Isolation of Vibrio tapetis from cultivated Atlantic halibut (Hippoglossus hippoglossus L.)

Reid, Helen I.; Duncan, Hazel; Laidler, L. A.; Hunter, Douglas B.; Birkbeck, T. H.

doi:10.1016/s0044-8486(03)00060-7

Cited by 41 publications

(46 citation statements)

References 12 publications

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“…This disease has been described in 2 different clam species -Manila clam Ruditapes philippinarum and carpet-shell clam R. decussatus -in France, England, Ireland, Italy, Spain, and Portugal (Paillard et al 1994, Figueras et al 1996, Allam et al 2000, Paillard 2004). However, V. tapetis has also been isolated from venus clam Venerupis aurea, common cockle Cerastoderma edule, Atlantic halibut Hippoglossus hippoglossus, and corkwing wrasse Symphodus melops from several European countries (Jensen et al 2003, Reid et al 2003.…”

Section: Introductionmentioning

confidence: 99%

Molecular fingerprinting of Vibrio tapetis strains using three PCR-based methods: ERIC-PCR, REP-PCR and RAPD

Rodrı́guez

López-Romalde²,

Beaz³

et al. 2006

Dis. Aquat. Org.

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Brown Ring Disease (BRD) is a bacterial disease caused by Vibrio tapetis which affects cultured clams and causes heavy economic losses. In this study, 28 V. tapetis strains isolated from 5 different hosts were intraspecifically characterized by 3 different polymerase chain reaction-(PCR-) based typing methods: enterobacteria repetitive intergenic consensus (ERIC)-PCR, repetitive extragenic palindromic (REP)-PCR and randomly amplified polymorphic DNA (RAPD)-PCR. Cluster analysis of genetic profiles obtained from these molecular techniques clearly showed the existence of 3 genetic groups strongly correlated to the host origin. The first group was formed by 23 V. tapetis strains isolated from Manila clam Ruditapes philippinarum, 1 isolated from venus clam Venerupis aurea, and 1 isolated from common cockle Cerastoderma edule, all collected from France and Spain. The second group was formed by 2 strains isolated from carpet-shell clam R. decussatus cultured in the northwest of Spain. The third group was composed of 1 strain isolated from Atlantic halibut Hippoglossus hippoglossus from the UK. We concluded that the 3 typing methods based on PCR were useful for the intraspecific typing of V. tapetis strains, and that they can potentially be used as a fast and reliable tool for epidemiological studies in the future. KEY WORDS: Vibrio tapetis · RAPD · ERIC-PCR · REP-PCR · Typing Resale or republication not permitted without written consent of the publisherDis Aquat Org 69: [175][176][177][178][179][180][181][182][183] 2006 demonstrated some genetic variations among V. tapetis strains (Castro et al. 1997, Romalde et al. 2002, Chevalier et al. 2003.Repetitive element PCR is a group of techniques that generates DNA fingerprints which can be utilized for the discrimination of bacterial species and/or strains (Versalovic et al. 1991(Versalovic et al. , 1994. These methods involve the application of oligonucleotide primers based on families of short, highly conserved extragenic repetitive sequences, including the repetitive extragenic palindromic (REP) and the enterobacterial repetitive intergenic consensus (ERIC) sequences (Stern et al. 1984, Hulton et al. 1991. REP and ERIC sequences are present in species throughout the Enterobacteriaceae family (Versalovic et al. 1991, Bachellier et al. 1999. The use of appropriate outward-facing PCR primers directed at these repeated sequences generates multiple amplification products, which reflect distance polymorphisms between adjacent DNA repeats. PCR with primers based on ERIC and REP sequences has been successfully used to differentiate bacterial strains from diverse species (de Bruijn 1992, Bennasar et al. 2002, Bruant et al. 2003, Hahm et al. 2003.The RAPD assay is based on the use of short random sequence primers, 9 to 10 bases in length, which hybridize with sufficient affinity to chromosomal DNA sequences to render a pattern of bands that is used for fingerprinting bacterial strains (Welsh & McClelland 1990, Williams et al. 1990. A number of studies have reported...

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Section: Introductionmentioning

confidence: 99%

Molecular fingerprinting of Vibrio tapetis strains using three PCR-based methods: ERIC-PCR, REP-PCR and RAPD

Rodrı́guez

López-Romalde²,

Beaz³

et al. 2006

Dis. Aquat. Org.

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“…comm.) and cultivated Atlantic halibut in Scotland (Reid et al 2003). As disease prevalence is seasonal and depends on the location of the clams, environmental conditions are thought to affect clam susceptibility to BRD.…”

Section: Introductionmentioning

confidence: 99%

Salinity effects on immune parameters of Ruditapes philippinarum challenged with Vibrio tapetis

Reid¹,

Soudant²,

Lambert³

et al. 2003

Dis. Aquat. Org.

103

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The occurrence of brown ring disease (BRD) in farmed Manila clams Ruditapes philippinarum is seasonal. Development of the disease is believed to require the presence of the infective agent Vibrio tapetis and particular environmental conditions. This paper studies the effect of salinity (20 to 40 ‰) on measurable immune parameters of Manila clams, and the progression of BRD in experimentally infected individuals. At 20 ‰ salinity, the total haemocyte count was reduced and disease prevalence was highest. At 40 ‰ salinity significantly fewer clams presented signs of BRD, and this was correlated with increases in the total haemocyte count, hyalinocyte count, phenoloxidase levels and phagocytic activity of haemocytes. Inoculation of clams with V. tapetis did not have a significant effect on the immune parameters measured. Thus, this laboratory-based study relates environmental stress to disease development.KEY WORDS: Vibrio tapetis · Brown ring disease · Aquaculture · Salinity · Immune response Resale or republication not permitted without written consent of the publisherDis Aquat Org 56: [249][250][251][252][253][254][255][256][257][258] 2003 To ascertain salinity tolerance, Strains CECT 4600 T and IS1 of Vibrio tapetis were cultured at a range of salinities from 5 to 60 ‰ in TSB; salinity was adjusted by addition of appropriate amounts of a stock solution of 20% (w/v) NaCl in TSB. The medium was dispensed in 200 µl vol in a 96-well microtitre plate, and at each salinity triplicate wells were inoculated with 10 µl of a standard suspension of V. tapetis to a final concentration of 10 6 bacteria ml -1. The plates were incubated at 20°C with gentle shaking, and growth was measured as the mean optical density at 600 nm.For the inoculation of clams, bacteria were cultured in 100 ml TSB at salinities of 20, 30 and 40 ‰, at 20°C with gentle shaking in an orbital incubator.Inoculation of clams. Adult Manila clams (35 to 40 mm) were obtained from Marennes, France. The clams were maintained in twelve 50 l aquaria with under-gravel filters containing aerated artificial seawater (ASW) at 20, 30 and 40 ‰. Clams were held at a density of 1 individual l -1 water and at a temperature of 15.5 ± 1.5°C. Upon arrival at the laboratory, the clams were initially held in ASW at 30 ‰ salinity for 2 d, after which time the salinity was adjusted as necessary over a 6 d period (3.3 ‰ every 2 d) from 30 ‰ to 20 or 40 ‰ (4 aquaria per salinity, each containing 50 clams). Clams were held for a further week before inoculation to allow acclimatisation.A third of the volume of water was exchanged with fresh ASW of the appropriate salinity 3 times a week. Water was removed from the tanks with a siphon, and faecal waste was removed from the bottom of the tanks during this procedure. It was established by measuring levels of nitrite and ammonia (TetraTests, TetraWerke) that this cleaning/water-replacement regime maintained low concentrations of these toxic compounds.For the duration of the experiment, clams were fed daily on a mixed alg...

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“…Furthermore, the bacterium has been detected (although not in Ireland to date) in other clam species, e.g. R. decussatus and Venerupis aurea Novoa et al 1998), and in other animals such as the cockle, Cerastoderma edule, , the wrasse, Symphodus melops (Jensen et al 2003) and the halibut, Hippoglossus hippoglossus (Reid et al 2003a), thereby suggesting that it is widespread in the environment in general.…”

Section: Discussionmentioning

confidence: 99%

“…Colonies, which fulfilled the V. tapetis characteristics, were suspected to be V. tapetis isolates. Their identity was confirmed by a slide agglutination test using anti-serum to V. tapetis, obtained from the Institute of Biomedical and Life Sciences, University of Glasgow (Reid et al 2003a). A positive control, V. tapetis CECT 4600, and a negative control, phosphate buffered saline (PBS), were tested concurrently, to ensure the efficacy of the anti-serum, and the validity of the slide agglutination results.…”

Section: Isolation and Characterisation Of Vibrio Tapetismentioning

confidence: 99%

A survey of the health status of the Manila clam Ruditapes philippinarum in Ireland with specific reference to brown ring disease

2009

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The present work investigated the health status of the Manila clam in Ireland, with particular reference to brown ring disease (BRD) caused by V. tapetis which has been responsible for high mortalities of this bivalve throughout Europe. BRD was diagnosed in Ireland in the 1990s, causing heavy mortalities in Manila clam stocks in the north-west coast. In the current study, samples of clams from an Irish hatchery were obtained and screened from two sites, Drumcliff Bay, Co. Sligo and Dungloe Bay, Co. Donegal. Turbellarians and trematodes with some minor infections were the only parasites observed. Clams were examined for BRD, by analysis of shell valves for brown ring signs, and for V. tapetis by microbiological methods and by polymerase chain reaction (PCR). BRD was not diagnosed in cultivated clams from either site. It was, however, diagnosed in residual clams, which had survived the initial outbreak of BRD in the 1990s in Dungloe Bay. V. tapetis, however was detected in clams from both sites. Additionally, BRD was diagnosed, and V. tapetis isolated, in a once-off sample of clams from Mulroy Bay, Co. Donegal, where clams had been on-grown from imported seed. Minor heterogeneity at the 16S rDNA gene was observed between some sequenced products from Mulroy Bay and from Drumcliff Bay and Dungloe Bay indicating that in addition to V. tapetis, a V. tapetis-like strain may have been present in Mulroy Bay. The detection of V. tapetis in asymptomatic clams in Drumcliff Bay and Dungloe Bay may indicate that disease development may only occur when a number of factors combine to induce disease symptoms.

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Isolation of Vibrio tapetis from cultivated Atlantic halibut (Hippoglossus hippoglossus L.)

Cited by 41 publications

References 12 publications

Molecular fingerprinting of Vibrio tapetis strains using three PCR-based methods: ERIC-PCR, REP-PCR and RAPD

Molecular fingerprinting of Vibrio tapetis strains using three PCR-based methods: ERIC-PCR, REP-PCR and RAPD

Salinity effects on immune parameters of Ruditapes philippinarum challenged with Vibrio tapetis

A survey of the health status of the Manila clam Ruditapes philippinarum in Ireland with specific reference to brown ring disease

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