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Microbial Pathogenesis Apr 2017Vibrio sp. are autochthonous to marine and estuarine waters. Several species of Vibrio are pathogens. It is of utmost importance to detect and discriminate the Vibrio...
Vibrio sp. are autochthonous to marine and estuarine waters. Several species of Vibrio are pathogens. It is of utmost importance to detect and discriminate the Vibrio sp. that are often involved in food and water borne infections. Since 16S rRNA based identification has limited utility in differentiating the closely related pathogenic species from non pathogenic species, we have evaluated the discriminatory power of groEL PCR-RFLP for identification of closely related Vibrio sp. Accordingly, in the current study, the efficiency of groEL PCR- RFLP for detection and accurate differentiation of known pathogens among Vibrio sp. such as V. cholerae, V. parahaemolyticus, V. vulnificus, V. mimicus, V. fluvialis, V. alginolyticus, V. anguillarum was evaluated. PCR amplified groEL gene fragment of each Vibrio sp. was digested separately using 5 restriction enzymes viz. Hha1, Rsa1, Alu1, Dde1 and Mbo1. The accuracy of the method was further validated by insilico restriction analysis of multiple strains of each species using NEBcutter. The method proved to be efficient for detection and differentiation of Vibrio species under study. Phylogenetic analysis also revealed groEL gene to be a better phylogenetic marker for Vibrio compared to 16S rRNA. Hence, the method can be employed for accurate detection of Vibrio sp. including the closely related species.
Topics: Amplified Fragment Length Polymorphism Analysis; Bacterial Proteins; Base Sequence; Chaperonin 60; Computer Simulation; Genes, Bacterial; Microbiological Techniques; Phylogeny; Polymorphism, Restriction Fragment Length; RNA, Ribosomal, 16S; Sensitivity and Specificity; Vibrio
PubMed: 28235640
DOI: 10.1016/j.micpath.2017.02.029 -
The Journal of the Louisiana State... 2016Vibrio mimicus, although named for having many of the same virulent factors as Vibrio cholera, is a rare cause of significant gastrointestinal illness. Like many of the...
Vibrio mimicus, although named for having many of the same virulent factors as Vibrio cholera, is a rare cause of significant gastrointestinal illness. Like many of the Vibrio species, the strongest risk factor for V. mimicus infection is seafood consumption. After consuming crabs, a 64-year-old male presented with a three day history of voluminous, non-bloody, water diarrhea. The severity of the diarrhea caused the patient to have orthostatic hypotension and acute kidney injury, which improved with fluid resuscitation. The diarrhea resolved in 24-hours, and the patient was discharged without medications. Stool studies later returned positive for V. mimicus. Clinicians, especially those in coastal regions, should consider this rare pathogen in the setting of refractory diarrhea and a history significant for seafood consumption. With early clinical suspicion, clinicians can focus on volume repletion while limiting the use of anti-microbials.
Topics: Diarrhea; Humans; Male; Middle Aged; Seafood; Vibrio Infections; Vibrio mimicus
PubMed: 28045686
DOI: No ID Found -
Veterinary Immunology and... Dec 2016Vibrio mimicus is the causative agent of ascites disease in fish. The heat-labile hemolytic toxin designated VMH is an immunoprotective antigen of V. mimicus. However,...
Vibrio mimicus is the causative agent of ascites disease in fish. The heat-labile hemolytic toxin designated VMH is an immunoprotective antigen of V. mimicus. However, its epitopes have not been well characterized. Here, a commercially available phage displayed 12-mer peptide library was used to screen epitopes of VMH protein using polyclonal rabbit anti-rVMH protein antibodies, and then five positive phage clones were identified by sandwich and competitive ELISA. Sequences analysis showed that the motif of DPTLL displayed on phage clone 15 and the consensus motif of SLDDDST displayed on the clone 4/11 corresponded to the residues 134-138 and 238-244 of VMH protein, respectively, and the synthetic motif peptides could also be recognized by anti-rVMH-HD antibody in peptide-ELISA. Thus, both motifs DPTLL and SLDDDST were identified as minimal linear B-cell epitopes of VMH protein. Although no similarity was found between VMH protein and the consensus motif of ADGLVPR displayed on the clone 2/6, the synthetic peptide ADGLVPR could absorb anti-rVMH-HD antibody and inhibit the antibody binding to rVMH protein in enhanced chemoluminescence Western blotting, whereas irrelevant control peptide did not affect the antibody binding with rVMH. These results revealed that the peptide ADGLVPR was a mimotope of VMH protein. Taken together, three novel B-cell epitopes of VMH protein were identified, which provide a foundation for developing epitope-based vaccine against V. mimicus infection in fish.
Topics: Amino Acid Sequence; Animals; Antibodies, Bacterial; Bacterial Proteins; Bacterial Vaccines; Enzyme-Linked Immunosorbent Assay; Epitopes, B-Lymphocyte; Fish Diseases; Fishes; Hemolysin Proteins; Peptide Library; Rabbits; Recombinant Fusion Proteins; Vibrio Infections; Vibrio mimicus
PubMed: 27863546
DOI: 10.1016/j.vetimm.2016.09.005 -
PloS One 2016Vibrio mimicus is a pathogen that causes ascites disease in fish. We have previously demonstrated that the outer membrane protein U (OmpU) is an important adhesin in V....
Vibrio mimicus is a pathogen that causes ascites disease in fish. We have previously demonstrated that the outer membrane protein U (OmpU) is an important adhesin in V. mimicus. Here eight specific OmpU-binding phage clones, which presented three different OmpU-binding peptides (designated P1, P2, P3), were screened from a commercially available phage displayed 12-mer peptide library using rOmpU protein as target. Then, synthetic OmpU-binding peptides were measured for their adhesion antagonistic activity and binding affinity via adhesion inhibition test and non-competitive ELISA, respectively. The results showed that after co-incubated with the mixture of rOmpU and P3, visible green fluorescence could be observed on the epithelioma papulosum cyprinidi (EPC) cells surface; while the EPC cells co-incubated with the mixture of rOmpU and P1/P2 exhibited little green fluorescence. The average adhesion number of V. mimicus 04-14 isolate before and after treatment with peptide was 21.4 ± 1.5, 20.8 ± 0.8 (irrelevant peptide), 20.2 ± 0.5 (P3), 5.1 ± 0.7 (P1) and 3.4 ± 0.8 (P2), respectively. There was a significant decrease in the adhesive level of 04-14 isolate treated with P1/ P2 compared to the untreated isolate (p<0.01). The affinity constants of P1 and P2 were (6.17 ± 0.19) × 108 L/mol and (1.24 ± 0.56) × 109 L/mol, respectively. Furthermore, protective effects of P1 and P2 on grass carps challenged with V. mimicus were preliminary detected. It was found there was delayed death of fish in the groups treated with P1/P2, and the survival rate of challenged fish improved with the increase of the dose of adhesion antagonistic peptide. Taken together, two novel OmpU-binding peptides, which possessed adhesion antagonistic activity, high affinity and a certain degree of antibacterial activity against V. mimicus, were screened and identified.
Topics: Adhesins, Bacterial; Animals; Anti-Bacterial Agents; Bacterial Adhesion; Carps; Cell Line; Fish Diseases; Fishes; Peptide Library; Peptides; Vibrio mimicus
PubMed: 27832083
DOI: 10.1371/journal.pone.0165092 -
Virulence Aug 2017
Topics: Animals; Bacterial Vaccines; Catfishes; Fish Diseases; Sequence Analysis, DNA; Vibrio Infections; Vibrio cholerae; Vibrio mimicus; Virulence; Whole Genome Sequencing
PubMed: 27763808
DOI: 10.1080/21505594.2016.1250996 -
Fish & Shellfish Immunology Aug 2016
Corrigendum to "Immunological responses of customised probiotics-fed marron, Cherax tenuimanus, (Smith 1912) when challenged with Vibrio mimicus" [Fish Shellfish Immunol. 35 (2) (2013) 262-270].
PubMed: 27475718
DOI: 10.1016/j.fsi.2016.07.028 -
Journal of Basic Microbiology Oct 2016Vibrio mimicus is an estuarine bacterium, while it can cause severe diarrhea, wound infection, and otitis media in humans. This pathogen secretes a relatively important...
Vibrio mimicus is an estuarine bacterium, while it can cause severe diarrhea, wound infection, and otitis media in humans. This pathogen secretes a relatively important toxin named V. mimicus metalloprotease (VMP). In this study, we clarified regulation of the VMP production according to the quorum-sensing master regulatory protein named LuxR. First, the full length of luxR gene, encoding LuxR, was detected in V. mimicus strain E-37, an environmental isolate. Next, the putative consensus binding sequence of LuxR protein could be detected in the upstream (promoter) region of VMP encoding gene, vmp. Finally, the effect of disruption of luxR gene on the expression of vmp and production of VMP was evaluated. Namely, the expression of vmp was significantly diminished by luxR disruption and the production of VMP was severely altered. Taken together, here we report that VMP production is under the positive regulation of the quorum-sensing master regulatory protein, LuxR.
Topics: Base Sequence; Binding Sites; DNA, Bacterial; Gene Expression Regulation, Bacterial; Metalloproteases; Promoter Regions, Genetic; Quorum Sensing; Repressor Proteins; Trans-Activators; Vibrio mimicus
PubMed: 27160384
DOI: 10.1002/jobm.201600002 -
Wei Sheng Wu Xue Bao = Acta... May 2016To study the immunological cross-reactivity and cross-protection characteristics of OmpU in Vibrio species.
OBJECTIVE
To study the immunological cross-reactivity and cross-protection characteristics of OmpU in Vibrio species.
METHODS
The ompU genes from 10 Vibrio strains were cloned, sequenced, followed by bioinformatics analysis. Western blot and whole-cell ELISA assay were used respectively to determine immunological cross-reaction feature and subcellular location of OmpU with rabbit serum against recombinant OmpU from V. parahaemolyticus ATCC17802, V. alginolyticus ATCC33787, V. vulnificus ATCC27562, V. mimicus ATCC33653 and V. cholera Vb0. Finally, the cross-protective property of recombinant OmpU (V. cholera-derived) was evaluated through vaccination and subsequent challenge with heterogeneous virulent Vibrio strains in mice.
RESULTS
The similarities of OmpU proteins of Vibrio ranged from 73.0 to 100% intra-species, and from 58.6 to 89.0% inter-species. Furthermore, homologous epitopes were found in OmpU and shared by different species of Vibrios. Western blot of rabbit serum against recombinant OmpU showed cross-recognition intra- and inter-species. Bands were observed ranging from 35 to 40 kDa. Whole-cell ELISA assay further confirmed that the antiserum of recombinant OmpU from V. parahaemolyticus ATCC17802, V. vulnifgicus ATCC27562 and V. mimicus ATCC33653 recognized the tested Vibrio species, implying that epitopes of OmpU were located on the cell surface. Recorded relative percent survival of the vaccinated group varied from 43.0 to 100%, showing that mice were protected from Vibrio infection after immunization with OmpU protein.
CONCLUSION
OmpU was a conserved antigen among tested Vibrio species and might be a universal vaccine candidate for the prevention of Vibriosis.
Topics: Animals; Antibodies, Bacterial; Bacterial Outer Membrane Proteins; Bacterial Vaccines; Cross Protection; Cross Reactions; Epitope Mapping; Epitopes; Female; Humans; Male; Mice; Rabbits; Vibrio; Vibrio Infections
PubMed: 29727148
DOI: No ID Found -
Asian Pacific Journal of Tropical... Jan 2016To investigate the isolation of enterobacteria associated with Macrobrachium amazonicum (M. amazonicum) farming and evaluate the in vitro antimicrobial susceptibility...
OBJECTIVE
To investigate the isolation of enterobacteria associated with Macrobrachium amazonicum (M. amazonicum) farming and evaluate the in vitro antimicrobial susceptibility of Vibrio strains.
METHODS
Strains were isolated from female M. amazonicum prawns and environmental and hatchery water. Biochemical assays were used to identify bacterial genera and those belonging to the genus Vibrio were submitted to further analyses for species identification, through Vitek 2 automated system and serotyping. Susceptibility test was performed according to Clinical Laboratory Standards Institute.
RESULTS
The following genera of enterobacteria were recovered: Enterobacter (n = 11), Citrobacter (n = 10), Proteus (n = 2), Serratia (n = 2), Kluyvera (n = 2), Providencia (n = 2), Cedecea (n = 1), Escherichia (n = 1), Edwardsiella (n = 1) and Buttiauxella (n = 1). As for Vibrio, three species were identified: Vibrio cholerae non-O1/non-O139 (n = 4), Vibrio vulnificus (V. vulnificus) (n = 1) and Vibrio mimicus (n = 1). Vibrio spp. showed minimum inhibitory concentrations values within the susceptibility range established by Clinical Laboratory Standards Institute for almost all antibiotics, except for V. vulnificus, which presented intermediate profile to ampicillin.
CONCLUSIONS
Enterobacteria do not seem to be the most important pathogens associated with M. amazonicum farming, whereas the recovery of Vibrio spp. from larviculture, with emphasis on Vibrio cholerae and V. vulnificus, deserves special attention due to their role as potentially zoonotic aquaculture-associated pathogens. Furthermore, the intermediate susceptibility of V. vulnificus to ampicillin reflects the importance of monitoring drug use in prawn farming.
PubMed: 26851782
DOI: 10.1016/j.apjtm.2015.12.006 -
PloS One 2016Vibrio mimicus is a gram-negative bacterium responsible for diseases in humans. Three strains of V. mimicus identified as V. mimicus 87, V. mimicus 92 and V. mimicus 93...
Vibrio mimicus is a gram-negative bacterium responsible for diseases in humans. Three strains of V. mimicus identified as V. mimicus 87, V. mimicus 92 and V. mimicus 93 were isolated from a shrimp processing facility in Guaymas, Sonora, Mexico. The strains were analyzed using several molecular techniques and according to the cluster analysis they were different, their similarities ranged between 51.3% and 71.6%. ERIC-PCR and RAPD (vmh390R) were the most discriminatory molecular techniques for the differentiation of these strains. The complete genomes of two strains (V. mimicus 87, renamed as CAIM 1882, and V. mimicus 92, renamed as CAIM 1883) were sequenced. The sizes of the genomes were 3.9 Mb in both strains, with 2.8 Mb in ChI and 1.1 Mb in ChII. A 12.7% difference was found in the proteome content (BLAST matrix). Several virulence genes were detected (e.g. capsular polysaccharide, an accessory colonization factor and genes involved in quorum-sensing) which were classified in 16 categories. Variations in the gene content between these genomes were observed, mainly in proteins and virulence genes (e.g., hemagglutinin, mobile elements and membrane proteins). According to these results, both strains were different, even when they came from the same source, giving an insight of the diversity of V. mimicus. The identification of various virulence genes, including a not previously reported V. mimicus gene (acfD) in ChI in all sequenced strains, supports the pathogenic potential of this species. Further analysis will help to fully understand their potential virulence, environmental impact and evolution.
Topics: Animals; Bacterial Proteins; Bacterial Typing Techniques; DNA Fingerprinting; DNA, Bacterial; Food Contamination; Food Handling; Food Microbiology; Freezing; Genes, Bacterial; Hemolysin Proteins; Mexico; Penaeidae; Random Amplified Polymorphic DNA Technique; Ribotyping; Sequence Alignment; Sequence Analysis, DNA; Species Specificity; Vibrio mimicus; Virulence; Water Microbiology
PubMed: 26730584
DOI: 10.1371/journal.pone.0144885