K6 Strains Research Articles

Production of S-methyl-methionine using engineered Saccharomyces cerevisiae sake K6.

Introducing a plant-derived MMT gene encoding methionine S-methyltransferase into engineered Saccharomyces cerevisiae sake K6 allowed microbial production of S-methyl-methionine (SMM).

Genomic Study of Chromosomally and Plasmid-Mediated Multidrug Resistance and Virulence Determinants in Klebsiella Pneumoniae Isolates Obtained from a Tertiary Hospital in Al-Kharj, KSA.

Klebsiella pneumoniae is an emergent pathogen causing respiratory tract, bloodstream, and urinary tract infections in humans. This study defines the genomic sequence data, genotypic and phenotypic characterization of K. pneumoniae clinically isolated from Al-Kharj, KSA. Whole-genome analysis of four K. pneumoniae strains was performed, including de novo assembly, functional annotation, whole-genome-phylogenetic analysis, antibiotic-resistant gene identification, prophage regions, virulent factor, and pan-genome analysis. The results showed that K6 and K7 strains were MDR and ESBL producers, K16 was an ESBL producer, and K8 was sensitive to all tested drugs except ampicillin. K6 and K7 were identified with sequence type (ST) 23, while K16 and K8 were identified with STs 353 and 592, respectively. K6 and K7 were identified with the K1 (wzi1 genotype) capsule and O1 serotype, while K8 was identified with the K57 (wzi206 genotype) capsule and O3b. K6 isolates harbored 10 antimicrobial resistance genes (ARGs) associated with four different plasmids; the chloramphenicol acetyltransferase (catB3), blaOXA-1 and aac(6')-Ib-cr genes were detected in plasmid pB-8922_OXA-48. K6 and K7 also carried a similar gene cassette in plasmid pC1K6P0122-2; the gene cassettes were the trimethoprim-resistant gene (dfrA14), integron integrase (IntI1), insertion sequence (IS1), transposase protein, and replication initiation protein (RepE). Two hypervirulent plasmids were reported in isolates K6 and K7 that carried synthesis genes (iucA, iucB, iucC, iucD, and iutA) and iron siderophore genes (iroB, iroC, iroD, and iroN). The presence of these plasmids in high-risk clones suggests their dissemination in our region, which represents a serious health problem.

Vibrio parahaemolyticus Is Associated with Diarrhea Cases in Mexico, with a Dominance of Pandemic O3:K6 Clones.

In the present study, we conducted surveillance of the V. parahaemolyticus strains present in clinical samples from six geographical regions of Mexico (22 states) from 2004 to 2011. The serotype dominance, virulence genes, presence of pandemic O3:K6 strains, and antibiotic resistance of the isolates were investigated. In total, 144 strains were isolated from the clinical samples. Seven different O serogroups and twenty-five serovars were identified. Most clinical isolates (66%, 95/144) belonged to the pandemic clone O3:K6 (tdh+, toxRS/new+ and/or orf8+) and were detected in 20 of the 22 states. Among the pandemic clones, approximately 17.8% (17/95) of the strains cross-reacted with the antisera for the K6 and K59 antigens (O3:K6, K59 serotype). Other pathogenic strains (tdh+ and/or trh+, toxRS/new−, orf8−) accounted for 26.3%, and the nonpathogenic strains (tdh− and/or trh−) accounted for 7.6%. Antimicrobial susceptibility testing showed that most of the strains were resistant to ampicillin (99.3%) but were sensitive to most tested antibiotics. The level of multidrug resistance was 1.3%. Our results indicate that pandemic O3:K6 is present in most Mexican states, thus, constant surveillance of V. parahaemolyticus strains in diarrhea patients is a public health priority and is useful for conducting risk assessments of foodborne illnesses to prevent V. parahaemolyticus outbreaks. Overall, our observations indicate that the pandemic O3:K6 clone of V. parahaemolyticus has become a relatively stable subpopulation and may be endemically established in Mexico; therefore, constant surveillance is needed to avoid new outbreaks of this pathogen.

Emergence and predominance of a new serotype of Vibrio parahaemolyticus in Huzhou, China

BackgroundDuring our surveillance period, we found a new type of Vibrio parahaemolyticus (V. parahaemolyticus) with serotype O10:K4, which had increased over the past 2 years in Huzhou and became the second most common serotype after O3:K6. MethodsStrains were isolated from stool samples of diarrhea patients in the First People's Hospital in Huzhou. The serotypes, virulence-associated genes, antimicrobial susceptibility, and pulsed field gel electrophoresis (PFGE) of these strains were analyzed. ResultsBetween January 2017 and December 2021, there were 598 (5.36%) V. parahaemolyticus-positive samples of 11,166 stool specimens. The V. parahaemolyticus detection rate was high in summer months. The O3:K6 was the dominant serotype in 2017–2020 and a new serotype, O10:K4, was the predominant serotype in 2021. The majority of isolates tested were resistant to ampicillin (86.8%). We randomly chose the strains with serotype O3:K6 and O10:K4 for PFGE to compare the genetic relationship between these two serotypes. The results showed that the PFGE profiles of V. parahaemolyticus O3:K6 and O10:K4 were genetically similar. The strains showed a tendency to cluster on the basis of their serotype profiles. However, some O3:K6 strains showed 100% similarity with O10:K4 strains. ConclusionA new serotype with pandemic potential of V. parahaemolyticus, O10:K4, was detected in 2020 and became dominant in 2021 in Huzhou.

Susceptibility of O3:K6 and environmental strains of Vibrio parahaemolyticus to acid inactivation, and survival competition between these strains and Pseudomonas fluorescens and indigenous bacteria

Vibrio parahaemolyticus is a common food poisoning pathogen associated with seafood. New O3:K6 strains originating from India in 1996 were the first pandemic strains and spread rapidly throughout many Asian countries. This study examines the characteristics of these new O3:K6 strains with respect to acid susceptibility and survival competition with Pseudomonas fluorescens and indigenous oyster spoilage microflora. Results revealed that the exponential phase cultures of new O3:K6 strains were significantly more tolerant of acid at pH 3.0 than the environmental strains. In these co-culture experiments, survival of the V. parahaemolyticus strains was influenced by the bacterial strains, incubation medium and incubation temperature. All V. parahaemolyticus strains showed similar survival ability when co-cultured with P. fluorescens in 1/10 tryptic soy broth. In the oyster medium, the environmental strain survived better than the new O3:K6 strains at 4 or 25°C. Both of the new O3:K6 strains (1121 and 1137) showed similar survivability in this study. These data offer hints on the survival of the new O3:K6 strains and their ability to spread.

Isolation, Molecular Characterization and Antibiotic Susceptibility Pattern of Vibrio parahaemolyticus from Aquatic Products in the Southern Fujian Coast, China.

Vibrio parahaemolyticus is a major gastroenteritis-causing pathogen in many Asian countries. Antimicrobial resistance in V. parahaemolyticus has been recognized as a critical threat to food safety. In this study, we determined the prevalence and incidence of antimicrobial resistance in V. parahaemolyticus in the southern Fujian coast, China. A total of 62 isolates were confirmed in retail aquatic products from June to October of 2018. The serotype O3:K6 strains, the virulence genes tdh and trh, antibiotic susceptibility and molecular typing were investigated. Then plasmid profiling analysis and curing experiment were performed for multidrug-resistant strains. The results showed that the total occurrence of V. parahaemolyticus was 31% out of 200 samples. Five strains (8.1%) out of 62 isolates were identified as the V. parahaemolyticus O3:K6 pandemic clone. A large majority of isolates exhibited higher resistance to penicillin (77.4%), oxacillin (71%), ampicillin (66.1%) and vancomycin (59.7%). Seventy-one percent (44/62) of the isolates exhibited multiple antimicrobial resistance. All 62 isolates were grouped into 7 clusters by randomly amplified polymorphic DNA, and most of the isolates (80.6%) were distributed within cluster A. Plasmids were detected in approximately 75% of the isolates, and seven different profiles were observed. Seventy-six percent (25/33) of the isolates carrying the plasmids were eliminated by 0.006% SDS incubated at 42°C, a sublethal condition. The occurrence of multidrug-resistant strains could be an indication of the excessive use of antibiotics in aquaculture farming. The rational use of antimicrobial agents and the surveillance of antibiotic administration may reduce the acquisition of resistance by microorganisms in aquatic ecosystems.

A new emerging serotype of Vibrio parahaemolyticus in China is rapidly becoming the main epidemic strain

ObjectivesDuring surveillance, we found a new type of Vibrio parahaemolyticus named ‘O4:KUT-recAin’ and studied the phenotypic, pathogenic and epidemiological characteristics of O4:KUT-recAin. MethodsV. parahaemolyticus were isolated from acute diarrhoeal patients in coastal hospitals of China. Serum agglutination test, specific PCR assay, growth curves under different conditions and rabbit diarrhoeal models were using to characterize O4:KUT-recAin. ResultsThe O4:KUT-recAin strain has a new type of K antigen and a 25 043-bp-large fragment encoding 20 proteins inserted in the housekeeping gene recA. Retrospective analysis found that only one O4:KUT-recAin strain was detected in 563 V. parahaemolyticus strains in 2014; then the proportion increased rapidly and reached 17.8% (105/590) in 2016 and 31.1% (224/721) in 2017, making O4:KUT-recAin the second dominant serotype following O3:K6. O4:KUT-recAin strains (100%, 14/14) exhibited increased acid resistance and could reproduce in medium at pH 4.9, while 92.9% (13/14) of the O3:K6 strains could not grow at this pH value. O4:KUT-recAin could cause diarrhoea and small intestinal tissue lesions in infant rabbits, but its diarrhoeal (93.1%, 27/29) and mortality (78.6%, 22/28) rates were slightly lower than those of O3:K6 (100% 16/16, 100% 16/16). Based on diarrhoea patients, there were no significant differences in the two groups for most clinical symptoms and laboratory results, except media age, haemoglobin and the number of red blood cells in stool samples. ConclusionsO4:KUT-recAin had enhanced acid resistance, was capable of causing infectious diarrhoea in both rabbits and humans, and has become widespread during a short period of time in China.

Alleviation of phytotoxic effects of cadmium on rice seedlings by cadmium resistant PGPR strain Enterobacter aerogenes MCC 3092

Heavy metal resistant PGPR mediated bioremediation, phytostimulation and stress alleviation is an eco-friendly method for sustainable agriculture in the metal contaminated soil. The isolation of such PGPR is highly demanding to reduce heavy metals in contaminated cultivated fields for agricultural benefit. The present study was successful to isolate a potent multi-heavy metal resistant PGPR strain, identified as Enterobacter aerogenes strain K6 based on MALDI-TOF MS, FAME analysis and 16S rDNA sequence homology, from rice rhizosphere contaminated with a variety of heavy metals/metalloid near industrial area. The strain exhibited high degree of resistance to Cd2+, Pb2+ and As3+ upto 4000 μg/mL, 3800 μg/mL and 1500 μg/mL respectively. Intracellular Cd accumulation of this strain was evidenced by AAS-SEM-TEM-EDX-XRF studies. Moreover, it showed several important PGP traits like IAA production, nitrogen fixation, phosphate solubilization, ACC deaminase activity even under high Cd stress (upto 3000 μg/mL). The combined effect of Cd resistance and PGP activities of this strain was manifested to the significant (p < 0.05) growth promotion of rice seedling under Cd stress by reducing oxidative stress (through antioxidants), stress ethylene and Cd uptake in seedlings. Thus K6 strain conferred Cd-tolerance in rice seedlings and could be applied as PGPR in contaminated fields.

A New Serotype of &lt;i&gt;Vibrio Parahaemolyticus&lt;/i&gt; Is Becoming the Main Epidemic Strain in China

Background: During surveillance, we found a new type of Vibrio parahaemolyticus that agglutinated with O4 serum but not with K serum, and its recA housekeeping gene was affected by a large insertion. We named this strain “O4:KUT-recAin”. Methods: We analyzed clinical data from acute diarrhea patients. By serum agglutination tests and sequence analyses, we identified a new type of V. parahaemolyticus strain. Through the establishment of a rabbit diarrhea model, we confirmed the pathogenicity of this strain. Findings: O4:KUT-recAin strain possessed a new type of K antigen, a 25,043 bp large fragment encoding 20 proteins inserted in the housekeeping gene recA. Retrospective analysis found that only one O4:KUT-recAin strain was detected in 2014; then, the proportion increased rapidly and reached 17·8% in 2016 and 31·1% in 2017, making O4:KUT-recAin the second dominant serotype following O3:K6. O4:KUT-recAin strains exhibited increased acid resistance and could reproduce in medium at pH= 4·9, while most of the O3:K6 strains could not grow at this pH. O4:KUT-recAin could cause diarrhea and small intestinal tissue lesions in infant rabbits, but its diarrhea and mortality rates were slightly lower than those of O3:K6. Based on diarrhea patients, most clinical symptoms and laboratory results were comparable in the two groups. Interpretation: O4:KUT-recAin had enhanced acid resistance, was capable of causing infectious diarrhea in both rabbits and humans, and became widespread during a short period of time in China. Therefore, it is necessary to raise global awareness and perform surveillance and molecular epidemiological studies to avoid further spread of O4:KUT-recAin. Funding Statement: This work was supported by the grants from the National Key Programs for Infectious Diseases of China (2017ZX10103008), the National Nature Science Foundation of China (81802067) and Sanming Project of Medicine in Shenzhen (SZSM201811071). Declaration of Interests: We declare no competing interests. Ethics Approval Statement: The study protocol was approved by the ethics committees of the First Affiliated Hospital, College of Medicine, Zhejiang University, and all participants provided written informed consent.

Prevalence of Vibrio parahaemolyticus in seafood and water environment in the Mekong Delta, Vietnam.

A total of 449 samples including 385 seafood and 64 water samples in the Mekong Delta of Vietnam collected in 2015 and 2016 were examined. Of 385 seafood samples, 332 (86.2%) samples were contaminated with Vibrio parahaemolyticus and 25 (6.5%) samples were pathogenic V. parahaemolyticus carrying tdh and/or trh genes. The tdh gene positive V. parahaemolyticus strains were detected in 22 (5.7%) samples and trh gene positive V. parahaemolyticus strains were found in 5 (1.3%) samples. Of 25 pathogenic V. parahaemolyticus strains, two strains harbored both tdh and trh genes and the other 23 strains carried either tdh or trh gene. Of 64 water samples at aquaculture farms, 50 (78.1%) samples were contaminated with V. parahaemolyticus. No tdh gene positive V. parahaemolyticus strains were detected; meanwhile, trh gene positive V. parahaemolyticus strain was detected in 1 (1.6%) sample. Twenty-six pathogenic V. parahaemolyticus strains isolated were classified into 6 types of O antigen, in which the serotype O3:K6 was detected in 4 strains. All pathogenic strains were group-specific PCR negative except for 4 O3:K6 strains. The result of antimicrobial susceptibility test indicated that pathogenic strains showed high resistance rates to streptomycin (84.6%), ampicillin (57.7%) and sulfisoxazole (57.7%). These findings can be used for understanding microbiological risk of seafood in the Mekong Delta, Vietnam.

Variation of genomic islands and flanking fragments in Vibrio parahaemolyticus isolates from environmental and clinical sources in Taiwan

Vibrio parahaemolyticus is a halophilic foodborne pathogenic bacterium that causes gastroenteritis; it has become an issue of global concern since the emergence and spread of pandemic O3:K6 strains. This study evaluated the role of Vibrio pathogenicity island (VPaI)-associated fragments in the genetic variation and grouping of this pathogen. Distribution of some VPaI fragments and flanking fragments (VPaI-1, VPaI-4, VPaI-5, VPaI-6 and VPaI-7) was determined in a total of 53 V. parahaemolyticus isolates from environmental and clinical sources in Taiwan, and supported by the sequences of seven fragments of VPaI-4 and its flanking fragment VP2145. As determined from the distribution of these VPaI-associated fragments, the clinical pandemic isolates were closely related in a single cluster; the clinical nonpandemic isolates were grouped into several clusters, while the environmental isolates were comparatively highly diversified. The profiles of virulence-associated genes of environmental pathogenic isolates varied, and were closer to those of clinical nonpandemic isolates than those of pandemic isolates. Isolates with atypical profiles of the VPaI-associated fragments and virulence-associated genes were identified. Sequences of VP2145 exhibited a close phylogenetic relationship among these local isolates, which were distinct from most V. parahaemolyticus strains from other geographic regions. This investigation demonstrated the application of VPaI-associated fragments in studying the genetic variation and clustering of V. parahaemolyticus isolates from different sources.

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

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

A pandemic Vibrio parahaemolyticus O3:K6 clone causing most associated diarrhea cases in the Pacific Northwest coast of Mexico.

Between September and October of 2004, more than 1230 cases of gastroenteritis due to pandemic O3:K6 strains of Vibrio parahaemolyticus (V. parahaemolyticus) were reported in the relatively small geographical area of Southern Sinaloa, a state located in Northwest Mexico. Since then, V. parahaemolyticus-associated gastroenteritis cases have gradually increased in prevalence spreading from south to north. The present study conducted an epidemiological surveillance of V. parahaemolyticus strains in both environmental and clinical samples along the Pacific coast of Sinaloa from 2011 to 2013. The genetic relatedness, serotype dominance and antibiotic resistance of isolates were investigated. A total of 46 strains were isolated from environmental samples (e.g., sediment, seawater and shrimp), whereas 249 strains were obtained from stools of patients with gastroenteritis. Nine different O serogroups and 16 serovars were identified. Serovars O3:K6 and O6:K46 were identified in both environmental and clinical strains. Whereas most environmental isolates carried the tdh gene (71.74%, 33/46), only three (6.52%) belonged to pandemic clones (O3:K6, O3:KUT and OUT:KUT). In contrast, 81.1% (202/249) of clinical isolates belonged to pandemic serotypes, with O3:K6 (tdh, toxRS/new, and/or orf8) representing the predominant serovar (97%, 196/202). This prevalence of pathogenic (tdh and/or trh positive) and O3:K6 pandemic V. parahaemolyticus isolates in this study were similar to those found from 2004 to 2010. As investigated by REP-PCR, genetic lineages of selected O3:K6 strains isolated in this study and some isolated earlier were nearly identical. Antimicrobial susceptibility testing showed that most strains (93.8%) were resistant to ampicillin but sensitive to chloramphenicol (98.8%). Multidrug resistance significantly increased from 8.6% (2004–2010) to 22.93% (2011–2013; p < 0.05). Our data indicate that pandemic O3:K6 clone has endemically established in the Pacific Coast of Mexico.

Pathogenic Vibrio parahaemolyticus isolated from biofouling on commercial vessels and harbor structures

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.

Genetic variation in Vibrio parahaemolyticus isolated from the aquacultural environments.

Environmental strains of Vibrio parahaemolyticus, a foodborne pathogen, were isolated from milkfish and grouper aquaculture facilities in southern Taiwan and characterized by pulsed-field gel electrophoresis (PFGE) and polymerase chain reaction targeting on the virulence-associated and genomic island (VPaI) marker genes. Analyses of 62 environmental isolates, including two putative pathogenic isolates, by NotI-PFGE revealed 11 pulse-type clusters with a similarity of 85%. Some of the T3SS2α-associated genes (vopB2, vopC and vopT) were not present in all of these two putative pathogenic isolates. Marker genes of VPaI-1 (MTase gene), VPaI-2 (VP0636) and VPaI-3 (VP1073 and VP1077) were detected in 14-100% of the environmental isolates examined, and the VP1073 and VP1077 of VPaI-3 marker genes were not detected. This study confirmed the high genetic variability of the environmental isolates including the putative pathogenic strains and identified some marker genes of VPaI in the environmental V. parahaemolyticus isolates. Vibrio parahaemolyticus is a prevalent seafood-borne enteropathogen with the appearance of pandemic O3:K6 strains in 1996. This study characterized the environmental nontoxigenic and toxigenic isolates by pulsed-field gel electrophoresis and the presence of marker genes of genomic islands. Results showed that the T3SS2α-associated genes are not present in all environmental tdh(+) isolates, and the presence of movable elements may contribute to genetic variation in the environmental V. parahaemolyticus isolates.

Predictive model of Vibrio parahaemolyticus O3:K6 growth on cooked Litopenaeus vannamei

Vibrio parahaemolyticus is the leading cause of seafood-derived illness in China and a possible mechanism leading to illness is cross contamination of cooked shrimp. A growth rate model of pandemic V. parahaemolyticus O3:K6 strain on cooked shrimp stored at isothermal temperatures was developed and validated. The maximum specific growth rate of this pandemic O3:K6 strain ranged from 0.11 to 1.47 (1/h) between 12 °C and 40 °C. A square root type model was then used to quantify the dependency between the maximum specific growth rate and temperature. The temperature at which the specific maximum growth rate = 0 was 1.0 °C. A goodness-of-fit measure, quantified by an average bias and accuracy factor indicated that the models of growth of V. parahaemolyticus O3:K6 on cooked shrimp under dynamic temperature regimes were reliable. Cooked shrimp also appears to be a better growth medium than TSB broth between 12–16 °C. Therefore relying on models developed in TSB broth for measuring the risk of V. parahaemolyticus will result in an underestimation of risk at these temperatures. Additionally, the growth models developed and validated in cooked shrimp gives fail-safe prediction of V. parahaemolyticus growth on fresh shrimp.

Vibrio parahaemolyticus, Southern Coastal Region of China, 2007–2012

We analyzed the prevalence and characteristics of Vibrio parahaemolyticus among patients with acute infectious diarrhea in the southern coastal region of China. V. parahaemolyticus was the leading cause of bacterial infectious diarrhea in this region during 2007–2012. Serotype O3:K6 strains were most common, followed by serotypes O4:K8 and O3:K29.

Pandemic Vibrio parahaemolyticus O3:K6 on the American continent

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.

Genome anatomy of the gastrointestinal pathogen, Vibrio parahaemolyticus of crustacean origin.

Vibrio parahaemolyticus, an important human pathogen, is associated with gastroenteritis and transmitted through partially cooked seafood. It has become a major concern in the production and trade of marine food products. The prevalence of potentially virulent and pathogenic V. parahaemolyticus in raw seafood is of public health significance. Here we describe the genome sequence of a V. parahaemolyticus isolate of crustacean origin which was cultured from prawns in 2008 in Selangor, Malaysia (isolate PCV08-7). The next generation sequencing and analysis revealed that the genome of isolate PCV08-7 has closest similarity to that of V. parahaemolyticus RIMD2210633. However, there are certain unique features of the PCV08-7 genome such as the absence of TDH-related hemolysin (TRH), and the presence of HU-alpha insertion. The genome of isolate PCV08-7 encodes a thermostable direct hemolysin (TDH), an important virulence factor that classifies PCV08-7 isolate to be a serovariant of O3:K6 strain. Apart from these, we observed that there is certain pattern of genetic rearrangements that makes V. parahaemolyticus PCV08-7 a non-pandemic clone. We present detailed genome statistics and important genetic features of this bacterium and discuss how its survival, adaptation and virulence in marine and terrestrial hosts can be understood through the genomic blueprint and that the availability of genome sequence entailing this important Malaysian isolate would likely enhance our understanding of the epidemiology, evolution and transmission of foodborne Vibrios in Malaysia and elsewhere.

Isolation and Characterization of Pandemic and Nonpandemic Strains ofVibrio parahaemolyticusfrom an Outbreak of Diarrhea in North 24 Parganas, West Bengal, India

Strains of the enteric pathogen Vibrio parahaemolyticus harboring the thermostable hemolysin (TDH) encoding gene tdh is known to cause epidemic and pandemic diarrhea. In industrialized countries, this pathogen causes sporadic or outbreaks of diarrheal illness associated with consumption of raw or improperly cooked seafood. This report describes a foodborne outbreak of gastroenteritis caused by V. parahaemolyticus in June 2011 following consumption of food served at a funeral reception held at Habra, North 24 Parganas, West Bengal, India. About 650 people attended the function, of whom 44 had acute watery diarrhea with other clinical symptoms; 35 of them were admitted to the District Hospital for the rehydration treatment. Stool specimens collected from three hospitalized cases were positive for V. parahaemolyticus, of which two strains were identified as an O4:K8 serovar and one was identified as O3:K6 serovar. The O3:K6 strain also possessed the pandemic group-specific toxRS gene target (GS), whereas the O4:K8 strains were negative. All strains were polymerase chain reaction-positive for tdh but were polymerase chain reaction-negative for trh. All of the strains were resistant to ampicillin but were pansensitive to other antimicrobials tested. Pulsed-field gel electrophoresis (PFGE) analysis using NotI showed that the O3:K6 strain was similar to that of a recent clinical strain from Kolkata, but had diverged from other strains during previous years. In contrast, PFGE analysis showed that the O4:K8 strains were closely related but differed from the Kolkata strain.