Vibrio Spp

Background and Aim: The use of bioactive compounds is a promising tool to improve shrimp health regarding vibriosis. This study aimed to determine the effect of the dietary Majapahit (Crescentia cujete L.) fruit powder on the cellular immune response of vannamei shrimp (Litopenaeus vannamei) infected with Vibrio harveyi, Vibrio alginolyticus, and Vibrio parahaemolyticus.

Materials and Methods: Twelve vannamei shrimp (aged 1 month) were randomly divided into four groups (n=3). Vannamei shrimp received experimental feed with different concentrations of Majapahit fruit powder for 20 days: Group A, 0%; Group B, 3.04%; Group C, 6.08%, and Group D, 9.12%. Subsequently, vannamei shrimp was infected with V. harveyi, V. alginolyticus, and V. parahaemolyticus. The total hemocytes, total differential hemocytes (hyaline, semi-granular, and granular cells), and phagocytic activity were assessed. Data were analyzed through analysis of variance (p<0.05) using SPSS ver. 24 for Windows.

Results: Majapahit fruit powder at a dose of 3.04% increased the number of hyaline cells of L. vannamei after infection with Vibrio spp. Supplementation of the feed formula with Majapahit fruit powder at a dose of 3.04% increased the number of semi-granular and granular cells compared with the control. Furthermore, Majapahit fruit powder at doses of 3.04% and 6.08% increased the hemocytes compared with a dose of 9.12%. The phagocytic activity of L. vannamei after infection with Vibrio spp. tends to increase after supplementation with 3.04% Majapahit fruit powder.

Conclusion: This study showed that the addition of C. cujete L. powder to the feed formula increased the cellular immune response. The most effective dose ranged from 3.04% to 6.08%.

The present study was carried out to evaluate the antibacterial activity of crude extracts of Cellana radiata collected from Visakhapatnam coast, Andhra Pradesh. The whole animal extracts from methanol and ethyl acetate were obtained by both dry and wet methods and tested for their activity against three pathogenic bacteria i.e. Escherichia coli, Staphylococcus aureus and Vibrio harveyi. The extracts obtained from methanol (wet), ethyl acetate (dry) and ethyl acetate (wet) showed inhibition zones of 12 mm, 13 mm and 18 mm respectively against V. harveyi.

Background and Aim: The use of bioactive compounds is a promising tool to improve shrimp health regarding vibriosis. This study aimed to determine the effect of the dietary Majapahit (Crescentia cujete L.) fruit powder on the cellular immune response of vannamei shrimp (Litopenaeus vannamei) infected with Vibrio harveyi, Vibrio alginolyticus, and Vibrio parahaemolyticus. Materials and Methods: Twelve vannamei shrimp (aged 1 month) were randomly divided into four groups (n=3). Vannamei shrimp received experimental feed with different concentrations of Majapahit fruit powder for 20 days: Group A, 0%; Group B, 3.04%; Group C, 6.08%, and Group D, 9.12%. Subsequently, vannamei shrimp was infected with V. harveyi, V. alginolyticus, and V. parahaemolyticus. The total hemocytes, total differential hemocytes (hyaline, semi-granular, and granular cells), and phagocytic activity were assessed. Data were analyzed through analysis of variance (p&lt;0.05) using SPSS ver. 24 for Windows. Results: ...

Background and Aim: The use of bioactive compounds is a promising tool to improve shrimp health regarding vibriosis. This study aimed to determine the effect of the dietary Majapahit (Crescentia cujete L.) fruit powder on the cellular immune response of vannamei shrimp (Litopenaeus vannamei) infected with Vibrio harveyi, Vibrio alginolyticus, and Vibrio parahaemolyticus. Materials and Methods: Twelve vannamei shrimp (aged 1 month) were randomly divided into four groups (n=3). Vannamei shrimp received experimental feed with different concentrations of Majapahit fruit powder for 20 days: Group A, 0%; Group B, 3.04%; Group C, 6.08%, and Group D, 9.12%. Subsequently, vannamei shrimp was infected with V. harveyi, V. alginolyticus, and V. parahaemolyticus. The total hemocytes, total differential hemocytes (hyaline, semi-granular, and granular cells), and phagocytic activity were assessed. Data were analyzed through analysis of variance (p&lt;0.05) using SPSS ver. 24 for Windows. Results: ...

Vibrio vulnificus, a bacterium ubiquitous in oysters and coastal water, is capable of causing ailments ranging from gastroenteritis to grievous wound infections or septicemia. The uptake of these bacteria into oysters is often examined in vitro by placing oysters in seawater amended with V. vulnificus. Multiple teams have obtained similar results in studies where laboratory-grown bacteria were observed to be rapidly taken up by oysters but quickly eliminated. This technique, along with suggested modifications, is reviewed here. In contrast, the natural microflora within oysters is notoriously difficult to eliminate via depuration. The reason for the transiency of exogenous bacteria is that those bacteria are competitively excluded by the oyster&#39;s preexisting microflora. Evidence of this phenomenon is shown using in vitro oyster studies and a multiyear in situ case study. Depuration of the endogenous oyster bacteria occurs naturally and can also be artificially induced, but both ...

The genus Vibrio, among the members of the γ-proteobacteria with the most rapid motility and most intricate series of quorum-sensing based transcriptional regulations, is also among the most transduction competent of the Vibrionaceae family due to a secondary chromosome encoding for a variety of virulence factors, to which its rapid pathogenesis and status as a source of high mortality in bivalve hatcheries is owed. This secondary chromosome is a source of great diversity for the 126 Vibrio species, encoding genes which are translated into metalloproteases, hemolysins, and cytolysins. V. tubiashii subsp. europaeus PP-638 T was originally isolated from a broodstock tank of flat oyster (Ostrea edulis) during an episode of larval mortality in a shellfish hatchery in Galicia, Spain. 1 Genomic DNA was isolated from PP-638 T grown overnight in YP30 using the Wizard purification kit (Promega), sequenced using an Illumina MiSeq at the Genomics and Sequencing Center at the University of Rhode Island, Kingston, RI, assembled and mapped to Vibrio tubiashi ATCC 19109 using CLC Genomics Workbench and Microbial Genome Finishing module. 2 The resulting contingent sequence of the second chromosome was mapped to a p251-type megaplasmid and a p57-type plasmid. Similarly, V. bivalvicida 605 T was originally isolated from an episode of larval mortality of carpet shell clam (Ruditapes decussatus) in a bivalve hatchery located in Galicia (northwest Spain). Genomic DNA was sequenced using an Illumina MiSeq sequencer by Sistemas Genómicos (Valencia, Spain). 3 Across the genomes of these two subspecies of the Orientalis clade were a series of genes encoding for proteases, phospholipases and hemolysins that shows similarity to those of various V. anguillarum serotypes. In V. bivalvicida 605 T , high sequence homology was found to the phospholipase Plp in V. anguillarum M93Sm and a hemolysin annotated as HlyA that shows similarity to Vah1 in V. anguillarum M93Sm. In the second chromosome of V. tubiashii subsp. europaeus PP-638 T , a protease was encoded with 71% similarity to Epp in Vibrio anguillarum M93Sm. As well, homology to M93Sm was found in V. tubiashii subsp. europaeus PP-638 T , in which the Plp homolog is divergently transcribed away from a pore-forming cytolysin, as has been previously sequenced. 4 The mature toxin binds and aggregates on eukaryotic cells, leading to permeability disruption and osmotic lysis. The phosphatidylcholine-specific lipase provides a mechanism for species specific hemolysis, providing possible indication of genes activated during the shift from zooplanktonic reservoir state in intermediate hosts and selective colonization, enabling persistence. Herein details attempts at formulating a rapid colony forming unit (CFU) assay per aqueous quantity (mL) by obtaining the optical density readings using a spectrophotometer for known concentrations to develop algorithms relating CFU/mL to optical density to rapidly assess viable cell number using biomass readings in shellfish hatcheries similar to the that from which the isolate was originally collected. This enabled elucidation of divergent virulence factors between V. anguillarum 1. Prado S, Dubert J, Barja JL. 2015. Characterization of pathogenic vibrios isolated from bivalve hatcheries in Galicia, NW Atlantic coast of Spain. Description of Vibrio tubiashii subsp. europaensis subsp. nov. Syst Appl Microbiol 38:26–

Vibrio species are an important and widespread component of marine microbial communities. Some Vibrio strains are potentially pathogenic to marine vertebrates and invertebrates. The aim of this study was to identify vibrios, in particular Vibrio splendidus and related species, isolated from clams (Chamelea gallina) collected along the coasts of the Abruzzi region from May to October 2007. The isolates obtained were phenotyped and classified as belonging to the genus Vibrio. The strains underwent biochemical testing in accordance with Alsina&#39;s scheme for V. splendidus identification. Molecular analysis of the 16S-23S intergenic space region and recA gene was used to identify V. splendidus and related species. All the samples examined were found to contain halophylic Vibrio species, with V. alginolyticus, V. splendidus-related species and V. mediterranei most commonly found. A polymerase chain reaction of the 16S-23S intergenic space region and sequencing of the recA gene from iso...

Vibrio species are an important and widespread component of marine microbial communities. Some Vibrio strains are potentially pathogenic to marine vertebrates and invertebrates. The aim of this study was to identify vibrios, in particular Vibrio splendidus and related species, isolated from clams (Chamelea gallina) collected along the coasts of the Abruzzi region from May to October 2007. The isolates obtained were phenotyped and classified as belonging to the genus Vibrio. The strains underwent biochemical testing in accordance with Alsina’s scheme for V. splendidus identification. Molecular analysis of the 16S-23S intergenic space region and recA gene was used to identify V. splendidus and related species. All the samples examined were found to contain halophylic Vibrio species, with V. alginolyticus, V. splendidus-related species and V. mediterranei most commonly found. A polymerase chain reaction of the 16S-23S intergenic space region and sequencing of the recA gene from isolates confirmed that phenotyping of Vibrio species is not sufficient to distinguish between different species. Differentiation of the highly related species among V. splendidus-related clusters remains an important issue. In this regard, our data suggests sequencing the recA genes was far more discriminatory than sequencing 16S rDNA for this purpose.

Vibrio vulnificus, a bacterium ubiquitous in oysters and coastal water, is capable of causing ailments ranging from gastroenteritis, to grievous wound infections or septicemia.  The uptake of these bacteria into oysters is often examined in vitro, by placing oysters in seawater amended with V. vulnificus.  Multiple teams have obtained similar results, wherein the laboratory-grown bacteria are observed to be rapidly taken up by oysters but quickly eliminated.  This technique, along with suggested modifications, is reviewed.  In contrast, the natural microflora within oysters is notoriously difficult to eliminate via depuration.  The reason for the transiency of exogenous bacteria is explained by competitive exclusion by from the oyster’s preexisting microflora.  Evidence of this phenomenon is shown using in vitro oyster studies and a multi-year in situ case study.  Depuration of the endogenous oyster bacteria occurs naturally and can be artificially induced, but both of these events require extreme conditions, natural or otherwise.  These are also presented here.  Finally, the “viable but non-culturable” (VBNC) state of Vibrio bacteria is discussed.  This bacterial torpor can easily be confused with a reduction in bacterial abundance as bacteria in this state fail to grow on culture media.  Thus, oysters collected from colder months may appear to be relatively Vibrio free, but in reality harbor VBNC cells that respond to exogenous bacteria and prevent colonization of oyster matrices.  Bacterial uptake experiments combined with studies involving cell-free spent media are detailed that demonstrate this occurrence which could explain why the microbial community in oysters does not always mirror that of the surrounding water.

The United States has federal regulations in place to reduce the risk of seafood-related infection caused by the estuarine bacteria Vibrio vulnificus and Vibrio parahaemolyticus. However, data to support the development of regulations have been generated in a very few specific regions of the nation. More regionally specific data are needed to further understand the dynamics of human infection relating to shellfish-harvesting conditions in other areas. In this study, oysters and water were collected from four oyster harvest sites in North Carolina over an 11-month period. Samples were analyzed for the abundances of total Vibrio spp., V. vulnificus, and V. parahaemolyticus; environmental parameters, including salinity, water temperature, wind velocity, and precipitation, were also measured simultaneously. By utilizing these data, preliminary predictive management tools for estimating the abundance of V. vulnificus bacteria in shellfish were developed. This work highlights the need for...

Bacterial evolution is due to the adaptive nature of the core bacterial genomes that plays critical role in diversification, fitness and adaptation of the species to different environment. Host-pathogen interactions and environmental settings are such forces that guide the adaptation of bacterial genomes. Since Vibrio cholerae represents an appropriate model organism for studying the interplay of environment and host driven factors shaping the microbial genome structure and function, the current study aims to identify genes that are under these strong forces in V. cholerae. Here, we employed a comparative genomics approach to identify genes that are under positive selection in ten strains of Vibrio sp. including four pathogenic V. cholerae strains. From the available genome sequence data, a total of 421 orthologous genes were identified by reciprocal BLAST best-hit method, recombination breakpoint frequency analysis and tree comparison method. These 421 genes, representing the core ...

This study aimed to isolate, define the genetic profile, assess potential pathogenicity and evaluate the seasonal distribution of Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus strains isolated from the Vibrata river (Abruzzo Region, Italy) during a monitoring period of one year. Detection was performed according to ISO/TS 21872-1-2:2007. Species identification and characterisation were achieved using molecular methods. Vibrio spp. were detected in 50% (23) of the water samples. In particular, V. cholerae, V. parahaemolyticus, and V. vulnificus were isolated in 18 (39.1%), 4 (8.7%), and 2 (4.3%) samples, respectively. All V. parahaemolyticus strains were tdh gene negative, 75% were positive for trh gene. In 30 V. cholerae isolates, the polymerase chain reaction (PCR) assay for detecting virulence and regulatory genes (ctxA, toxR, tcpA, ompU, hlyA, tcpI, zot, and stn/sto) revealed 6 genotypes associated to different levels of pathogenicity. Pulsed-field gel electrophoresis (PFGE) characterisation of the V. cholerae strains identified 13 different pulsotypes. A greater degree of similarity was shown for strains isolated in the same period of the year. Results of our study reveal a potential health risk associated with the waters of the Vibrata river, which are used for irrigation and next to the swimming areas of Abruzzo coastline.

Vibrio species are an important and widespread component of marine microbial communities. Some Vibrio strains are potentially pathogenic to marine vertebrates and invertebrates. The aim of this study was to identify vibrios, in particular Vibrio splendidus and related species, isolated from clams (Chamelea gallina) collected along the coasts of the Abruzzi region from May to October 2007. The isolates obtained were phenotyped and classified as belonging to the genus Vibrio. The strains underwent biochemical testing in accordance with Alsina&#39;s scheme for V. splendidus identification. Molecular analysis of the 16S-23S intergenic space region and recA gene was used to identify V. splendidus and related species. All the samples examined were found to contain halophylic Vibrio species, with V. alginolyticus, V. splendidus-related species and V. mediterranei most commonly found. A polymerase chain reaction of the 16S-23S intergenic space region and sequencing of the recA gene from iso...

This study aimed to isolate, define the genetic profile, assess potential pathogenicity and evaluate the seasonal distribution of Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus strains isolated from the Vibrata river (Abruzzo Region, Italy) during a monitoring period of one year. Detection was performed according to ISO/TS 21872-1-2:2007. Species identification and characterisation were achieved using molecular methods. Vibrio spp. were detected in 50% (23) of the water samples. In particular, V. cholerae, V. parahaemolyticus, and V. vulnificus were isolated in 18 (39.1%), 4 (8.7%), and 2 (4.3%) samples, respectively. All V. parahaemolyticus strains were tdh gene negative, 75% were positive for trh gene. In 30 V. cholerae isolates, the polymerase chain reaction (PCR) assay for detecting virulence and regulatory genes (ctxA, toxR, tcpA, ompU, hlyA, tcpI, zot, and stn/sto) revealed 6 genotypes associated to different levels of pathogenicity. Pulsed-field gel electroph...

Vibrio vulnificus, a bacterium ubiquitous in oysters and coastal water, is capable of causing ailments ranging from gastroenteritis to grievous wound infections or septicemia. The uptake of these bacteria into oysters is often examined in vitro by placing oysters in seawater amended with V. vulnificus. Multiple teams have obtained similar results in studies where laboratory-grown bacteria were observed to be rapidly taken up by oysters but quickly eliminated. This technique, along with suggested modifications, is reviewed here. In contrast, the natural microflora within oysters is notoriously difficult to eliminate via depuration. The reason for the transiency of exogenous bacteria is that those bacteria are competitively excluded by the oyster&#39;s preexisting microflora. Evidence of this phenomenon is shown using in vitro oyster studies and a multiyear in situ case study. Depuration of the endogenous oyster bacteria occurs naturally and can also be artificially induced, but both ...

The United States has federal regulations in place to reduce the risk of seafood-related infection caused by the estuarine bacteria Vibrio vulnificus and Vibrio parahaemolyticus. However, data to support the development of regulations have been generated in a very few specific regions of the nation. More regionally specific data are needed to further understand the dynamics of human infection relating to shellfish-harvesting conditions in other areas. In this study, oysters and water were collected from four oyster harvest sites in North Carolina over an 11-month period. Samples were analyzed for the abundances of total Vibrio spp., V. vulnificus, and V. parahaemolyticus; environmental parameters, including salinity, water temperature, wind velocity, and precipitation, were also measured simultaneously. By utilizing these data, preliminary predictive management tools for estimating the abundance of V. vulnificus bacteria in shellfish were developed. This work highlights the need for...

This study aimed to isolate, define the genetic profile, assess potential pathogenicity and evaluate the seasonal distribution of Vibrio cholerae, Vibrio parahaemolyticus, and Vibrio vulnificus strains isolated from the Vibrata river (Abruzzo Region, Italy) during a monitoring period of one year. Detection was performed according to ISO/TS 21872-1-2:2007. Species identification and characterisation were achieved using molecular methods. Vibrio spp. were detected in 50% (23) of the water samples. In particular, V. cholerae, V. parahaemolyticus, and V. vulnificus were isolated in 18 (39.1%), 4 (8.7%), and 2 (4.3%) samples, respectively. All V. parahaemolyticus strains were tdh gene negative, 75% were positive for trh gene. In 30 V. cholerae isolates, the polymerase chain reaction (PCR) assay for detecting virulence and regulatory genes (ctxA, toxR, tcpA, ompU, hlyA, tcpI, zot, and stn/sto) revealed 6 genotypes associated to different levels of pathogenicity. Pulsed-field gel electroph...

In this study, we show the induction of lateral flagella by the action of the sodium channel blocker phenamil, in the marine bacterium Vibrio shilonii, a coral pathogen that causes bleaching. We analyzed the growth and morphology of cells treated with phenamil. A time course analysis showed that after 30 min of exposure to the sodium channel blocker, lateral flagella were present and could be detected by electron microscopy. Detection of the mRNA of the master regulator (lafK) and lateral flagellin (lafA) by RT-PCR confirmed the expression of lateral flagellar genes. We show the simultaneous isolation of polar and, for the first time, lateral flagellar hook-basal bodies. This allowed us to compare the dimensions and morphological characteristics of the 2 structures.

The use of effective biocides as disinfectants is essential in aquaculture facilities. However, while most biocides act effectively on free-living planktonic pathogens, they are seldom useful against biofilms. In this study, we evaluate the biocidal efficacy and antimicrobial specific contact time of three disinfectants, Virkon™Aquatic (VirA), peracetic acid (PerA) and hydrogen peroxide (HydP), on Vibrio anguillarum, V. harveyi, V. alginolyticus, and Photobacterium damselae subspecies piscicida against their both life phases. By using the minimum inhibitory, bactericidal, and eradication concentrations of disinfectants acting on the free-living planktonic state (MIC; MBC) and biofilms (MBIC; MBEC), we determined the in vitro susceptibility of each bacterial strain against three different individual concentrations of VirA, PerA, and HydP added at 1, 5, and 10 min intervals. PerA and VirA had the highest bactericidal efficacies against the free-living planktonic state and biofilm of all bacteria. Kinetically, PerA gave a positive result more quickly in both cases regardless of the strain in question, while the weakest HydP required longer than 10 min to act effectively. Moreover, we conducted a short in vivo safety trial by pouring the suggested MIC of each disinfectant into tanks containing juvenile Gilthead seabream (Sparus aurata). Significant mortality after 24 h was observed pointing to the potential risk a mishap of these chemicals might cause to fish. Nevertheless, collectively, our results support the inclusion of biocides within biosecurity protocols in aquaculture facilities and highlight PerA as the most effective disinfectant for fighting against biofilms produced by V. anguillarum, V. harveyi, V. alginolyticus, or P. damselae subsp. piscicida.