Oysters can accumulate potentially pathogenic water bacteria. The objective of this study was to compare two procedures to quantify Vibrio species present in oysters to determine the most sensitive method. We analyzed oyster samples from the Gulf of Mexico, commercialized in Mexico City. The samples were inoculated in tubes with alkaline peptone water (APW), based on three tubes and four dilutions (10 to 10). From these tubes, the first quantification of Vibrio species was performed (most probable number (MPN) from tubes) and bacteria were inoculated by streaking on thiosulfate-citrate-bile salts-sucrose (TCBS) petri dishes. Colonies were isolated for a second quantification (MPN from dishes). Polymerase chain reaction (PCR) was used to determine species with specific primers: ompW for Vibrio cholerae, tlh for Vibrio parahaemolyticus, and VvhA for Vibrio vulnificus. Simultaneously, the sanitary quality of oysters was determined. The quantification of V. parahaemolyticus was significantly higher in APW tubes than in TCBS dishes. Regarding V. vulnificus counts, the differences among both approaches were not significant. In contrast, the MPNs of V. cholerae obtained from dishes were higher than from tubes. The quantification of MPNs through PCR of V. parahaemolyticus and V. vulnificus obtained from APW was sensitive and recommendable for the detection of both species. In contrast, to quantify V. cholerae, it was necessary to isolate colonies on TCBS prior PCR. Culturing in APW at 42 °C could be an alternative to avoid colony isolation. The MPNs of V. cholerae from dishes was associated with the bad sanitary quality of the samples.
The need to separate bacteria from sediment in order to appropriately count them has led to test the efficacy of different techniques. In this research, traditional techniques such as manual shaking, homogenization, ultrasonication, and surfactant are compared. Moreover, the possibility of using a set of enzymes (pancreatine) and an antibiotic (ampicillin) for sediment coliform extraction is proposed. Samples were obtained from Xochimilco Lake in Mexico City. The most probable number of coliform bacteria was determined after applying the appropriate separation procedure. Most of the techniques tested led to numbers similar to those of the control (manual shaking). Only with the use of ampicillin, a greater total coliform concentration was observed (Mann-Whitney, z = 2.09; p = 0.03). It is possible to propose the use of ampicillin as a technique for total coliform extraction; however, it is necessary to consider sensitivity of bacteria to the antibiotic.
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