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Letters in Applied Microbiology Sep 2020In this study, the prevalence of Vibrio cholerae, Vibrio parahaemolyticus, Vibrio mimicus, Vibrio vulnificus and Vibrio spp. in shrimp from retail markets in Reynosa,...
In this study, the prevalence of Vibrio cholerae, Vibrio parahaemolyticus, Vibrio mimicus, Vibrio vulnificus and Vibrio spp. in shrimp from retail markets in Reynosa, Mexico was determined. A total of 765 isolates, identified as Vibrio spp. (59·1%), V. cholerae (17·8%), V. mimicus (6·7%) and V. parahaemolyticus (4·6%), were obtained; V. vulnificus was not detected. Most of the strains were isolated from supermarkets (48·1%), followed by street vendors (37·3%) and retail stores (14·6%). Moreover, several virulence genes were identified in V. cholerae: toxR (100%), OmpU (76·5%), hlyA (76·5%), VPI (19·9%) and tcpA (5·1%); in V. mimicus: vmh (100%), wzb (74·5%), pilF (54·9%), VPI (43·1%), OmpU (29·4%) and tdh (9·8%); and in V. parahaemolyticus: toxR (100%), tlh (100%), VP1680 (51·4%) and VPI (11·4%). These results show the low safety of this food and the potential risk to consumers' health, since this product in Mexican cuisine is sometimes served raw or semi-cooked. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows the prevalence of pathogenic Vibrio cholerae, Vibrio mimicus and Vibrio parahaemolyticus isolated from shrimp that is commercialized in Reynosa city. This could represent a risk to consumers' health, since outbreaks related to shrimp contaminated with Vibrio have been previously reported. Additionally, shrimp fishing has a major role in Mexico's economy.
Topics: Animals; Food Contamination; Food Safety; Mexico; Penaeidae; Prevalence; Raw Foods; Seafood; Supermarkets; Vibrio cholerae; Vibrio mimicus; Vibrio parahaemolyticus; Vibrio vulnificus; Virulence
PubMed: 32408383
DOI: 10.1111/lam.13315 -
Journal of Microbiological Methods Jun 2020Vibrio parahaemolyticus is a major foodborne pathogen worldwide. Contamination of V. parahaemolyticus in foods must be detected as quickly as possible because raw...
Vibrio parahaemolyticus is a major foodborne pathogen worldwide. Contamination of V. parahaemolyticus in foods must be detected as quickly as possible because raw seafood, a major source of V. parahaemolyticus infection, is shipped immediately after production due to its short expiration date. In this study, we generated monoclonal antibodies (mAbs) against V. parahaemolyticus to develop a rapid and specific detection assay. Obtained mAbs were categorized into four groups according to their specificity. Of the groups, Group 1 (mAb VP7, VP11, and VP24) reacted to O1-O12 of V. parahaemolyticus without cross-reaction with human pathogenic Vibrio spp. (V. alginolyticus, V. cholerae, V. fluvialis, V. furnissii, V. mimicus, and V. vulnificus). We developed an immunochromatographic (IC) strip for the rapid detection of V. parahaemolyticus in the field using VP7 as a membrane-immobilized antibody and VP24 as a colloidal gold-conjugated antibody. The IC strip detected any and all serogroups (O1 to O12) or isolates (clinical, food, and environmental strains) of V. parahaemolyticus, regardless of the presence of virulence factors thermostable direct hemolysin (TDH) or TDH-related hemolysin (TRH). It did not cross-react with any other non-V. parahaemolyticus strains tested. To elucidate the target of the IC strip, we analyzed the antigen recognized by these mAbs. Group 1 mAbs showed two specific bands at molecular masses of approximately 11 and 16 kDa by western blotting analysis. Nano liquid chromatography mass spectrometry (LC-MS)/MS analysis revealed that the candidate antigen recognized by these mAbs was outer membrane (OM) lipoprotein Q87G48. We verified that mAb VP7 detected His-tagged OM lipoprotein synthesized by reconstituted cell-free protein synthesis reagent. Reactivity to an N-terminus deletion form and protease digestion form of the OM lipoprotein showed that the extent of epitope recognized by VP mAbs was 22nd-41st amino acids (AAs) from N-terminus of the OM lipoprotein, with the sequence "SDDAATANAAKLDEL." This region was also confirmed to be a V. parahaemolyticus-specific sequence by comparing putative orthologs of OM lipoprotein among Vibrio spp. The C-terminus deletion form (1st-39th AAs) including the sequence primarily recognized by VP mAbs (22nd-36th AAs) showed poor reactivity, indicating that the sequence after 40 residues of OM lipoprotein is also important for recognition by VP mAbs and VP mAbs recognize a conformational epitope. Bioinformatics research demonstrated that the OM lipoprotein is an ortholog of the lpp protein conserved throughout many bacteria. Lpp is an abundant and constitutively expressed protein and exists on the bacterial surface, suggesting it may be a good target for detection of V. parahaemolyticus.
Topics: Antibodies, Monoclonal; Antigens, Bacterial; Bacterial Toxins; Bacteriological Techniques; Blotting, Western; Chromatography, Affinity; Cross Reactions; Epitopes; Hemolysin Proteins; Humans; Immunoprecipitation; Species Specificity; Vibrio cholerae; Vibrio parahaemolyticus; Virulence Factors
PubMed: 32289368
DOI: 10.1016/j.mimet.2020.105919 -
Frontiers in Physiology 2019A feeding trial was carried out to evaluate the effects of substitution of fishmeal (FM) by dietary poultry by-product meal, fermented by and on growth, intestinal...
A feeding trial was carried out to evaluate the effects of substitution of fishmeal (FM) by dietary poultry by-product meal, fermented by and on growth, intestinal health, microbial composition, immune related cytokines and disease resistance of freshwater crayfish, marron () against Two isonitrogenous and isocaloric diets were formulated by replacing FM protein with fermented poultry by-product meal (FPBM) protein at 0% (Control) and 75% (FPBM), and fed marron for 70 days. The results indicated no significant difference ( > 0.05) in final body weights between two groups of marron, whilst intestinal microvilli number per fold was increased in marron fed FPBM than the control. The 16S rRNA sequences revealed an increased number of and , and decreased number of at genus level in the distal intestine of marron fed FPBM. Marron fed FPBM showed up-regulated expression of IL-8, IL-10, and IL-17F genes in the distal intestine. Significantly ( < 0.05) increased lysozyme and phagocytic activity, and higher survival was found in marron fed FPBM following a bacterial challenge with . Therefore, it is concluded that FPBM is beneficial to marron in terms of microbial community, immune-related cytokines and disease resistance against
PubMed: 32082185
DOI: 10.3389/fphys.2019.01635 -
Journal of Applied Microbiology Jul 2020The current study was conducted to determine the incidence, antibiotic resistance and virulence genes of Vibrio strains isolated from ready-to-eat shrimps in Edo and...
AIM
The current study was conducted to determine the incidence, antibiotic resistance and virulence genes of Vibrio strains isolated from ready-to-eat shrimps in Edo and Delta States, Nigeria.
METHODS AND RESULTS
A total of 1440 ready-to-eat shrimp samples were obtained from open markets from November 2016 to October 2017 and analysed using standard culture-based procedures. Overall, our result showed that the ready-to-eat shrimp samples had high mean aerobic mesophilic bacterial count between 3·543 and 7·489 log CFU per gram. Vibrio cell densities ranged between 0·663 and 6·761 log CFU per gram. From the total samples, 1343/1440 (93·3%) were positive for Vibrio species where 120 Vibrio isolates were randomly selected and confirmed using genus- and species-specific PCR approach. The PCR identification revealed the presence of Vibrio parahaemolyticus 46 (38·33%), Vibrio vulnificus 14 (11·67%), Vibrio fluvialis 12 (10%), Vibrio alginolyticus 8 (6·67%), Vibrio cholerae 2 (1·67%), Vibrio mimicus 10 (8·33%), Vibrio harveyi 3 (2·5%) and other Vibrio sp. 25 (20·83%). All Vibrio isolates were sensitive to colistin and gentamycin with varying percentage of resistance to other antibiotics used in the study. Multiple antibiotic-resistant (MAR) index ranged from 0·08 to 0·83. The tcp, tdh and trl virulence genes were identified in 95 (79·2%), 92 (76·7%) and 95 (79·2%) of the examined isolates respectively. Antibiotic-resistant genes also revealed the presence of class 1 integrase 75 (62·5%), sul2 87 (72·5%), strB 94 (78·3%) and catB3 68 (56·7%).
CONCLUSION
Our findings revealed that the ready-to-eat shrimps may serve as potential reservoirs and medium in the dissemination of prospective MAR pathogens to the consumers and thus constitute a potential risk to public health.
SIGNIFICANCE AND IMPACT OF THE STUDY
The findings from this study represent the first comprehensive report of Vibrio isolates from ready-to-eat shrimps in Edo and Delta States, Nigeria. Incessant monitoring of Vibrio strains and their predisposition to antimicrobials is a necessity to guarantee seafood safety and guarantee the best treatment regimen for patients with gastroenteritis.
Topics: Animals; Anti-Bacterial Agents; Drug Resistance, Microbial; Genes, Bacterial; Microbial Sensitivity Tests; Nigeria; Shellfish; Vibrio; Virulence Factors
PubMed: 31977129
DOI: 10.1111/jam.14590 -
Virulence Dec 2020is a foodborne pathogen, which is widely distributed in the aquatic environment. Moreover, it is often involved in aquatic animal diseases. In recent years, is an...
is a foodborne pathogen, which is widely distributed in the aquatic environment. Moreover, it is often involved in aquatic animal diseases. In recent years, is an emerging pathogen in some species of Siluriformes. The strain SCCF01 was isolated from yellow catfish (). In this study, we aimed to perform genomic analysis of strain SCCF01 to identify genetic features and evolutionary relationships. Information on gene function and classification was obtained by functional annotation, and circular graph of strain SCCF01 genome, which was created by Circos v0.64. Information on virulence genes (adhesion, flagellum system, exotoxin, and secretory system, etc.) was obtained by virulence genes annotation. Genome element prediction showed that most of the mobile elements were distributed in chromosome I. Therefore, chromosome I of SCCF01 genome has more plasticity than chromosome II and might be larger in size. Genomic linear relationship between the strain of s and strain SCCF01 was analyzed by linear pairwise comparison but was unable to determine the relationship. Gene family analysis predicted that the evolutionary direction of strain SCCF01 was: clinical strain → environmental strain → SCCF01 strain. Phylogenetic analysis showed that the strain SCCF01 was more closely related to environmental strains. According to gene family analysis and phylogenetic analysis, we speculated that strain SCCF01 has probably diverged from environmental strains.
Topics: Animals; Bacterial Adhesion; Catfishes; Exotoxins; Flagella; Fresh Water; Genes, Bacterial; Genomics; Interspersed Repetitive Sequences; Phylogeny; Vibrio mimicus; Virulence; Virulence Factors
PubMed: 31826705
DOI: 10.1080/21505594.2019.1702797 -
Scientific Reports Nov 2019A potential mechanism for the global distribution of waterborne pathogens is through carriage by the migratory waterbirds. However, this mode of transmission has yet...
A potential mechanism for the global distribution of waterborne pathogens is through carriage by the migratory waterbirds. However, this mode of transmission has yet been confirmed epidemiologically. Here, we conducted whole genome sequencing of Vibrio spp. collected from waterbirds, sediments, and mollusks in the estuary of the Liaohe River in China to investigate this transmission mode. We found that a V. parahaemolyticus strain isolated from a waterbird was clonally related to the other V. parahaemolyticus strains obtained from the sediments and mollusks, and three V. mimicus strains isolated from bird feces were genomically related to those found in the mollusks and upstream groundwater, suggesting that the bird-carried Vibrio strains were acquired through the direct predation of the local mollusks. Surprisingly, two bird-carried V. parahaemolyticus strains belonging to the same clone were identified in Panjin and Shanghai, which are over 1,150 km apart, and another two were found at two locations 50 km apart, further supporting that waterbirds are capable of carrying and disseminating these pathogens over long distances. Our results provide the first evidence of direct transmission from mollusks to waterbirds and confirm that waterbirds act as disseminating vehicles of waterborne pathogens. Effective surveillance of migratory waterbirds along their routes will be valuable for predicting future epidemics of infectious diseases.
Topics: Animal Migration; Animals; Birds; Fisheries; Food Contamination; Genome, Bacterial; Genomics; Geography; Phylogeny; Phylogeography; Public Health Surveillance; Rivers; Vibrio; Vibrio Infections
PubMed: 31704994
DOI: 10.1038/s41598-019-52791-5 -
Fish & Shellfish Immunology Mar 2020Intestinal mucosal immunity plays a vital role against Vibrio mimicus infection because it is an enteric pathogen causing serious vibriosis in fish. In the previous...
Intestinal mucosal immunity plays a vital role against Vibrio mimicus infection because it is an enteric pathogen causing serious vibriosis in fish. In the previous studies, we developed an oral double-targeted DNA vaccine of V. mimicus and demonstrated that the vaccine could elicit significantly higher intestinal mucosal immune response than did naked DNA vaccine. But, little is known underlying regulatory molecular mechanisms of the enhanced intestinal mucosal immunity. Here the transcriptome and proteome in the intestines of the grass carps immunized or not with the double-targeted DNA vaccine were investigated by using RNA-seq and iTRAQ-coupled LC-MS/MS. Compared with the control group, a total of 5339 differentially expressed genes (DEGs) and 1173 differentially expressed proteins (DEPs) were identified in the immunized fish intestines. Subsequently, the integrated analysis between transcriptome and proteome data revealed that 250 DEPs were matched with the corresponding DEGs (named associated DEPs/DEGs) at both transcriptome and proteome levels. Fifty of all the associated DEPs/DEGs were immune-related and mainly enriched in phagosome, antigen-processing and presentation, complement and coagulation cascades, NLRs and MAPK signaling pathways via Gene Ontology and KEGG pathway analyses, which suggested the coordination of the five activated pathways was essential to the enhanced intestinal mucosal immune response in the immunized fish. The protein-protein interaction analysis showed that 60 of the 63 immune-related DEPs to form an integrated network. Additionally, randomly selected DEGs and DEPs were respectively validated by quantitative real-time RT-PCR and multiple reaction monitoring (MRM) assay, indicating that the both RNA-Seq and iTRAQ results in the study were reliable. Overall, our comprehensive transcriptome and proteome data provide some key genes and their protein products for further research on the regulatory molecular mechanisms underlying the enhanced intestinal mucosal immunity.
Topics: Animals; Bacterial Vaccines; Carps; Gene Expression Profiling; Intestines; Proteome; Transcriptome; Vaccination; Vaccines, DNA; Vibrio Infections; Vibrio mimicus
PubMed: 31678536
DOI: 10.1016/j.fsi.2019.10.045 -
PeerJ 2019This study aimed to investigate the effects of as a dietary probiotic supplement in fishmeal based diet on growth, gut microbiota and immune performance of marron ()....
This study aimed to investigate the effects of as a dietary probiotic supplement in fishmeal based diet on growth, gut microbiota and immune performance of marron (). Marron were randomly distributed into two different treatment groups, control and probiotic fed group. After 42 days of feeding trial, the results revealed a significant ( < 0.05) increase in growth due to increase in number of moults in marron fed probiotics. The probiotic diet also significantly enhanced the total haemocyte counts (THC), lysozyme activity in the haemolymph and protein content of the tail muscle in marron. Compared to control, the 16S rRNA sequences data demonstrated an enrichment of bacterial diversity in the probiotic fed marron where significant increase of abundance was observed. The abundance for crayfish pathogen and were found to be significantly reduced post feeding with probiotic diet. Predicted metabolic pathway revealed an increased activity for the metabolism and absorption of carbohydrate, degradation of amino acid, fatty acid and toxic compounds, and biosynthesis of secondary metabolites. supplementation also significantly modulated the expression level of immune-responsive genes of marron post challenged with . The overall results suggest that could be used as dietary probiotic supplement in marron aquaculture.
PubMed: 31523510
DOI: 10.7717/peerj.7553 -
Fish & Shellfish Immunology Sep 2019Vibrio mimicus (V. mimicus) is a significant pathogen in freshwater catfish, though knowledge of virulence determinants and effective vaccine is lacking. Multiplex...
Vibrio mimicus (V. mimicus) is a significant pathogen in freshwater catfish, though knowledge of virulence determinants and effective vaccine is lacking. Multiplex genome editing by natural transformation (MuGENT) is an easy knockout method, which has successfully used in various bacteria except for V. mimicus. Here, we found V. mimicus strain SCCF01 can uptake exogenous DNA and insert it into genome by natural transformation assay. Subsequently, we exploited this property to make five mutants (△Hem, △TS1, △TS2, △TS1△TS2, and △II), and removed the antibiotic resistance marker by Flp-recombination. Finally, all of the mutants were identified by PCR and RT-PCR. The results showed that combination of natural transformation and FLP-recombination can be applied successfully to generate targeted gene disruptions without the antibiotic resistance marker in V. mimicus. In addition, the five mutants showed mutant could be inherited after several subcultures and a 668-fold decrease in the virulence to yellow catfish (Pelteobagrus fulvidraco). This study provides a convenient method for the genetic manipulation of V. mimicus. It will facilitate the identification and characterization of V. mimicus virulence factors and eventually contribute to a better understanding of V. mimicus pathogenicity and development of attenuated vaccine.
Topics: Animals; Bacterial Vaccines; Catfishes; Fish Diseases; Gene Editing; Gene Knockout Techniques; Vaccines, Attenuated; Vibrio Infections; Vibrio mimicus
PubMed: 31202969
DOI: 10.1016/j.fsi.2019.06.025 -
Marine Pollution Bulletin Apr 2019Analyses of thermotolerant coliform and heterotrophic bacteria as well as Escherichia coli and Vibrio species were carried out on plastic samples and in the surrounding...
Analyses of thermotolerant coliform and heterotrophic bacteria as well as Escherichia coli and Vibrio species were carried out on plastic samples and in the surrounding waters of Guanabara Bay to evaluate plastic debris as vehicles of bacterial dispersal. Chemical characterizations of plastics were performed using Fourier transform infrared spectroscopy (FTIR). Plastic debris with high coliform contents were found, while their respective water samples had only low titers. No correlations were observed, however, between the amounts of bacteria and the chemical compositions of the plastic debris. Forty-four bacterial strains were PCR-confirmed as E. coli pathotypes, and 59 strains of Vibrio spp. (with 12 being identified as Vibrio cholerae [6], Vibrio vulnificus [5], and Vibrio mimicus [1]). These findings suggest these plastics can function as a substrate for bacterial biofilms (including pathogens). These debris, in turn, can be dispersed in aquatic environments not otherwise showing recent fecal bacterial contamination.
Topics: Bays; Biofilms; Brazil; Enterobacteriaceae; Escherichia coli; Heterotrophic Processes; Plastics; Polymerase Chain Reaction; Spectroscopy, Fourier Transform Infrared; Vibrio; Vibrio vulnificus; Waste Products; Wastewater; Water Pollutants, Chemical
PubMed: 30955768
DOI: 10.1016/j.marpolbul.2019.02.064