Haplosporidiosis in the Pacific oyster Crassostrea gigas from the French Atlantic coast

Renault, Tristan; Stokes, Nancy A.; Chollet, Bruno; Cochennec, Nathalie; Berthe, Franck; Gérard, André; Burreson, Eugene M.

doi:10.3354/dao042207

Cited by 76 publications

(78 citation statements)

References 20 publications

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“…The extracted DNA of 463 sampled M. arenaria was subjected to PCR amplification using Ostreid Herpes Virus (OSHV)-specific primers C2/C6 and OSHV-For/OSHV-3 Rev (Renault & Arzul, 2001), as the clams were obtained from an area (Bannow Bay, Wexford, Ireland) where this virus is present in Pacific Oysters, Crassostrea gigas. Both Mya arenaria (n=463) and E. siliqua (n=484) samples were screened for the presence of bacterial DNA using the generic forward primer 18Scom-F and the reverse primer 18Scom-R (Zhang & Lin, 2005), and for detection of haplosporidian species using the forward primer (Hap-F1) and reverse primer (Hap-R3) (Renault et al, 2000). …”

Section: Softmentioning

confidence: 99%

Withdrawn: “A health status survey of clams, Mya arenaria and Ensis siliqua, in the Irish Sea”

Cross

Lynch

O’Riordan

et al. 2014

Journal of Invertebrate Pathology

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Section: Softmentioning

confidence: 99%

Withdrawn: “A health status survey of clams, Mya arenaria and Ensis siliqua, in the Irish Sea”

Cross

Lynch

O’Riordan

et al. 2014

Journal of Invertebrate Pathology

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“…Primer MSX A' is the same as MSX A originally designed by Stokes et al (1995) and used by Ulrich et al (2007), but has an extra 8 bases on the 5'-end and amplifies a 573 bp, rather than a 564 bp, section of the same region of the small subunit (SSU) rDNA (Renault et al 2000). Primer MSX A is much shorter than MSX B, resulting in a melting temperature (T m ) difference of 16.3°C.…”

Section: Methodsmentioning

confidence: 99%

“…Cycling parameters were 2 min at 94°C for initial denaturation, then 35 cycles at 94°C for 30 s, 59°C for 30 s, and 72°C for 1.5 min for the denaturation, annealing, and extension steps, respectively, and a final elongation at 72°C for 5 min (Renault et al 2000). A total of 10 µl of the amplification product were applied to a 2% agarose gel using SYBR Green (5 µl in 40 ml of gel buffer, pH 8.0) to stain the product, which was then photo-documented as a digital image.…”

Section: Methodsmentioning

confidence: 99%

Widespread survey finds no evidence of Haplosporidium nelsoni (MSX) in Gulf of Mexico oysters

Ford

Paterno²,

Scarpa³

et al. 2011

Dis. Aquat. Org.

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The advent of molecular detection assays has provided a set of very sensitive tools for the detection of pathogens in marine organisms, but it has also raised problems of how to interpret positive signals that are not accompanied by visual confirmation. PCR-positive results have recently been reported for Haplosporidium nelsoni (MSX), a pathogen of the oyster Crassostrea virginica in 31 of 40 oysters from 6 sites in the Gulf of Mexico and the Caribbean Sea. Histological confirmation of the PCR results was not undertaken, and no haplosporidian has been reported from the numerous histological studies and surveys of oysters in the region. To further investigate the possibility that H. nelsoni is present in this region, we sampled 210 oysters from 40 sites around the Gulf of Mexico and Puerto Rico using PCR and 180 of these using tissue-section histology also. None of the oysters showed evidence of H. nelsoni by PCR or of any haplosporidian by histology. We cannot, therefore, confirm that H. nelsoni is present and widespread in the Gulf of Mexico and the Caribbean Sea. Our results do not prove that H. nelsoni is absent from the region, but taken together with results from previous histological surveys, they suggest that for the purposes of controlling oyster importation, the region should continue to be considered free of the parasite. KEY WORDS: PCR · Oyster · Crassostrea virginica · Molecular detection · Infection · Shellfish importation · Haplosporidian Resale or republication not permitted without written consent of the publisherDis Aquat Org 93: [251][252][253][254][255][256] 2011 C. virginica from Nova Scotia, Canada, to Biscayne Bay, Florida, USA (Haskin & Andrews 1988, Barber et al. 1997, Stephenson et al. 2003, but none of the several histological studies of oysters in the Gulf of Mexico have reported H. nelsoni or any other haplosporidian (Couch 1985, Gauthier et al. 1990, Fisher et al. 1996, Kim et al. 1998, Winstead et al. 2004, Kim & Powell 2006. The most comprehensive of these is the annual status and trends (S&T) MATERIALS AND METHODSCrassostrea virginica were collected between December 2007 and February 2008 at 32 sites representing 14 bay systems from lower Laguna Madre, Texas, to Florida Bay, Florida, USA, and C. rhizophorae were collected from 2 bays in Puerto Rico (Fig. 1, Table 1). In May and July 2010, C. virginica were sampled from 6 additional sites, representing 2 bay systems in Texas and 1 in Florida (Fig. 1, Table 1). All samples were express shipped directly to HSRL. Upon receipt, 12 oysters from each sample were opened using a shucking knife, which was rinsed in 10% bleach between each oyster. A section of gill about 4 mm wide and encompassing all 4 demibranchs approximately 6 to 7 mm posterior to the gill-palp junction was excised and preserved in 70% ethanol (EtOH) for PCR analysis. Dissecting instruments were scraped clean in a bleach-sand mixture, dipped in EtOH and flamed between each oyster. Between the PCR section and the palps, and adjacent to the piece of gill ta...

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“…Only PCR assays for the Australian abalone herpesvirus [62] and oyster parasites M. mackini [63] and B. ostreae and B. exitiosa [52] are noteworthy for the degree of assessment applied. Although a long history of use does provide confidence that important tools such as the conventional PCR assays for H. nelsoni [64,65], M. refringens [34], B. ostreae [66] and P. marinus [49] perform reliably despite limited formal assessment, a 'long history of use' is no substitute for the formal validation that is essential to demonstrate that these molecular assays perform as intended. Lack of validation has opened the door to a proliferation of redundant assays that complicate the matter of formally determining which assays are suitable for which application.…”

Section: The Paradox Of Advanced Diagnosticsmentioning

confidence: 99%

Managing marine mollusc diseases in the context of regional and international commerce: policy issues and emerging concerns

Carnegie

Arzul

Bushek

2016

Phil. Trans. R. Soc. B

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Marine mollusc production contributes to food and economic security worldwide and provides valuable ecological services, yet diseases threaten these industries and wild populations. Although the infrastructure for mollusc aquaculture health management is well characterized, its foundations are not without flaws. Use of notifiable pathogen lists can leave blind spots with regard to detection of unlisted and emerging pathogens. Increased reliance on molecular tools has come without similar attention to diagnostic validation, raising questions about assay performance, and has been accompanied by a reduced emphasis on microscopic diagnostic expertise that could weaken pathogen detection capabilities. Persistent questions concerning pathogen biology and ecology promote regulatory paralysis that impedes trade and which could weaken biosecurity by driving commerce to surreptitious channels. Solutions that might be pursued to improve shellfish aquaculture health management include the establishment of more broad-based surveillance programmes, wider training and use of general methods like histopathology to ensure alertness to emerging diseases, an increased focus on assay assessment and validation as fundamental to assay development, investment in basic research, and application of risk analyses to improve regulation. A continual sharpening of diagnostic tools and approaches and deepening of scientific knowledge is necessary to manage diseases and promote sustainable molluscan shellfish industries.

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Haplosporidiosis in the Pacific oyster Crassostrea gigas from the French Atlantic coast

Cited by 76 publications

References 20 publications

Withdrawn: “A health status survey of clams, Mya arenaria and Ensis siliqua, in the Irish Sea”

Withdrawn: “A health status survey of clams, Mya arenaria and Ensis siliqua, in the Irish Sea”

Widespread survey finds no evidence of Haplosporidium nelsoni (MSX) in Gulf of Mexico oysters

Managing marine mollusc diseases in the context of regional and international commerce: policy issues and emerging concerns

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