Occurrence of Helicobacter spp. and Fecal Bacterial Contamination in High-altitude Aquatic Environments from the Andes.

The occurrence of Helicobacter spp. and fecal bacterial contamination was investigated in high-altitude environments from the Northeastern Andes of Venezuela. Helicobacter DNA was detected by PCR in streams, drinking and irrigation waters, and one culture from drinking water by the HP enrichment medium for selection of Helicobacter pylori, which displayed 98.98% homology to this pathogen based on 16S rRNA gene sequencing. FISH demonstrated predominant coccoid cells of the target bacteria indicative of the viable but nonculturable (VBNC) state in all water samples and HP cultures. Our work reveals for the first time Helicobacter spp. in waters from one of the highest places in the world. These results, together with the presence of fecal coliforms (2-160 MPN/100 mL) from the headwaters of rivers to drinking and irrigation waters, alert fecal contamination and epidemiological implications in this area of ecological and economic importance for the region.

Occurrence and virulence properties of Vibrio and Salinivibrio isolates from tropical lagoons of the southern Caribbean Sea.

The Vibrionaceae are Gram-negative bacteria present in marine and estuarine environments worldwide, including several species known as important pathogens to humans and aquatic organisms. The aim of this research was to investigate the occurrence and virulence properties of Vibrio and Salinivibrio isolated from lagoons at Cuare Wildlife Refuge and Margarita Island in the southern Caribbean Sea. Water, plankton and oyster samples were collected during October 2011 and March 2012 and examined by specific PCR and culture methods. Vibrio genus DNA was detected in 95% of samples, while the intergenic spacer region (ISR) of Vibrio cholerae and the genes that code for the thermolabile direct haemolysin (tl) of Vibrio parahaemolyticus and the haemolysin/cytolysin (vvhA) of Vibrio vulnificus were absent or amplified in low proportions (23, 5, and 0%, respectively). Nine isolates from water and plankton were confirmed as Vibrio or Salinivibrio by phenotypic tests, 16S rRNA gene sequencing and phylogenetic analysis. All the isolates presented similar patterns of virulence factors, in which the genes ctxA (encoding for cholera toxin), tl and vvhA were lacking, whereas seven isolates displayed antibiotic resistance against ampicillin and cephalosporins. The 16S rRNA phylogenetic analysis showed the clustering of Vibrio isolates in three main clades: the plankton isolate from Cuare Wildlife Refuge formed a group with V. cholerae and Vibrio mimicus while the Margarita isolates clustered with sequences from the harveyi clade and Salinivibrio. This is the first time that Salinivibrio species are reported in tropical lagoons of the Caribbean Sea with antibiotic resistance.

Differential distribution and biochemical characteristics of hydrolases among developmental stages of Schistosoma mansoni may offer new anti-parasite targets.

Schistosoma mansoni enzymes play important roles in host-parasite interactions and are potential targets for immunological and/or pharmacological attack. The aim of this study was to comparatively assess the presence of hydrolytic activities (phosphatases, glycosidases, aminopeptidases) in soluble (SF) and membrane (MF) fractions from different S. mansoni developmental stages (schistosomula 0 and 3h, juveniles, and adult worms of 28 and 45days-old, respectively), by using simple enzyme-substrate microassays. Our results show and confirm the prominent presence of alkaline phosphatase (AlP) activity in the MF of all the above parasite stages, highlighting also the relevant presence of MF-associated α-mannosidase (α-MAN) activity in juveniles. A soluble AlP activity, together with ß-N-D-acetylglucosaminidase (ß-NAG), and α-MAN activities, was detected in SF of schistosomulum 0h. Soluble ß-NAG, α-MAN, acid phosphatase (AcP), leucin (LAP) and alanine (AAP) aminopeptidase activities were also seen in the SF of the other different developmental stages. This work shows different soluble and membrane-associated hydrolytic capacities in each S. mansoni developmental stage from schistosomula to adults that might be exploitable as potential new targets for immune and/or chemoprophylactic strategies.

Multiple cag genotypes of Helicobacter pylori isolates colonize the oesophagus in individual hosts in a Venezuelan population.

PURPOSE: Multiple Helicobacter pylori strains colonize and coexist in the stomach of one single patient, carrying heterogeneous distributions of cag genotypes. The oesophagus provides a niche for H. pylori colonization; however, little is known about its adaptive role. METHODOLOGY: Using PCR for cagA, cagE and virB11 genes from cag-pathogenicity island (PAI) and Etest for antimicrobial susceptibility test, we determined cag-PAI genotypes associated with H. pylori virulence, when positive cultures were matching in both the stomach and the oesophagus (96 isolates; 8 out of 80 dyspeptic patients). RESULTS: The stomach showed complete cag-PAI islands in 77 % of the isolates, whereas the oesophagus showed complete cag-PAI islands only in 44 % of the isolates. Expression of CagA and interleukin 8 correlated with inflammatory processes and histopathological changes in the stomach, but not in the oesophagus. Different cag-PAI profiles were found in both mucosae of an individual host, and at least one oesophagus profile corresponded to one profile identified in stomach. The antibiotic resistance profiles showed variability in the colonization by single or mixed H. pylori isolates in the gastric and oesophageal mucosa both intra- and inter-individuals. CONCLUSION: These results demonstrate colonization with multiple H. pylori isolates in the oesophageal mucosa, like those found in the stomach of individual hosts. H. pylori was characterized by a dominant partial island, low interleukin 8 induction with lower histopathological damage and lower antibiotic resistance, suggesting that the microenvironmental changes in individual hosts select less virulent isolates in the oesophagus than in the stomach. New approaches to ensure effective eradication therapy in multi-resistant H. pylori strains must be developed.

BIOFILM FORMATION OF Vibrio cholerae ON STAINLESS STEEL USED IN FOOD PROCESSING.

Vibrio cholerae represents a significant threat to human health in developing countries. This pathogen forms biofilms which favors its attachment to surfaces and its survival and transmission by water or food. This work evaluated the in vitro biofilm formation of V. cholerae isolated from clinical and environmental sources on stainless steel of the type used in food processing by using the environmental scanning electron microscopy (ESEM). Results showed no cell adhesion at 4 h and scarce surface colonization at 24 h. Biofilms from the environmental strain were observed at 48 h with high cellular aggregations embedded in Vibrio exopolysaccharide (VPS), while less confluence and VPS production with microcolonies of elongated cells were observed in biofilms produced by the clinical strain. At 96 h the biofilms of the environmental strain were released from the surface leaving coccoid cells and residual structures, whereas biofilms of the clinical strain formed highly organized structures such as channels, mushroom-like and pillars. This is the first study that has shown the in vitro ability of V. cholerae to colonize and form biofilms on stainless steel used in food processing.

Prevalence and Distribution of Vibrio spp. in Wild Aquatic Birds of the Southern Caribbean Sea, Venezuela, 2011-12.

Vibrio spp. are associated with waterbirds mainly in temperate latitudes. We evaluated the prevalence and distribution of Vibrio spp. from fecal samples of resident and migratory aquatic birds collected during October 2011 and March 2012 at two coastal sites in the tropical southern Caribbean Sea. We amplified DNA by PCR in 40% of samples, resulting in 47% and 36% estimated prevalence for resident and migratory birds in Cuare Wildlife Refuge, and 33% and 44% in Margarita Island, respectively. We found nontoxigenic Vibrio cholerae in Cuare Wildlife Refuge with a higher prevalence in resident birds (18%). Our PCR results for Vibrio and V. cholerae were not significantly different between sites or bird migratory status. The 16S rRNA phylogenetic analysis sequences from fecal samples from Cuare Wildlife Refuge were highly similar to V. cholerae and Vibrio vulnificus , whereas sequences from Margarita Island samples formed clusters with species related to the Harveyi clade. Our findings indicate that several species of Vibrio are common in aquatic birds along the southern Caribbean Sea and contribute to our understanding of the role of birds as possible reservoirs of potentially pathogenic bacteria.

Evidence of Helicobacter spp. in freshwaters from Roraima Tepui, Guayana Shield, South America.

Helicobacter presence and viability in waters is not well characterized. The identification of natural reservoirs and infection sources may provide novel insights into its waterborne transmission. The goal of this study was to investigated the occurrence of Helicobacter spp. in natural freshwaters from Roraima Tepui, a little studied and unique ecosystem of the Guayana Shield. Freshwaters collected from two localities at Roraima Tepui were cultured in HP selective broth and agar for Helicobacter pylori and analysed by fluorescent in situ hybridization (FISH), specific PCR assays, 16S rRNA gene sequencing and phylogenetic analysis. The presence of other bacteria in freshwater enrichments was determined using clone library sequencing of the 16S rRNA gene and phylogenetic inferences. Helicobacter spp. were detected by semi-nested PCR and FISH in freshwater enrichments from both sites. Coccoid viable but nonculturable (VBNC) cells were evidenced using 16S rRNA gene Helicobacter species and H. pylori-specific probes. Partial 16S rRNA gene sequences of two HP enrichments showed high similarity to H. pylori and Helicobacter nemestrinae (99-100 %). Other bacteria such as Serratia, Aquitalea, Chromobacterium, Mycobacterium, Acinetobacter, Curvibacter and Dysgonomonas were also detected using complete 16S rRNA gene sequences, with Serratia, Aquitalea and Chromobacterium the most common genera (40.9, 18.2 and 15.2 %, respectively). This is the first time that Helicobacter spp. have been reported in freshwaters of a tepui ecosystem. Our results contribute to the current knowledge of these bacteria in the aquatic environment and expand their known/potential sites outside the human host.

Environmental scanning electron microscopy analysis of Proteus mirabilis biofilms grown on chitin and stainless steel.

Proteus mirabilis is a human pathogen able to form biofilms on the surface of urinary catheters. Little is known about P. mirabilis biofilms on natural or industrial surfaces and the potential consequences for these settings. The main aim of this work was to assess and compare the adhesion and biofilm formation of P. mirabilis strains from different origins on chitin and stainless steel surfaces within 4 to 96 h. Using environmental scanning electron microscopy, the biofilms of a clinical strain grown on chitin at 4 h showed greater adhesion, aggregation, thickness, and extracellular matrix production than those grown on stainless steel, whereas biofilms of an environmental strain had less aggregation on both surfaces. Biofilms of both P. mirabilis strains developed different structures on chitin, such as pillars, mushrooms, channels, and crystalline-like precipitates between 24 and 96 h, in contrast with flat-layer biofilms produced on stainless steel. Significant differences (p < 0.05) were found in the frequency of pillars and channels. Images of transmission electron microscopy demonstrated abundant fimbriae in 100 % of cells from both strains, which could be related to surface adherence and biofilm formation. This represents the first study of P. mirabilis showing adhesion, biofilm formation, and development of different structures on surfaces found outside the human host.

Helicobacter pylori Infection in Rural and Urban Dyspeptic Patients from Venezuela.

The goal of this work was to assess the Helicobacter pylori prevalence in a rural mestizo population and compare it to an urban population from Venezuela. The study was performed in gastric juice samples of 71 dyspeptic patients from Caracas (urban) and 39 from Tucupita (rural), in the Orinoco Delta region. Helicobacter pylori was detected by amplification of 16S rRNA, glmM, and ureA genes in 55.0% patients from urban and 87.2% from rural populations. cagA was found positive in 51% and 62% urban and rural patients, respectively. Non-H. pylori Helicobacter species were not detected in the urban population, but was found in 7.7% of patients in the rural study site. Frequency values of the 16S rRNA, glmM, and ureA genes were higher in the rural population. The odds ratio for each gene was 15.18 for 16S rRNA, 2.34 for glmM, 2.89 for ureA, and 1.53 cagA, showing significant differences except for cagA when gene frequency was compared in both populations. These results demonstrate a higher frequency of H. pylori and gastric non-H. pylori Helicobacter infection in a rural mestizo population with low hygienic standards as compared with city dwellers, representing a potential risk for the development of gastroduodenal diseases.

Survival, induction and resuscitation of Vibrio cholerae from the viable but non-culturable state in the Southern Caribbean Sea.

The causative agent of cholera, Vibrio cholerae, can enter into a viable but non-culturable (VBNC) state in response to unfavorable conditions. The aim of this study was to evaluate the in situ survival of V. cholerae in an aquatic environment of the Southern Caribbean Sea, and its induction and resuscitation from the VBNC state. V. cholerae non-O1, non-O139 was inoculated into diffusion chambers placed at the Cuare Wildlife Refuge, Venezuela, and monitored for plate, total and viable cells counts. At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state. Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells. Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season. Resuscitation studies showed significant recovery of cell cultivability with spent media addition (p < 0.05). These results suggest that V. cholerae can persist in the VBNC state in this Caribbean environment and revert to a cultivable form under favorable conditions. The VBNC state might represent a critical step in cholera transmission in susceptible areas.

Survival, induction and resuscitation of vibrio cholerae from the viable but non-culturable state in the Southern Caribbean sea / Supervivencia, inducción y resucitación de Vibrio cholerae del estado viable no cultivable en el sur del Mar Caribe

The causative agent of cholera, Vibrio cholerae, can enter into a viable but non-culturable (VBNC) state in response to unfavorable conditions. The aim of this study was to evaluate the in situ survival of V. cholerae in an aquatic environment of the Southern Caribbean Sea, and its induction and resuscitation from the VBNC state. V. cholerae non-O1, non-O139 was inoculated into diffusion chambers placed at the Cuare Wildlife Refuge, Venezuela, and monitored for plate, total and viable cells counts. At 119 days of exposure to the environment, the colony count was < 10 CFU/mL and a portion of the bacterial population entered the VBNC state. Additionally, the viability decreased two orders of magnitude and morphological changes occurred from rod to coccoid cells. Among the aquatic environmental variables, the salinity had negative correlation with the colony counts in the dry season. Resuscitation studies showed significant recovery of cell cultivability with spent media addition (p < 0.05). These results suggest that V. cholerae can persist in the VBNC state in this Caribbean environment and revert to a cultivable form under favorable conditions. The VBNC state might represent a critical step in cholera transmission in susceptible areas.

El agente causal del cólera, Vibrio cholerae, puede entrar a un estado viable no cultivable (VNC) en respuesta a condiciones desfavorables. El objetivo de este estudio fue evaluar la supervivencia in situ de V. cholerae en un ambiente acuático al sur del Mar Caribe y su inducción y resucitación del estado VBNC. V. cholerae no-O1, no-O139 fue inoculado en cámaras de difusión ubicadas en el Refugio de Fauna Cuare, Venezuela, y monitoreado para contaje de colonias, células totales y viables. En 119 días de exposición al ambiente, el contaje de colonias fue < 10 UFC/mL y una fracción de la población bacteriana entró al estado VBNC. Adicionalmente, la viabilidad disminuyó dos órdenes de magnitud y ocurrieron cambios morfológicos de células bacilares a cocoides. Entre las variables del ambiente acuático, la salinidad presentó correlación negativa con el contaje de colonias. Los estudios de resucitación mostraron recuperación significativa de la cultivabilidad celular con adición de sobrenadantes de cultivos en crecimiento activo (p < 0.05). Estos resultados sugieren que V. cholerae puede persistir en estado VBNC en este ambiente de Caribe y revertir a una forma cultivable bajo condiciones favorables. El estado VBNC podría representar un paso crítico en la transmisión del cólera en áreas susceptibles.

Heterogeneity of cag genotypes of Helicobacter pylori in the esophageal mucosa of dyspeptic patients and its relation to histopathological outcomes.

BACKGROUND: A high prevalence of Helicobacter pylori in the esophageal mucosa of dyspeptic Venezuelan patients has been reported. We aimed to assess the genetic composition of the cag genotypes of H. pylori and its relation to histopathological outcomes in the gastroesophageal mucosa. METHODS: The presence of cagA, cagE, and virB11 cag pathogenicity island (PAI) genes was detected by PCR in 80 of 150 H. pylori-positive dyspeptic patients in both mucosae. Alterations of the gastroesophageal mucosa were assessed by histological techniques. RESULTS: The frequency of intact, partial, and deleted cag-PAI genes in the stomach of dyspeptic patients was found to be 57.5%, 21.3%, and 21.3%, respectively, whereas in the esophagus, frequencies were 33.8%, 33.8%, and 32.5% respectively. The genetic composition in the stomach was 57.5% cagA-positive, 20.0% cagA-negative, 75.0% cagE, and 77.5% virB11, whereas in the esophagus the distribution was 36.3% cagA-positive, 30.0% cagA-negative, 61.3% cagE, and 63.8% virB11. The gene with the largest difference between the two mucosae was cagA, with 58.8% in the stomach and 37.5% in the esophagus; cagE and virB11 were less variable. The correlation among single and/or mixed cag genotypes with histopathological outcomes in both mucosae from the same patient was higher for intact single cag-PAI genotypes, showing severe alterations. CONCLUSIONS: H. pylori may coexist in similar proportions without dominance of one cag genotype, suggesting a heterogeneous distribution in the esophagus. The cagE and virB11 genes can be used as markers of cag-PAI in the esophagus. The single cag-PAI genotype in both mucosae confers an increased risk of developing histological damage.

Detection of Helicobacter in the digestive tract of an Atlantic spotted dolphin (Stenella frontalis).

Helicobacter spp. occur in the digestive system of a broad range of animal taxa, including marine mammals. Only one formally recognized species, Helicobacter cetorum, has been described in marine mammals. Helicobacter has not been reported in the Atlantic spotted dolphin (Stenella frontalis). The purpose of our study was to examine the digestive tract of a stranded spotted dolphin for Helicobacter. Tissue and content samples were collected at necropsy and examined by histopathology and molecular analyses using Helicobacter genus-specific 16S rDNA polymerase chain reaction (PCR) and DNA sequencing. Helicobacter was detected in all stomach divisions and the duodenal ampulla. A sequence type of the 16S rRNA gene shared a 98-99% identity to sequences from H. cetorum. This study reports for the first time Helicobacter in S. frontalis.

Vibrio cholerae non-O1, non-O139 associated with seawater and plankton from coastal marine areas of the Caribbean Sea.

The aim of this study was to characterize the virulence properties and the antimicrobial resistance of Vibrio cholerae isolates from a coastal area of the Caribbean Sea. Three V. cholerae isolates were obtained from seawater and plankton using the HP selective medium for Helicobacter pylori. These V. cholerae isolates belonged to the non-O1, non-O139 serogroups and they did not have cholera toxin genes. They were resistant to penicillins and some cephalosporins and were sensitive to netilmicin, tetracyclines, sulfamethoxazole-trimethoprim and quinolones. This is the first study that provides biochemical and molecular evidence of non-O1, non-O139 V. cholerae isolates, non-toxigenic, carrying antibiotic resistance in seawater and plankton from a coastal area of the Caribbean Sea.

Evidencias de la transmisión acuática de Helicobacter Pylori / Evidences of aquatic transmission of Helicobacter Pylori / Evidências da transmisão aquática de Helicobacter Pylori

Helicobacter pylori es agente causal de gastritis y úlcera duodenal y juega un rol importante en el desarrollo del cáncer gástrico. Más de la mitad de la población mundial está infectada con este microorganismo, considerándose en la actualidad uno de los patógenos humanos de mayor importancia. En vista de la incertidumbre existente acerca de su transmisión ambiental y a la dificultad de su detección en fuentes no humanas, en este trabajo se realiza una revisión acerca de la presencia de H. pylori en distintas fuentes de agua y sus estrategias de supervivencia en ambientes acuáticos. Estudios epidemiológicos han establecido que el agua contaminada con materia fecal es una fuente de infección, principalmente en países en vías de desarrollo. Diferentes métodos han sido evaluados para su detección en muestras de agua. Se han desarrollado técnicas basadas en el cultivo en medios selectivos, inmunofluorescencia, hibridación in situ por fluorescencia y PCR. A pesar del éxito obtenido con algunos de estos métodos, hasta el presente no existe un procedimiento estándar para la detección de este microorganismo en el agua. La formación de biopelículas y la persistencia bajo el estado viable no cultivable (VNC) en el agua han sido propuestas como estrategias de supervivencia de H. pylori en el ambiente. Futuros trabajos deberían dirigirse a optimizar la detección de H. pylori en muestras ambientales para esclarecer la vía de transmisión asociada al medio acuático.

Helicobacter pylori is a causing agent of gastritis and duodenal ulcer, and plays an important role in the etiology of stomach cancer. This organism infects over one half of the world population and it is currently considered as one of the most important human pathogens. In view of the existing uncertainty about its transmission from the environment and the difficulty of detecting it in non-human sources, a review is made in the present paper about the presence of H. pylori in different water sources and its survival strategies in aquatic environments. Epidemiological studies have established that, mainly in developing countries, water contaminated with feces is a source of infection. Different methods for detection in water have been evaluated. Techniques have been developed based on culture in selective media, immunofluorescence, in situ hybridization by fluorescence and PCR. Despite the success obtained with some of these methods, to date no standard procedure exists for the detection of this microorganism in water. Bio-film formation and the persistence in a viable non-cultivable (VNC) state in water have been proposed as survival strategies of H. pylori in the environment. Future work should focus on optimizing the detection of H. pylori in environmental samples so as to clarify the transmission path associated with the aquatic medium.

Helicobacter pylori é agente causal de gastrite e úlcera duodenal, e tem um papel importante no desenvolvimento do câncer gástrico. Mais da metade da população mundial está infectada com este microorganismo, considerando-se na atualidade um dos patógenos humanos de maior importância. Em vista da incertidumbre existente sobre sua transmissão ambiental e a dificuldade de sua detecção em fontes não humanas, neste trabalho se realiza uma revisão sobre a presença de H. pylori em distintas fontes de água e suas estratégias de sobrevivência em ambientes aquáticos. Estudos epidemiológicos têm estabelecido que a água contaminada com matéria fecal é uma fonte de infecção, principalmente em países em vias de desenvolvimento. Diferentes métodos têm sido avaliados para sua detecção em amostras de água. Tem-se desenvolvido técnicas baseadas no cultivo em meios seletivos, imunofluorescência, hibridização in situ por fluorescência e PCR. A pesar do sucesso obtido com alguns destes métodos, até o presente não existe um procedimento estándar para a detecção de este microorganismo na água. A formação de biopelículas e a persistência sob o estado viável não cultivável (VNC) na água tem sido propostas como estratégias de sobrevivência de H. pylori no ambiente. Futuros trabalhos deveriam dirigir-se a aperfeiçoar a detecção de H. pylori em amostras ambientais para esclarecer a via de transmissão associada ao meio aquático.

Occurrence of Proteus mirabilis associated with two species of venezuelan oysters.

The fecal contamination of raw seafood by indicators and opportunistic pathogenic microorganisms represents a public health concern. The objective of this study was to investigate the presence of enteric bacteria colonizing oysters collected from a Venezuelan touristic area. Oyster samples were collected at the northwestern coast of Venezuela and local salinity, pH, temperature, and dissolved oxygen of seawater were recorded. Total and fecal coliforms were measured for the assessment of the microbiological quality of water and oysters, using the Multiple Tube Fermentation technique. Analyses were made using cultures and 16S rRNA gene sequencing. Diverse enrichment and selective culture methods were used to isolate enteric bacteria. We obtained pure cultures of Gram-negative straight rods with fimbriae from Isognomon alatus and Crassostrea rhizophorae. Our results show that P. mirabilis was predominant under our culture conditions. We confirmed the identity of the cultures by biochemical tests, 16S rRNA gene sequencing, and data analysis. Other enterobacteria such as Escherichia coli, Morganella morganii and Klebsiella pneumoniae were also isolated from seawater and oysters. The presence of pathogenic bacteria in oysters could have serious epidemiological implications and a potential human health risk associated with consumption of raw seafood.

Occurrence of Proteus mirabilis associated with two species of venezuelan oysters

The fecal contamination of raw seafood by indicators and opportunistic pathogenic microorganisms represents a public health concern. The objective of this study was to investigate the presence of enteric bacteria colonizing oysters collected from a Venezuelan touristic area. Oyster samples were collected at the northwestern coast of Venezuela and local salinity, pH, temperature, and dissolved oxygen of seawater were recorded. Total and fecal coliforms were measured for the assessment of the microbiological quality of water and oysters, using the Multiple Tube Fermentation technique. Analyses were made using cultures and 16S rRNA gene sequencing. Diverse enrichment and selective culture methods were used to isolate enteric bacteria. We obtained pure cultures of Gram-negative straight rods with fimbriae from Isognomon alatus and Crassostrea rhizophorae. Our results show that P. mirabilis was predominant under our culture conditions. We confirmed the identity of the cultures by biochemical tests, 16S rRNA gene sequencing, and data analysis. Other enterobacteria such as Escherichia coli, Morganella morganii and Klebsiella pneumoniae were also isolated from seawater and oysters. The presence of pathogenic bacteria in oysters could have serious epidemiological implications and a potential human health risk associated with consumption of raw seafood.

A contaminação fecal de frutos do mar crus por microrganismos oportunistas patogênicos representa problema de saúde pública. O objetivo deste estudo é investigar a presença de bactérias entéricas que colonizam ostras coletadas em área turística da Venezuela. Amostras de ostras foram coletadas na costa noroeste da Venezuela e foram registrados a salinidade local, pH, temperatura e o oxigênio dissolvido na água do mar. O total de coliformes fecais foi medido para a avaliação da qualidade microbiológica da água e das ostras, usando a técnica de fermentação em tubos múltiplos. Análises foram feitas usando culturas e seqüência do gene 16S rRNA. Enriquecimento diversificado e métodos de cultura seletivos foram usados para isolar a bactéria entérica. Obtivemos culturas puras de bastões retos Gram negativos com fímbrias de Isognomon alatus e Crassostrea rhizophorae. Nossos resultados mostram que P. mirabilis foi predominante nas nossas condições de cultura. Confirmamos a identidade das culturas por testes bioquímicos, seqüência do gene 16rRNA e a análise de dados. Outras enterobactérias como Escherichia coli, Morganella morganii e Klebsiella pneumoniae foram também isoladas da água do mar e ostras. A presença de bactérias patogênicas em ostras podem ter implicações epidemiológicas e potencial risco para a saúde humana quando do consumo de frutos do mar crus.