Isolation ofVibrio parahaemolyticus andVibrio cholerae (Non-O1 and O139) from moribund shrimp (Penaeus monodon) and experimental challenge study against post larvae and juveniles

Haldar, Soumya; Chatterjee, Shruti; Asakura, Masahiro; Vijayakumaran, M; Yamasaki, Shinji

doi:10.1007/bf03175050

Cited by 36 publications

(15 citation statements)

References 26 publications

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“…Genomic DNA was prepared according to the method of Ausubel et al (2002). Partial sequencing of the 16S rRNA and uridylate kinase (pyrH) genes was done for all 28 strains as described previously (Haldar et al, 2007;Thompson et al, 2007). The sequence of the pyrH gene from all experimental strains was compared with published sequence data in GenBank using BLAST (http://www.ncbi.nih.gov/BLAST).…”

Section: Methodsmentioning

confidence: 99%

Identification of Vibrio campbellii isolated from diseased farm-shrimps from south India and establishment of its pathogenic potential in an Artemia model

et al. 2011

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Shrimp diseases are frequently reported to be caused by closely related vibrios, and in many cases they are tentatively but inaccurately identified as Vibrio harveyi and related vibrios. In the present study, 28 biochemically identified V. harveyi-related strains isolated from diseased shrimps were randomly selected for further characterization by molecular tools. Twenty-six strains were identified as Vibrio campbellii and two as V. harveyi by sequence analysis of 16S rRNA and uridylate kinase genes. Haemolysin-gene-based species-specific multiplex PCR also confirmed these results. Experimental challenge studies using Artemia as a model showed that eight isolates were highly pathogenic, three were moderately pathogenic and the remaining 17 were non-pathogenic. Ribotyping with BglI clearly distinguished V. campbellii from V. harveyi, but it failed to separate pathogenic and non-pathogenic clusters. Artemia nauplii challenged with a fluorescently labelled highly pathogenic strain (IPEY54) showed patches in the digestive tract. However, no patches were observed for a non-pathogenic strain (IPEY41). Direct bacterial counts also supported colonization potential for the highly pathogenic strain. To our knowledge, this is the first report on the isolation and accurate identification of large numbers of V. campbellii associated with shrimp disease in aquacultural farms. V. campbellii has long been considered to be non-pathogenic and classified with V. harveyi-related bacteria. However, we show that this species may be an emerging aquaculture pathogen. This study will help to formulate suitable strategies to combat this newly identified pathogen. INTRODUCTIONHalophilic vibrios such as Vibrio harveyi are ubiquitous in the marine environment and are implicated as the causes of several diseases in wild and cultured aquatic organisms. Due to the plasticity of Vibrio genomes, with frequent horizontal gene transfer events, species boundaries are very narrow in the marine environment (Fraser et al., 2007). Hence, the identification of V. harveyi and related species isolated from the marine environment is sometimes difficult. Luminous vibrios including V. harveyi have been implicated principally with disease outbreaks in shrimp larval culture facilities and grow-out ponds worldwide. Due to the high level of phylogenetic similarity among marine vibrios, bacteria associated with disease outbreaks have often been misidentified. For example, although strain LMG 19703 T (or AK1 T ) showed 99.4 % sequence similarity in 16S rRNA with Vibrio mediterranei (ATCC 43341 T ), Kushmaro et al. (2001) initially classified it as a new species, Vibrio shiloi, due to large differences in phenotypic properties. Later, Thompson et al. (2001), on the basis of genotypic features such as fluorescent amplified fragment length polymorphism and DNA-DNA hybridization, as Abbreviations: ASW, artificial seawater; DOC, days of culture; DTAF, 5-(4,6-dichlorotriazin-2-yl)aminofluorescein; NJ, neighbour joining. The GenBank/EMBL/DDBJ accession numbers for the se...

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Section: Methodsmentioning

confidence: 99%

Identification of Vibrio campbellii isolated from diseased farm-shrimps from south India and establishment of its pathogenic potential in an Artemia model

et al. 2011

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“…Sequencing of genes like 16S rRNA, 23S rRNA, 16S‐23S intergenic spacer region or gyrB cannot differentiate these closely related species (Gomez‐Gill et al. 2004; Haldar et al. 2007).…”

Section: Introductionmentioning

confidence: 99%

Development of a haemolysin gene-based multiplex PCR for simultaneous detection ofVibrio campbellii,Vibrio harveyiandVibrio parahaemolyticus

Haldar

Neogi

Kogure

et al. 2010

Letters in Applied Microbiology

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Aim: To develop a haemolysin (hly) gene‐based species‐specific multiplex PCR for simple and rapid detection of Vibrio campbellii, V. harveyi and V. parahaemolyticus. Methods and Results: The complete hly genes of three V. campbellii strains isolated from diseased shrimps were sequenced and species‐specific PCR primers were designed based on these sequences and the registered hly gene sequences of Vibrio harveyi and Vibrio parahaemolyticus. Specificity and sensitivity of the multiplex PCR was validated with 27 V. campbellii, 16 V. harveyi, and 69 V. parahaemolyticus, 18 other Vibrio species, one Photobacterium damselae and nine other bacterial species. The detection limits of all the three target species were in between 10 and 100 cells per PCR tube. Conclusions: Specificity and sensitivity of the multiplex PCR is 100% each and sufficient to be considered as an effective tool in a prediction system to prevent potential disease outbreak by these Vibrio species. Significance and Impact of the Study: Because there is lack of simple, rapid and cost‐effective method to differentiate these closely related V. campbellii, V. harveyi and V. parahaemolyticus species, the multiplex PCR developed in this study will be very effective in epidemiological, ecological and economical points of view.

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“…Furthermore, V. cholerae (20), V. vulnificus (7), and V. parahaemolyticus (2) are pathogenic to shrimp and other aquacultured animals. Therefore, the monitoring of these pathogenic Vibrio species in the aquacultural environment is important for both the aqua culture industry and human health.…”

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confidence: 99%

Occurrence of Vibrio parahaemolyticus, Vibrio cholerae, and Vibrio vulnificus in the Aquacultural Environments of Taiwan

Tey

Jong

Fen

et al. 2015

Journal of Food Protection

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The occurrence of Vibrio parahaemolyticus, Vibrio vulnificus, and Vibrio cholerae in a total of 72 samples from six aquaculture ponds for groupers, milk fish, and tilapia in southern Taiwan was examined by the membrane filtration and colony hybridization method. The halophilic V. parahaemolyticus was only recovered in seawater ponds, with a high isolation frequency of 86.1% and a mean density of 2.6 log CFU/g. V. cholerae was found in both the seawater and freshwater ponds but preferentially in freshwater ponds, with a frequency of 72.2% and a mean density of 1.65 log CFU/g. V. vulnificus was identified mainly in seawater ponds, with an isolation frequency of 27.8%. The density of V. parahaemolyticus in seawater ponds was positively related to water temperature (Pearson correlation coefficient, r = 0.555) and negatively related to salinity (r = 2 0.333). The density of V. cholerae in all six ponds was positively related to water temperature (r = 0.342) and negatively related to salinity (r = 2 0.432). Two putatively pathogenic tdh(+) V. parahaemolyticus isolates (1.4% of the samples) and no ctx(+) V. cholerae isolates were identified. The experimental results may facilitate assessments of the risk posed by these pathogenic Vibrio species in Taiwan, where aquaculture provides a large part of the seafood supply.

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Isolation ofVibrio parahaemolyticus andVibrio cholerae (Non-O1 and O139) from moribund shrimp (Penaeus monodon) and experimental challenge study against post larvae and juveniles

Cited by 36 publications

References 26 publications

Identification of Vibrio campbellii isolated from diseased farm-shrimps from south India and establishment of its pathogenic potential in an Artemia model

Identification of Vibrio campbellii isolated from diseased farm-shrimps from south India and establishment of its pathogenic potential in an Artemia model

Development of a haemolysin gene-based multiplex PCR for simultaneous detection ofVibrio campbellii,Vibrio harveyiandVibrio parahaemolyticus

Occurrence of Vibrio parahaemolyticus, Vibrio cholerae, and Vibrio vulnificus in the Aquacultural Environments of Taiwan

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