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Evaluation of enrichment media for improved PCR‐based detection of V. cholerae and V. vulnificus from estuarine water and plankton
Author(s) -
Malayil L.,
Turner J.W.,
Mote B.L.,
Howe K.,
Lipp E.K.
Publication year - 2011
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/j.1365-2672.2011.04996.x
Subject(s) - vibrio cholerae , plankton , estuary , biology , vibrionaceae , microbiology and biotechnology , oceanography , environmental science , bacteria , ecology , geology , genetics
Aims: Pathogenic Vibrio spp., including V. cholerae and V. vulnificus , are commonly found along the estuaries of the south‐east United States; however, it is often difficult to recover these species directly from environmental samples. Pre‐enrichment assays are commonly used to improve the detection of pathogenic vibrios from environmental sources. Here, we evaluated a novel enrichment procedure using freshly collected and autoclaved natural estuarine water amended with 1% peptone (designated as estuarine peptone water, EPW) and compared it to traditional alkaline peptone water (APW) for detection by PCR of V. cholerae and V. vulnificus . Methods and Results: Of the 50 samples collected in total, V. cholerae DNA was detected in APW 10% of the time and in EPW 40% of the time. Likewise, the cholera toxin gene ( ctxA ) was detected in 4 vs 18% of the samples using APW and EPW, respectively. Conversely, APW showed improved recovery for V. vulnificus relative to EPW with respective detection frequencies of 46 and 20%. Results showed similar patterns across different sample types (water and plankton). Conclusions: While enrichment in traditional APW was adequate for the recovery of Vibrio vulnificius , use of sterile estuarine water amended with peptone significantly improved the detection of V. cholerae and the virulence gene ctxA from estuarine sources.