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1.
BMC Genom Data ; 22(1): 38, 2021 09 27.
Artigo em Inglês | MEDLINE | ID: mdl-34579653

RESUMO

BACKGROUND: The V. parahaemolyticus pandemic clone, results in the development of gastrointestinal illness in humans. Toxigenic strains of this species are frequently isolated from aquatic habitats and organisms such as mollusks and crustaceans. Reports on the isolation of the pandemic clone started in 1996, when a new O3:K6 clone was identified in Asia, that rapidly spread worldwide, becoming the predominant clone isolated from clinical cases. In this study whole genome sequencing was accomplished with an Illumina MiniSeq platform, upon six novel V. parahaemolyticus strains, that have been isolated in Mexico since 1998 and three representative genomes of strains that were isolated from reported outbreaks in other American countries, and were deposited in the GenBank. These nine genomes were compared against the reference sequence of the O3:K6 pandemic strain (RIMD 2210633), which was isolated in 1996, to determine sequence differences within American isolates and between years of isolation. RESULTS: The results indicated that strains that were isolated at different times and from different countries, were highly genetically similar, among them as well as to the reference strain RIMD 2210633, indicating a high level of genetic stability among the strains from American countries between 1996 to 2012, without significant genetic changes relative to the reference strain RIMD 2210633, which was isolated in 1996 and was considered to be representative of a novel O3:K6 pandemic strain. CONCLUSIONS: The genomes of V. parahaemolyticus strains isolated from clinical and environmental sources in Mexico and other American countries, presented common characteristics that have been reported for RIMD 2210633 O3:K6 pandemic strain. The major variations that were registered in this study corresponded to genes non associated to virulence factors, which could be the result of adaptations to different environmental conditions. Nevertheless, results do not show a clear pattern with the year or locality where the strains were isolated, which is an indication of a genomic stability of the studied strains.


Assuntos
Instabilidade Genômica , Pandemias , Vibrioses/epidemiologia , Vibrioses/virologia , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/genética , América/epidemiologia , Surtos de Doenças/estatística & dados numéricos , Humanos , México/epidemiologia , Vibrio parahaemolyticus/isolamento & purificação
2.
Emerg Infect Dis ; 22(7): 1235-7, 2016 07.
Artigo em Inglês | MEDLINE | ID: mdl-27315090

RESUMO

In 2009, an outbreak of Vibrio parahaemolyticus occurred in Piura, Cajamarca, Lambayeque, and Lima, Peru. Whole-genome sequencing of clinical and environmental samples from the outbreak revealed a new V. parahaemolyticus clone. All the isolates identified belonged to a single clonal complex described exclusively in Asia before its emergence in Peru.


Assuntos
Surtos de Doenças , Vibrioses/epidemiologia , Vibrioses/microbiologia , Vibrio parahaemolyticus/classificação , Genoma Bacteriano , Humanos , Epidemiologia Molecular/métodos , Tipagem de Sequências Multilocus , Peru/epidemiologia , Software , Vibrio parahaemolyticus/genética
3.
BMC Genomics ; 16: 176, 2015 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-25880192

RESUMO

BACKGROUND: New strains of Vibrio parahaemolyticus that cause diarrhea in humans by seafood ingestion periodically emerge through continuous evolution in the ocean. Influx and expansion in the Southern Chilean ocean of a highly clonal V. parahaemolyticus (serotype O3:K6) population from South East Asia caused one of the largest seafood-related diarrhea outbreaks in the world. Here, genomics analyses of isolates from this rapidly expanding clonal population offered an opportunity to observe the molecular evolutionary changes often obscured in more diverse populations. RESULTS: Whole genome sequence comparison of eight independent isolates of this population from mussels or clinical cases (from different years) was performed. Differences of 1366 to 217,729 bp genome length and 13 to 164 bp single nucleotide variants (SNVs) were found. Most genomic differences corresponded to the presence of regions unique to only one or two isolates, and were probably acquired by horizontal gene transfer (HGT). Some DNA gain was chromosomal but most was in plasmids. One isolate had a large region (8,644 bp) missing, which was probably caused by excision of a prophage. Genome innovation by the presence of unique DNA, attributable to HGT from related bacteria, varied greatly among the isolates, with values of 1,366 (ten times the number of highest number of SNVs) to 217,729 (a thousand times more than the number of highest number of SNVs). CONCLUSIONS: The evolutionary forces (SNVs, HGT) acting on each isolate of the same population were found to differ to an extent that probably depended on the ecological scenario and life circumstances of each bacterium.


Assuntos
Variação Genética , Genoma Bacteriano , Vibrio parahaemolyticus/genética , Animais , Bivalves/microbiologia , Diarreia/epidemiologia , Diarreia/microbiologia , Transferência Genética Horizontal , Humanos , Pandemias , Plasmídeos/genética , Plasmídeos/metabolismo , Polimorfismo de Nucleotídeo Único , Vibrioses/epidemiologia , Vibrioses/microbiologia , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/isolamento & purificação
4.
Biofouling ; 31(3): 275-82, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25921866

RESUMO

Ballast water is a significant vector of microbial dissemination; however, biofouling on commercial vessel hulls has been poorly studied with regard to pathogenic bacteria transport. Biofouling on three commercial vessels and seven port structures in Ensenada, Baja California, Mexico, was examined by qPCR to identify and quantify Vibrio parahaemolyticus, a worldwide recognized food-borne human pathogen. Pathogenic variants (trh+, tdh+) of V. parahaemolyticus were detected in biofouling homogenates samples from several docks in Ensenada and on the hulls of ships with Japanese and South Korean homeports, but not in reference sampling stations. A total of 26 tdh+ V. parahaemolyticus colonies and 1 ORF8+/O3:K6 strain were also isolated from enriched biofouling homogenate samples confirming the qPCR analysis. Our results suggest that biofouling is an important reservoir of pathogenic vibrios. Thus, ship biofouling might be an overlooked vector with regard to the dissemination of pathogens, primarily pathogenic V. parahaemolyticus.


Assuntos
Incrustação Biológica , Navios , Vibrio parahaemolyticus/isolamento & purificação , Biofilmes , DNA Bacteriano/isolamento & purificação , Reservatórios de Doenças , Genes Bacterianos , México , Reação em Cadeia da Polimerase em Tempo Real , Sorotipagem , Vibrio parahaemolyticus/classificação
5.
PLoS One ; 10(2): e0117485, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-25679989

RESUMO

Vibrio parahaemolyticus is an important human pathogen whose transmission is associated with the consumption of contaminated seafood. Consistent multilocus sequence typing for V. parahaemolyticus has shown difficulties in the amplification of the recA gene by PCR associated with a lack of amplification or a larger PCR product than expected. In one strain (090-96, Peru, 1996), the produced PCR product was determined to be composed of two recA fragments derived from different Vibrio species. To better understand this phenomenon, we sequenced the whole genome of this strain. The hybrid recA gene was found to be the result of a fragmentation of the original lineage-specific recA gene resulting from a DNA insertion of approximately 30 kb in length. This insert had a G+C content of 38.8%, lower than that of the average G+C content of V. parahaemolyticus (45.2%), and contained 19 ORFs, including a complete recA gene. This new acquired recA gene deviated 24% in sequence from the original recA and was distantly related to recA genes from bacteria of the Vibrionaceae family. The reconstruction of the original recA gene (recA3) identified the precursor as belonging to ST189, a sequence type reported previously only in Asian countries. The identification of this singular genetic feature in strains from Asia reveals new evidence for genetic connectivity between V. parahaemolyticus populations at both sides of the Pacific Ocean that, in addition to the previously described pandemic clone, supports the existence of a recurrent transoceanic spreading of pathogenic V. parahaemolyticus with the corresponding potential risk of pandemic expansion.


Assuntos
Recombinases Rec A/genética , Vibrioses/microbiologia , Vibrioses/transmissão , Vibrio parahaemolyticus/genética , Biologia Computacional , Ordem dos Genes , Variação Genética , Genoma Bacteriano , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Tipagem de Sequências Multilocus , Oceano Pacífico , Peru , Filogenia , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/isolamento & purificação
6.
Mol Biol Evol ; 32(6): 1396-410, 2015 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-25605790

RESUMO

We investigated global patterns of variation in 157 whole-genome sequences of Vibrio parahaemolyticus, a free-living and seafood associated marine bacterium. Pandemic clones, responsible for recent outbreaks of gastroenteritis in humans, have spread globally. However, there are oceanic gene pools, one located in the oceans surrounding Asia and another in the Mexican Gulf. Frequent recombination means that most isolates have acquired the genetic profile of their current location. We investigated the genetic structure in the Asian gene pool by calculating the effective population size in two different ways. Under standard neutral models, the two estimates should give similar answers but we found a 27-fold difference. We propose that this discrepancy is caused by the subdivision of the species into a hundred or more ecotypes which are maintained stably in the population. To investigate the genetic factors involved, we used 51 unrelated isolates to conduct a genome-wide scan for epistatically interacting loci. We found a single example of strong epistasis between distant genome regions. A majority of strains had a type VI secretion system associated with bacterial killing. The remaining strains had genes associated with biofilm formation and regulated by cyclic dimeric GMP signaling. All strains had one or other of the two systems and none of isolate had complete complements of both systems, although several strains had remnants. Further "top down" analysis of patterns of linkage disequilibrium within frequently recombining species will allow a detailed understanding of how selection acts to structure the pattern of variation within natural bacterial populations.


Assuntos
Pool Gênico , Genética Populacional , Genoma Bacteriano , Vibrio parahaemolyticus/genética , Vibrio parahaemolyticus/isolamento & purificação , Ásia , Biofilmes , Cromossomos Bacterianos/genética , Epistasia Genética , Loci Gênicos , México , Oceanos e Mares , Filogenia , Filogeografia , Polimorfismo de Nucleotídeo Único , Recombinação Genética , Água do Mar/microbiologia , Análise de Sequência de DNA , Vibrio parahaemolyticus/classificação
7.
Appl Environ Microbiol ; 81(5): 1689-99, 2015 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-25548045

RESUMO

Moribund shrimp affected by acute hepatopancreatic necrosis disease (AHPND) from farms in northwestern Mexico were sampled for bacteriological and histological analysis. Bacterial isolates were molecularly identified as Vibrio parahaemolyticus by the presence of the tlh gene. The tdh-negative, trh-negative, and tlh-positive V. parahaemolyticus strains were further characterized by repetitive extragenic palindromic element-PCR (rep-PCR), and primers AP1, AP2, AP3, and AP and an ems2 IQ2000 detection kit (GeneReach, Taiwan) were used in the diagnostic tests for AHPND. The V. parahaemolyticus strains were used in immersion challenges with shrimp, and farmed and challenged shrimp presented the same clinical and pathological symptoms: lethargy, empty gut, pale and aqueous hepatopancreas, and expanded chromatophores. Using histological analysis and bacterial density count, three stages of AHNPD (initial, acute, and terminal) were identified in the affected shrimp. The pathognomonic lesions indicating severe desquamation of tubular epithelial cells of the hepatopancreas were observed in both challenged and pond-infected shrimp. The results showed that different V. parahaemolyticus strains have different virulences; some of the less virulent strains do not induce 100% mortality, and mortality rates also rise more slowly than they do for the more virulent strains. The virulence of V. parahaemolyticus strains was dose dependent, where the threshold infective density was 10(4) CFU ml(-1); below that density, no mortality was observed. The AP3 primer set had the best sensitivity and specificity. Field and experimental results showed that the V. parahaemolyticus strain that causes AHPND acts as a primary pathogen for shrimp in Mexico compared with the V. parahaemolyticus strains reported to date.


Assuntos
Penaeidae/microbiologia , Vibrioses/veterinária , Vibrio parahaemolyticus/isolamento & purificação , Estruturas Animais/microbiologia , Estruturas Animais/patologia , Animais , Técnicas Bacteriológicas , México , Técnicas de Diagnóstico Molecular , Reação em Cadeia da Polimerase , Vibrioses/microbiologia , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/genética
8.
J Food Prot ; 77(7): 1069-77, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24988011

RESUMO

The abundance of total and pathogenic Vibrio parahaemolyticus (Vp) strains in American oysters (Crassostrea virginica) harvested in two different harvest sites from the Mandinga lagoon System was evaluated monthly for 1 year (January through December 2012). Frequencies of species-specific genes and pathogenic genes exhibited a seasonal distribution. The annual occurrence of Vp with the species-specific tlh gene (tlh(+)) was significantly higher during the winter windy season (32.50%) and spring dry season (15.0%), with the highest densities observed during spring dry season at 283.50 most probable number (MPN)/g (lagoon bank A, near human settlements), indicating the highest risk of infection during warmer months. Pathogenic Vp tlh(+)/tdh(+) frequency was significantly higher during the winter windy and the spring dry seasons at 22.50 and 10.00%, respectively, with highest densities of 16.22 and 41.05 MPN/g (bank A), respectively. The tlh/trh and tdh/trh gene combinations were also found in Vp isolates during the spring dry season at 1.25 and 1.3%, respectively, with densities of 1.79 and 0.4 MPN/g (bank A), respectively. The orf8 genes were detected during the winter windy season (1.25%) with highest densities of 5.96 MPN/g (bank A) and 3.21 MPN/g (bank B, near mangrove islands and a heron nesting area). Densities of Vp tdh(+) were correlated (R(2) = 0.245, P < 0.015) with those of Vp orf8(+). The seasonal dynamics of Vp harboring pathogenic genes varied with seasonal changes, with very high proportions of Vp tdh(+) and Vp orf8(+) isolates in the winter windy season at 46.2 and 17.0%, respectively, which suggests that environmental factors may differentially affect the abundance of pathogenic subpopulations. Although all densities of total Vp (Vp tlh(+)) were lower than 10(4) MPN/g, thus complying with Mexican regulations, the presence of pathogenic strains is a public health concern. Our results suggest that total Vp densities may not be appropriate for assessing oyster contamination and predicting the risk of infection. Evaluation of the presence of pathogenic strains would be a better approach to protecting public health.


Assuntos
Crassostrea/microbiologia , Contaminação de Alimentos/análise , Frutos do Mar/microbiologia , Vibrio parahaemolyticus/isolamento & purificação , Animais , Proteínas de Bactérias/genética , Crassostrea/crescimento & desenvolvimento , Inocuidade dos Alimentos , Humanos , México , Estações do Ano , Estados Unidos , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/genética
9.
Front Cell Infect Microbiol ; 3: 110, 2014 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-24427744

RESUMO

Vibrio parahaemolyticus is one of the most important seafood-borne bacterial in recent years and is the leading causal agent of human acute gastroenteritis, primarily following the consumption of raw, undercooked or mishandled marine products. Until 1996, infections caused by V. parahaemolyticus were generally associated with diverse serovars. However, in February 1996, a unique serovar (O3:K6) of V. parahaemolyticus with specific genetic markers (tdh, toxRS/New and/or orf8) appeared abruptly in Kolkata, India. In subsequent years, O3:K6 isolates similar to those isolated in Kolkata have been reported from food borne outbreaks in Southeast Asia, as well as in the Atlantic and Gulf coasts of the United States (U.S). More recently, there have been reports in Europe, Africa and Central and South America. Specifically, in the American continent, some countries have reported cases of gastroenteritis due to the pandemic O3:K6 strain and its serovariants; the pandemic strain was first detected in Peru (1996, >100 cases), subsequently spreading to Chile in 1998 (>16,804 human cases), to the U.S. in 1998 (>700 cases), to Brazil in 2001 (>18 cases) and to Mexico in 2004 (>1200 cases). The arrival of the pandemic clone on the American continent may have resulted in a significant shift on the epidemic dynamics of V. parahaemolyticus. However, although O3:K6 is the predominant serovar of the recognized clinical strains in some countries in the Americas, a decrease in clinical cases caused by O3:K6 and an increase in cases associated with a new serotype (O3:K59, Chile) have been recently reported. The emergence and worldwide dissemination of O3:K6 and other pandemic strains since 1996 have come to represent a threat to public health and should concern health authorities. This review focuses on the presence, distribution and virulence factors of the V. parahaemolyticus O3:K6 pandemic clone and its serovariants in clinical and environmental strains on the American continent.


Assuntos
Pandemias , Vibrioses/epidemiologia , Vibrioses/microbiologia , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/isolamento & purificação , América/epidemiologia , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/microbiologia , Gastroenterite/epidemiologia , Gastroenterite/microbiologia , Humanos , Sorotipagem
10.
Rev. argent. microbiol ; Rev. argent. microbiol;45(4): 267-70, dic. 2013.
Artigo em Espanhol | LILACS, BINACIS | ID: biblio-1171794

RESUMO

The enzymatic characterization of vibrios has been used as a virulence indicator of sanitary interest. The objective of this study was to determine the enzymatic profile of Vibrio parahaemolyticus strains (n=70) isolated from Crassostrea rhizophorae oysters. The strains were examined for the presence of gelatinase (GEL), caseinase (CAS), elastase (ELAS), phospholipase (PHOS), lipase (LIP), amilase (AML) and DNase. All enzymes, except elastase, were detected in more than 60


of the strains. The most recurrent enzymatic profiles were AML + DNase + PHOS + GEL + LIP (n=16; 22.9


) and AML + CAS + DNase + PHOS + GEL + LIP (n=21; 30


). Considering the fact that exoenzyme production by vibrios is closely related to virulence, one must be aware of the bacteriological risk posed to human health by the consumption of raw or undercooked oysters.


Assuntos
Crassostrea/microbiologia , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/enzimologia , Animais , Vibrio parahaemolyticus/isolamento & purificação
11.
PLoS Negl Trop Dis ; 7(5): e2210, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23696906

RESUMO

Vibrio parahaemolyticus is a foodborne pathogen that has become a public health concern at the global scale. The epidemiological significance of V. parahaemolyticus infections in Latin America received little attention until the winter of 1997 when cases related to the pandemic clone were detected in the region, changing the epidemic dynamics of this pathogen in Peru. With the aim to assess the impact of the arrival of the pandemic clone on local populations of pathogenic V. parahaemolyticus in Peru, we investigated the population genetics and genomic variation in a complete collection of non-pandemic strains recovered from clinical sources in Peru during the pre- and post-emergence periods of the pandemic clone. A total of 56 clinical strains isolated in Peru during the period 1994 to 2007, 13 strains from Chile and 20 strains from Asia were characterized by Multilocus Sequence Typing (MLST) and checked for the presence of Variable Genomic Regions (VGRs). The emergence of O3:K6 cases in Peru implied a drastic disruption of the seasonal dynamics of infections and a shift in the serotype dominance of pathogenic V. parahaemolyticus. After the arrival of the pandemic clone, a great diversity of serovars not previously reported was detected in the country, which supports the introduction of additional populations cohabitating with the pandemic group. Moreover, the presence of genomic regions characteristic of the pandemic clone in other non-pandemic strains may represent early evidence of genetic transfer from the introduced population to the local communities. Finally, the results of this study stress the importance of population admixture, horizontal genetic transfer and homologous recombination as major events shaping the structure and diversity of pathogenic V. parahaemolyticus.


Assuntos
Variação Genética , Vibrioses/epidemiologia , Vibrioses/microbiologia , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/genética , Análise por Conglomerados , DNA Bacteriano/química , DNA Bacteriano/genética , Doenças Transmitidas por Alimentos/epidemiologia , Doenças Transmitidas por Alimentos/microbiologia , Humanos , Epidemiologia Molecular , Dados de Sequência Molecular , Tipagem de Sequências Multilocus , Peru/epidemiologia , Sorotipagem , Vibrio parahaemolyticus/isolamento & purificação
12.
Rev Argent Microbiol ; 45(4): 267-70, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-24401782

RESUMO

The enzymatic characterization of vibrios has been used as a virulence indicator of sanitary interest. The objective of this study was to determine the enzymatic profile of Vibrio parahaemolyticus strains (n=70) isolated from Crassostrea rhizophorae oysters. The strains were examined for the presence of gelatinase (GEL), caseinase (CAS), elastase (ELAS), phospholipase (PHOS), lipase (LIP), amilase (AML) and DNase. All enzymes, except elastase, were detected in more than 60% of the strains. The most recurrent enzymatic profiles were AML + DNase + PHOS + GEL + LIP (n=16; 22.9%) and AML + CAS + DNase + PHOS + GEL + LIP (n=21; 30%). Considering the fact that exoenzyme production by vibrios is closely related to virulence, one must be aware of the bacteriological risk posed to human health by the consumption of raw or undercooked oysters.


Assuntos
Crassostrea/microbiologia , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/enzimologia , Animais , Vibrio parahaemolyticus/isolamento & purificação
13.
Rev. argent. microbiol ; 45(4): 267-70, 2013 Oct-Dec.
Artigo em Espanhol | BINACIS | ID: bin-132750

RESUMO

The enzymatic characterization of vibrios has been used as a virulence indicator of sanitary interest. The objective of this study was to determine the enzymatic profile of Vibrio parahaemolyticus strains (n=70) isolated from Crassostrea rhizophorae oysters. The strains were examined for the presence of gelatinase (GEL), caseinase (CAS), elastase (ELAS), phospholipase (PHOS), lipase (LIP), amilase (AML) and DNase. All enzymes, except elastase, were detected in more than 60


of the strains. The most recurrent enzymatic profiles were AML + DNase + PHOS + GEL + LIP (n=16; 22.9


) and AML + CAS + DNase + PHOS + GEL + LIP (n=21; 30


). Considering the fact that exoenzyme production by vibrios is closely related to virulence, one must be aware of the bacteriological risk posed to human health by the consumption of raw or undercooked oysters.


Assuntos
Crassostrea/microbiologia , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/enzimologia , Animais , Vibrio parahaemolyticus/isolamento & purificação
14.
PLoS One ; 7(1): e30823, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22292049

RESUMO

The emergence of the pandemic strain Vibrio parahaemolyticus O3:K6 in 1996 caused a large increase of diarrhea outbreaks related to seafood consumption in Southeast Asia, and later worldwide. Isolates of this strain constitutes a clonal complex, and their effectual differentiation is possible by comparison of their variable number tandem repeats (VNTRs). The differentiation of the isolates by the differences in VNTRs will allow inferring the population dynamics and microevolution of this strain but this requires knowing the rate and mechanism of VNTRs' variation. Our study of mutants obtained after serial cultivation of clones showed that mutation rates of the six VNTRs examined are on the order of 10(-4) mutant per generation and that difference increases by stepwise addition of single mutations. The single stepwise mutation (SSM) was deduced because mutants with 1, 2, 3, or more repeat unit deletions or insertions follow a geometric distribution. Plausible phylogenetic trees are obtained when, according to SSM, the genetic distance between clusters with different number of repeats is assessed by the absolute differences in repeats. Using this approach, mutants originated from different isolates of pandemic V. parahaemolyticus after serial cultivation are clustered with their parental isolates. Additionally, isolates of pandemic V. parahaemolyticus from Southeast Asia, Tokyo, and northern and southern Chile are clustered according their geographical origin. The deepest split in these four populations is observed between the Tokyo and southern Chile populations. We conclude that proper phylogenetic relations and successful tracing of pandemic V. parahaemolyticus requires measuring the differences between isolates by the absolute number of repeats in the VNTRs considered.


Assuntos
Evolução Molecular , Repetições Minissatélites/genética , Vibrioses/epidemiologia , Vibrioses/microbiologia , Vibrio parahaemolyticus/genética , Variação Antigênica/genética , Técnicas de Tipagem Bacteriana/métodos , DNA Bacteriano/análise , DNA Bacteriano/genética , Variação Genética , Humanos , Repetições de Microssatélites/genética , Taxa de Mutação , Pandemias , Filogenia , Vibrio parahaemolyticus/classificação
15.
Appl Environ Microbiol ; 78(6): 1794-803, 2012 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-22247160

RESUMO

In 2004, more than 1,230 cases of gastroenteritis due to pandemic O3:K6 strains of Vibrio parahaemolyticus were reported in southern Sinaloa, a state in Northwestern Mexico. Recurrent sporadic cases arose from 2004 to 2010, spreading from the south to the north. In the present study, Vibrio parahaemolyticus was detected in both environmental samples and clinical cases along the Pacific coast of Sinaloa during 2004 to 2010. An evaluation was made of the serotypes, distribution of virulence genes, and presence of pandemic O3:K6 strains. A total of 144 strains were isolated from environmental samples (from sediment, seawater, and shrimp), and 154 clinical strains were isolated. A total of 10 O serogroups and 30 serovars were identified in the strains. Environmental strains (n = 144) belonged to 10 O serogroups and 28 serovars, while clinical strains (n = 154) belonged to 8 O serogroups and 14 serovars. Ten serovars were shared by both environmental and clinical strains. Among 144 environmental isolates, 4.1% (6/144) belonged to the pandemic clone, with 83.3% containing the orf8 gene and with O3:K6 accounting for 67%. On the other hand, pathogenic strains (tdh and/or trh) accounted for 52% (75/144) of the environmental isolates. Interestingly, among 154 clinical isolates, 80.5% (124/154) were pandemic strains, with O3:K6 (tdh, toxRS(new), and orf8) representing the predominant serovar (99.2%, 123/124). Overall, our results indicate that in spite of a high serodiversity and prevalence of pathogenic Vibrio parahaemolyticus in the environment, the pandemic strain O3:K6 caused >79% of reported cases between 2004 and 2010 in Sinaloa, Mexico.


Assuntos
Microbiologia Ambiental , Gastroenterite/epidemiologia , Gastroenterite/microbiologia , Vibrioses/epidemiologia , Vibrioses/microbiologia , Vibrio parahaemolyticus/isolamento & purificação , Análise por Conglomerados , Diarreia/epidemiologia , Diarreia/microbiologia , México/epidemiologia , Recidiva , Sorotipagem , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/genética , Vibrio parahaemolyticus/imunologia , Fatores de Virulência/genética
16.
J Infect Dev Ctries ; 5(7): 502-10, 2011 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-21795818

RESUMO

INTRODUCTION: Vibrio (V.) parahaemolyticus has endemically established in Chilean sea shores, causing outbreaks every year, with an important number of cases. In order to know the genetic relationship, genotype dominance and antibiotic resistance of isolates obtained from two outbreaks, this study characterized 110 strains isolated from environmental and clinical samples in years 2005 and 2007 in Chile. METHODOLOGY: Genotyping was performed by determination of PFGE profiles, and pandemic group and integrons were screened by PCR. Antimicrobial susceptibility was studied by the disk diffusion method. RESULTS: High antibiotic susceptibility frequency was found, mainly among 2007 isolates, except to ampicillin, cephalothin, cefoxitin, cefpodoxime, amikacin, streptomycin and kanamycin. Strains belonging to the pandemic group in clinical isolates account for 88% in 2005, decreasing to 66% in 2007 and among environmental isolates were detected in 20% of the strains from 2005, rising to 36% in 2007. In 2005, nine different PFGE profiles were identified, with 78% of the strains corresponding to a single clone. In 2007, sixteen different PFGE profiles were detected, with 61% of the strains included into a sole clone. The same clone was prevalent in both years. None of class 1, 2, 3 and SXT integrases genes was detected; however, the superintegron integrase gene (intIA) was present in almost all strains. CONCLUSIONS: These results suggest the persistence and dominance of a unique PFGE clone of V. parahaemolyticus during 2005 and 2007, and the absence of genetic elements that capture antibiotic resistance genes described in other species of Vibrio.


Assuntos
Surtos de Doenças , Doenças Endêmicas , Microbiologia Ambiental , Vibrioses/epidemiologia , Vibrioses/microbiologia , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/efeitos dos fármacos , Antibacterianos/farmacologia , Chile/epidemiologia , Análise por Conglomerados , DNA Bacteriano/genética , Eletroforese em Gel de Campo Pulsado , Genótipo , Humanos , Integrons , Testes de Sensibilidade Microbiana , Epidemiologia Molecular , Tipagem Molecular , Fenótipo , Reação em Cadeia da Polimerase , Vibrio parahaemolyticus/genética , Vibrio parahaemolyticus/isolamento & purificação
17.
Mol Ecol ; 19(18): 3924-37, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20735744

RESUMO

The dynamics of dissemination of the environmental human pathogen Vibrio parahaemolyticus are uncertain. The O3:K6 clone was restricted to Asia until its detection along the Peruvian coasts and in northern Chile in 1997 in phase with the arrival of El Niño waters. A subsequent emergence of O3:K6 strains was detected in austral Chile in 2004. The origin of these 1997 and 2004 population radiations has not yet been conclusively determined. Multiple loci VNTR analysis using seven polymorphic loci was carried out with a number of representative strains from Asia, Peru and Chile to determine their genetic characteristics and population structure. Asian and Chilean subpopulations were the most genetically distant groups with an intermediate subpopulation in Peru. Population structure inferred from a minimum-spanning tree and Bayesian analysis divided the populations into two genetically distinct groups, consistent with the epidemic dynamics of the O3:K6 clone in South America. One group comprised strains from the original Asiatic population and strains arriving in Peru and Chile in 1997. The second group included the remaining Peruvian Strains and Chilean strains obtained from Puerto Montt in 2004. The analysis of the arrival of the O3:K6 clone at the Pacific coasts of South America has provided novel insights linking the origin of the invasion in 1997 to Asian populations and describing the successful establishment of the O3:K6 populations, first in Peru and subsequently in the South of Chile owing to a possible radiation of Peruvian populations.


Assuntos
Variação Genética , Repetições Minissatélites , Vibrio parahaemolyticus/genética , Algoritmos , Ásia , Teorema de Bayes , Chile , Análise por Conglomerados , DNA Bacteriano/genética , Genética Populacional , Genótipo , Desequilíbrio de Ligação , Repetições de Microssatélites , Modelos Genéticos , Peru , Filogenia , Análise de Sequência de DNA , Vibrio parahaemolyticus/classificação
18.
Appl Environ Microbiol ; 75(23): 7482-7, 2009 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-19801458

RESUMO

Seafood consumption-related diarrhea became prevalent in Chile when the pandemic strain of Vibrio parahaemolyticus serotype O3:K6 reached a region in the south of Chile (Region de los Lagos) where approximately 80% of the country's seafood is produced. In spite of the large outbreaks of clinical infection, the load of V. parahaemolyticus in shellfish of this region is relatively low. The pandemic strain constitutes a small but relatively stable group of a diverse V. parahaemolyticus population, composed of at least 28 genetic groups. Outbreaks in Region de los Lagos began in 2004 and reached a peak in 2005 with 3,725 clinical cases, all associated with the pandemic strain. After 2005, reported cases steadily decreased to a total of 477 cases in 2007. At that time, 40% of the clinical cases were associated with a pandemic strain of a different serotype (O3:K59), and 27% were related to V. parahaemolyticus isolates unrelated to the pandemic strain. In the results published here, we report that in the summer of 2008, when reported cases unexpectedly increased from 477 to 1,143, 98% of the clinical cases were associated with the pandemic strain serotype O3:K6, a change from 2007. Nevertheless, in 2009, when clinical cases decreased to 441, only 64% were related to the pandemic strain; the remaining cases were related to a nonpandemic tdh- and trh-negative strain first identified in shellfish in 2006. Overall, our observations indicate that the pandemic strain has become a relatively stable subpopulation and that when the number of diarrhea cases related to the pandemic strain is low, previously undetected V. parahaemolyticus pathogenic strains become evident.


Assuntos
Diarreia/epidemiologia , Surtos de Doenças , Microbiologia Ambiental , Doenças Transmitidas por Alimentos/epidemiologia , Alimentos Marinhos/microbiologia , Vibrioses/epidemiologia , Vibrio parahaemolyticus/isolamento & purificação , Técnicas de Tipagem Bacteriana , Chile/epidemiologia , Diarreia/microbiologia , Doenças Transmitidas por Alimentos/microbiologia , Incidência , Sorotipagem , Vibrioses/microbiologia , Vibrio parahaemolyticus/classificação
19.
Appl Environ Microbiol ; 75(12): 4079-88, 2009 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-19376898

RESUMO

Epidemics of Vibrio parahaemolyticus in Chile have occurred since 1998. Direct genome restriction enzyme analysis (DGREA) using conventional gel electrophoresis permitted discrimination of different V. parahaemolyticus isolates obtained from these outbreaks and showed that this species consists of a highly diverse population. A multiple-locus variable-number tandem-repeat (VNTR) analysis (MLVA) approach was developed and applied to 22 clinical and 91 environmental V. parahaemolyticus isolates from Chile to understand their clonal structures. To this end, an advanced molecular technique was developed by applying multiplex PCR, fluorescent primers, and capillary electrophoresis, resulting in a high-resolution and high-throughput (HRHT) genotyping method. The genomic basis of this HRHT method was eight VNTR loci described previously by Kimura et al. (J. Microbiol. Methods 72:313-320, 2008) and two new loci which were identified by a detailed molecular study of 24 potential VNTR loci on both chromosomes. The isolates of V. parahaemolyticus belonging to the same DGREA pattern were distinguishable by the size variations in the indicative 10 VNTRs. This assay showed that these 10 VNTR loci were useful for distinguishing isolates of V. parahaemolyticus that had different DGREA patterns and also isolates that belong to the same group. Isolates that differed in their DGREA patterns showed polymorphism in their VNTR profiles. A total of 81 isolates was associated with 59 MLVA groups, providing fine-scale differentiation, even among very closely related isolates. The developed approach enables rapid and high-resolution analysis of V. parahaemolyticus with pandemic potential and provides a new surveillance tool for food-borne pathogens.


Assuntos
Técnicas de Tipagem Bacteriana/métodos , Eletroforese Capilar/métodos , Repetições Minissatélites , Vibrioses/microbiologia , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/genética , Chile/epidemiologia , Análise por Conglomerados , Impressões Digitais de DNA , DNA Bacteriano/genética , Genótipo , Humanos , Epidemiologia Molecular/métodos , Polimorfismo Genético , Sensibilidade e Especificidade , Vibrioses/epidemiologia , Vibrio parahaemolyticus/isolamento & purificação
20.
Emerg Infect Dis ; 15(2): 163-8, 2009 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-19193258

RESUMO

Disease outbreaks caused by Vibrio parahaemolyticus in Puerto Montt, Chile, began in 2004 and reached a peak in 2005 at 3,600 clinical cases. Until 2006, every analyzed case was caused by the serovar O3:K6 pandemic strain. In the summer of 2007, only 475 cases were reported; 73% corresponded to the pandemic strain. This decrease was associated with a change in serotype of many pandemic isolates to O3:K59 and the emergence of new clinical strains. One of these strains, associated with 11% of the cases, was genotypically different from the pandemic strain but contained genes that were identical to those found on its pathogenicity island. These findings suggest that pathogenicity-related genes were laterally transferred from the pandemic strain to one of the different V. parahaemolyticus groups comprising the diverse and shifting bacterial population in shellfish in this region.


Assuntos
Surtos de Doenças , Frutos do Mar/microbiologia , Vibrioses/epidemiologia , Animais , Chile/epidemiologia , Humanos , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Sorotipagem , Vibrioses/microbiologia , Vibrio parahaemolyticus/classificação , Vibrio parahaemolyticus/genética , Vibrio parahaemolyticus/isolamento & purificação , Vibrio parahaemolyticus/patogenicidade
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