Detection of Haplosporidium nelsoni (Haplosporidia:Haplosporidiidae) in oysters by PCR amplification

Na, Stokes; Me, Siddall; Burreson, EM

doi:10.3354/dao023145

Cited by 41 publications

(32 citation statements)

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“…These results may have been due to the permissive conditions used in their PCR assay. Ulrich et al (2007) amended the reaction published by Stokes et al (1995), increasing the MgCl 2 concentration from 1.5 to 3.5 mM. They claimed that they increased stringency of the PCR by increasing the annealing temperature from 50 to 55°C, which would be true if that were the only change made to the published protocol.…”

Section: Discussionmentioning

confidence: 99%

“…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%

“…The PCR assay for detection of genomic DNA from Haplosporidium nelsoni was modified from that published by Stokes et al (1995) using the primers MSX A' (5'-CGA CTT TGG CAT TAG GTT TCA GAC C-3') and MSX B (5'-ATG TGT TGG TGA CGC TAA CCG-3'). 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).…”

Section: Methodsmentioning

confidence: 99%

“…In enzootic locations along the mid-Atlantic and northeastern United States where seasonal infection patterns have been described, Haplosporidium nelsoni prevalence can be high in the summer, depending on when in the infection cycle oysters are collected, but infection prevalence and intensity are universally elevated in the winter (Andrews 1966, Ford & Haskin 1982, Matthiessen et al 1990). The southern-most location for which seasonal infection prevalence is available from histological analyses comes from the entrance to Charleston Harbor, South Carolina, USA, where sites were sampled in 1994and 1995(Bobo et al 1997. At the site with the highest prevalence, infections predominated from fall into spring, and, at another site, they were more abundant in the fall and spring than in the winter or summer, but, when all sites were considered together, infections were found throughout the year.…”

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confidence: 99%

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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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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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

Ford

Paterno²,

Scarpa³

et al. 2011

Dis. Aquat. Org.

Self Cite

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“…Transmission occurs over large distances and can be rapid under favorable climatic conditions, resulting in rapid parasite proliferation and elevated adult mortalities (Andrews and Wood 1967;Lafferty and Kuris 1993;Burreson and Ragone-Calvo 1996;Ford and Tripp 1996). Throughout the region in which the disease organism can exist, most hosts are exposed to the disease organism and probably become infected, even though infections may be undetected (Bushek et al 1994;Stokes et al 1995;Culloty et al 2003). The spread and intensification of infections can result in epizootics, which are disease epidemics in nonhuman animal populations.…”

Section: Systematic Positionmentioning

confidence: 99%

First record of protozoan parasites, Tetrahymena rostrata and Callimastix equi from the edible oyster in Sundarbans region of West Bengal, India

Biswas

Bandyopadhyay

2014

J Parasit Dis

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Several protozoan parasites have been found infecting the edible oysters, hence deteriorating the meat quality. Protozoan parasites such as, Tetrahymena rostrata and Callimastix equi infested the edible oyster in Sundarbans region, West Bengal, India, are first record from this region. Due to filter feeding habit of the organisms, oysters provides excellent ecological services in regard to efficient cleaning of infectious agents from surrounding water as a potential measure to improve water quality. However, these environmental benefits are associated with public heath risks from contaminated oysters intended for human consumption.

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Genetic by environmental variation but no local adaptation in oysters (Crassostrea virginica)

Hughes

Hanley

Byers

et al. 2016

Ecology and Evolution

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Functional trait variation within and across populations can strongly influence population, community, and ecosystem processes, but the relative contributions of genetic vs. environmental factors to this variation are often not clear, potentially complicating conservation and restoration efforts. For example, local adaptation, a particular type of genetic by environmental (G*E) interaction in which the fitness of a population in its own habitat is greater than in other habitats, is often invoked in management practices, even in the absence of supporting evidence. Despite increasing attention to the potential for G*E interactions, few studies have tested multiple populations and environments simultaneously, limiting our understanding of the spatial consistency in patterns of adaptive genetic variation. In addition, few studies explicitly differentiate adaptation in response to predation from other biological and environmental factors. We conducted a reciprocal transplant experiment of first‐generation eastern oyster (Crassostrea virginica) juveniles from six populations across three field sites spanning 1000 km in the southeastern Atlantic Bight in both the presence and absence of predation to test for G*E variation in this economically valuable and ecologically important species. We documented significant G*E variation in survival and growth, yet there was no evidence for local adaptation. Condition varied across oyster cohorts: Offspring of northern populations had better condition than offspring from the center of our region. Oyster populations in the southeastern Atlantic Bight differ in juvenile survival, growth, and condition, yet offspring from local broodstock do not have higher survival or growth than those from farther away. In the absence of population‐specific performance information, oyster restoration and aquaculture may benefit from incorporating multiple populations into their practices.

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Detection of Haplosporidium nelsoni (Haplosporidia:Haplosporidiidae) in oysters by PCR amplification

Cited by 41 publications

References 1 publication

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

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

First record of protozoan parasites, Tetrahymena rostrata and Callimastix equi from the edible oyster in Sundarbans region of West Bengal, India

Genetic by environmental variation but no local adaptation in oysters (Crassostrea virginica)

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