Virulent properties and genomic diversity of Vibrio vulnificus isolated from environment, human, diseased fish.

The incidence of Vibrio vulnificus infections, with high mortality rates in humans and aquatic animals, has escalated, highlighting a significant public health challenge. Currently, reliable markers to identify strains with high virulence potential are lacking, and the understanding of evolutionary drivers behind the emergence of pathogenic strains is limited. In this study, we analyzed the distribution of virulent genotypes and phenotypes to discern the infectious potential of V. vulnificus strains isolated from three distinct sources. Most isolates, traditionally classified as biotype 1, possessed the virulence-correlated gene-C type. Environmental isolates predominantly exhibited YJ-like alleles, while clinical and diseased fish isolates were significantly associated with the nanA gene and pathogenicity region XII. Hemolytic activity was primarily observed in the culture supernatants of clinical and diseased fish isolates. Genetic relationships, as determined by multiple-locus variable-number tandem repeat analysis, suggested that strains originating from the same source tended to cluster together. However, multilocus sequence typing revealed considerable genetic diversity across clusters and sources. A phylogenetic analysis using single nucleotide polymorphisms of diseased fish strains alongside publicly available genomes demonstrated a high degree of evolutionary relatedness within and across different isolation sources. Notably, our findings reveal no direct correlation between phylogenetic patterns, isolation sources, and virulence capabilities. This underscores the necessity for proactive risk management strategies to address pathogenic V. vulnificus strains emerging from environmental reservoirs.IMPORTANCEAs the global incidence of Vibrio vulnificus infections rises, impacting human health and marine aquacultures, understanding the pathogenicity of environmental strains remains critical yet underexplored. This study addresses this gap by evaluating the virulence potential and genetic relatedness of V. vulnificus strains, focusing on environmental origins. We conduct an extensive genotypic analysis and phenotypic assessment, including virulence testing in a wax moth model. Our findings aim to uncover genetic and evolutionary factors that drive pathogenic strain emergence in the environment. This research advances our ability to identify reliable virulence markers and understand the distribution of pathogenic strains, offering significant insights for public health and environmental risk management.

Potential applications of Rhodomyrtus tomentosa leaf extract as natural anti-staphylococcal additive in food systems: Efficacy and in vivo safety evaluation.

This work aimed to explore the potential use of Rhodomyrtus tomentosa ethanol leaf extract (RTEL) as an alternative food preservative agent for controlling the growth of Staphylococcus aureus. Antibacterial activities against food-isolated S. aureus were performed using disc diffusion and broth microdilution assays, followed by evaluating in vivo subacute oral toxicity of the extract. Salad dressing was used as a food model to study bactericidal properties and consumer acceptability. RTEL remarkably inhibited S. aureus with minimum inhibitory concentrations (MICs) ranging from 7.81-62.5 µg/mL. Repeated oral doses (5, 50, and 300 mg/kg RTEL) for 28 days did not affect any of the measured toxicity parameters. The no-observed-adverse-effect-level (NOAEL) of RTEL was noted as more than 300 mg/kg body weight/day. The utilization of RTEL (12.5 mg/mL) in the vinaigrette salad dressing did not affect the consumer acceptability of the product, remarkably killed the pathogen within 3-9 h of exposure. The results indicated that RTEL is safe and effective as a natural anti-staphylococcal controlling agent that could be utilized in food systems. Further work is required on the effects of enterotoxin production, an important virulence factor of S. aureus responsible for food-borne disease.

Thai herbal formulation 'Ya-Pit-Samut-Noi': Its antibacterial activities, effects on bacterial virulence factors and in vivo acute toxicity.

ETHNOPHARMACOLOGICAL RELEVANCE: A Thai herbal formulation 'Ya-Pit-Samut-Noi' containing Nigella sativa (seed), Piper retrofractum (fruit), Punica granatum (pericarp), and Quercus infectoria (nutgall) has long been traditionally used to treat diarrhea or bloody mucous diarrhea. Scientific information is very important to support its therapeutic effects and traditional drug development. AIM OF THE STUDY: This study aimed to evaluate the antibacterial activities of Ya-Pit-Samut-Noi against diarrhea-causing bacteria and determine its effects on bacterial virulence factors and in vivo acute toxicity. MATERIALS AND METHODS: Ethanol and water extracts of Ya-Pit-Samut-Noi and its plant components were prepared. The agar diffusion method was used for preliminary screening of antibacterial activity of the extracts against diarrhea-causing bacteria including Staphylococcus aureus, Vibrio cholerae, and Vibrio parahaemolyticus. The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) were assessed using broth microdilution method. The effects on bactericidal activity, bacterial cell wall, and cell membrane were examined by time-kill, lysis, and leakage assays, respectively. The effects on bacterial virulence factors including quorum-sensing system, biofilm production, and swarming motility were determined. Phytochemical screening was carried out to identify the group of chemical compounds present in the formulation extracts. Acute toxicity study was conducted by a single oral dose of 2000 mg/kg body weight in Wistar albino rats. RESULTS: Ethanol and water extracts of Ya-Pit-Samut-Noi and Quercus infectoria demonstrated antibacterial efficacy against all bacterial strains as revealed by zones of inhibition ranging from 7.0 to 24.5 mm. The ethanol and water extracts of Ya-Pit-Samut-Noi and Quercus infectoria produced strong bacteriostatic activity against V. parahaemolyticus (n = 11) with an MIC range of 7.81-250 µg/ml. Only the ethanol extract of Ya-Pit-Samut-Noi produced MBC values less than or equal to 1000 µg/ml against all V. parahaemolyticus. Based on time-kill study, no surviving V. parahaemolyticus (ATCC 17802 and 5268) cells were detected within 6-12 h after treatment with the ethanol extract of Ya-Pit-Samut-Noi at MBC-4MBC concentrations. Vibrioparahaemolyticus ATCC 17802 cells treated with the ethanol extract of Ya-Pit-Samut-Noi demonstrated no lysis or leakage through the bacterial membrane was not observed. At low concentrations (0.125-0.25 µg/ml) the ethanol extract of Ya-Pit-Samut-Noi inhibited violacein production by Chromobacterium violaceum DMST 21761 without affecting the bacterial growth. The ethanol (31.25-62.5 µg/ml) and water (31.25-250 µg/ml) extracts of Ya-Pit-Samut-Noi inhibited biofilm production by S. aureus. The ethanol and water extracts of Ya-Pit-Samut-Noi at 1000 µg/ml reduced the swarming motility of Escherichia coli O157: H7 by 74.98% and 52.65%, respectively. Tannins and terpenoids were detected in both the ethanol and water extracts. Flavonoids were present only in the ethanol extract. Alkaloids and antraquinones were not noticed in either extract. In the acute toxicity study, there were no significant changes in hematological and biochemical parameters nor were adverse effects on mortality, general behaviors, body weight, or organ weights detected. CONCLUSIONS: The scientific evidence from this study supported the therapeutic effects and safety of the traditional Thai herbal formulation 'Ya-Pit-Samut-Noi' which has been used as an alternative treatment for gastrointestinal infections in Thailand.