knopp
Received: 19 November 2014 / Revised: 18 February 2015 / Accepted: 25 February 2015 / Published: 2 March 2015
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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. View Full-Text
Keywords: Campylobacter; urban effluents; isolation protocols; antimicrobial resistance Campylobacter; urban effluents; isolation protocols; antimicrobial resistance
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).

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MDPI and ACS Style

Ugarte-Ruiz, M.; Florez-Cuadrado, D.; Wassenaar, T.M.; Porrero, M.C.; Domínguez, L. Method Comparison for Enhanced Recovery, Isolation and Qualitative Detection of C. jejuni and C. coli from Wastewater Effluent Samples. Int. J. Environ. Res. Public Health 2015, 12, 2749-2764.

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