Vibrio Isolates Research Articles

Potential role of alginate in marine bacteria-yeast interactions.

The ability of microorganisms to decompose brown algae has attracted attention. This study aims to clarify the characteristics of marine microbial communities in which prokaryotic and eukaryotic microorganisms interact via the metabolism of brown algae carbohydrates. Amplicon-based microbiome analysis revealed the predominance of the genera Marinomonas and Vibrio in seawater and seaweed samples mixed with alginate and mannitol, which are the primary carbohydrates in brown algae. Three Vibrio species and Candida intermedia were isolated via alginate enrichment culture. Although C. intermedia did not utilize alginate as a nutrient source, the yeast grew in the spent alginate medium in which Vibrio algivorus had been cultured. Coculture with C. intermedia and the Vibrio isolates, especially V. algivorus, also enhanced the growth of the yeast on alginate. These results suggested that C. intermedia grew because of the supply of nutrients via alginate metabolism by Vibrio species. In the coculture medium, the amount of phosphatidylserine increased in the early phase but decreased with the growth of C. intermedia, indicating that phosphatidylserine secreted by Vibrio is involved in the putative mutualistic interaction. We examined whether such interaction is applicable to the production of useful substances and succeeded in lipid production by oleaginous marine yeast Yarrowia lipolytica through coculture with V. algivorus. Our study suggested the potential of mutualistic interaction via degradation of alginate by marine Vibrio for production of industrially useful substances in yeast cells.IMPORTANCEIn this study, we analyzed the microbiome of seawater and seaweed in the presence of brown algae carbohydrates and reconstructed the putative mutualistic relationship of marine Vibrio and Candida intermedia mediated by metabolism of brown algae in the ocean.

Polyphasic identification of Vibrio species from aquatic sources using mass spectrometry, housekeeping gene sequencing and whole genome analysis

Accurate bacterial identification is essential for determining the causative agent of an infection, thus facilitating appropriate treatment and management strategies in both human and animal health contexts. Some species in the Vibrio genus are recognized pathogens, associated with a variety of infections. However, identification of these bacteria is oftentimes controversial. Therefore, we aimed to evaluate different identification approaches in terms of their reliability in distinguishing Vibrio species. To achieve this, we selected a set of 40 Vibrio isolates previously recovered from water and floating plastic samples in a large bay environment and identified them employing MALDI-TOF mass spectrometry, and rrs and pyrH gene sequencing. A subset of isolates was also submitted to whole genome sequencing. Overall, MALDI-TOF was found to be a fast-screening methodology for identification, notably at genus-level. However, for better species discrimination, pyrH gene sequencing stood out as a more reliable tool in contrast to rrs gene sequencing and MALDI-TOF, as corroborated by whole genome sequencing analysis.

Further Evidence of Anthropogenic Impact: High Levels of Multiple-Antimicrobial-Resistant Bacteria Found in Neritic-Stage Sea Turtles

Background/Objectives: Marine turtles are globally threatened and face daily anthropogenic threats, including pollution. Water pollution from emerging contaminants such as antimicrobials is a major and current environmental concern. Methods: This study investigated the phenotypic antimicrobial resistance and heavy metal resistance genes of 47 Vibrio isolates from different stages of sea turtles (oceanic stage vs neritic stage) from the Taiwanese coast. Results: The results show that a high proportion (48.9%; 23/47) of the Vibrio species isolated from sea turtles in our study had a multiple antimicrobial resistance (MAR) pattern. It was found that Vibrio spp. isolates with a MAR pattern and those with a MAR index value greater than 0.2 were both more likely to be observed in neritic-stage sea turtles. Furthermore, isolates from neritic-stage sea turtles exhibited greater resistance to the majority of antimicrobials tested (with the exception of beta-lactams and macrolides) than isolates from the oceanic-stage groups. Isolates from neritic sea turtles were found to be more resistant to nitrofurans and aminoglycosides than isolates from oceanic sea turtles. Furthermore, isolates with a MAR pattern (p = 0.010) and those with a MAR index value greater than 0.2 (p = 0.027) were both found to be significantly positively associated with the mercury reductase (merA) gene. Conclusions: The findings of our study indicate that co-selection of heavy metals and antimicrobial resistance may occur in aquatic bacteria in the coastal foraging habitats of sea turtles in Taiwan.

Prophages in Vibrio.

Although tailed bacteriophages (phages) of the class Caudoviricetes are thought to constitute the most abundant and ecologically relevant group of phages that can integrate their genome into the host chromosome, it is becoming increasingly clear that other prophages are widespread. Here, we show that prophages derived from filamentous and tailless phages with genome sizes below 16 kb make up the majority of prophages in marine bacteria of the genus Vibrio. To estimate prophage prevalence unaffected by database biases, we combined comparative genomics and chemical induction of 58 diverse Vibrio cyclitrophicus isolates, resulting in 107 well-curated prophages. Complemented with computationally predicted prophages, we obtained 1,158 prophages from 931 naturally co-existing strains of the family Vibrionaceae. Prophages resembling tailless and filamentous phages predominated, accounting for 80% of all prophages in V. cyclitrophicus and 60% across the Vibrionaceae. In our experimental model, prophages of all three viral realms actively replicated upon induction indicating their ability to transfer to new hosts. Indeed, prophages were rapidly gained and lost, as suggested by variable prophage content between closely related V. cyclitrophicus. Prophages related to filamentous and tailless phages were integrated into only three genomic locations and restored the function of their integration site. Despite their small size, they contained highly diverse accessory genes that may contribute to host fitness, such as phage defense systems. We propose that, like their well-studied tailed equivalent, tailless and filamentous temperate phages are active and highly abundant drivers of host ecology and evolution in marine Vibrio, which have been largely overlooked.

Antimicrobial resistance and genotypic attributes of virulence among Vibrio spp. isolated from Japanese retail seafood

Bacterial diseases caused by Vibrio spp. are a significant impediment to the aquaculture industry owing to difficult-to-treat infections and mortality in farmed animals, affecting socioeconomic development. This study aimed to evaluate the role of Japanese retail seafood in the production of antimicrobial-resistant Vibrio spp. and pathogenic factors. One hundred seafood samples purchased from various retail supermarkets in Hiroshima, Japan, were examined using microbiological, phenotypic, and molecular techniques. One hundred and twenty-eight Vibrio isolates belonging to more than 11 species were identified and characterized. Among the 100 samples, 26 (26 %) tested positive for Vibrio spp. with antimicrobial resistance, while 44 (44 %) were positive for those carrying virulence determinants. Out of 128 isolates, V. alginolyticus was predominant (38 %), followed by V. parahaemolyticus (29 %), V. neocaledonicus (7.0 %), V. cholerae (6.3 %), among others. For virulence gene assessment, only six of the 10 virulence attributes, tlh (34.4 %), VPI (29.7 %), ompW (7.0 %), toxR (5.5 %), hlyA (3.9 %), and ompU (3.1 %), were detected in the isolates. tcpA, ctxA, ctxB, and tdh are absent. Eight V. cholerae isolates of international origin were VPI-positive, seven of which harbored the toxR regulon and other virulence genes. The blaTEM resistance gene was identified in V. alginolyticus (22 isolates). All 46 antimicrobial resistance gene-harboring isolates were phenotypically susceptible to cephalosporins, aztreonam, meropenem, tetracycline, chloramphenicol, and ciprofloxacin. Ampicillin resistance was the highest (91.3 %), followed by colistin (30.4 %) and fosfomycin (15.2 %). The multiple-antibiotic resistance index ranged from 0.2 to 0.47, suggesting that some isolates originate from high-risk contamination sources. Our results shed light on the incidence of potentially pathogenic Vibrio spp. and highlighted the importance of continuous monitoring to improve food safety in the seafood supply chain. These findings will be useful for developing microbiological risk assessments for retail seafood management.

Genomic evidence of two-staged transmission of the early seventh cholera pandemic

The seventh cholera pandemic started in 1961 in Indonesia and spread across the world in three waves in the decades that followed. Here, we utilised genomic evidence to detail the first wave of the seventh pandemic. Genomes of 22 seventh pandemic Vibrio cholerae isolates from 1961 to 1979 were completely sequenced. Together with 152 publicly available genomes from the same period, they fell into seven phylogenetic clusters (CL1–CL7). By multilevel genome typing (MGT), all were assigned to MGT2 ST1 (Wave 1) except three isolates in CL7 which were typed as MGT2 ST2 (Wave 2). The Wave 1 seventh pandemic expanded in two stages, with Stage 1 (CL1–CL5) spread across Asia and Stage 2 (CL6 and CL7) spread to the Middle East and Africa. Three non-synonymous mutations, one each, in three regulatory genes, csrD (global regulator), acfB (chemotaxis), and luxO (quorum sensing) may have critically contributed to its pandemicity. The three MGT2 ST2 isolates in CL7 were the progenitors of Wave 2 and evolved from within Wave 1 with acquisition of a novel IncA/C plasmid. Our findings provide new insight into the evolution and transmission of the early seventh pandemic, which may aid future cholera prevention and control.

Thiolated chitosan nanoparticles encapsulated nisin and selenium: antimicrobial/antibiofilm/anti-attachment/immunomodulatory multi-functional agent

BackgroundThe increase in the resistance of bacterial strains to antibiotics has led to research into the bactericidal potential of non-antibiotic compounds. This study aimed to evaluate in vitro antibacterial/ antibiofilm properties of nisin and selenium encapsulated in thiolated chitosan nanoparticles (N/Se@TCsNPs) against prevalent enteric pathogens including standard isolates of Vibrio (V.) cholerae O1 El Tor ATCC 14,035, Campylobacter (C.) jejuni ATCC 29,428, Salmonella (S.) enterica subsp. enterica ATCC 19,430, Shigella (S.) dysenteriae PTCC 1188, Escherichia (E.) coli O157:H7 ATCC 25,922, Listeria (L.) monocytogenes ATCC 19,115, and Staphylococcus (S.) aureus ATCC 29,733.MethodsThe synthesis and comprehensive analysis of N/Se@TCsNPs have been completed. Antibacterial and antibiofilm capabilities of N/Se@TCsNPs were evaluated through broth microdilution and crystal violet assays. Furthermore, the study included examining the cytotoxic effects on Caco-2 cells and exploring the immunomodulatory effects of N/Se@TCsNPs. This included assessing the levels of both pro-inflammatory (IL-6 and TNFα) and anti-inflammatory (IL-10 and TGFβ) cytokines and determining the gene expression of TLR2 and TLR4.ResultsThe N/Se@TCsNPs showed an average diameter of 136.26 ± 43.17 nm and a zeta potential of 0.27 ± 0.07 mV. FTIR spectroscopy validated the structural features of N/Se@TCsNPs. Scanning electron microscopy (SEM) images confirmed their spherical shape and uniform distribution. Thermogravimetric Analysis (TGA)/Differential Scanning Calorimetry (DSC) tests demonstrated the thermal stability of N/Se@TCsNPs, showing minimal weight loss of 0.03%±0.06 up to 80 °C. The prepared N/Se@TCsNPs showed a thiol content of 512.66 ± 7.33 µmol/g (p < 0.05), an encapsulation efficiency (EE) of 69.83%±0.04 (p ≤ 0.001), and a drug release rate of 74.32%±3.45 at pH = 7.2 (p ≤ 0.004). The synthesized nanostructure demonstrated potent antibacterial activity against various isolates, with effective concentrations ranging from 1.5 ± 0.08 to 25 ± 4.04 mg/mL. The ability of N/Se@TCsNPs to reduce bacterial adhesion and internalization in Caco-2 cells underscored their antibiofilm properties (p ≤ 0.0001). Immunological studies indicated that treatment with N/Se@TCsNPs led to decreased levels of inflammatory cytokines IL-6 (14.33 ± 2.33 pg/mL) and TNFα (25 ± 0.5 pg/mL) (p ≤ 0.0001), alongside increased levels of anti-inflammatory cytokines IL-10 (46.00 ± 0.57 pg/mL) and TGFβ (42.58 ± 2.10 pg/mL) in infected Caco-2 cells (p ≤ 0.0001). Moreover, N/Se@TCsNPs significantly reduced the expression of TLR2 (0.22 ± 0.09) and TLR4 (0.16 ± 0.05) (p < 0.0001).ConclusionIn conclusion, N/Se@TCsNPs exhibited significant antibacterial/antibiofilm/anti-attachment/immunomodulatory effectiveness against selected Gram-positive and Gram-negative enteric pathogens. However, additional ex-vivo and in-vivo investigations are needed to fully assess the performance of nanostructured N/Se@TCsNPs.

Vibrio neptunius-ULV11 cell-free supernatant as a promising antifouling approach in reverse osmosis systems

One of the most common problems observed in continuous operation of desalination plants, is the presence of biofouling in ultrafiltration (UF) modules and reverse osmosis membranes (ROMs), decreasing permeated water obtention and increasing energy consumption among other adverse effects. To solve this problem, desalination industry applied UF modules and ROMs cleaning procedures that aims to increase half-life operation time, however, many of the actual cleaning procedures are focused on the addition of chemical compounds that provokes secondary effects such as UF modules and ROMs disintegration leading to structural changes and a decrease in plant performance. Thus, new approaches to replace current cleaning compounds with new ecofriendly and less secondary effects technologies are necessary.In this study, we present a Vibrio neptunius strain ULV11 isolate, with a high capacity to remove fouling in RO systems. In detail, ULV11 cell-free supernatant (CFS) was used in replace of chemical agents as a cleaning solution according to standard operational protocol on 3-year-old membranes of continuous operation. The results showed that CFS can be used under operational conditions (such high pressure and flux) allowed us to recover permeate volumes similar to those observed in traditional chemical treatment with better conductivity. This recovery was accompanied by a reestablishment of membrane topology observed by atomic force microscopy and EPS remotion since concanavalin A signal trough confocal microscopy was decrease as compared with biofouling control.

Study on Antibiotic Resistance Profiles Exhibited by Vibrio Species Isolated from Landfill Soils in Zaria Metropolis, Northern Nigeria

Study’s Novelty/Excerpt This study investigates the occurrence and antibiotic resistance profiles of Vibrio species isolated from landfill soils in Zaria Metropolis, with a focus on environmental reservoirs of antibiotic resistance. The research uniquely identifies a high prevalence of Vibrio cholerae non-O1 among the isolates and reveals significant multidrug resistance (MDR), particularly against Ampicillin, while also showing complete susceptibility to Gentamicin and Chloramphenicol. These findings highlight the potential for landfills to serve as environmental reservoirs of antibiotic resistance, underscoring the need for further research to understand the implications for public health and environmental management. Full Abstract Antibiotic resistance in bacteria presents a major risk to public health and the environment. Infections caused by Vibrio species continue to be a serious public health concern. This study investigated the occurrence and antibiotic resistance profiles of Vibrio species isolates from landfill soils in Zaria Metropolis. A total of one hundred and twenty (120) soil samples were collected from designated landfills in four locations in Sabon-Gari, Samaru, Tudun-Wada, and Zaria City. Vibrio species were isolated using Thiosulphate citrate bile salt sucrose (TCBS) agar. Bacteriological analysis of the soil samples revealed 9(7.50%) Vibrio species isolates with Vibrio cholerae non-O1 exhibiting the highest prevalence of 4 (3.33%) among all isolates. Using the Kirby-Bauer method, the isolates were tested for susceptibility against ten commonly used antibiotics belonging to three different classes. The highest resistance was to Ampicillin (88. 89%), while all the isolates (100%) showed susceptibility to Gentamicin and Chloramphenicol. Five isolates (55.56%) were Multidrug Resistance (MDR). The highest Multidrug Resistance (MDR, 60%) was observed in Vibrio cholerae non-O1. The isolate resistant to the highest number of antibiotics was obtained from the Tudun-Wada sample location. Three isolates (33.33%) showed resistance to 4 antibiotics, while 2 isolates, 1(11.11%) each were resistant to 5 and 6 antibiotics. The isolates were Multiple Antibiotic Resistance (MAR) to 4-6 antibiotics, and four different phenotypic resistance profiles were observed among them. The origin and varying levels of resistance to multiple antibiotics indicated could be traced to the faecal constituent of the waste in landfills produced by people or animals that have been treated indiscriminately with various antibiotics or items containing residual antimicrobial agents disposed of in dump soils, highlighting the potential environmental reservoirs of antibiotic resistance and call for further research to understand the implications for public health and environmental management.

Antibiotic resistance trends among Vibrio vulnificus and Vibrio parahaemolyticus isolated from the Chesapeake Bay, Maryland: a longitudinal study.

Antibiotics are often used to treat severe Vibrio infections, with third-generation cephalosporins and tetracyclines combined or fluoroquinolones alone being recommended by the US Centers for Disease Control and Prevention. Increases in antibiotic resistance of both environmental and clinical vibrios are of concern; however, limited longitudinal data have been generated among environmental isolates to inform how resistance patterns may be changing over time. Hence, we evaluated long-term trends in antibiotic resistance of vibrios isolated from Chesapeake Bay waters (Maryland) across two 3-year sampling periods (2009-2012 and 2019-2022). Vibrio parahaemolyticus (n = 134) and Vibrio vulnificus (n = 94) toxR-confirmed isolates were randomly selected from both sampling periods and tested for antimicrobial susceptibility against eight antibiotics using the Kirby-Bauer disk diffusion method. A high percentage (94%-96%) of V. parahaemolyticus isolates from both sampling periods were resistant to ampicillin and only 2%-6% of these isolates expressed intermediate resistance or resistance to third-generation cephalosporins, amikacin, tetracycline, and trimethoprim-sulfamethoxazole. Even lower percentages of resistant V. vulnificus isolates were observed and those were mostly recovered from 2009 to 2012, however, the presence of multiple virulence factors was observed. The frequency of multi-drug resistance was relatively low (6%-8%) but included resistance against antibiotics used to treat severe vibriosis in adults and children. All isolates were susceptible to ciprofloxacin, a fluoroquinolone, indicating its sustained efficacy as a first-line agent in the treatment of severe vibriosis. Overall, our data indicate that antibiotic resistance patterns among V. parahaemolyticus and V. vulnificus recovered from the lower Chesapeake Bay have remained relatively stable since 2009.IMPORTANCEVibrio spp. have historically been susceptible to most clinically relevant antibiotics; however, resistance and intermediate-resistance have been increasingly recorded in both environmental and clinical isolates. Our data showed that while the percentage of multi-drug resistance and resistance to antibiotics was relatively low and stable across time, some Vibrio isolates displayed resistance and intermediate resistance to antibiotics typically used to treat severe vibriosis (e.g., third-generation cephalosporins, tetracyclines, sulfamethoxazole-trimethoprim, and aminoglycosides). Also, given the high case fatality rates observed with Vibrio vulnificus infections, the presence of multiple virulence factors in the tested isolates is concerning. Nevertheless, the continued susceptibility of all tested isolates against ciprofloxacin, a fluoroquinolone, is indicative of its use as an effective first-line treatment of severe Vibrio spp. infections stemming from exposure to Chesapeake Bay waters or contaminated seafood ingestion.

Antibiotics resistance and phenotypic virulence factors of Vibrio species isolated from poultry litters at Obafemi Awolowo University, research farm.

The study investigates the antimicrobial resistance and phenotypic virulence factors of Vibrio isolates in poultry litter. Samples were enriched in 5 mL alkaline peptone water 1:10 (w/v) for 6 hours and cultured on Thiosulphate Citrate Bile salt (TCBS) agar. Vibrio isolates were identified through biochemical tests assay and were subjected to antibiotic susceptibility tests using standards disc diffusion technique. The phenotypic virulence factors, including hemolytic and proteolytic activity were determined. A total of 100 Vibrio isolates were identified as Vibrio parahaemolyticus (49%), V. cholerae (26%), V. vulnificus (11%), V. alginolyticus (9%), and V. damsela (5%). The Vibrio isolates showed a significant resistance to the selected antibiotics tested with 99% resistant to Gentamycin, 95% Cefixime, 96% Ofloxacin, 98% Augmentin, 96% Nitrofurantoin, 100% Ciprofloxacin, and 100% Cefuroxime. The Vibrio isolates showed varying levels of haemolysis with 17% alpha haemolysis, 61% beta haemolysis, 22% gamma haemolysis, and 29% proteolytic activity. The study reveals that poultry litter contains Vibrio isolates with antibiotic resistance and virulence factors, increasing their risk of severe infections. The high beta haemolysis prevalence highlights the interconnected relationship between animal, environmental, and human health, underscoring the need for effective management.

Complete genome of Vibrio harveyi isolate K2014767 from the hepatopancreas of captive Caribbean spiny lobster (Panulirus argus).

The complete genome sequence is reported for Vibrio harveyi isolate K2014767, isolated from a captive Caribbean spiny lobster (Panulirus argus) during a species-specific mortality event in a public display aquarium in the United States.

Prevalence, genetic diversity, and antimicrobial susceptibility of Vibrio spp. infected gilthead sea breams from coastal farms at Damietta, Egypt

BackgroundVibriosis is one of the most serious bacterial diseases and causes high morbidity and mortality among cultured sea breams. This study was undertaken to track the surveillance of Vibrio infection and its correlation to environmental factors. A total of 115 gilthead sea breams were collected seasonally from a private earthen pond fish farm in the Shatta area of Damietta, Egypt from September 2022 to July 2023. Physicochemical parameters of water were analyzed, and heavy metal levels were measured. The fish samples were subjected to clinical, bacteriological, Enterobacterial Repetitive Intergenic Consensus (ERIC) fingerprinting, and hematoxylin and Eosin histopathological staining.ResultsThe results revealed significant variations in the water quality parameters over different seasons, in addition to an increase in heavy metals. Naturally infected fish showed external signs and postmortem lesions that were relevant to bacterial infection. Two dominant Vibrio subspecies of bacteria were identified: V. alginolyticus (205 isolates) and V. fluvialis (87 isolates). PCR confirmed the presence of V. alginolyticus using the species-specific primer collagenase at 737 bp. The highest prevalence of V. alginolyticus was detected during the summer season (57.72%), and the lowest prevalence was observed in autumn (39.75%). The correlation analysis revealed a positive relationship between V. alginolyticus and water temperature (r = 0.69). On the other hand, V. fluvialis showed a high prevalence during the autumn season (25.30%) and the lowest prevalence during the summer season (10.56%), where it was negatively correlated with water temperatures (r =—0.03). ERIC fingerprinting showed genetic variation within the Vibrio isolates. Antimicrobial susceptibility testing revealed sensitivity to ciprofloxacin and doxycycline, and resistance to amoxicillin and erythromycin. The multiple antibiotic resistance (MAR) index values for V. alginolyticus and V. fluvialis ranged from 0.3 to 0.7, with a multi-drug resistance pattern to at least three antibiotics. Histopathological alterations in the affected tissues revealed marked hemorrhage, vascular congestion, and hemosiderosis infiltration.ConclusionThis study provides insights into the potential propagation of waterborne diseases and antibiotic resistance in the environment. Ensuring that the environment does not serve as a reservoir for virulent and contagious Vibrio species is a critical concern for regional aquaculture industries. Therefore, we recommend implementing environmental context-specific monitoring and surveillance tools for microbial resistance.

Detection and quantitative microbial risk assessment of pathogenic Vibrio cholerae in a river used for drinking, domestic, fresh produce irrigation and recreational purposes

Cholera infection results from the ingestion of water or food contaminated with toxigenic Vibrio cholerae. This study evaluated the occurrence of toxigenic V. cholerae in Asata River, Enugu Metropolis, Nigeria, and estimated V. cholerae infection risks from use of the River water for drinking, domestic and recreational purposes. Vibrio was detected and quantified using membrane filtration technique and thiosulfate–citrate–bile salts–sucrose agar. Isolates were screened by PCR, using specific primers targeting the internal transcribed spacer (its) region between 16 and 23S rRNA and the cholera toxin (ctx) gene. Sequenced 16SrRNA gene amplicons of two selected isolates were used for phylogenetic analysis. Quantitative microbial risk assessment (QMRA) was conducted using the β-Poisson dose–response model. About 81% (58/72) of Asata River samples recorded Vibrio counts above 1.0 ×\\documentclass[12pt]{minimal} \\usepackage{amsmath} \\usepackage{wasysym} \\usepackage{amsfonts} \\usepackage{amssymb} \\usepackage{amsbsy} \\usepackage{mathrsfs} \\usepackage{upgreek} \\setlength{\\oddsidemargin}{-69pt} \\begin{document}$$\ imes$$\\end{document} 103 cfu/100 mL. Of the fifty Vibrio isolates screened, its was detected in 54% (27/50), out of which 74% (20/27) had the ctx gene of toxigenic V. cholerae. Evolutionary relatedness of the sequenced isolates to V. cholerae was revealed. The estimated risks of cholera infection in persons exposed to untreated Asata River water were above 0.5 for all the exposure scenarios, for both the rainy and dry seasons. The risks were highest (~ 0.9) for exposure via drinking water and annual risk of infection was deduced to have a probability of 1.0. Therefore, dependence on the untreated Asata River water for drinking, recreational, domestic and irrigation purposes may present a potential public health risk of cholera outbreak. We recommend increased monitoring and surveillance of River water for Vibrio abundance and that Asata River be protected from further degradation.

Biofilm inhibitor Con A lectin feed additive protects shrimp against pathogenic Vibrio harveyi and V. parahaemolyticus

Extracellular matrix (ECM) is a major component of all bacterial biofilms that protect the bacterial cells living within. It consists mainly of carbohydrates, proteins, lipids, and nucleic acids. We characterized the ECM of an acute hepatopancreatic necrosis disease (AHPND) causing Vibrio parahaemolyticus isolate 3HP and two luminescent shrimp disease-causing V. harveyi isolates (1114GL or VH0 and its Siphoviridae phage-lysogenized isolate, VH1) using 11 FITC-labeled plant lectins with different terminal sugar specificities. Canavalia ensiformis lectin (Con A), wheat germ lectin (Triticum vulgaris, WGA), Griffonia simplicifolia lectin II (GSL-II), and Maclura pomifera Lectin (MPL) were found to react with the biofilms of all three isolates. Based on these results and availability, Con A and WGA and their respective specific sugars (D-mannose or glucose and N-acetyl-D-glucosamine) were tested for effects on V. harveyi and V. parahaemolyticus biofilm formations. Biofilms of these bacteria were evaluated by thickness after growth with and without the two lectins and with and without their respective sugar targets at different concentrations. Only Con A at 0.1 mg/ml and 0.5 mg/ml (with and without D-mannose or d-glucose) were found to inhibit biofilms of all three Vibrio isolates in a dose-dependent manner. Con A was then tested for efficacy in protecting shrimp against V. harveyi (VH1) and V. parahaemolyticus 3-HP by its addition to shrimp feed at 0.05 mg/g and 0.5 mg/g and fed for seven days before they were immersion challenged with either VH1 or 3HP on day 8. After the challenge, the shrimp continued to be fed with their respective feed, and their health and survival were monitored for another 11 and 12 days for the VH1 and 3HP experiments, respectively. From both experiments, the groups fed with Con A-supplemented feed gave significantly higher survival rates [51.9% and 53.6% (VH1) and 52.8% and 62.9% (3HP)] than those fed with un-supplemented feed [38.3% (VH1) and 21.2% (3HP)] (p < 0.05). These results suggested that D-mannose/d-glucose plays an essential role in V. harveyi and V. parahaemolyticus biofilm formations, and Con A lectin might be helpful in monitoring and controlling biofilms to prevent Vibrio diseases.

Recovering high-quality bacterial genomes from cross-contaminated cultures: a case study of marine Vibrio campbellii

BackgroundEnvironmental monitoring of bacterial pathogens is critical for disease control in coastal marine ecosystems to maintain animal welfare and ecosystem function and to prevent significant economic losses. This requires accurate taxonomic identification of environmental bacterial pathogens, which often cannot be achieved by commonly used genetic markers (e.g., 16S rRNA gene), and an understanding of their pathogenic potential based on the information encoded in their genomes. The decreasing costs of whole genome sequencing (WGS), combined with newly developed bioinformatics tools, now make it possible to unravel the full potential of environmental pathogens, beyond traditional microbiological approaches. However, obtaining a high-quality bacterial genome, requires initial cultivation in an axenic culture, which is a bottleneck in environmental microbiology due to cross-contamination in the laboratory or isolation of non-axenic strains.ResultsWe applied WGS to determine the pathogenic potential of two Vibrio isolates from coastal seawater. During the analysis, we identified cross-contamination of one of the isolates and decided to use this dataset to evaluate the possibility of bioinformatic contaminant removal and recovery of bacterial genomes from a contaminated culture. Despite the contamination, using an appropriate bioinformatics workflow, we were able to obtain high quality and highly identical genomes (Average Nucleotide Identity value 99.98%) of one of the Vibrio isolates from both the axenic and the contaminated culture. Using the assembled genome, we were able to determine that this isolate belongs to a sub-lineage of Vibrio campbellii associated with several diseases in marine organisms. We also found that the genome of the isolate contains a novel Vibrio plasmid associated with bacterial defense mechanisms and horizontal gene transfer, which may offer a competitive advantage to this putative pathogen.ConclusionsOur study shows that, using state-of-the-art bioinformatics tools and a sufficient sequencing effort, it is possible to obtain high quality genomes of the bacteria of interest and perform in-depth genomic analyses even in the case of a contaminated culture. With the new isolate and its complete genome, we are providing new insights into the genomic characteristics and functional potential of this sub-lineage of V. campbellii. The approach described here also highlights the possibility of recovering complete bacterial genomes in the case of non-axenic cultures or obligatory co-cultures.

Unveiling potential virulence determinants in Vibrio isolates from Anadara tuberculosa through whole genome analyses.

The genus Vibrio includes pathogenic bacteria able to cause disease in humans and aquatic organisms, leading to disease outbreaks and significant economic losses in the fishery industry. Despite much work on Vibrio in several marine organisms, no specific studies have been conducted on Anadara tuberculosa. This is a commercially important bivalve species, known as "piangua hembra," along Colombia's Pacific coast. Therefore, this study aimed to identify and characterize the genomes of Vibrio isolates obtained from A. tuberculosa. Bacterial isolates were obtained from 14 A. tuberculosa specimens collected from two locations along the Colombian Pacific coast, of which 17 strains were identified as Vibrio: V. parahaemolyticus (n = 12), V. alginolyticus (n = 3), V. fluvialis (n = 1), and V. natriegens (n = 1). Whole genome sequence of these isolates was done using Oxford Nanopore Technologies (ONT). The analysis revealed the presence of genes conferring resistance to β-lactams, tetracyclines, chloramphenicol, and macrolides, indicating potential resistance to these antimicrobial agents. Genes associated with virulence were also found, suggesting the potential pathogenicity of these Vibrio isolates, as well as genes for Type III Secretion Systems (T3SS) and Type VI Secretion Systems (T6SS), which play crucial roles in delivering virulence factors and in interbacterial competition. This study represents the first genomic analysis of bacteria within A. tuberculosa, shedding light on Vibrio genetic factors and contributing to a comprehensive understanding of the pathogenic potential of these Vibrio isolates.IMPORTANCEThis study presents the first comprehensive report on the whole genome analysis of Vibrio isolates obtained from Anadara tuberculosa, a bivalve species of great significance for social and economic matters on the Pacific coast of Colombia. Research findings have significant implications for the field, as they provide crucial information on the genetic factors and possible pathogenicity of Vibrio isolates associated with A. tuberculosa. The identification of antimicrobial resistance genes and virulence factors within these isolates emphasizes the potential risks they pose to both human and animal health. Furthermore, the presence of genes associated with Type III and Type VI Secretion Systems suggests their critical role in virulence and interbacterial competition. Understanding the genetic factors that contribute to Vibrio bacterial virulence and survival strategies within their ecological niche is of utmost importance for the effective prevention and management of diseases in aquaculture practices.

Genomic and functional insights into antibiotic resistance genes floR and strA linked with the SXT element of Vibrio cholerae non-O1/non-O139.

The emergence and spread of antibiotic-resistant bacterial pathogens are a critical public health concern across the globe. Mobile genetic elements (MGEs) play an important role in the horizontal acquisition of antimicrobial resistance genes (ARGs) in bacteria. In this study, we have decoded the whole genome sequences of multidrug-resistant Vibrio cholerae clinical isolates carrying the ARG-linked SXT, an integrative and conjugative element, in their large chromosomes. As in others, the SXT element has been found integrated into the 5'-end of the prfC gene (which encodes peptide chain release factor 3 involved in translational regulation) on the large chromosome of V. cholerae non-O1/non-O139 strains. Further, we demonstrate the functionality of SXT-linked floR and strAB genes, which confer resistance to chloramphenicol and streptomycin, respectively. The floR gene-encoded protein FloR belongs to the major facilitator superfamily efflux transporter containing 12 transmembrane domains (TMDs). Deletion analysis confirmed that even a single TMD of FloR is critical for the export function of chloramphenicol. The floR gene has two putative promoters, P1 and P2. Sequential deletions reveal that P2 is responsible for the expression of the floR. Deletion analysis of the N- and/or C-terminal coding regions of strA established their importance for conferring resistance against streptomycin. Interestingly, qPCR analysis of the floR and strA genes indicated that both of the genes are constitutively expressed in V. cholerae cells. Further, whole genome-based global phylogeography confirmed the presence of the integrative and conjugative element SXT in non-O1/non-O139 strains despite being non-multidrug resistant by lacking antimicrobial resistance (AMR) gene cassettes, which needs monitoring.

Antimicrobial resistance and genomic analysis of Vibrio parahaemolyticus isolates from foodborne outbreaks, Huzhou, China, 2019-2023.

The purpose of this study was to investigate the epidemiological and genomic characteristics of Vibrio parahaemolyticus (V. parahaemolyticus) isolates from outbreaks in Huzhou, China. This study aims to analyze the epidemiological data on V. parahaemolyticus outbreaks reported in Huzhou from 2019 to 2023. A total of 70 V. parahaemolyticus outbreak isolates were collected. The antibiotic resistance, serotypes, molecular typing, and genomic characteristics of these isolates were analyzed. Most outbreaks of V. parahaemolyticus infection occurred in the summer, and the majority of outbreaks occurred in restaurants and rural banquets. High resistance rates were observed for ampicillin (AMP, 24.29%), followed by tetracycline (TET, 15.71%) and trimethoprim-sulfamethoxazole (SXT, 15.71%). The newly emerged serotype O10:K4 became dominant from 2021 to 2023, with most isolates belonging to ST3. The resistance gene blaCARB was frequently detected among these isolates. The pulsed-field gel electrophoresis (PFGE) and whole-genome single-nucleotide polymorphisms (wgSNPs) effectively differentiated the nine outbreaks. The newly emerged serotype O10:K4 became dominant from 2021 to 2023, with most isolates being ST3. PFGE and WGS technologies provided reliable methods for typing and identifying V. parahaemolyticus for outbreaks.

Assessment of Vibrionaceae prevalence in seafood from Qidong market and analysis of Vibrio parahaemolyticus strains.

The aim of this study was to investigate the prevalence of Vibrionaceae family in retail seafood products available in the Qidong market during the summer of 2023 and to characterize Vibrio parahaemolyticus isolates, given that this bacterium is the leading cause of seafood-associated food poisoning. We successfully isolated a total of 240 Vibrionaceae strains from a pool of 718 seafood samples. The breakdown of the isolates included 146 Photobacterium damselae, 59 V. parahaemolyticus, 18 V. campbellii, and 11 V. alginolyticus. Among these, P. damselae and V. parahaemolyticus were the predominant species, with respective prevalence rates of 20.3% and 8.2%. Interestingly, all 59 isolates of V. parahaemolyticus were identified as non-pathogenic. They demonstrated proficiency in swimming and swarming motility and were capable of forming biofilms across a range of temperatures. In terms of antibiotic resistance, the V. parahaemolyticus isolates showed high resistance to ampicillin, intermediate resistance to cefuroxime and cefazolin, and were sensitive to the other antibiotics evaluated. The findings of this study may offer valuable insights and theoretical support for enhancing seafood safety measures in Qidong City.