Toxigenic V. cholerae, V. parahaemolyticus, and V. vulnificus in oysters from the Gulf of Mexico and sold in Mexico City.

Oysters can accumulate potentially pathogenic bacteria, such as Vibrio cholerae, V. parahaemolyticus, and V. vulnificus. The aim of this study was to detect the presence of these Vibrio species and their toxigenic variants in oysters from the Gulf of Mexico sold in Mexico City. Oyster samples were studied using traditional culture and molecular polymerase chain reaction analysis. V. cholerae was present in 30.4% of the samples and its toxigenic variant chxA+ in 26.1%. It was isolated only in deshelled oysters, mainly in the dry season. V. parahaemolyticus was present in 95.7% of the samples and the toxigenic variant was found in 17.4%. V. vulnificus was identified in 60.9% of the samples, 38% of which corresponded to the environmental genotype and 21.7% to the clinical genotype, mainly in the cold season. Consumption of the oysters analyzed poses health risks due to the presence of Vibrio species, especially in deshelled oysters.

Genetic Analysis of Vibrio parahaemolyticus O3:K6 Strains That Have Been Isolated in Mexico Since 1998.

Vibrio parahaemolyticus is an important human pathogen that has been isolated worldwide from clinical cases, most of which have been associated with seafood consumption. Environmental and clinical toxigenic strains of V. parahaemolyticus that were isolated in Mexico from 1998 to 2012, including those from the only outbreak that has been reported in this country, were characterized genetically to assess the presence of the O3:K6 pandemic clone, and their genetic relationship to strains that are related to the pandemic clonal complex (CC3). Pathogenic tdh+ and tdh+/trh+ strains were analyzed by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Also, the entire genome of a Mexican O3:K6 strain was sequenced. Most of the strains were tdh/ORF8-positive and corresponded to the O3:K6 serotype. By PFGE and MLST, there was very close genetic relationship between ORF8/O3:K6 strains, and very high genetic diversities from non-pandemic strains. The genetic relationship is very close among O3:K6 strains that were isolated in Mexico and sequences that were available for strains in the CC3, based on the PubMLST database. The whole-genome sequence of CICESE-170 strain had high similarity with that of the reference RIMD 2210633 strain, and harbored 7 pathogenicity islands, including the 4 that denote O3:K6 pandemic strains. These results indicate that pandemic strains that have been isolated in Mexico show very close genetic relationship among them and with those isolated worldwide.

Quantification of Vibrio species in oysters from the Gulf of Mexico with two procedures based on MPN and PCR.

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 (10-1 to 10-4). 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.

Genetic characterization of Vibrio vulnificus strains isolated from oyster samples in Mexico.

Vibrio vulnificus strains were isolated from oysters that were collected at the main seafood market in Mexico City. Strains were characterized with regard to vvhA, vcg genotype, PFGE, multilocus sequence typing (MLST), and rtxA1. Analyses included a comparison with rtxA1 reference sequences. Environmental (vcgE) and clinical (vcgC) genotypes were isolated at nearly equal percentages. PFGE had high heterogeneity, but the strains clustered by vcgE or vcgC genotype. Select housekeeping genes for MLST and primers that were designed for rtxA1 domains divided the strains into two clusters according to the E or C genotype. Reference rtxA1 sequences and those from this study were also clustered according to genotype. These results confirm that this genetic dimorphism is not limited to vcg genotyping, as other studies have reported. Some environmental C genotype strains had high similarity to reference strains, which have been reported to be virulent, indicating a potential risk for oyster consumers in Mexico City.

Filtration rate, assimilation and assimilation efficiency in Crassostrea virginica (Gmelin) fed with Tetraselmis suecica under cadmium exposure.

Crassostrea virginica is an epibentic filter-feeding bivalve of economical importance in coastal lagoons of the Gulf of Mexico, locations with increasing inputs of heavy metals such as cadmium that have become environmental stressors. In this study, feeding and assimilation of the species were evaluated as physiological indicators of cadmium exposure. For this purpose, the filtration rate (FR), food assimilation (A) and assimilation efficiency (AE) of oysters from the Mandinga Lagoon, Veracruz, Mexico, were examined under sublethal and environmentally realistic cadmium concentrations (95 and 170 micro gCd L(-1)). Semi-static, 12-day bioassays were conducted with organisms placed into individual chambers and fed daily with Tetraselmis suecica. FR was calculated by measuring the depletion in algal density. Caloric contents of food and feces produced were also obtained. Condition Index (CI) and morphometric parameters were evaluated at the beginning and at the end of the assay. Total cadmium concentrations were quantified in water and tissue, and the metal bioconcentration factor (BCF) was calculated. Cadmium exposure significantly reduced FR in oysters (mean value: 0.64 L h(-1) and 0.44 L h(-1)) from control values (1.17 L h(-1)). Extreme values among results demonstrate the existence of a high FR (over 4 L h(-1)) mainly in control oysters, and this was associated with a better physiological condition; a low FR (under 2.5 L h(-1)) indicated metabolic stress as a consequence of Cd exposure. A and AE were significantly modified due to cadmium external levels, and time of exposure. FR and A were linearly related, and both decreased as metal BCF increased. Cadmium bioaccumulation was linearly related with external metal levels. The physiological deterioration of native C. virginica from Mandinga Lagoon was reflected in the alteration of FR, A and AE due to cadmium exposure in concentrations considered sublethal, lowering the feeding and assimilation capability of the organisms. The weight loss and mortality recorded in the oysters exposed to the highest metal concentration, was the final consequence of the overall adverse effect of cadmium exposure.