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Acute hepatopancreatic necrosis disease (AHPND) is a severe condition that has caused significant damage and substantial financial losses to the global shrimp industry. Consequently, developing a convenient, sensitive, and specific analytical assay for accurately detecting AHPND is crucial for controlling the spread of the disease. The etiologic agent of AHPND is a strain of Vibrio species that carries a plasmid encoding the virulent binary toxins PirA and PirB. In this study, a pair of novel high-affinity monoclonal antibodies (MAbs) targeting the PirB toxin was generated and employed to develop a highly sensitive lateral flow immunochromatographic assay (LFIA). The LFIA results are visible to an unaided eye within 15 min of applying the bacterial colonies or pre-enrichment sample prepared in the application buffer to the test strip. This assay exhibited high specificity, successfully identifying AHPND-causing Vibrio species (Vibrio parahaemolyticus and Vibrio campbellii) without cross-reacting with non-AHPND Vibrio species or other bacterial isolates examined. The assay demonstrated a detection limit of approximately 40 ng/ml for PirB toxin, representing a 100-fold improvement over the sensitivity of the previous test strip for PirB toxin. Notably, the detection limit was further enhanced after a 6-h pre-enrichment process, enabling the identification of toxin-producing bacteria at concentrations as low as 1 colony-forming unit (CFU)/ml. Thus, it achieves a detection limit comparable to that of one-step PCR assays. Due to its high specificity, ease of use, rapid results, and no requirement for specialized equipment, the LFIA strip is an ideal tool for monitoring AHPND outbreaks and accurately detecting AHPND infections in shrimp cultures. [ABSTRACT FROM AUTHOR]

The most important threatening disease problem facing freshwater aquaculture is Vibriosis that cause water-borne diseases and foodborne gastroenteritis resulted from ingesting of uncooked or undercooked fish products. Also, it is one of the most zoonotic disease which transmitted to human from fish and another aquaculture. This study aimed to isolate different vibrio species from freshwater fish. Microbiological identification of Vibrio species were made using colonial morphology, biochemical characteristics and Identification system for Gram negative bacteria (GN 24) (Diagnostic sro kits) from total of 100 clinically diseased fish samples (Oreochromis niloticus) were collected alive from two various fish farms ( each 50 fish from one farm) from Kafrelsheik Governorate., the prevalence of Vibrio species were noticed in samples tilabia (23/100) 23% which V. parahaemolyticus was found in 6/100 (6%) and V. alginolyticus was detected in 16/100 (16%) and in V. cholera 1/100 (1%), Also we used PCR using 16S rRNA gene-specific for the genus Vibrio, detection of virulence and antibacterial resistance associated genes in v. parahaemolyticus and V. alginolyticus which the most pathogenic vibrio species .The present study also focused on the emergence of Multiple Antibiotic Resistance (MAR) index with the aim of developing strategies to mitigate the potential for infection and ensure effective treatment in the near future [ABSTRACT FROM AUTHOR]

Synthetic biology aims to establish engineering principles for biological systems by leveraging the design-build-test-learn (DBTL) cycle. Central to the success of the DBTL cycle is the selection of a suitable chassis, which is the environment in which biological designs are tested. Every step of this cycle is strongly influenced by the properties of chassis. A successful chassis must meet various criteria, prompting ongoing research regarding new candidates. Recently, species within the Vibrio genus, notably Vibrio natriegens and related strains, have emerged as promising next-generation chassis due to their rapid growth rates, versatile substrate utilization, and biosafety level 1 classification. These properties make them highly attractive for accelerating the DBTL cycle with the potential for efficient protein and metabolite production. This review emphasizes the foundational requirements for efficient engineering in synthetic biology, including genetic parts, vectors, and genome engineering technologies tailored to Vibrio species. Practical applications, such as metabolic engineering and protein expression, have been discussed, offering a comprehensive summary of recent advances. This paper also outlines the future directions and suggestions for fully unlocking the potential of Vibrio species as next-generation chassis. [ABSTRACT FROM AUTHOR]

Background: Vibrio species bloodstream infections have been associated with significant mortality and morbidity. Limited information is available regarding the epidemiology of bloodstream infections because of Vibrio species in the Australian context. Aims: The objective of this study was to define the incidence and risk factors for developing Vibrio species bloodstream infections and compare differences between different species. Methods: All patients with Vibrio spp. isolated from positive blood cultures between 1 January 2000 and 31 December 2019 were identified by the state‐wide Pathology Queensland laboratory. Demographics, clinical foci of infections and comorbid conditions were collected in addition to antimicrobial susceptibility results. Results: About 100 cases were identified between 2000 and 2019 with an incidence of 1.2 cases/1 million person‐years. Seasonal and geographical variation occurred with the highest incidence in the summer months and in the tropical north. Increasing age, male sex and multiple comorbidities were identified as risk factors. Vibrio vulnificus was isolated most frequently and associated with the most severe disease. Overall case fatality was 19%. Conclusions: There is potential for increasing cases of Vibrio species infections globally with ageing populations and climate change. Ongoing clinical awareness is required to ensure optimal patient outcomes. [ABSTRACT FROM AUTHOR]

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Inhibitory effects of water-soluble fraction (WSF) of crude oil, diesel, spent engine oil, and their composite mixture on luminescent Vibrio species isolated from a tropical marine fish were assessed to determine the acute toxicity and suitability of the bacteria. This was evaluated with a slight modification of the procedure for freshly prepared luminescent marine bacteria according to the International Organization for Standardization. Toxicity was assessed following bacterial exposure to different concentrations of the WSFs. Water quality parameters and total polycyclic aromatic hydrocarbons (PAHs) were determined using standard methods. Chemical analysis revealed cumulative loads of PAHs in the WSFs of crude oil (7.22 mg L-1) > diesel (6.92 mg L-1) > composite (6.23 mg L-1) > spent engine oil (4.58 mg L-1). The WSFs of crude oil and diesel showed rapid inhibition of bacterial light yield (11–81%), with maximum inhibition at 100% treatment level. However, luminescence inhibition was evident at 45 mins following exposure to spent engine oil (41–80%) for most of the treatment level tested, even though the 100% treatment level showed rapid (T0) and elevated percent inhibition at 75%. Exposure to composite mixtures of the hydrocarbons (1:1:1) revealed a gradual rise in percent inhibition with time. The EC50 values noted for the entire test samples during the period of exposure (0–60 mins) were between 14.79 and 257.57 mg L-1. Results obtained from this study revealed low toxic impacts of the WSFs and the observed sensitivity of the freshly isolated bioluminescent bacteria from our tropical waters. More so, it further demonstrates the potential for application of the bioluminescent Vibrio campbellii in standardized toxicity testing. Vibrio campbellii can be utilized to assess toxicity outcome in the event of bioremediation of oil residues but equally as a simple and generic indicator of oil spill and consequences on coastal waterways.

Abstract Background Virulence determinants are crucial to the risk assessment of pathogens in an environment. This study investigated the presence of eleven key virulence-associated genes in Vibrio cholerae (n = 111) and Vibrio mimicus (n = 22) and eight virulence determinants in Vibrio alginolyticus (n = 65) and Vibrio parahaemolyticus (n = 17) isolated from six important water resources in Eastern Cape, South Africa, using PCR techniques. The multiple virulence gene indexes (MVGI) for sampling sites and isolates as well as hotspots for potential vibriosis outbreaks among sampling sites were determined statistically based on the comparison of MVGI. Result The PCR assay showed that all the V. cholerae isolates belong to non-O1/non-O139 serogroups. Of the isolates, Vibrio Cholera (84%), V. mimicus (73%), V. alginolyticus (91%) and V. parahaemolyticus (100%) isolates harboured at least one of the virulence-associated genes investigated. The virulence gene combinations detected in isolates varied at sampling site and across sites. Typical virulence-associated determinants of V. cholerae were detected in V. mimicus while that of V. parahaemolyticus were detected in V. alginolyticus. The isolates with the highest MVGI were recovered from three estuaries (Sunday river, Swartkopps river, buffalo river) and a freshwater resource (Lashinton river). The cumulative MVGI for V. cholerae, V. mimicus, V. alginolyticus and V. parahaemolyticus isolates were 0.34, 0.20, 0.45, and 0.40 respectively. The targeted Vibrio spp. in increasing order of the public health risk posed in our study areas based on the MVGI is V. alginolyticus > V. parahaemolyticus > V. cholerae > V. mimicus. Five (sites SR, PA5, PA6, EL4 and EL6) out of the seventeen sampling sites were detected as the hotspots for potential cholera-like infection and vibriosis outbreaks. Conclusions Our findings suggest that humans having contact with water resources in our study areas are exposed to potential public health risks owing to the detection of virulent determinants in human pathogenic Vibrio spp. recovered from the water resources. The study affirms the relevancy of environmental Vibrio species to the epidemiology of vibriosis, cholera and cholera-like infections. Hence we suggest a monitoring program for human pathogenic Vibrio spp. in the environment most especially surface water that humans have contact with regularly.

Abstract Background Vibrio spp. are a diverse group of ecologically important marine bacteria responsible for several foodborne outbreaks of gastroenteritis around the world. Their detection and characterization are moving away from conventional culture-based methods towards next generation sequencing (NGS)-based approaches. However, genomic methods are relative in nature and suffer from technical biases arising from library preparation and sequencing. Here, we introduce a quantitative NGS-based method that enables the quantitation of Vibrio spp. at the limit of quantification (LOQ) through artificial DNA standards and their absolute quantification via digital PCR (dPCR). Results We developed six DNA standards, called Vibrio-Sequins, together with optimized TaqMan assays for their quantification in individually sequenced DNA libraries via dPCR. To enable Vibrio-Sequin quantification, we validated three duplex dPCR methods to quantify the six targets. LOQs were ranging from 20 to 120 cp/µl for the six standards, whereas the limit of detection (LOD) was ~ 10 cp/µl for all six assays. Subsequently, a quantitative genomics approach was applied to quantify Vibrio-DNA in a pooled DNA mixture derived from several Vibrio species in a proof-of-concept study, demonstrating the increased power of our quantitative genomic pipeline through the coupling of NGS and dPCR. Conclusions We significantly advance existing quantitative (meta)genomic methods by ensuring metrological traceability of NGS-based DNA quantification. Our method represents a useful tool for future metagenomic studies aiming at quantifying microbial DNA in an absolute manner. The inclusion of dPCR into sequencing-based methods supports the development of statistical approaches for the estimation of measurement uncertainties (MU) for NGS, which is still in its infancy.

Background: Virulence determinants are crucial to the risk assessment of pathogens in an environment. This study investigated the presence of eleven key virulence-associated genes in Vibrio cholerae (n = 111) and Vibrio mimicus (n = 22) and eight virulence determinants in Vibrio alginolyticus (n = 65) and Vibrio parahaemolyticus (n = 17) isolated from six important water resources in Eastern Cape, South Africa, using PCR techniques. The multiple virulence gene indexes (MVGI) for sampling sites and isolates as well as hotspots for potential vibriosis outbreaks among sampling sites were determined statistically based on the comparison of MVGI. Result: The PCR assay showed that all the V. cholerae isolates belong to non-O1/non-O139 serogroups. Of the isolates, Vibrio Cholera (84%), V. mimicus (73%), V. alginolyticus (91%) and V. parahaemolyticus (100%) isolates harboured at least one of the virulence-associated genes investigated. The virulence gene combinations detected in isolates varied at sampling site and across sites. Typical virulence-associated determinants of V. cholerae were detected in V. mimicus while that of V. parahaemolyticus were detected in V. alginolyticus. The isolates with the highest MVGI were recovered from three estuaries (Sunday river, Swartkopps river, buffalo river) and a freshwater resource (Lashinton river). The cumulative MVGI for V. cholerae, V. mimicus, V. alginolyticus and V. parahaemolyticus isolates were 0.34, 0.20, 0.45, and 0.40 respectively. The targeted Vibrio spp. in increasing order of the public health risk posed in our study areas based on the MVGI is V. alginolyticus > V. parahaemolyticus > V. cholerae > V. mimicus. Five (sites SR, PA5, PA6, EL4 and EL6) out of the seventeen sampling sites were detected as the hotspots for potential cholera-like infection and vibriosis outbreaks. Conclusions: Our findings suggest that humans having contact with water resources in our study areas are exposed to potential public health risks owing to the detection of virulent determinants in human pathogenic Vibrio spp. recovered from the water resources. The study affirms the relevancy of environmental Vibrio species to the epidemiology of vibriosis, cholera and cholera-like infections. Hence we suggest a monitoring program for human pathogenic Vibrio spp. in the environment most especially surface water that humans have contact with regularly. [ABSTRACT FROM AUTHOR]

Background Vibrio species are among the autochthonous bacterial populations found in surface waters and associated with various life-threatening extraintestinal diseases, especially in human populations with underlying illnesses and wound infections. Presently, very diminutive information exists regarding these species' mutational diversity of virulence and resistance genes. This study evaluated variations in endonucleases and mutational diversity of the virulence and resistance genes of Vibrio isolates, harboring virulence-correlated gene (vcgCPI), dihydropteroate synthase type 1 and type II genes (Sul 1 and 11), (aadA) aminoglycoside (3'') (9) adenylyltransferase gene, (aac(3)-IIa, (aacC2)a, aminoglycoside N(3)-acetyltransferase III, and (strA) aminoglycoside 3'-phosphotransferase resistance genes. Methods Using combinations of molecular biology techniques, bioinformatics tools, and sequence analysis. Results Our result revealed various nucleotide variations in virulence determinants of V. vulnificus (vcgCPI) at nucleotide positions (codon) 73-75 (A → G) and 300--302 (N → S). The aminoglycosides resistance gene (aadA) of Vibrio species depicts a nucleotide difference at position 482 (A → G), while the aminoglycosides resistance gene (sul 1 and 11) showed two variable regions of nucleotide polymorphism (102 and 140). The amino acid differences exist with the nucleotide polymorphism at position 140 (A → E). The banding patterns produced by the restriction enzymes HinP1I, MwoI, and StyD4I showed significant variations. Also, the restriction enzyme digestion of protein dihydropteroate synthase type 1 and type II genes (Sul 1 and 11) differed significantly, while enzymes DpnI and Hinf1 indicate no significant differences. The restriction enzyme NlaIV showed no band compared to reference isolates from the GenBank. However, the resistant determinants show significant point nucleotide mutation, which does not produce any amino acid change with diverse polymorphic regions, as revealed in the restriction digest profile. Conclusion The described virulence and resistance determinants possess specific polymorphic locus relevant to pathogenomics studies, pharmacogenomic, and control of such water-associated strains. [ABSTRACT FROM AUTHOR]

This study aimed to determine the incidence and virulence factor profiling of Vibrio species from hospital wastewater (HWW) and community wastewater effluents. Wastewater samples from selected sites were collected, processed, and analysed presumptively by the culture dependent methods and molecular techniques. A total of 270 isolates were confirmed as Vibrio genus delineating into V. cholerae (27%), V. parahaemolyticus (9.1%), V. vulnificus (4.1%), and V. fluvialis (3%). The remainder (>50%) may account for other Vibrio species not identified in the study. The four Vibrio species were isolated from secondary hospital wastewater effluent (SHWE), while V. cholerae was the sole specie isolated from Limbede community wastewater effluent (LCWE) and none of the four Vibrio species was recovered from tertiary hospital wastewater effluent (THWE). However, several virulence genes were identified among V. cholerae isolates from SHWE: ToxR (88%), hylA (81%), tcpA (64%), VPI (58%), ctx (44%), and ompU (34%). Virulence genes factors among V. cholerae isolates from LCWE were: ToxR (78%), ctx (67%), tcpA (44%), and hylA (44%). Two different genes (vfh and hupO) were identified in all confirmed V. fluvialis isolates. Among V. vulnificus, vcgA (50%) and vcgB (67%) were detected. In V. parahaemolyticus, tdh (56%) and tlh (100%) were also identified. This finding reveals that the studied aquatic niches pose serious potential health risk with Vibrio species harbouring virulence signatures. The distribution of virulence genes is valuable for ecological site quality, as well as epidemiological marker in the control and management of diseases caused by Vibrio species. Regular monitoring of HWW and communal wastewater effluent would allow relevant establishments to forecast, detect, and mitigate any public health threats in advance. [ABSTRACT FROM AUTHOR]

This study evaluated the inhibitory effects and adhesion properties of the isolated Bacillus safensis strain VQV8 (abbreviated as VQV8) against pathogenic Vibrio species under different culture conditions. VQV8 was cultured under specific conditions of temperature, pH, and NaCl (%) before testing its antibacterial activities using the dot-spotting method. In addition, a medium from agriculture by-products was screened for replacing Marine broth (MB) medium to culture VQV8. The bacteriostatic activities of VQV8 against pathogenic Vibrio bacteria were also determined. The adhesion properties of this bacterium were evaluated based on three main attributes: hydrophobicity, aggregation, and biofilm formation. The results indicated that VQV8 had the highest antibacterial activities against the tested Vibrio species when cultured in a medium containing 2% NaCl and incubation temperature ranged from 25 to 37 °C. However, the antibacterial activities against V. parahaemolyticus, V. haveyi, and V. alginolyticus were not found at 45 °C. Additionally, the pH from 6 to 9 did not demonstrate a significant effect on the antibacterial activities of VQV8. On the other hand, the use of pineapple peel extract (20–30%) medium demonstrated no significant difference in either growth or antibacterial activities of VQV8 compared to those found in MB medium. These results suggest the potential ability of this bacterial strain to strongly inhibit the growth of all tested Vibrio spp. High adhesion ability was also confirmed through the results of the hydrophobicity, aggregation, and biofilm formation assays. Thus, our results suggest that VQV8 could be used as a potential probiotic candidate for the prevention of Vibrio spp. infections in aquaculture. [ABSTRACT FROM AUTHOR]

The protective effects of a phage cocktail composed of vB_Vc_SrVc2 and vB_Vc_SrVc9 were tested in Pacific white shrimp (Litopenaeus vannamei) postlarvae, which were originally isolated from diseased shrimps and selected due to their broad-host-range properties against several pathogenic Vibsrio species. We used culture-dependent and culture-independent approaches to explore its effect on bacterial communities associated with shrimp postlarvae. Both methods revealed that the levels of Vibrio species were significantly reduced after phage cocktail administration. Phage-treated shrimp also exhibisuppted lesser damage and higher lipid accumulation in B cells of the hepatopancreas, as revealed by histopathological examination. Taken together, this study provides clear evidence that phage therapy can selectively and effectively reduce Vibrio species, thereby providing an environmentally safe alternative to the prophylactic use of antibiotics in shrimp aquaculture. [ABSTRACT FROM AUTHOR]

The outbreak of waterborne diseases such as cholera and non-cholera (vibriosis) is continuously increasing in the environment due to fecal and sewage discharge in water sources. Cholera and vibriosis are caused by different species of Vibrio genus which are responsible for acute diarrheal disease and soft tissue damage. Although incidences of cholera and vibriosis have been reported from the Vaishali district of Bihar, India, clinical or environmental strains have not been characterized in this region. Out of fifty environmental water samples, twelve different biochemical test results confirmed the presence of twenty Vibrio isolates. The isolates were found to belong to five different Vibrio species, namely V. proteolyticus, V. campbellii, V. nereis, V. cincinnatiensis, and V. harveyi. From the identified isolates, 65% and 45% isolates were found to be resistant to ampicillin and cephalexin, respectively. Additionally, two isolates were found to be resistant against six and four separately selected antibiotics. Furthermore, virulent hlyA and ompW genes were detected by PCR in two different isolates. Additionally, phage induction was also noticed in two different isolates which carry lysogenic phage in their genome. Overall, the results reported the identification of five different Vibrio species in environmental water samples. The isolates showed multiple antibacterial resistance, phage induction, and virulence gene profile in their genome. [ABSTRACT FROM AUTHOR]

The Greenland shark (Somniosus microcephalus) is a large species of shark found in the North Atlantic and Arctic Oceans and is believed to be the longest living vertebrate. Relatively little is known about its biology, abundance, health or diseases. In March 2022, only the third reported UK stranding of this species occurred and it was the first to undergo post-mortem examination. The animal was a sexually immature female, measuring 3.96 m in length and 285 kg in weight, and was in poor nutritional state. Gross findings included haemorrhages in the skin and soft tissues, particularly of the head, and silt in the stomach suggestive of live stranding, bilateral corneal opacity, slightly turbid cerebrospinal fluid (CSF) and patchy congestion of the brain. Histopathological findings included keratitis and anterior uveitis, fibrinonecrotic and lymphohistiocytic meningitis of the brain and proximal spinal cord and fibrinonecrotizing choroid plexitis. A near pure growth of a Vibrio organism was isolated from CSF. This is believed to be the first report of meningitis in this species. [ABSTRACT FROM AUTHOR]

Abstract Marine Vibrio species are natural degraders of chitin and usually secrete high levels of chitinolytic enzymes to digest recalcitrant chitin to chitooligosaccharides. This study used an endochitinase (VhChiA) from Vibrio campbellii to produce high-quality chitobiose from crustacean chitins. The enzyme was shown to be fully active and stable over 24 h when BSA was used as an additive. When different chitin sources were tested, VhChiA preferentially digested shrimp and squid (α) chitins compared to crab (β) chitin and did not utilize non-chitin substrates. The overall yields of chitobiose obtained from small-scale production using a single-step reaction was 96% from shrimp, and 91% from squid pen and crab-shell chitins. Larger-scale production yielded 200 mg of chitobiose, with > 99% purity after a desalting and purification step using preparative HPLC. In conclusion, we report the employment of an in-house produced chitinase as an effective biocatalyst to rapidly convert chitin food wastes to chitobiose, in a quantity and quality suitable for use in research and commercial purposes. Chitobiose production by this economical and eco-friendly approach can be easily scaled up to obtain multi-gram quantities of chitobiose for chemo-enzymic synthesis of rare chitooligosaccharide derivatives and long chain chitooligosaccharides, as well as preparation of sugar-based functionalized nanomaterials. Graphical Abstract

Vibrio spp. is the primary source of human infections linked to seafood. Vibrio parahaemolyticus is one of the species that is most frequently reported. It is found in tilapia, a fishery product that has a sizable global market. This study investigated the antibacterial activity of Brazilian peppertree and Cassia glauca extracts against Vibrio parahaemolyticus in tilapia fillets. In a number of fish products, including fried oysters, grilled oysters, tilapia fillets, smoked herring, and salted sardine. We also measured the total phenolic content, flavonoid content, antioxidant effect, and antibacterial and sensory evaluation scores of Brazilian peppertree and Cassia glauca mixture extract on tilapia fillets. The results also showed that Vibrio parahaemolyticus is present in many fish products, with Tilapia Fillets showing the highest positive percentage (76%) among all types of fish. The Brazilian peppertree and cassia glauca have an antioxidant effect, especially when mixed, that contains a high concentration of total phenolic content (750.88 mg GAE/g) and flavonoid contents (162.9 mg catechol/g), and the mixture has more effective antibacterial activity with inhibition zone 29.1 mm against Vibrio parahaemolyticus in tilapia fillet, especially with the high concentration of the mixture extract. Additionally, we found an improved sensory score for tilapia fillet with different extracts of Brazilian peppertree and Cassia glauca mixture extract. • The detection of V. parahaemolyticus in fish products and tilapia fillets had the highest positive percentage (76%). • Phytochemical Analysis of Plant Extracts (Phenolic Content, Total Flavonoid Content, and Antioxidant Activity) • Antibacterial activity of plant extracts, antibiotics, and minimum inhibitory concentrations against V. parahaemolyticus. • Antibacterial effect of mix extract against Vibrio parahaemolyticus experimentally inoculated into Tilapia Fillets • Sensory evaluation of Tilapia Fillets fortified with Mix Extract (Brazilian Peppertree and Cassia glauca). [ABSTRACT FROM AUTHOR]

Vibrio spp. are ubiquitous in marine areas worldwide and can cause diseases in humans. Aqueous environments favor the transfer of genes between strains, resulting in the loss of strain-specific target genes. This study aimed to explore new molecular targets for the detection of Vibrio strains at the species level using pan-genome analysis for nucleic acid detection and differentiation of major pathogenic Vibrio in aquatic products. Pan-genome analysis and PCR validation revealed a total of 10 specific targets: three for Vibrio parahaemolyticus , one for Vibrio alginolyticus , three for Vibrio vulnificus , three for Vibrio fluvialis. We constructed primers for the selected genes, and we verified their specificity using various Vibrio species and non- Vibrio strains. After systematic optimization, a novel molecular target-based 4-multiplex PCR (4-mPCR) method was developed, exhibiting a detection limit of 103 CFU/mL for pure cultures and 104 CFU/g for fish samples. The initial inoculum was approximately 1 Log 10 CFU/g, and all samples were positive after 4 h of enrichment. Not only that, the feasibility of the mined new targets in the field of nucleic acid detection was further validated by the 4-mqPCR and LAMP techniques. These results suggest that novel targets mined by pan-genomic have good generalizability for use in a variety of nucleic acid detection techniques and can help improve the accuracy of nucleic acid-based detection techniques in the detection of pathogenic Vibrio species in aquatic samples, with potential applications for food safety investigations and disease diagnosis. • Three specific genes for Vibrio parahaemolyticus , one for Vibrio alginolyticus , three for Vibrio vulnificus , and three for Vibrio fluvialis were obtained by pan-genomic analysis, respectively. • The function of most specific genes is unknown. • 196 strains proved that new molecular targets are more specific to the species. • 4-mqPCR and LAMP techniques demonstrated that novel targets have general applicability across multiple nucleic acid detection technologies. • We demonstrate the potential application of the novel target for the rapid detection of Vibrio in food samples. [ABSTRACT FROM AUTHOR]

White feces syndrome (WFS) is a multifactorial disease that affects global shrimp production. The diagnostic approach to identify WFS involves traditional and molecular scientific methods by examining histopathology, bioassays, PCR (polymerase chain reaction), and calorimetric estimation. The pathogenesis of WFS is closely associated with Vibrio spp., intestinal microbiota (IM) dysbiosis, and Enterocytozoon hepatopenaei (EHP). It also has caused over 10–15 % loss in the aquaculture industry and is also known to cause retardation, lethargy and slowly leading to high mortality in shrimp farms. Therefore, it is necessary to understand the molecular mechanisms processed under the association of IM dysbiosis, Vibrio spp., and EHP to analyze the impact of disease on the innate immune system of shrimp. However, only very few reviews have described the molecular pathways involved in WFS. Hence, this review aims to elucidate an in-depth analysis of molecular pathways involved in the innate immune system of shrimp and their response to pathogens. The analysis and understanding of the impact of shrimp's innate immune system on WFS would help in developing treatments to prevent the spread of disease, thereby improving the economic condition of shrimp farms worldwide. [Display omitted] • White feces syndrome (WFS) causes mortality and affects global shrimp production. • WFS is mainly caused by Vibrio spp., along with other causative agents. • The innate immune system in shrimp plays the main role in the pathogenesis of WFS. • In-depth analysis of innate immune response aids in disease prevention. • This review is providing up to date comprehensive knowledge about WFS. [ABSTRACT FROM AUTHOR]

This study aimed to determine the incidence and virulence factor profiling of Vibrio species from hospital wastewater (HWW) and community wastewater effluents. Wastewater samples from selected sites were collected, processed, and analysed presumptively by the culture dependent methods and molecular techniques. A total of 270 isolates were confirmed as Vibrio genus delineating into V. cholerae (27%), V. parahaemolyticus (9.1%), V. vulnificus (4.1%), and V. fluvialis (3%). The remainder (>50%) may account for other Vibrio species not identified in the study. The four Vibrio species were isolated from secondary hospital wastewater effluent (SHWE), while V. cholerae was the sole specie isolated from Limbede community wastewater effluent (LCWE) and none of the four Vibrio species was recovered from tertiary hospital wastewater effluent (THWE). However, several virulence genes were identified among V. cholerae isolates from SHWE: ToxR (88%), hylA (81%), tcpA (64%), VPI (58%), ctx (44%), and ompU (34%). Virulence genes factors among V. cholerae isolates from LCWE were: ToxR (78%), ctx (67%), tcpA (44%), and hylA (44%). Two different genes (vfh and hupO) were identified in all confirmed V. fluvialis isolates. Among V. vulnificus, vcgA (50%) and vcgB (67%) were detected. In V. parahaemolyticus, tdh (56%) and tlh (100%) were also identified. This finding reveals that the studied aquatic niches pose serious potential health risk with Vibrio species harbouring virulence signatures. The distribution of virulence genes is valuable for ecological site quality, as well as epidemiological marker in the control and management of diseases caused by Vibrio species. Regular monitoring of HWW and communal wastewater effluent would allow relevant establishments to forecast, detect, and mitigate any public health threats in advance.

To understand the characteristics of Vibrio isolates in aquatic environments in China and their public health significance, this study investigated water samples in six cities in China in 2020. A total of 88 sampling locations were included and Vibrio isolates were identified in 81 of them. A total of 143 Vibrio isolates belonging to 16 species were selected for characterization. The population structure of Vibrio species showed great differences among the six cities, indicating regional specificity. The presence of virulence genes was examined for the isolates (n = 78) of five pathogenic Vibrio species. All isolates except one (n = 77) contained at least one virulence gene and isolates belonging to the same species showed very similar virulence gene profiles. Then, 26 isolates from 12 species were examined by multilocus sequence typing and were assigned to 25 STs, of which 24 STs were new. Also, the presence of antibiotic-resistant genes was investigated for all 143 isolates and only three isolates were found to contain genes from aminoglycosides, phenicols, beta-lactams or the tetracycline family. Our results provide valuable insights into the Vibrio community in Chinese aquatic environments and can be applied as guidance for the environmental surveillance of the risk of Vibrio isolates. [ABSTRACT FROM AUTHOR]

Vibrio species are significant pathogens affecting aquatic species. Around 12 species of Vibrio can cause a gastrointestinal illness (gastroenteritis) in humans resulting from eating contaminated food such as raw or undercooked shellfish. The indiscriminate use of antibiotics accelerates the development of resistance representing a severe challenge for controlling Vibrio outbreaks. In this study, the antibiotic resistance profile and the prevalence of pathogenic Vibrio species of apparently healthy and diseased fishes isolated from different types of fish in Kafr ELSheikh Governorate in Egypt during 2018 were determined. Samples obtained from fishes were inoculated onto a Vibrio-selective medium (TCBS) and phenotypically identified using the biochemical characteristics and representative cultures were checked by PCR to confirm the identified isolates. In the present study, V. alginolyticus (16.00%) was the predominant species followed by V. cholerae (7.33%) and V. parahaemolyticus (5.33%). The tested isolates were resistant to ampicillin (80.00%) and sensitive to ciprofloxacin and norfloxacin (100%). A total number of 15 Vibrio isolates (five Vibrio parahaemolyticus, five V. alginolyticus, and five V. cholerae) were screened for five housekeeping genes and pathogenic virulence markers by PCR. Results showed that 100% of the V. parahaemolyticus isolates carried the tlh gene and 60.00% carried the tdh gene. In V. alginolyticus, 100% of the isolates carried the collagenase gene 0.00% carry the tdh gene; and 80.00% of V. cholerae isolates carried the ctx gene. The results showed that many isolates in this study had virulence characteristics that might correspond with the potential of infections and diseases. [ABSTRACT FROM AUTHOR]

Marine Vibrio species are natural degraders of chitin and usually secrete high levels of chitinolytic enzymes to digest recalcitrant chitin to chitooligosaccharides. This study used an endochitinase (VhChiA) from Vibrio campbellii to produce high-quality chitobiose from crustacean chitins. The enzyme was shown to be fully active and stable over 24 h when BSA was used as an additive. When different chitin sources were tested, VhChiA preferentially digested shrimp and squid (α) chitins compared to crab (β) chitin and did not utilize non-chitin substrates. The overall yields of chitobiose obtained from small-scale production using a single-step reaction was 96% from shrimp, and 91% from squid pen and crab-shell chitins. Larger-scale production yielded 200 mg of chitobiose, with > 99% purity after a desalting and purification step using preparative HPLC. In conclusion, we report the employment of an in-house produced chitinase as an effective biocatalyst to rapidly convert chitin food wastes to chitobiose, in a quantity and quality suitable for use in research and commercial purposes. Chitobiose production by this economical and eco-friendly approach can be easily scaled up to obtain multi-gram quantities of chitobiose for chemo-enzymic synthesis of rare chitooligosaccharide derivatives and long chain chitooligosaccharides, as well as preparation of sugar-based functionalized nanomaterials. [ABSTRACT FROM AUTHOR]

This study investigated the bacteriological quality of ready-to-eat (RTE) African salads in Enugu metropolis, Enugu, Nigeria. A total of 10 samples of African salad were purchased from 10 different vendors in Enugu metropolis. The samples were purchased from Agbani Road, Ogbete, Mayor, Uwani, Kenyatta, Achara Layout, Obiagu and Timber. Isolation and enumeration of bacterial isolates were done using Nutrient agar, Eosin Methylene Blue (EMB) agar, Thiosulphate-citrate-bile salts-sucrose (TCBS) agar, Salmonella-Shigella Agar (SSA) and MacConkey agar, following standard methods. Identification of the bacterial isolates were done through biochemical tests and the Analytical Profile Index (API 20E) test kit. The antibiotic sensitivity of the bacterial isolates was also done using the Kirby Bauer disc diffusion method. Total culturable heterotrophic count was above 300 colonies across the samples. The highest bacterial counts recorded on EMB, SSA and TCBS across the samples were 6.3 × 106 CFU/g, 7.4 × 106 CFU/g and 1.21 × 107 CFU/g respectively. The identities of the organisms were; Salmonella spp., Staphylococcus aureus, Escherichia coli, Vibrio mimicus, Vibrio fluvialis, Vibrio cholerae, Vibrio parahaemolyticus and Vibrio hollisae. The prevalent organism across the samples was Vibrio spp. The antibiotic sensitivity test suggested that Vibrio spp. was resistant to Ampiclox and Amoxycillin but sensitive to Erythromycin, Pefloxacin and Septrin. From this study, it was discovered that consumers of RTE African salad from majority of the vendors across Enugu metropolis are at risk of severe food poisoning.

The outbreak of waterborne diseases such as cholera and non-cholera (vibriosis) is continuously increasing in the environment due to fecal and sewage discharge in water sources. Cholera and vibriosis are caused by different species of Vibrio genus which are responsible for acute diarrheal disease and soft tissue damage. Although incidences of cholera and vibriosis have been reported from the Vaishali district of Bihar, India, clinical or environmental strains have not been characterized in this region. Out of fifty environmental water samples, twelve different biochemical test results confirmed the presence of twenty Vibrio isolates. The isolates were found to belong to five different Vibrio species, namely V. proteolyticus, V. campbellii, V. nereis, V. cincinnatiensis, and V. harveyi. From the identified isolates, 65% and 45% isolates were found to be resistant to ampicillin and cephalexin, respectively. Additionally, two isolates were found to be resistant against six and four separately selected antibiotics. Furthermore, virulent hlyA and ompW genes were detected by PCR in two different isolates. Additionally, phage induction was also noticed in two different isolates which carry lysogenic phage in their genome. Overall, the results reported the identification of five different Vibrio species in environmental water samples. The isolates showed multiple antibacterial resistance, phage induction, and virulence gene profile in their genome.

The potentially pathogenic species of the genus Vibrio pose a threat to both humans and animals, creating medical burdens and economic losses to the mariculture industry. Improvements in surveillance and diagnosis are needed to successfully manage vibriosis outbreaks. Matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) can provide rapid diagnosis and has been widely used in the identification of Vibrio spp. The main weakness of this technology is the limited number of strains and species of Vibrio in the existing commercial database. Here, we develop a new in-house database named PVBase containing 790 main spectra projections (MSP) of ten Vibrio species that come from various regions of China and include abundant clinical and environmental strains. PVBase was validated through a blind test of 65 Vibrio strains. The identification accuracy and scoring of Vibrio strains was greatly improved through the addition of PVBase. Identification accuracy increased from 73.4 to 100%. The number of strains with identification scores above 2.2 increased from 53.1% to 96.9% and 53.1% of strains had an identification score above 2.59. Moreover, perfect discrimination was obtained when using all of the MSPs created for the Vibrio species, even for very closely related species such as V. cholerae , V. albensis , and V. mimicus or V. alginolyticus , V. parahaemolyticus , and V. harveyi. In addition, we used phyloproteomic analysis to study whether there are differences in protein fingerprints of different regions or pathogenic strains. We found that MSP characteristics of Vibrio species were not related to their region or source. With the construction of PVBase, the identification efficiency of potentially pathogenic Vibrio species has been greatly improved, which is an important advance for epidemic prevention and control, and aquaculture disease detection. [ABSTRACT FROM AUTHOR]

Outbreaks of Vibrio infections have a long history of global public health concern and threat to the aquaculture industry. This 3-decade (1990-2019) meta-synthesis of global research progress in Vibrio species and associated disease outbreaks was undertaken to generate the knowledge needed to design effective interventions with policy implications. Using PRISMA protocol, we obtained data on the online version of the Institute for Scientific Information (ISI), Web of Science (WOS), and Scopus from January 1990 to September 2021 by title search of the keywords " Vibrio species OR Vibrio spp. OR vibriosis." On the 3-decade survey, the result has shown that a total of 776 publications document types were published on the subject, with an average of 24.25 ± 13.6 published documents per year with an annual growth rate of 4.71%. The year 2020 recorded the highest output of 52 published documents accounting for 6.70% of the total. The most prolific author, Blanch A., published 12 articles on the subject and has received citations of 1003 with an h-index of 10. While the most global cited paper author is the journal of J. Bacteriol (Bassler et al), receiving total citation (TC) (550) and per Year (22). The top active corresponding authors country is the United States of America with (92) articles, freq. 12.40%; TC of 3103. The observations in this study, such as the collaborations network map, and index, which have outlined a big difference between countries based on economic status, have underscored the need for a sustained research mentorship program that can define future policies. [ABSTRACT FROM AUTHOR]

Background:Vibrio infections have increased in Benin, and this phenomenon is expected to increase due to climate change, increased consumption of contaminated water and the number of people who are immunocompromised. The aim of the work: The objective of this study was to evaluate the risk of Vibrio transmission linked to the use of contaminated water in Benin. Methodology: Water samples [n = 220] were analyzed to isolate Vibrio strains using their biochemical and cultural characteristics. The species were identified by the Polymerase Chain Reaction technique by monitoring the search for genes encoding the cholera toxin of Vibrio cholerae [ctxA and ctxB] and the direct thermostable and thermostable hemolysins linked to Vibrio parahaemolyticus [tdh and trh]. Results: Among the 220 collected samples, the biochemical tests revealed 86 strains of Vibrio species; Vibrio cholerae [35%], Vibrio parahaemolyticus [18.60%] and Vibrio alginolyticus [13.95%] were identified using molecular tool. The presence of genes encoding the main virulence factors of the strains studied. Thus 6.67%, 10% and 3.33% of the strains of Vibrio cholerae respectively contain the toxins ctxA, ctxB and the couple ctxA and ctxB. Likewise, the Vibrio parahaemolyticus strains contain 12.5% tdh toxins and 31.25% [tdh and trh]. The search for genes [tdh and trh] in Vibrio alginolyticus was also negative. Conclusion: Epidemics can be triggered by natural or fabricated events that contaminate drinking water or compromise access to safe drinking water and sanitation. The incidence of vibriosis is increasing, perhaps in part because of the spread of Vibrio species promoted by climate change and increasing water temperature.

The potentially pathogenic species of the genus Vibrio pose a threat to both humans and animals, creating medical burdens and economic losses to the mariculture industry. Improvements in surveillance and diagnosis are needed to successfully manage vibriosis outbreaks. Matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) can provide rapid diagnosis and has been widely used in the identification of Vibrio spp. The main weakness of this technology is the limited number of strains and species of Vibrio in the existing commercial database. Here, we develop a new in-house database named PVBase containing 790 main spectra projections (MSP) of ten Vibrio species that come from various regions of China and include abundant clinical and environmental strains. PVBase was validated through a blind test of 65 Vibrio strains. The identification accuracy and scoring of Vibrio strains was greatly improved through the addition of PVBase. Identification accuracy increased from 73.4 to 100%. The number of strains with identification scores above 2.2 increased from 53.1% to 96.9% and 53.1% of strains had an identification score above 2.59. Moreover, perfect discrimination was obtained when using all of the MSPs created for the Vibrio species, even for very closely related species such as V. cholerae, V. albensis, and V. mimicus or V. alginolyticus, V. parahaemolyticus, and V. harveyi. In addition, we used phyloproteomic analysis to study whether there are differences in protein fingerprints of different regions or pathogenic strains. We found that MSP characteristics of Vibrio species were not related to their region or source. With the construction of PVBase, the identification efficiency of potentially pathogenic Vibrio species has been greatly improved, which is an important advance for epidemic prevention and control, and aquaculture disease detection.

Vibrio species are important pathogens of marine animals and aquaculture populations and some of them can cause serious infections in humans through consumption of contaminated seafood and aquaculture products. Lytic bacteriophages can potentially alleviate Vibrio contamination in the aquaculture organisms and in the processing of aquatic products and have gained significant scientific attention in recent years. In the present study, bacteriophages were isolated from sewage of local aquatic products markets and grown using Vibrio mimicus CICC 21613 as host cells. The lytic vibriophage OY1 belonging to the newly proposed family Autographiviridae and the genus Maculvirus was identified by observation under electron microscope and comparative genomic analysis. The phage OY1 showed lytic activity against 24 among 32 tested strains belonging to eight Vibrio species. The complete phage OY1 genome consists of a single circular double-stranded DNA of 43,479 bp with a total GC content of 49.27% and was predicted to encode 40 open reading frames (ORFs). To evaluate its potential against vibrios, the one-step growth curve, thermal and pH stability, host range, and lytic activity of the OY1 phage against Vibrio species were evaluated. The results showed that phage OY1 had a range of thermal and pH tolerance, and exhibited a significant inhibitory effect on the growth of tested Vibrio species. Bacterial growth in the fish muscle extract juice (FMEJ) inoculated with Vibrio mimicus CICC 21613, Vibrio parahaemolyticus CICC 21617, Vibrio alginolyticus VJ14, and the mixed bacterial culture was reduced by 2.65 log CFU/ml, 2.42 log CFU/ml, 1.93 log CFU/ml, and 2.01 log CFU/ml, respectively, by incubation with phage OY1 at 25°C for 36 h. Phage OY1 also showed a strong ability to prevent biofilm formation and destroy formed Vibrio species biofilms. These results indicate that phage OY1 is a potential biocontrol agent against Vibrio species in the aquaculture industry and in food safety control. [ABSTRACT FROM AUTHOR]

Shrimp farming has experienced rising costs as a result of disease outbreaks associated with Vibrio spp. Suitable strategies for disease prevention and control are therefore urgently needed. This study aimed to evaluate the antimicrobial effect of Moringa oleifera seed powder against Vibrio cholerae in the rearing water of Pacific white shrimp (Penaeus vannamei) postlarvae. In vitro assays included the determination of minimum inhibitory concentration (MIC) of M. oleifera seed powder against V. cholerae, whereas in vivo assays included the effect of M. oleifera seed powder on bacterial load and water quality parameters in the rearing tanks, as well as its effect on shrimp postlarvae survival. M. oleifera seed powder inhibited the growth of V. cholerae with MIC values of 62·5 µg ml−1. Moreover, seawater pH of treated tanks (8·66) was significantly lower (P < 0·01) than pH of the control tanks (9·02), whereas the visibility of treated tanks (37·08 cm) was significantly higher (P < 0·01) as compared to control tanks (35·37 cm). Likewise, V. cholerae load was significantly reduced (P < 0·01) from 4·7 × 104 to 3·1 × 103 CFU per ml in tanks treated with M. oleifera seed powder. Altogether, this study demonstrates the antimicrobial activity of M. oleifera against V. cholerae in shrimp culture. [ABSTRACT FROM AUTHOR]

Vibrio species are important pathogens of marine animals and aquaculture populations and some of them can cause serious infections in humans through consumption of contaminated seafood and aquaculture products. Lytic bacteriophages can potentially alleviate Vibrio contamination in the aquaculture organisms and in the processing of aquatic products and have gained significant scientific attention in recent years. In the present study, bacteriophages were isolated from sewage of local aquatic products markets and grown using Vibrio mimicus CICC 21613 as host cells. The lytic vibriophage OY1 belonging to the newly proposed family Autographiviridae and the genus Maculvirus was identified by observation under electron microscope and comparative genomic analysis. The phage OY1 showed lytic activity against 24 among 32 tested strains belonging to eight Vibrio species. The complete phage OY1 genome consists of a single circular double-stranded DNA of 43,479 bp with a total GC content of 49.27% and was predicted to encode 40 open reading frames (ORFs). To evaluate its potential against vibrios, the one-step growth curve, thermal and pH stability, host range, and lytic activity of the OY1 phage against Vibrio species were evaluated. The results showed that phage OY1 had a range of thermal and pH tolerance, and exhibited a significant inhibitory effect on the growth of tested Vibrio species. Bacterial growth in the fish muscle extract juice (FMEJ) inoculated with Vibrio mimicus CICC 21613, Vibrio parahaemolyticus CICC 21617, Vibrio alginolyticus VJ14, and the mixed bacterial culture was reduced by 2.65 log CFU/ml, 2.42 log CFU/ml, 1.93 log CFU/ml, and 2.01 log CFU/ml, respectively, by incubation with phage OY1 at 25°C for 36 h. Phage OY1 also showed a strong ability to prevent biofilm formation and destroy formed Vibrio species biofilms. These results indicate that phage OY1 is a potential biocontrol agent against Vibrio species in the aquaculture industry and in food safety control.

Adequate comprehension of the genomics of microbial resistance to an antimicrobial agent will advance knowledge on the management of associated pathologies and public health safety. However, continued emergences and reemergence of pathogens, including Vibrio species, hallmarks a potential knowledge gap. A clear understanding of the process and forecast of the next trend should be in place to nip in the bud, microbial acquisition of resistance to antibiotics. Therefore, this two-decade (1 January 2000 to 31 December 2019) systematic review and meta-analytical study articulated the prevalence and incidence of antibiotics resistance genes in Vibrio species isolated from environmental samples. Articles from the Web of Science and PubMed electronic databases was engaged. Heterogeneity of the data and bias were analyzed with random effect model meta-analysis and funnel plot. A total of 1920 Vibrio sp. were reported by the ten selected articles included in this study; out of which 32.39% of identified isolates displayed antimicrobial resistance and associated genes. The distribution of antibiotics resistance genes in Vibrio sp., reported within six countries was 21% tetracycline (tet), and 20% sulphonamide (sul) and β-lactamase (bla) respectively. The quinolone, tetracycline and sulfonamide resistance genes showed 32.97% (95% CI 0.18–0.53) prevalence while chloramphenicol, macrolides and aminoglycoside resistance genes are expressed in percentages as 28.67% (95% CI 0.15–0.47) and β-lactamase resistance genes 27.93% (95% CI 0.11–0.56) respectively. The Vibrio antibiotics resistance genes (V-ARG) distribution depicts no regular trend or pattern from the analyzed data. Consequently, more studies would be required to articulate the structure of cohesion in the distribution of the resistance determinants in microbes.

Bacteria have developed diverse strategies to counteract virus predation, one of which is the clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR associated (Cas) proteins immune defense system. In this study, the structure and function of the CRISPR-Cas system in 120 Vibrio strains were analyzed by bioinformatics methods, as well as the correlation between CRISPR and mobile genetic elements (MEGs). Only 61 Vibrio strains contained one or more CRISPR structures, and finally 102 CRISPRs were identified. The typical repeat size was 28 bp, and the total length of CRISPRs is nearly 60 bp, which was the most stable length of CRISPR in Vibrio strains. The types of CRISPR-Cas present in 61 strains were I-C, I-E, I-F, II-B, III-B, III-D and the rare type IV systems. Through principal component analysis, we found that Cas gene was most closely related to CRISPR. In addition, phages and plasmids were also highly correlated, showing negative correlation with CRISPR-Cas system. CRISPR-Cas predominantly present on chromosome within Vibrio while rarely in plasmids. Comparing the structural characteristics of plasmids containing CRISPR and without CRISPR, we found plasmid pMBL287 with CRISPR contained a bacteriophage f237, with more MGES, suggesting the diversity was greater. In addition, the same mobile genetic elements IS256 and ISL3 were found in the upstream and downstream of CRISPR. This study provides the prevalence, diversity and phylogenetic distribution of CRISPR-Cas in Vibrio, revealing which type of CRISPR-Cas system is predominant, and the factors affecting its function, as well as its relationship with mobile genetic elements. [ABSTRACT FROM AUTHOR]

Some Vibrio species are important human pathogens owing to they cause infectious diseases such as gastroenteritis, wound infections, septicemia or even death. Many of these illnesses are associated with consumption of contaminated seafood. In the present study, we evaluated the presence of pathogenic Vibrio species, their virulence and antimicrobial susceptibility from 285 different kind of seafood samples from "La Nueva Viga" market in Mexico City. The PCR assay was used for amplification the vppC (collagenase), vmh (hemolysin), tlh (thermolabile hemolysin), and vvhA (hemolytic cytolysin) genes that are specific to Vibrio alginolyticus (detected in 27%), Vibrio mimicus (23.2%), Vibrio parahaemolyticus (28.8%) and Vibrio vulnificus (21.1%), respectively. Several genes encoding virulence factors were amplified. These included V. alginolyticus: pvuA (17.9%), pvsA (50%), wza and lafA (100%); V. mimicus: iut A (60%), toxR (100%); V. parahaemolyticus: pvuA (58.7%), pvsA (26.1%), wza (2.2%), and lafA (100%); and V. vulnificus: wcrA (77.5%), gmhD (57.5%), lafA (100%) and motA (30%). The antibiotic susceptibility of the Vibrio species isolates revealed that most of them were resistant to ampicillin, cephalothin and carbenicillin but susceptible to pefloxacin and trimethoprim-sulfamethoxazole. Our results indicated a high prevalence of pathogenic Vibrio species in seafood, a high presence of virulence genes and that Vibrio species continuously exposed to antibiotics, therefore, consumption of these kind of seafood carries a potential risk for foodborne illness. [ABSTRACT FROM AUTHOR]

Vibrio species, widely distributed in water environments, has emerged as a prominent cause of water and food-related disease outbreaks posing significant risk to human and animal health worldwide. About 40% of presumptive isolates recovered from four selected rivers in Southwest Nigeria and, established as Vibrio species genus through polymerase chain reaction techniques., were subjected to antibiotic susceptibility testing against a panel of 18 commonly used antibiotics. The relative prevalence of key Vibrio species (V. parahaemolyticus, V. vulnificus, V. mimicus, V. harveyi, and V. cholerae) was in the order 17%, 13.3%, 4.4%, 2.2%, and 2.2% respectively. Antibiotic resistance by all Vibrio species was mostly observed against doxycycline (71–89%), erythromycin (86–100%), tetracycline (71–89%), rifampicin (86–100%), and sulfamethoxazole (87–100%), though susceptibility to meropenem (86–100%), cephalothin (60–100%), norfloxacin (93–100%), ciprofloxacin (88–100%), amikacin (64–100%), gentamicin (57–74%), and trimethoprim/sulfamethoxazole (57–81%) was equally observed in all species. Vibrio mimicus expressed highest resistance against streptomycin and chloramphenicol (64%), while V. vulnificus (52%) and V. cholerae (57%) had the highest resistance against cephalothin. High resistance against ampicillin (57%) and amoxicillin (50%) was exhibited by V. cholerae and V. mimicus respectively. Indexes of multiple antibiotic resistances (MARI) among Vibrio species ranged between 0.11 and 0.72 with the highest MAR index of 0.72 observed in one isolate of V. vulnificus. This study reveals high prevalence of Vibrio species in the selected rivers as well as elevated resistance against some first-line antibiotics, which suggests possible inappropriate antimicrobial usage around study communities. We conclude that the freshwater resources investigated are unfit for domestic, industrial, and recreational uses without treatment prior to use and are potential reservoirs of antibiotic-resistant Vibrio species in this environment. [ABSTRACT FROM AUTHOR]

To understand the characteristics of Vibrio isolates in aquatic environments in China and their public health significance, this study investigated water samples in six cities in China in 2020. A total of 88 sampling locations were included and Vibrio isolates were identified in 81 of them. A total of 143 Vibrio isolates belonging to 16 species were selected for characterization. The population structure of Vibrio species showed great differences among the six cities, indicating regional specificity. The presence of virulence genes was examined for the isolates (n = 78) of five pathogenic Vibrio species. All isolates except one (n = 77) contained at least one virulence gene and isolates belonging to the same species showed very similar virulence gene profiles. Then, 26 isolates from 12 species were examined by multilocus sequence typing and were assigned to 25 STs, of which 24 STs were new. Also, the presence of antibiotic-resistant genes was investigated for all 143 isolates and only three isolates were found to contain genes from aminoglycosides, phenicols, beta-lactams or the tetracycline family. Our results provide valuable insights into the Vibrio community in Chinese aquatic environments and can be applied as guidance for the environmental surveillance of the risk of Vibrio isolates.

Vibriosis, an illness caused by the Vibrio bacteria species, results in significant economic loss in olive flounder farms. Here we present a novel anti- Vibrio feed vaccine protecting multiple strains of Vibrio pathogens, a universal vaccine effect. The vaccine was generated by engineering Lactococcus lactis BFE920 to express the fusion antigens of Vibrio outer membrane protein K (OmpK) and flagellin B subunit (FlaB). These antigen genes are highly conserved among Vibrio species. Olive flounder (7.1 ± 0.8 g and 140 ± 10 g) were fed the vaccine adsorbed to a regular feed (1 × 107 CFU/g) for one week with a 1-week interval, repeating three times (a triple boost). The vaccinated fish increased the significant levels of antigen-specific antibodies, T cell numbers (CD4-1 , CD4-2, and CD8α), cytokine production (T-bet and IFN-γ), and innate immune responses (TLR5M , IL-1β, and IL-12p40). Also, the survival rates of adult and juvenile fish fed the vaccine were significantly elevated when challenged with V. anguillarum , V. alginolyticus , and V. harveyi. In addition, weight gain rate and feed conversion ratio were improved in vaccinated fish. The feed vaccine protected multiple Vibrio pathogens, a universal vaccine effect, by activating innate and adaptive immune responses. This oral vaccine may be developed as an anti- Vibrio vaccine to protect against a broad spectrum of Vibrio pathogens. • Lactococcus lactis BFE920 was engineered to express the fusion protein that are highly conserved among Vibrio pathogens. • LAB vaccine fed olive flounder were pretected from multiple Vibrio pathogens: a universal vaccine effect. • The vaccine-fed fish increased both innate immunity and adaptive immunity. • The vaccinated olive flounder improved the weight gain rate and feed conversion ratio. [ABSTRACT FROM AUTHOR]

The contamination of seafood with Vibrio species can have severe repercussions in the seafood industry. Vibrio species can form mature biofilms and persist on the surface of several seafoods such as crabs, oysters, mussels, and shrimp, for extended duration. Several conventional approaches have been employed to inhibit the growth of planktonic cells and prevent the formation of Vibrio biofilms. Since Vibrio biofilms are mostly resistant to these control measures, novel alternative methods need to be urgently developed. In this review, we propose environmentally friendly approaches to suppress Vibrio biofilm formation using a hypothesized mechanism of action. [ABSTRACT FROM AUTHOR]

Abstract Background Bacteria are prey for many viruses that hijack the bacterial cell in order to propagate, which can result in bacterial cell lysis and death. Bacteria have developed diverse strategies to counteract virus predation, one of which is the clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR associated (Cas) proteins immune defense system. Species within the bacterial family Vibrionaceae are marine organisms that encounter large numbers of phages. Our goal was to determine the significance of CRISPR-Cas systems as a mechanism of defense in this group by investigating their prevalence, phylogenetic distribution, and genome context. Results Herein, we describe all the CRISPR-Cas system types and their distribution within the family Vibrionaceae. In Vibrio cholerae genomes, we identified multiple variant type I-F systems, which were also present in 41 additional species. In a large number of Vibrio species, we identified a mini type I-F system comprised of tniQcas5cas7cas6f, which was always associated with Tn7-like transposons. The Tn7-like elements, in addition to the CRISPR-Cas system, also contained additional cargo genes such as restriction modification systems and type three secretion systems. A putative hybrid CRISPR-Cas system was identified containing type III-B genes followed by a type I-F cas6f and a type I-F CRISPR that was associated with a prophage in V. cholerae and V. metoecus strains. Our analysis identified CRISPR-Cas types I-C, I-E, I-F, II-B, III-A, III-B, III-D, and the rare type IV systems as well as cas loci architectural variants among 70 species. All systems described contained a CRISPR array that ranged in size from 3 to 179 spacers. The systems identified were present predominantly within mobile genetic elements (MGEs) such as genomic islands, plasmids, and transposon-like elements. Phylogenetic analysis of Cas proteins indicated that the CRISPR-Cas systems were acquired by horizontal gene transfer. Conclusions Our data show that CRISPR-Cas systems are phylogenetically widespread but sporadic in occurrence, actively evolving, and present on MGEs within Vibrionaceae.

A total of 100 fish samples (Oreochromis niloticus) were collected as 25 fish apparently health and 75 fish appeared to be clinically diseased fish. Samples of kidney, heart, liver, gills and skin lesions are collected and cultured on TCBS medium. Eighty-three samples show biochemically positive to vibrio species.10 random samples amplified to 16srRNA gene by PCR technique for more accurate identification resulting in 8 isolates were positive to 16srRNA.The 8 isolates amplified to suspected species specific primers. Resulted in 4 V.parahaemolyticus (50%), 2 V. harveyi (25%),1 V. anguillarum (12.5%) and 1 V. alginolyticus (12.5%). Each species amplified to PCR technique for more identification to virulence genes. V.parahaemolyticus amplified to tdh and trh; V. harveyi to Partial hly and Vhh; V. alginolyticus amplified to trh and tdh and V.anguillarum amplified to virA and angM. This study proved that PCR protocols could be useful tools for rapid and simultaneous detection of different Vibrio species virulence factors. Also using of 16srRNA gene in molecular characterization (PCR) of Vibrio is the fastest and most accurate method to identify all isolates of Vibrio. [ABSTRACT FROM AUTHOR]

Background: New strategies are needed to combat multidrug-resistant bacteria. The restriction of iron uptake by bacteria is a promising way to inhibit their growth. We aimed to suppress the growth of Vibrio bacterial species by inhibiting their ferric ion-binding protein (FbpA) using food components. Methods: Twenty spices were selected for the screening of FbpA inhibitors. The candidate was applied to antibacterial tests, and the mechanism was further studied. Results: An active compound, rosmarinic acid (RA), was screened out. RA binds competitively and more tightly than Fe3+ to VmFbpA, the FbpA from V. metschnikovii, with apparent KD values of 8 μM vs. 17 μM. Moreover, RA can inhibit the growth of V. metschnikovii to one-third of the control at 1000 μM. Interestingly, sodium citrate (SC) enhances the growth inhibition effect of RA, although SC only does not inhibit the growth. The combination of RA/SC completely inhibits the growth of not only V. metschnikovii at 100/100 μM but also the vibriosis-causative pathogens V. vulnificus and V. parahaemolyticus, at 100/100 and 1000/100 μM, respectively. However, RA/SC does not affect the growth of Escherichia coli. Conclusions: RA/SC is a potential bacteriostatic agent against Vibrio species while causing little damage to indigenous gastrointestinal bacteria.

Flagellar motility in bacteria is a highly regulated and complex cellular process that requires high energy investment for movement and host colonization. Motility plays an important role in the lifestyle of Vibrio spp. in the aquatic environment and during host colonization. Flagellar motility in vibrios is associated with several cellular processes, such as movement, colonization, adhesion, biofilm formation, and virulence. The transcription of all flagella-related genes occurs hierarchically and is regulated positively or negatively by several transcription factors and regulatory proteins. The flagellar regulatory hierarchy is well studied in Vibrio cholerae and Vibrio parahaemolyticus. Here, we compared the regulatory cascade and molecules involved in the flagellar motility of V. cholerae and V. parahaemolyticus in detail. The evolutionary relatedness of the master regulator of the polar and lateral flagella in different Vibrio species is also discussed. Although they can form symbiotic associations of some Vibrio species with humans and aquatic organisms can be harmed by several species of Vibrio as a result of surface contact, characterized by flagellar movement. Thus, targeting flagellar motility in pathogenic Vibrio species is considered a promising approach to control Vibrio infections. This approach, along with the strategies for controlling flagellar motility in different species of Vibrio using naturally derived and chemically synthesized compounds, is discussed in this review. Key points: • Vibrio species are ubiquitous and distributed across the aquatic environments. • The flagellar motility is responsible for the chemotactic movement and initial colonization to the host. • The transition from the motile into the biofilm stage is one of the crucial events in the infection. • Several signaling pathways are involved in the motility and formation of biofilm. • Attenuation of motility by naturally derived or chemically synthesized compounds could be a potential treatment for preventing Vibrio biofilm-associated infections. [ABSTRACT FROM AUTHOR]

False yam (Icacina oliviformis) is a drought-resistant root crop found in Northern Ghana. Research confirms it to contain some levels of antimicrobial, antifungal, antidiarrheal, antioxidant, and antidiabetic activities. This study was carried out to determine the effectiveness of False yam extracts incorporated in liquid soap against selected microorganisms. Polar solvent (methanol, ethanol, and water) extracts from the leaves, tuber (root), peels (root bark), and seeds were diluted with liquid soap to give concentrations of 50 mg/mL, 25 mg/mL, and 12.5 mg/mL. Test organisms used were Staphylococcus aureus, Escherichia coli, and Vibrio species. The results revealed significant differences (P < 0.05) among the different solvents and concentrations of the plant materials used. The ethanol seed extract at a concentration of 50 mg/mL was most inhibitive on Vibrio species with an inhibition diameter of 13 mm. The methanol peel extract also had the greatest inhibition on Escherichia coli with average inhibition zone of 7 mm to 12.5 mm followed closely by the tuber with maximum inhibition zone of 9 mm. The leaf extract generally exhibited higher values of inhibition on Staphylococcus aureus with zone range of 10 mm to 15 mm. The highest activity occurred with an inhibition zone of 15 mm diameter. The tuber and peel extracts recorded the highest antibacterial activity. Staphylococcus aureus appeared to be the most susceptible organism and the methanol extracts exhibited a relatively high frequency of inhibition. Further research efforts should combine the tuber and peel extracts to determine the possible synergistic and/or antagonistic effect. [ABSTRACT FROM AUTHOR]

Vibriosis is a globally threatening bacterial disease affecting mariculture with high mortalities and severe economic losses. Isolation and Identification of different vibrio species were performed to a total number of one hundred moribund and freshly dead Solea aegyptiaca, Epinephelus marginatus and Mugil cephalus collected from Lake Qarun and Gulf of Suez. Phenotypic picture and molecular identification based on use of 16SrRNA gene sequence confirmed 44 strains as vibrio species. Further molecular identification of retrieved vibrio spp. using species specific primers for collagenase, ToxR and Vvh genes categorized 10 isolates belong to V. alginolyticus, 8 isolates belong to V. parahaemolyticus and 6 isolates belong to V. vulnificus. The total prevalence of vibriosis was 44% where the highest prevalence was recorded in Lake Qarun examined fishes.

Treated sewage harbours pathogenic microbes, such as enteric bacteria and protozoa, are capable of causing several diseases. Some of these are emerging pathogens sometimes recovered in the absence of common water quality indicator organisms. The possibility of selected treatments plants serving as momentary reservoirs of Vibrio pathogens during a non-outbreak period was assessed. The occurrence and diversity of Vibrio pathogens were monitored for one year (December 2016 to November 2017) in the treated effluents and upstream and downstream areas of the receiving water bodies of two wastewater treatment plants (WWTPs), designated AL and TS. Physicochemical parameters of TS and AL WWTPs’ water samples were analysed using a multi-parameter meter (Hanna, model HI 9828, Padova, Italy) and a turbidimeter (HACH, model 2100P, Johannesburg, South Africa). Water samples were augmented with alkaline peptone water and cultured on thiosulfate citrate bile salts sucrose agar at 37 °C for 24 h. The recovered probable pathogens were confirmed via PCR amplification, using primers specific for Vibrio species of public health significance. The distribution of Vibrio species positively and significantly (p < 0.01) correlated with turbidity (r = 0.630), temperature (r = 0.615), dissolved oxygen (r = 0.615), pH (r = 0.607), biological oxygen demand (r = 0.573), total dissolved solid (r = 0.543), total suspended solid (r = 0.511), electrical conductivity (r = 0.499), residual chlorine (r = 0.463) and salinity (r = 0.459). The densities of Vibrio species were found to be significantly higher (p < 0.05) in effluents from both AL and TS WWTPs than upstream and downstream of the receiving rivers across the sampling regime. Furthermore, the maximum Vibrio species density across the sampling regime were observed during the warmer Summer and Spring season. Moreover, six medically important Vibrio species were detected in the water samples, indicating that the methods employed were efficient in revealing that WWTPs are potential reservoirs of Vibrio pathogens, which could pose a substantial public health risk if the receiving water is used for domestic purposes. Our findings further strengthen existing calls for the inclusion of emerging bacterial pathogens, including Vibrio species, as water quality indicators by the South African Department of Water Affairs. Hence, we recommend regular monitoring of treated effluents and receiving water bodies to ensure early control of potential outbreaks of vibriosis and cholera.

Background: Bacteria are prey for many viruses that hijack the bacterial cell in order to propagate, which can result in bacterial cell lysis and death. Bacteria have developed diverse strategies to counteract virus predation, one of which is the clustered regularly interspaced short palindromic repeat (CRISPR) and CRISPR associated (Cas) proteins immune defense system. Species within the bacterial family Vibrionaceae are marine organisms that encounter large numbers of phages. Our goal was to determine the significance of CRISPR-Cas systems as a mechanism of defense in this group by investigating their prevalence, phylogenetic distribution, and genome context. Results: Herein, we describe all the CRISPR-Cas system types and their distribution within the family Vibrionaceae. In Vibrio cholerae genomes, we identified multiple variant type I-F systems, which were also present in 41 additional species. In a large number of Vibrio species, we identified a mini type I-F system comprised of tniQcas5cas7cas6f, which was always associated with Tn7-like transposons. The Tn7-like elements, in addition to the CRISPR-Cas system, also contained additional cargo genes such as restriction modification systems and type three secretion systems. A putative hybrid CRISPR-Cas system was identified containing type III-B genes followed by a type I-F cas6f and a type I-F CRISPR that was associated with a prophage in V. cholerae and V. metoecus strains. Our analysis identified CRISPR-Cas types I-C, I-E, I-F, II-B, III-A, III-B, III-D, and the rare type IV systems as well as cas loci architectural variants among 70 species. All systems described contained a CRISPR array that ranged in size from 3 to 179 spacers. The systems identified were present predominantly within mobile genetic elements (MGEs) such as genomic islands, plasmids, and transposon-like elements. Phylogenetic analysis of Cas proteins indicated that the CRISPR-Cas systems were acquired by horizontal gene transfer. Conclusions: Our data show that CRISPR-Cas systems are phylogenetically widespread but sporadic in occurrence, actively evolving, and present on MGEs within Vibrionaceae. [ABSTRACT FROM AUTHOR]