ABSTRACT
The mariculture industry has seen a rapid expansion in recent years due to the increasing global demand for seafood. However, the industry faces challenges from climate change and increased pathogen pressure. Additionally, the chemicals used to enhance mariculture productivity are changing ocean ecosystems. This study analyzed 36 surface-water metagenomes from South Korean mussel, oyster, scallop, and shrimp farms to expand our understanding of aquaculture microbial genetic resources and the potential impacts of these anthropogenic inputs. We recovered 240 non-redundant species-level metagenome-assembled genomes (MAGs), comprising 224 bacteria, 13 archaea, and three eukaryotes. Most MAGs were assigned to Proteobacteria, Bacteroidota, and Actinobacteriota, with 40.7% remaining unclassified at the species level. Among the three eukaryotic MAGs, one was identified as a novel lineage of green algae, highlighting the uncharacterized genetic diversity in mariculture environments. Additionally, 22 prokaryotic MAGs harbored 26 antibiotic and metal resistance genes, with MAGs carrying beta-lactamases being particularly prevalent in most farms. The obtained microbiome data from mariculture environments can be utilized in future studies to foster healthy, sustainable mariculture practices.
Subject(s)
Aquaculture , Metagenome , Republic of Korea , Animals , Bacteria/genetics , Bacteria/classification , Microbiota , Ostreidae/microbiology , Archaea/genetics , Pectinidae/microbiology , Pectinidae/genetics , Penaeidae/microbiology , Penaeidae/geneticsABSTRACT
INTRODUCTION: Vibrio parahaemolyticus is a common pathogen that can cause seafood-borne gastroenteritis in humans. We determined the prevalence and characteristics of V. parahaemolyticus isolated from clinical specimens and oysters in Thailand. METHODOLOGY: Isolates of V. parahaemolyticus from clinical specimens (n = 77) and oysters (n = 224) were identified by biochemical testing, polymerase chain reaction (PCR) assays, and serotyping. The toxin genes, antimicrobial resistance, and ß-lactamase production were determined. RESULTS: A total of 301 isolates were confirmed as V. parahaemolyticus by PCR using specific primers for the toxR gene. The majority of clinical isolates carried the tdh+/trh- genotype (82.1%), and one of each isolate was tdh-/trh+ and tdh+/trh+ genotypes. One isolate from oyster contained the tdh gene and another had the trh gene. Twenty-six serotypes were characterized among these isolates, and O3:K6 was the most common (37.7%), followed by OUT:KUT, and O4:K9. In 2010, most clinical and oyster isolates were susceptible to antibiotics, with the exception of ampicillin. In 2012, clinical isolates were not susceptible to cephalothin (52.4%), streptomycin (95.2%), amikacin (66.6%), kanamycin (61.9%), and erythromycin (95.2%), significantly more frequently than in 2010. More than 95% of isolates that were not susceptible to ampicillin produced ß-lactamase enzymes. CONCLUSIONS: We found toxin genes in two oyster isolates, and the clinical isolates that were initially determined to be resistant to several antibiotics. Toxin genes and antimicrobial susceptibility profiles of V. parahaemolyticus from seafood and environment should be continually monitored to determine the spread of toxin and antimicrobial resistance genes.
Subject(s)
Ostreidae , Vibrio Infections , Vibrio parahaemolyticus , Vibrio parahaemolyticus/genetics , Vibrio parahaemolyticus/isolation & purification , Vibrio parahaemolyticus/drug effects , Vibrio parahaemolyticus/classification , Thailand/epidemiology , Ostreidae/microbiology , Humans , Animals , Vibrio Infections/microbiology , Vibrio Infections/epidemiology , beta-Lactamases/genetics , Anti-Bacterial Agents/pharmacology , Microbial Sensitivity Tests , Serotyping , Polymerase Chain Reaction , Prevalence , Genotype , Drug Resistance, Bacterial/genetics , Bacterial Toxins/genetics , Male , Adult , Female , Middle AgedABSTRACT
This study aimed to determine the prevalence of V. parahaemolyticus in oysters from the northwestern coast of Mexico and to identify the serotypes, virulence factors, and antibiotic resistance of the strains. Oyster samples were collected from 2012 to 2020 from the northwest coast of Mexico; biochemical and molecular methods were used to identify V. parahaemolyticus from oysters; antiserum reaction to determine V. parahaemolyticus serotypes, and PCR assays were performed to identify pathogenic (tdh and/or trh) or pandemic (toxRS/new, and/or orf8) strains and antibiotic resistance testing. A total of 441 oyster samples were collected and tested for V. parahaemolyticus. Forty-seven percent of oyster samples were positive for V. parahaemolyticus. Ten different O serogroups and 72 serovars were identified, predominantly serotype O1:KUT with 22.2% and OUT:KUT with 17.3%. Twenty new serotypes that had not been previously reported in our region were identified. We detected 4.3% of pathogenic clones but no pandemic strains. About 73.5% of strains were resistant to at least one antibiotic, mainly ampicillin and ciprofloxacin; 25% were multi-drug resistant. In conclusion, the pathogenic strains in oysters and antibiotic resistance are of public health concern, as the potential for outbreaks throughout northwestern Mexico is well established.
Subject(s)
Anti-Bacterial Agents , Ostreidae , Shellfish , Vibrio parahaemolyticus , Virulence Factors , Animals , Vibrio parahaemolyticus/genetics , Vibrio parahaemolyticus/drug effects , Vibrio parahaemolyticus/isolation & purification , Mexico/epidemiology , Ostreidae/microbiology , Virulence Factors/genetics , Anti-Bacterial Agents/pharmacology , Shellfish/microbiology , Drug Resistance, Bacterial , Serogroup , Virulence/genetics , Microbial Sensitivity TestsABSTRACT
To evaluate the antibiotic susceptibility of Vibrio parahaemolyticus from prawns and oysters marketed in Zhanjiang, Guangdong, China. 84 strains of V. parahaemolyticus were isolated from prawns and oysters sampled from 9 major markets. The results showed that 84 V. parahaemolyticus strains had the highest rate of antibiotic resistance to oxytetracycline (69.05 %, 58/84) and the lowest rate of antibiotic resistance to enrofloxacin (1.19 %, 1/84), ciprofloxacin (4.76 %, 4/84) and norfloxacin (7.14 %, 6/84) in quinolone. Meanwhile, 96.42 % of the strains showed multiple antibiotic resistance (MAR). PCR results showed that the resistance phenotype was closely related to the antibiotic resistance genes and efflux pump genes (p < 0.01), and the efflux pump gene was the key causing MAR. The combination of antibiotics significantly eliminated multidrug resistance. In addition, efflux pump inhibitors also reduce MAR. This study may provide information on antibiotic susceptibility, antibiotic resistance and strategies for the control of V. parahaemolyticus.
Subject(s)
Anti-Bacterial Agents , Microbial Sensitivity Tests , Ostreidae , Vibrio parahaemolyticus , Vibrio parahaemolyticus/drug effects , Anti-Bacterial Agents/pharmacology , China , Animals , Ostreidae/microbiology , Drug Resistance, Bacterial/genetics , Oxytetracycline/pharmacologyABSTRACT
Vibrio parahaemolyticus and Vibrio vulnificus are bacteria with a significant public health impact. Identifying factors impacting their presence and concentrations in food sources could enable the identification of significant risk factors and prevent incidences of foodborne illness. In recent years, machine learning has shown promise in modeling microbial presence based on prevalent external and internal variables, such as environmental variables and gene presence/absence, respectively, particularly with the generation and availability of large amounts and diverse sources of data. Such analyses can prove useful in predicting microbial behavior in food systems, particularly under the influence of the constant changes in environmental variables. In this study, we tested the efficacy of six machine learning regression models (random forest, support vector machine, elastic net, neural network, k-nearest neighbors, and extreme gradient boosting) in predicting the relationship between environmental variables and total and pathogenic V. parahaemolyticus and V. vulnificus concentrations in seawater and oysters. In general, environmental variables were found to be reliable predictors of total and pathogenic V. parahaemolyticus and V. vulnificus concentrations in seawater, and pathogenic V. parahaemolyticus in oysters (Acceptable Prediction Zone >70 %) when analyzed using our machine learning models. SHapley Additive exPlanations, which was used to identify variables influencing Vibrio concentrations, identified chlorophyll a content, seawater salinity, seawater temperature, and turbidity as influential variables. It is important to note that different strains were differentially impacted by the same environmental variable, indicating the need for further research to study the causes and potential mechanisms of these variations. In conclusion, environmental variables could be important predictors of Vibrio growth and behavior in seafood. Moreover, the models developed in this study could prove invaluable in assessing and managing the risks associated with V. parahaemolyticus and V. vulnificus, particularly in the face of a changing environment.
Subject(s)
Machine Learning , Ostreidae , Seawater , Vibrio parahaemolyticus , Vibrio vulnificus , Ostreidae/microbiology , Seawater/microbiology , Vibrio parahaemolyticus/isolation & purification , Vibrio parahaemolyticus/growth & development , Animals , Vibrio vulnificus/isolation & purification , Vibrio vulnificus/growth & development , Food Microbiology , Food Contamination/analysis , Shellfish/microbiology , Seafood/microbiology , Temperature , Vibrio/isolation & purificationABSTRACT
BACKGROUND: The global dissemination of critical-priority carbapenem-resistant hypervirulent Klebsiella pneumoniae (CR-hvKp) via food sources represents a significant public health concern. Epidemiological data on CR-hvKp in oysters in Egypt is limited. This study aimed to investigate the potential role of oysters sold in Egypt as a source for carbapenem-resistant K. pneumoniae (CRKP), hypervirulent K. pneumoniae (hvKp), and CR-hvKp and assess associated zoonotic risks. METHODS: A sample of 330 fresh oysters was randomly purchased from various retail fish markets in Egypt and divided into 33 pools. Bacteriological examination and the identification of Klebsiella pneumoniae were performed. Carbapenem resistance in K. pneumoniae isolates was determined by phenotypic and molecular methods. Additionally, the presence of hypervirulent K. pneumoniae was identified based on virulence gene markers (peg-344, rmpA, rmpA2, iucA, and iroB), followed by a string test. The clustering of CR-hvKp strains was carried out using R with the pheatmap package. RESULTS: The overall prevalence of K. pneumoniae was 48.5% (16 out of 33), with 13 isolates displaying carbapenem resistance, one intermediate resistance, and two sensitive. Both carbapenem-resistant K. pneumoniae and carbapenem-intermediate-resistant K. pneumoniae strains exhibited carbapenemase production, predominantly linked to the blaVIM gene (68.8%). HvKp strains were identified at a rate of 62.5% (10/16); notably, peg-344 was the most prevalent gene. Significantly, 10 of the 13 CRKP isolates possessed hypervirulence genes, contributing to the emergence of CR-hvKp. Moreover, cluster analysis revealed the clustering of two CR-hvKp isolates from the same retail fish market. CONCLUSION: This study provides the first insight into the emergence of CR-hvKp among oysters in Egypt. It underscores the potential role of oysters as a source for disseminating CR-hvKp within aquatic ecosystems, presenting a possible threat to public health.
Subject(s)
Anti-Bacterial Agents , Carbapenems , Klebsiella Infections , Klebsiella pneumoniae , Microbial Sensitivity Tests , Ostreidae , Klebsiella pneumoniae/drug effects , Klebsiella pneumoniae/genetics , Klebsiella pneumoniae/pathogenicity , Klebsiella pneumoniae/isolation & purification , Animals , Egypt/epidemiology , Carbapenems/pharmacology , Klebsiella Infections/microbiology , Klebsiella Infections/epidemiology , Ostreidae/microbiology , Anti-Bacterial Agents/pharmacology , Humans , Virulence , Public Health , Virulence Factors/genetics , Prevalence , Carbapenem-Resistant Enterobacteriaceae/genetics , Carbapenem-Resistant Enterobacteriaceae/isolation & purification , Carbapenem-Resistant Enterobacteriaceae/drug effects , Carbapenem-Resistant Enterobacteriaceae/pathogenicityABSTRACT
Marine bacteria experience fluctuations in osmolarity that they must adapt to, and most bacteria respond to high osmolarity by accumulating compatible solutes also known as osmolytes. The osmotic stress response and compatible solutes used by the coral and oyster pathogen Vibrio coralliilyticus were unknown. In this study, we showed that to alleviate osmotic stress V. coralliilyticus biosynthesized glycine betaine (GB) and transported into the cell choline, GB, ectoine, dimethylglycine, and dimethylsulfoniopropionate, but not myo-inositol. Myo-inositol is a stress protectant and a signaling molecule that is biosynthesized and used by algae. Bioinformatics identified myo-inositol (iol) catabolism clusters in V. coralliilyticus and other Vibrio, Photobacterium, Grimontia, and Enterovibrio species. Growth pattern analysis demonstrated that V. coralliilyticus utilized myo-inositol as a sole carbon source, with a short lag time of 3 h. An iolG deletion mutant, which encodes an inositol dehydrogenase, was unable to grow on myo-inositol. Within the iol clusters were an MFS-type (iolT1) and an ABC-type (iolXYZ) transporter and analyses showed that both transported myo-inositol. IolG and IolA phylogeny among Vibrionaceae species showed different evolutionary histories indicating multiple acquisition events. Outside of Vibrionaceae, IolG was most closely related to IolG from a small group of Aeromonas fish and human pathogens and Providencia species. However, IolG from hypervirulent A. hydrophila strains clustered with IolG from Enterobacter, and divergently from Pectobacterium, Brenneria, and Dickeya plant pathogens. The iol cluster was also present within Aliiroseovarius, Burkholderia, Endozoicomonas, Halomonas, Labrenzia, Marinomonas, Marinobacterium, Cobetia, Pantoea, and Pseudomonas, of which many species were associated with marine flora and fauna.IMPORTANCEHost associated bacteria such as Vibrio coralliilyticus encounter competition for nutrients and have evolved metabolic strategies to better compete for food. Emerging studies show that myo-inositol is exchanged in the coral-algae symbiosis, is likely involved in signaling, but is also an osmolyte in algae. The bacterial consumption of myo-inositol could contribute to a breakdown of the coral-algae symbiosis during thermal stress or disrupt the coral microbiome. Phylogenetic analyses showed that the evolutionary history of myo-inositol metabolism is complex, acquired multiple times in Vibrio, but acquired once in many bacterial plant pathogens. Further analysis also showed that a conserved iol cluster is prevalent among many marine species (commensals, mutualists, and pathogens) associated with marine flora and fauna, algae, sponges, corals, molluscs, crustaceans, and fish.
Subject(s)
Inositol , Multigene Family , Osmotic Pressure , Vibrio , Inositol/metabolism , Animals , Vibrio/metabolism , Vibrio/genetics , Vibrio/physiology , Anthozoa/microbiology , Ostreidae/microbiology , Betaine/metabolism , Bacterial Proteins/genetics , Bacterial Proteins/metabolismABSTRACT
BACKGROUND: Oyster polypeptide (OP) is a mixture of oligopeptides extracted from oysters through enzyme lysis, separation, and purification. It is associated with immunomodulatory effects, but the underlying mechanisms are not known. This study therefore combined proton nuclear magnetic resonance (1H-NMR) urinary metabolomics and 16S rRNA gene sequencing of the gut microbiome to determine the immunoprotective mechanisms of OP in rats subjected to cyclophosphamide-induced immunosuppression. RESULTS: Oyster polypeptide restored the body weight and the structure of spleen and thymus in rats with cyclophosphamide-induced immunosuppression. It upregulated the levels of white blood cells (WBCs), hemoglobin (HGB), platelets (PLT), red blood cells (RBCs), immunoglobulin G (IgG), immunoglobulin M (IgM), cytokines such as interleukin6 (IL-6) and tumor necrosis factor-α (TNF-α), and increased the numbers of CD3+ and CD4+ T cells in the immunosuppressed rats. The 1H-NMR metabolomics results showed that OP significantly reversed the levels of ten metabolites in urine, including 2-oxoglutarate, citrate, dimethylamine, taurine, N-phenylacetylglycine, alanine, betaine, creatinine, uracil, and benzoate. The 16S rRNA gene sequencing results showed that OP restored the gut microbiome homeostasis by increasing the abundance of beneficial bacteria and reducing the abundance of pathogenic bacteria. Finally, a combination of metabolomics and microbiomics found that the metabolism of taurine and hypotaurine, and the metabolism of alanine, aspartate, and glutamate were disturbed, but these metabolic pathways were restored by OP. CONCLUSION: This study demonstrated that OP had immunoprotective effects in rats with cyclophosphamide-induced immunosuppression by restoring key metabolic pathways and the gut microbiome homeostasis. Our findings provide a framework for further research into the immunoregulatory mechanisms of OP and its potential use in drugs and nutritional supplements. © 2024 Society of Chemical Industry.
Subject(s)
Cyclophosphamide , Gastrointestinal Microbiome , Ostreidae , Peptides , Rats, Sprague-Dawley , Animals , Rats , Male , Gastrointestinal Microbiome/drug effects , Peptides/pharmacology , Ostreidae/microbiology , Ostreidae/chemistry , Bacteria/classification , Bacteria/isolation & purification , Bacteria/genetics , Bacteria/drug effects , RNA, Ribosomal, 16S/genetics , Humans , Spleen/drug effects , Spleen/metabolism , Protective Agents/pharmacology , Protective Agents/administration & dosage , Tumor Necrosis Factor-alpha/genetics , Tumor Necrosis Factor-alpha/metabolism , Tumor Necrosis Factor-alpha/immunology , Immunosuppressive Agents/pharmacology , Immunoglobulin G , Interleukin-6/genetics , Interleukin-6/metabolism , Interleukin-6/immunology , Cytokines/metabolism , Cytokines/geneticsABSTRACT
Com o objetivo de avaliar o grau de contaminação por coliformes presentes na água e em ostras e isolar cepas de Escherichia coli para testar seu perfil de resistência aos antibióticos β-lactâmicos, foram coletadas amostras de água e ostras nas regiões de Baixão de Guaí e Capanema, Estuário da Baía do Iguape, BA. A partir das análises microbiológicas de contagem de coliformes à 35ºC e 45ºC, cepas de E. coli foram isoladas e identificadas para realização dos testes de suscetibilidade aos antimicrobianos pertencentes a família dos β-lactâmicos amoxicilina (10μg), ampicilina (10μg), azetronan (30μg), imipenem (10μg), cefalotina (30μg), ceftriaxona (30μg), oxacilina (1μg), utilizando a técnica de difusão de disco em placas. Foi verificado que todos os isolados apresentaram resistência à oxacilina, em contrapartida 100% das cepas de E. coli avaliadas demonstraram perfil de sensibilidade à azetronam, seguidas por 97% ao imepenem e ceftriaxona e 90% à ampicilina.
Subject(s)
Escherichia coli/isolation & purification , Escherichia coli/pathogenicity , Water Microbiology , Ostreidae/microbiology , Drug Resistance, Microbial , Microbiological Techniques/methodsABSTRACT
Vibrio alginolyticus has acquired increasing importance because this microorganism may be pathogenic to aquatic animals and humans. It has been reported that some V. alginolyticus strains carry virulence genes derived from pathogenic V. cholerae and V. parahaemolyticus strains. In this work V. alginolyticus was isolated from oyster samples acquired from a food-market in Mexico City. Thirty isolates were identified as V. alginolitycus. Strains showed β-haemolysis and proteolytic activity and produced a capsule. Strains displayed swimming and swarming motility and 93.3% of them produced siderophores. Several genes encoding virulence factors were detected using PCR amplification. These included proA, wza, vopD, vopB, hcp, vasH and vgrG genes, which were present in all strains. Other genes had a variable representation: tdh (86.6%), lafA (96.6%), pvsA (62%) and pvuA (16%). The trh gene could not be amplified from any of the strains. The antimicrobial resistance profile revealed that more than 90% of the strains were resistant to beta-lactams antibiotics, 60% to cephalotin, 45% to amikacin, 16% to cephotaxime, and 10% to pefloxacin, while 100% were susceptible to ceftriaxone. The V. alginolyticus strains isolated from oysters showed multiple resistance to antibiotics and several virulence factors described in well-characterized pathogenic vibrios (AU)
No disponible
Subject(s)
Vibrio alginolyticus/pathogenicity , Vibrio Infections/drug therapy , Anti-Bacterial Agents/therapeutic use , Ostreidae/microbiology , Drug Resistance, Bacterial , Polysaccharides, Bacterial/immunology , Virulence Factors/analysis , Microbial Sensitivity Tests/statistics & numerical data , Mollusca/microbiology , Food ContaminationABSTRACT
The following study aimed to determine the antimicrobial susceptibility profile of Vibrio parahaemolyticus strains from fresh and frozen oysters Crassostrea rhizophorae sold in Fortaleza-Brazil. An antibiogram was performed on 87 isolates using nine antibiotics: gentamicin (Gen 10 µg), ampicillin (Amp 10 µg), penicillin G (Pen 10U), ciprofloxacin (Cip 5 µg), chloramphenicol (Chl 30 µg), nalidixic acid (Nal 30 µg), tetracycline (Tet 30 µg), vancomycin (Van 30 µg) and erythromycin (Ery 15 µg). All strains were resistant to at least one antibiotic, and 85 (97.7%) were multi-resistant, with predominance of the Van+ Pen+Amp resistance profile (n = 46). Plasmid resistance to Pen, Amp and Ery was detected. Thus, the risk that raw oyster consumption poses to the health of consumers is highlighted, due to the fact that these bivalves may host antibacterial-resistant microorganisms.
O presente estudo objetivou determinar o perfil de suscetibilidade a antimicrobianos de cepas de Vibrio parahaemolyticus oriundas de ostras “in natura” e congeladas comercializadas em Fortaleza-Brasil. Oitenta e sete (87) cepas foram submetidas ao antibiograma com emprego de nove antibióticos: gentamicina (Gen 10 µg), ampicilina (Amp 10 µg), penicilina G (Pen 10U), ciprofloxacin (Cip 5 µg), cloranfenicol (Clo 30 µg), ácido nalidíxico (Nal 30 µg), tetraciclina (Tet 30 µg), vancomicina (Van 30 µg) e eritromicina (Eri 15 µg). Todas as cepas mostram-se resistentes a pelo menos um antibiótico, e 85 (97,7%) apresentaram multirresistência, com predomínio do perfil Van+ Pen+Amp (n = 46). Foi detectada resistência plasmidial a Pen, Amp e Eri. Dessa forma, o risco que o consumo de ostras cruas representa para a saúde dos consumidores merece ser destacado, uma vez que esses bivalves podem ser veículos de transmissão de micro organismos multirresistentes a fármacos antibacterianos.
Subject(s)
Animals , Humans , Anti-Bacterial Agents/pharmacology , Food Microbiology , Ostreidae/microbiology , Shellfish/microbiology , Vibrio parahaemolyticus/drug effects , Brazil , Colony Count, Microbial , Drug Resistance, Multiple, Bacterial , Microbial Sensitivity Tests , Vibrio parahaemolyticus/isolation & purificationABSTRACT
This study investigated the health of natural stocks of the oyster Crassostrea rhizophorae on the southern coast of Bahia in northeastern Brazil, during summer and winter 2010, at three localities (sampling points) in the estuaries of the Maraú (Camamu Bay) and Graciosa rivers. A total of 180 oysters (30/sampling point/season) were examined macroscopically for the presence of pathogens and anatomical changes. The specimens were subsequently fixed in Davidson solution, processed for paraffin embedding, sectioned and stained with Harris' hematoxylin and eosin. Histological analysis revealed the presence of Rickettsia-like organisms (RLOs), Ancistrocoma, Trichodina, Sphenophrya, Nematopsis, Urastoma, Bucephalus in the sporocyst phase, a nonspecific metacercaria, and a metacestode of genus Tylocephalum. The prevalence of infection was low except for parasitism by Nematopsis sp. which also caused histopathological changes. The presence of Bucephalus sp. caused parasitic castration. These two pathogens significantly affect the health of C. rhizophorae.
Este estudo investigou a saúde de ostras da espécie Crassostrea rhizophorae de estoques naturais do Litoral Sul do Estado da Bahia, Nordeste do Brasil, durante o verão e o inverno de 2010, em três pontos amostrais distribuídos nos estuários dos rios Maraú (Baía de Camamu) e Graciosa. Um total de 180 ostras (30/ponto amostral/período) foram examinadas macroscopicamente para a presença de patógenos e alterações anatômicas e posteriormente fixadas em solução de Davidson, processadas para inclusão em parafina, seccionadas e coradas com hematoxilina de Harris e eosina. A análise histológica evidenciou a presença de organismos com características similares a Rickettsia (RLOs), Ancistrocoma, Trichodina, Sphenophrya, Nematopsis, Urastoma, Bucephalus em fase esporocística, metacercária inespecífica e metacestóide de Tylocephalum. As prevalências de infecção foram baixas, com exceção do parasitismo por Nematopsis sp., o qual também causou alterações histopatológicas. A presença de Bucephalus sp. causou castração parasitária. Esses dois patógenos têm interferência significativa na saúde de C. rhizophorae.
Subject(s)
Animals , Ostreidae/microbiology , Ostreidae/parasitology , Brazil , SeasonsABSTRACT
INTRODUCTION: The present study aimed to survey the Vibrio microbiota of oysters (Crassostrea rhizophorae) obtained from restaurants in Fortaleza, State of Ceará, Brazil, and to identify virulence factors. METHODS: The isolated vibrios were submitted to biochemical identification and were tested for hemolytic and urease activities. RESULTS: The isolated strains belonged to 13 species, with predominance of Vibrio mimicus. Of the strain isolates only from fresh samples, 20.5% and 2.8% showed hemolytic and urease activities, respectively. CONCLUSIONS: The findings support the little-publicized claim that Vibrio species other than V. parahaemolyticus and V. vulnificus can represent a health risk to public health.
Subject(s)
Animals , Food Microbiology , Hemolysis , Ostreidae/microbiology , Urease/metabolism , Vibrio/metabolism , Food, Preserved/microbiology , Virulence Factors , Vibrio/classification , Vibrio/isolation & purificationABSTRACT
Vibrio parahaemolyticus is a marine bacterium, responsible for gastroenteritis in humans. Most of the clinical isolates produce thermostable direct hemolysin (TDH) and TDH-related hemolysin (TRH) encoded by tdh and trh genes respectively. In this study, twenty-three V. parahaemolyticus, previously isolated from oysters and mussels were analyzed by PCR using specific primers for the 16S rRNA and virulence genes (tdh, trh and tlh) and for resistance to different classes of antibiotics and PFGE. Nineteen isolates were confirmed by PCR as V. parahaemolyticus. The tlh gene was present in 100 percent of isolates, the tdh gene was identified in two (10.5 percent) isolates, whereas the gene trh was not detected. Each isolate was resistant to at least one of the nine antimicrobials tested. Additionally, all isolates possessed the blaTEM-116 gene. The presence of this gene in V. parahaemolyticus indicates the possibility of spreading this gene in the environment. Atypical strains of V. parahaemolyticus were also detected in this study.
Vibrio parahaemolyticus é uma bactéria marinha, responsável por gastroenterite em humanos. A maioria dos isolados clínicos produzem hemolisina termoestável direta (TDH) e hemolisina TDH-relacionada (TRH) codificadas por genes tdh e trh, respectivamente. Neste estudo, vinte e três V. parahaemolyticus, previamente isolados de ostras e mexilhões foram analisados por PCR utilizando indicadores específicos para o gene 16S rRNA, genes de virulência (tdh, trh e tlh), resistência a diferentes classes de antibióticos, e PFGE. Dezenove isolados foram confirmados por PCR, como V. parahaemolyticus. O gene tlh estava presente em 100 por cento dos isolados, o gene tdh foi identificado em dois (10,5 por cento) dos isolados, enquanto que o gene trh não foi detectado. Cada isolado foi resistente a pelo menos um dos nove antibióticos testados. Além disso, todos os isolados apresentaram resultado positivo para o gene blaTEM-116. A presença deste gene em V. parahaemolyticus indica a possibilidade de propagação desse gene no ambiente. Cepas atípicas de V. parahaemolyticus foram também detectadas neste estudo.
Subject(s)
Animals , Ostreidae/microbiology , Shellfish/microbiology , Vibrio parahaemolyticus/isolation & purification , Anti-Bacterial Agents/pharmacology , Brazil , Bacterial Proteins/genetics , Hemolysin Proteins/genetics , Microbial Sensitivity Tests , Polymerase Chain Reaction , Vibrio parahaemolyticus/drug effects , Vibrio parahaemolyticus/pathogenicity , Virulence Factors/geneticsABSTRACT
La ostra Crassostrea rhizophorae es consumida comúnmente en diversas playas del estado Sucre, sin conocer la carga microbiana que éstas presentan. Por tal motivo se evaluó la calidad microbiológica de la misma y de muestras de agua proveniente de tres estaciones del banco natural de Laguna Grande del Obispo, con el propósito de verificar el cumplimiento de los requisitos microbiológicos establecidos por la Legislación Sanitaria Venezolana y la Administración de Alimentos y Drogas de los Estados Unidos (FDA). Se realizaron muestreos mensuales entre agosto 2005 y julio 2006, determinándose el NMP/100 mL de coliformes totales (CT) y coliformes fecales (CF) en muestras de agua, mientras que en los bivalvos se cuantificó el NMP de CT, CF y Escherichia coli (EC) por gramo de ostra, además de la detección de Salmonella spp. Los resultados obtenidos de CT y CF en muestras de agua de las tres estaciones, cumplieron en la mayoría de los meses, con las exigencias permitidas (70 NMP/100 mL de CT y 14 NMP/100 mL de CF). Durante los meses de muestreo y entre las estaciones analizadas, se observaron variaciones considerables en los valores de CF y EC que estuvieron entre 0 y 2,4 × 10(4) NMP/g de ostra. Los límites permitidos de CF y EC (230 NMP/g de ostra) fueron excedidos por los CF en todas las estaciones durante agosto y diciembre 2005, y también en marzo, mayo y junio 2006, mientras que los de EC se excedieron sólo en diciembre 2005, y marzo y mayo 2006. Salmonella spp. fue detectada en la primera y segunda estación para el mes de septiembre 2005, en la tercera estación para el mes de diciembre del mismo año y en la segunda y tercera estación, para los meses de mayo y julio 2006. Los resultados obtenidos reflejan un alto riesgo de consumir estos moluscos sin haber sido previamente sometidos a una depuración.
In beaches of Sucre State commonly the oyster Crassostrea rhizophorae is consumed without knowing the microbial load microbial that may be presenting. For such motive, the microbiology quality of C. rhizophorae and water samples of three stations from natural bank in Laguna Grande of Obispo was evaluated, to check the compliance of microbiological requirements established by the Food and Drugs Administration of United States (FDA) and Sanitary Venezuelan Legislation. Monthly samplings between August 2005 and July 2006 was realized, determining the MPN/100 mL of coliforms totals (TC) and faecal coliforms (FC) in water samples, whereas in bivalves MPN of TC, FC and Escherichia coli (EC) was quantified for gram of oyster, further to detection Salmonella spp. The results obtained of TC and FC, in water samples of the three stations, fulfilled in majority of the months, with the requirements mentioned above. During the months sampling and between analyzed stations, considerable variations was observed in values of FC and EC that were between 0 and 2.4 × 10(4) MPN/g oyster. The limits allowed for FC and EC (230 MPN/g oyster) were exceeded by the FC on all stations during August, December 2005, and also in March, May and June 2006, whereas EC exceded only in December 2005, and March and May 2006. Salmonella spp. was detected on first and second station in September 2005, on the third station for December of the same year, and second and third station for May and July 2006. The obtained results reflect high risk of consuming these mollusks without having being before submitted to depuration.
Subject(s)
Coliforms/analysis , Crassostrea/microbiology , Escherichia coli , Ostreidae/microbiology , MicrobiologyABSTRACT
A ocorrência de toxinfecções alimentares em virtude do consumo de ostras contaminadas representa risco à saúde pública. Este estudo foi elaborado com o objetivo de avaliar os níveis temporais e espaciais de contaminação por coliformes totais (CT), coliformes termotolerantes (CTM), Staphylococcus coagulase positiva e Salmonella sp. na carne de ostras coletadas em bancos naturais. Durante o período de dezembro de 2005 a abril de 2006, foram analisados quatro pontos amostrais na Baía de Guaratuba, Paraná, perfazendo um total de 12 coletas por ponto. Constatou-se que a contaminação por CT e CTM nos bancos naturais de ostras teve relação direta com o aumento da população flutuante do litoral paranaense durante o verão, quando há um aumento na descarga de esgoto doméstico no estuário. Entre os pontos analisados, a maior contaminação registrada foi próxima da área urbana de Guaratuba, na Marina do Sol. Por outro lado, constatou-se que, no período de uma semana, as ostras passaram por um rápido processo de depuração natural, com redução significativa dos níveis de coliforme.
Subject(s)
Humans , Animals , Coliforms , Food Contamination , Food Microbiology , Food Samples , Ostreidae/microbiology , Shellfish , Salmonella/isolation & purification , Staphylococcus/isolation & purification , BrazilABSTRACT
Este estudo objetivou avaliar a qualidade microbiológica da água do cultivo de ostras, do Estuário do Rio Pacoti, assim como a qualidade das ostras, através do Número Mais Provável (NMP) de coliformes totais (Ct) e termotolerantes (CT). Foram realizadas 15 colheitas, entre junho e novembro de 2006. A água do cultivo se manteve dentro dos limites permitidos pela Legislação vingente. O NMP de Ct/100 mL variou de < 1,8 a 18.000 e de CT/100 mL de < 1,8 a 2.000, enquanto as ostras variaram os Ct e CT/g de < 1,8 a 3.500 e < 1,8 a 2.800, respectivamente. Vinte e cinco cepas de Escherichia coli isoladas da água de cultivo testadas quanto a susceptibilidade a alguns antimicrobianos se mostraram resistentes a ampicilina, nitrofurantoína, tetraciclina, sulfazotrin, ácido nalidíxico, ciprofloxacim, e a imipenem. Das ostras, somente quatro cepas, identificadas como E.coli mostraram-se resistentes a tetraciclina e a imipenem. Baseado nos valores obtidos de Ct e CT da amostra de água e ostras, foi possível constatar que: as águas do Rio Pacoti estão em boas condições segundo a legislação nacional; que a maioria das cepas de E. coli (59,43%), isoladas da água do Rio Pacoti foi sensível aos antimicrobianos testados, exceção do imipenem para o qual as cepas de E.coli apresentaram alto percentual de resistência (80%); a sensibilidade das cepas de E.coli isoladas das amostras de ostras apresentou-se alta à maioria dos antibióticos testados.
This study has been designed to assess the microbiological quality of oyster Crassostrea rhizophorae and of the water used in farms located at the Pacoti River estuary, Ceará State, by means of the MPN of total coliforms (Ct) and thermtolerant coliforms (CT). The database consisted of 15 samples collected from June to November, 2006. The water was kept within the limits set up by the current legislation. MPN values were in the ranges of <1.8 - 18,000 Ct per 100 ml and <1.8 - 2,000 CT per 100 ml, for the water, and <1.8 - 3,500 Ct per gram and <1.8 - 2,800 CT per gram for the oyster itself. Twenty-five strains isolated from the cultivation water and identified as Escherichia coli were tested for susceptibility to a few antimicrobians and proved resistant to ampicillin, nitrofurantoin, tetracycline, sulfazotrin, nalidixic acid, ciprofloxacin and imipenem. From the oysters, four strains were identified as E. coli and proved resistant to tetracycline and imipenem. From the afore-mentioned results, it can be concluded that: Pacoti River waters have received a clean bill of health according to the national legislation; the majority of water-isolated E.coli strains (59.43%) were sensitive to the currently used antimicrobians, except for imipenem to which 80% of the identified E. coli strains proved resistant; sensitivity of the oyster-isolated E. coli strains was found to be high to most of the tested antimicrobians. In all, it is suggested that more clear management measures be made available so as to allow for microbiological quality evaluation of raw-consumed mollusks.
Subject(s)
Animals , Escherichia coli/isolation & purification , Ostreidae/microbiology , Microbial Sensitivity Tests/methodsABSTRACT
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.
Subject(s)
Animals , Humans , Enterobacteriaceae/isolation & purification , Food Microbiology , Ostreidae/microbiology , Seawater/microbiology , Anti-Bacterial Agents/pharmacology , Bacterial Typing Techniques , Enterobacteriaceae/drug effects , Enterobacteriaceae/genetics , Microbial Sensitivity Tests , Proteus mirabilis/drug effects , Proteus mirabilis/genetics , Proteus mirabilis/isolation & purification , RNA, Bacterial , VenezuelaABSTRACT
Vibrio parahaemolyticus (Vp) é uma bactéria naturalmente presente em regiões estuarinas, sendo a principal causa de gastrenterite de origem bacteriana associada a pescados, principalmente ostras cruas. Nesta pesquisa, foi desenvolvida uma avaliação quantitativa de risco para avaliar a probabilidade de Vp causar doença após o consumo de ostra crua, produzida e comercializada no Estado de São Paulo. O estudo incluiu a identificação e caracterização do perigo, a avaliação da exposição e a caracterização do risco. Um modelo matemático foi desenvolvido. Este modelo leva em consideração o comportamento de Vp em ostras na cadeia produtiva, em cada estação do ano, além da relação entre a dose de Vp ingerida e a probabilidade de desenvolver a doença. A avaliação da exposição foi desenvolvida em três etapas: cultivo, pós-coleta e consumo. Na etapa de cultivo foram considerados os fatores que influenciam a prevalência e o número de Vp em ostras no momento da coleta. Na etapa pós-coleta, foram descritas as práticas da indústria e foram considerados os fatores associados ao processamento, transporte e manipulação. Já na etapa de consumo foram considerados os fatores como a quantidade de ostras consumidas por porção, o peso médio por ostra consumida e a população de Vp patogênico no momento do consumo. O resultado do modelo quantitativo da avaliação da exposição foi, então, integrado ao modelo dose-resposta, Beta-Poisson, para se obter uma estimativa do risco. Esta estimativa expressa o impacto da exposição humana a Vp, sobre a saúde pública, associada ao consumo de ostras. A simulação de Monte Carlo foi utilizada para avaliar o efeito da variabilidade e incerteza das variáveis do modelo sobre a estimativa do risco. O modelo prediz uma probabilidade de ocorrência de doença de 4,6x`10 POT.`MENOS`4`, por porção de ostra, consumida ao longo do ano...
Subject(s)
Crassostrea , Food Microbiology , Gastroenteritis/etiology , Ostreidae/microbiology , Vibrio parahaemolyticus , Risk AssessmentABSTRACT
A ostra é um filtrador capaz de ingerir partículas em suspensão, as quais podem carrear microrganismos patogênicos. Desta forma, o hábito de consumir ostras cruas pode causar toxinfecções alimentares em humanos. O presente trabalho teve como objetivo avaliar a qualidade microbiológica de ostras da espécie Crassostrea gigas cultivadas e comercializadas na região litorânea de Florianópolis, através da contagem de coliformes a 35ºC e 45ºC, Escherichia coli e estafilococos coagulase positiva e da pesquisa de Salmonella sp, Víbrio cholerae e Víbrio parahaemolyticus. Foram analisadas 90 (noventa) amostras das quais 45 foram coletadas em estabelecimentos comerciais destinados à venda de frutos do mar e 45 coletadas diretamente do local de cultivo. Todas as análises foram realizadas de acordo com métodos da American Public Health Association. Vibrio cholerae, Víbrio parahaemolyticus e Salmonella sp. não foram encontrados em nenhuma das amostras. Apenas uma amostra apresentou 80 UFC/g de estafilococos coagulase positiva, as demais amostras apresentaram <10 UFC/g. Com o resultado das contagens de coliformes a 35ºC e a 45ºC, evidencia-se contaminação tanto no local de cultivo quanto no local de venda. Escherichia coli foi encontrada em 4 (9 por cento) das amostras provenientes do local de cultivo e em 16 (35,5 por cento) amostras coletadas nos estabelecimentos comerciais. Estes resultados indicam a necessidade de se monitorar a qualidade de ostras cruas, com a implantação de programas de boas práticas de manipulação e manejo dos moluscos.