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PloS One 2017The morphology and the drug sensitivity of the strain GYX2014-1 isolated from the hepatic pancreatic tissue of moribund Litopenaeus vannamei were evaluated by...
The morphology and the drug sensitivity of the strain GYX2014-1 isolated from the hepatic pancreatic tissue of moribund Litopenaeus vannamei were evaluated by conventional culture characteristics, physical and chemical characteristics, and molecular biology methods. Detection of extracellulase and hemolysin activity shows that the isolated GYX2014-1 has protease, lipase, gelatinase activity, but none of amylase, or lecithinase activity. The 16S rRNA gene (GenBank accession number: KT781675) was analyzed, and a phylogenetic tree analysis showed that the isolated pathogen was most closely related to V. vulnificus (GenBank accession number: NR 118570)-a match of more than 99%. The phenotypic traits and molecular biology of isolated bacteria, determined their identity as Vibrio vulnificus (V. vulnificus). In addition, artificially infected L. vannamei with Vibrio vulnificus appeared with the same disease symptoms as those of naturally infected shrimp. Drug sensitivity tests showed that V. vulnificus is highly sensitive to fosfomycin, cefradine and sinomin, and was resistant to penicillin, amikacin and kanamycin. This experiment is the first to separate V. vulnificus from L. vannamei, and the findings of this study can be used as a reference for disease control and health management.
Topics: Animals; Base Sequence; Colony Count, Microbial; Extracellular Space; Microbial Sensitivity Tests; Penaeidae; Phenotype; Phylogeny; RNA, Ribosomal, 16S; Vibrio Infections; Vibrio vulnificus
PubMed: 29045415
DOI: 10.1371/journal.pone.0186135 -
Journal of Food Protection Oct 2013Vibrio vulnificus is a highly invasive human pathogen that exists naturally in estuarine environment and coastal waters. In this study, we used different PCR assays to...
Vibrio vulnificus is a highly invasive human pathogen that exists naturally in estuarine environment and coastal waters. In this study, we used different PCR assays to detect V. vulnificus in 260 seafood and 80 seawater samples. V. vulnificus was present in about 34 (13%) of the 260 seafood samples and 18 (23%) of the 80 seawater samples. Repetitive extragenic palindromic PCR (REP-PCR) and enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) were applied to subtype the V. vulnificus isolates. Twenty-five REP profiles and 45 ERIC profiles were observed, and the isolates were categorized into 9 and 10 distinct clusters at the similarity of 80%, by REP-PCR and ERIC-PCR, respectively. ERIC-PCR is more discriminative than REP-PCR in subtyping V. vulnificus, demonstrating high genetic diversity among the isolates.
Topics: Consumer Product Safety; DNA, Bacterial; Food Contamination; Genetic Variation; Humans; Malaysia; Polymerase Chain Reaction; Prevalence; Seafood; Seawater; Vibrio vulnificus
PubMed: 24112583
DOI: 10.4315/0362-028X.JFP-13-141 -
Journal of Clinical Microbiology Jan 2011We determined the association between DNA load and mortality in patients with Vibrio vulnificus infection. Real-time PCR performed on sera of 27 culture-positive...
We determined the association between DNA load and mortality in patients with Vibrio vulnificus infection. Real-time PCR performed on sera of 27 culture-positive patients showed a significantly higher median DNA load in nonsurvivors than in survivors. Hence, real-time PCR can be used as an early prognostic factor in V. vulnificus septicemia.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Bacterial Load; DNA, Bacterial; Female; Humans; Male; Middle Aged; Polymerase Chain Reaction; Prognosis; Sepsis; Vibrio Infections; Vibrio vulnificus; Young Adult
PubMed: 21068289
DOI: 10.1128/JCM.01913-09 -
BMC Microbiology Mar 2020Vibrio vulnificus hemolysin (VVH) is a pore-forming toxin secreted by Vibrio vulnificus. Cellular cholesterol was believed to be the receptor for VVH, because...
BACKGROUND
Vibrio vulnificus hemolysin (VVH) is a pore-forming toxin secreted by Vibrio vulnificus. Cellular cholesterol was believed to be the receptor for VVH, because cholesterol could bind to VVH and preincubation with cholesterol inhibited cytotoxicity. It has been reported that specific glycans such as N-acetyl-D-galactosamine and N-acetyl-D-lactosamine bind to VVH, however, it has not been known whether these glycans could inhibit the cytotoxicity of VVH without oligomer formation. Thus, to date, binding mechanisms of VVH to cellular membrane, including specific receptors have not been elucidated.
RESULTS
We show here that VVH associates with ganglioside GM1a, Fucosyl-GM1, GD1a, GT1c, and GD1b by glycan array. Among them, GM1a could pulldown VVH. Moreover, the GD1a inhibited the cytotoxicity of VVH without the formation of oligomers.
CONCLUSION
This is the first report of a molecule able to inhibit the binding of VVH to target cells without oligomerization of VVH.
Topics: Animals; Bacterial Proteins; Binding Sites; CHO Cells; Cell Membrane; Cholesterol; Cricetulus; Gangliosides; Glycomics; Hemolysin Proteins; Microarray Analysis; Protein Binding; Protein Conformation; Protein Multimerization; Vibrio vulnificus
PubMed: 32228455
DOI: 10.1186/s12866-020-01755-1 -
Characterization of temperature-dependent hemin uptake receptors HupA and HvtA in Vibrio vulnificus.MicrobiologyOpen Oct 2019The Gram-negative pathogen Vibrio vulnificus produces several iron-sequestration systems including a hemin uptake system in response to iron limitation as a means to...
The Gram-negative pathogen Vibrio vulnificus produces several iron-sequestration systems including a hemin uptake system in response to iron limitation as a means to acquire this essential element. Strains of this organism are capable of causing serious septicemia in humans and eels, where hemin is abundant and an advantageous source of iron. Vibrio vulnificus hemin uptake systems consist of HupA, a well studied outer membrane protein, and a recently identified HvtA protein receptor. In this study, we confirmed that the expression of the hvtA gene is iron-regulated in a fur-dependent manner. When analyzed for virulence in a hemin-overloaded murine model system, the hupA gene was more important for establishing infection than the hvtA gene. Transcriptional profiling of these genes using strains of two different biotypes, biotype 1 (human pathogen) and biotype 2 (eel pathogen), showed that the expression of the two receptors was also regulated in response to temperature. The expression of hupA was highly induced in elevated temperatures in the human pathogenic strain when tested in iron-depleted conditions. Conversely, hvtA expression was induced significantly in the eel pathogenic strain at a lower temperature, a condition where the hupA locus was relatively repressed. Our results indicate that although both hupA and hvtA are involved for optimal hemin uptake in V. vulnificus, their expression is dually regulated by the environmental cues of iron concentration and temperature. Together, these data suggest that the virulence genes hupA and hvtA are tightly regulated and strictly induced during iron limitation combined with the physiological temperature of the host organism.
Topics: Animals; Bacterial Outer Membrane Proteins; Cold Temperature; Disease Models, Animal; Gene Expression Profiling; Gene Expression Regulation, Bacterial; Hemin; Iron; Membrane Transport Proteins; Mice; Protein Transport; Vibrio Infections; Vibrio vulnificus; Virulence Factors
PubMed: 31290613
DOI: 10.1002/mbo3.905 -
Whole-genome comparison between reference sequences and oyster Vibrio vulnificus C-genotype strains.PloS One 2019Whole-genome sequences of Vibrio vulnificus clinical genotype (C-genotype) from the CICESE Culture Collection, isolated from oysters, were compared with reference... (Comparative Study)
Comparative Study
Whole-genome sequences of Vibrio vulnificus clinical genotype (C-genotype) from the CICESE Culture Collection, isolated from oysters, were compared with reference sequences of CMCP6 and YJ016 V. vulnificus C-genotype strains of clinical origin. The RAST web server estimated the whole genome to be ~4.8 Mb in CICESE strain 316 and ~4.7 Mb in CICESE strain 325. No plasmids were detected in the CICESE strains. Based on a phylogenetic tree that was constructed with the whole-genome results, we observed high similarity between the reference sequences and oyster C-genotype isolates and a sharp contrast with environmental genotype (E-genotype) reference sequences, indicating that the differences between the C- and E-genotypes do not necessarily correspond to their isolation origin. The CICESE strains share 3488 genes (63.2%) with the YJ016 strain and 3500 genes (63.9%) with the CMCP6 strain. A total of 237 pathogenicity associated genes were selected from reference clinical strains, where-92 genes were from CMCP6, 126 genes from YJ016, and 19 from MO6-24/O; the presence or absence of these genes was recorded for the CICESE strains. Of the 92 genes that were selected for CMCP6, 67 were present in both CICESE strains, as were as 86 of the 126 YJ016 genes and 13 of the 19 MO6-24/O genes. The detection of elements that are related to virulence in CICESE strains-such as the RTX gene cluster, vvhA and vvpE, the type IV pili cluster, the XII genomic island, and the viuB genes, suggests that environmental isolates with the C-genotype, have significant potential for infection.
Topics: Animals; Evolution, Molecular; Genome, Bacterial; Genotype; Ostreidae; Phylogeny; Vibrio vulnificus; Virulence Factors; Whole Genome Sequencing
PubMed: 31361763
DOI: 10.1371/journal.pone.0220385 -
Diseases of Aquatic Organisms Apr 2014Vibrio vulnificus is a potentially zoonotic bacterial pathogen of fish, which can infect humans (causing necrotic fasciitis). We analysed 24 V. vulnificus isolates (from...
Vibrio vulnificus is a potentially zoonotic bacterial pathogen of fish, which can infect humans (causing necrotic fasciitis). We analysed 24 V. vulnificus isolates (from 23 severe eel disease outbreaks in 8 Dutch eel farms during 1996 to 2009, and 1 clinical strain from an eel farmer) for genetic correlation and zoonotic potential. Strains were typed using biotyping and molecular typing by high-throughput multilocus sequence typing (hiMLST) and REP-PCR (Diversilab®). We identified 19 strains of biotype 1 and 5 of biotype 2 (4 from eels, 1 from the eel farmer), that were subdivided into 8 MLST types (ST) according to the international standard method. This is the first report of V. vulnificus biotype 1 outbreaks in Dutch eel farms. Seven of the 8 STs, of unknown zoonotic potential, were newly identified and were deposited in the MLST database. The REP-PCR and the MLST were highly concordant, indicating that the REP-PCR is a useful alternative for MLST. The strains isolated from the farmer and his eels were ST 112, a known potential zoonotic strain. Antimicrobial resistance to cefoxitin was found in most of the V. vulnificus strains, and an increasing resistance to quinolones, trimethoprim + sulphonamide and tetracycline was found over time in strain ST 140. Virulence testing of isolates from diseased eels is recommended, and medical practitioners should be informed about the potential risk of zoonotic infections by V. vulnificus from eels for the prevention of infection especially among high-risk individuals. Additional use of molecular typing methods such as hiMLST and Diversilab® is recommended for epidemiological purposes during V. vulnificus outbreaks.
Topics: Anguilla; Animals; Anti-Bacterial Agents; Aquaculture; Disease Outbreaks; Drug Resistance, Bacterial; Fish Diseases; Genetic Variation; Netherlands; Vibrio Infections; Vibrio vulnificus
PubMed: 24695233
DOI: 10.3354/dao02703 -
Microbiology Spectrum May 2024is a genus of halophilic, gram-negative bacteria found in estuaries around the globe. Integral parts of coastal cultures often involve contact with vectors of...
UNLABELLED
is a genus of halophilic, gram-negative bacteria found in estuaries around the globe. Integral parts of coastal cultures often involve contact with vectors of pathogenic spp. (e.g., consuming raw shellfish). High rates of mortality from certain spp. infections demonstrate the need for an improved understanding of spp. dynamics in estuarine regions. Our study assessed meteorological, hydrographic, and biological correlates of and at 10 sites in the Eastern Mississippi Sound System (EMSS) from April to October 2019. During the sampling period, median abundances of and were 2.31 log MPN/L and 2.90 log MPN/L, respectively. spp. dynamics were largely driven by site-based variation, with sites closest to freshwater inputs having the highest abundances. The E-W wind scalar, which affects Ekman transport, was a novel spp. correlate observed. A potential salinity effect on bacterial-particle associations was identified, where was associated with larger particles in conditions outside of their optimal salinity. Additionally, abundances were correlated to those of harmful algal species that did not dominate community chlorophyll. Correlates from this study may be used to inform the next iteration of regionally predictive models and may lend additional insight to spp. ecology in similar systems.
IMPORTANCE
spp. are bacteria found in estuaries worldwide; some species can cause illness and infections in humans. Relationships between spp. abundance, salinity, and temperature are well documented, but correlations to other environmental parameters are less understood. This study identifies unique correlates (e.g., E-W wind scalar and harmful algal species) that could potentially inform the next iteration of predictive models for the EMSS region. Additionally, these correlates may allow existing environmental monitoring efforts to be leveraged in providing data inputs for future Vibrio risk models. An observed correlation between salinity and /particle-size associations suggests that predicted environmental changes may affect the abundance of spp. in certain reservoirs, which may alter which vectors present the greatest vibrio risk.
Topics: Vibrio parahaemolyticus; Vibrio vulnificus; Estuaries; Alabama; Population Dynamics; Salinity; Vibrio Infections; Seawater; Water Microbiology
PubMed: 38578091
DOI: 10.1128/spectrum.03674-23 -
International Journal of Food... Sep 2022Vibrio vulnificus is a zoonotic pathogen linked to aquaculture that is spreading due to climate change. The pathogen can be transmitted to humans and animals by...
Vibrio vulnificus is a zoonotic pathogen linked to aquaculture that is spreading due to climate change. The pathogen can be transmitted to humans and animals by ingestion of raw shellfish or seafood feed, respectively. The aim of this work was to design and test a new procedure to detect V. vulnificus hazardous to human and/or animal health in food/feed samples. For this purpose, we combined a pre-enrichment step with multiplex PCR using primers for the species and for human and animal virulence markers. In vitro assays with mixed DNA from different Vibrio species and Vibrio cultures showed that the new protocol was 100 % specific with a detection limit of 10 cfu/mL. The protocol was successfully validated in seafood using artificially contaminated live shrimp and proved useful also in pathogen isolation from animals and their ecosystem. In conclusion, this novel protocol could be applied in health risk studies associated with food/feed consumption, as well as in the routine identification and subtyping of V. vulnificus from environmental or clinical samples.
Topics: Animals; Ecosystem; Humans; Multiplex Polymerase Chain Reaction; Seafood; Shellfish; Vibrio; Vibrio vulnificus
PubMed: 35696749
DOI: 10.1016/j.ijfoodmicro.2022.109778 -
Frontiers in Cellular and Infection... 2018Antimicrobial resistance (AMR) in pathogens is the result of indiscriminate use of antibiotics and consequent metabolic/genetic modulation to evolve survival strategies...
Antimicrobial resistance (AMR) in pathogens is the result of indiscriminate use of antibiotics and consequent metabolic/genetic modulation to evolve survival strategies and clonal-selection in AMR strains. As an alternative to antibiotic treatment, antivirulence strategies are being developed, not only to combat bacterial pathogenesis, but also to avoid emerging antibiotic resistance. is a foodborne pathogen that causes gastroenteritis, necrotizing wound infections, and sepsis with a high rate of mortality. Here, we developed an inhibitor-screening reporter platform to target HlyU, a master transcriptional regulator of virulence factors in by assessing transcription under its control. The inhibitor-screening platform includes wild type and Δ mutant strains of harboring the reporter construct P for desired luminescence signal detection and control background luminescence, respectively. Using the inhibitor-screening platform, we identified a small molecule, fursultiamine hydrochloride (FTH), that inhibits the transcription of the highly invasive repeat-in-toxin () and hemolysin () along with other HlyU regulated virulence genes. FTH has no cytotoxic effects on either host cells or pathogen at the tested concentrations. FTH rescues host cells from the necrotic cell-death induced by RtxA1 and decreases the hemolytic activity under conditions. The most important point is that FTH treatment does not induce the antivirulence resistance. Current study validated the antivirulence strategy targeting the HlyU virulence transcription factor and toxin-network of and demonstrated that FTH, exhibits a potential to inhibit the pathogenesis of deadly, opportunistic human pathogen, without inducing AMR.
Topics: Anti-Bacterial Agents; Bacterial Proteins; Drug Evaluation, Preclinical; Drug Resistance, Bacterial; Fursultiamin; Gene Expression Regulation, Bacterial; HeLa Cells; Hemolysin Proteins; Humans; Transcription Factors; Vibrio vulnificus; Virulence; Virulence Factors
PubMed: 29868508
DOI: 10.3389/fcimb.2018.00152