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Detection of the oyster parasite Bonamia ostreae by fluorescent in situ hybridization
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Bonamia ostreae is an economically significant protistan parasite of the flat oyster Ostrea edulis in Europe and North America. Management of this parasite depends partly upon its reliable identification in wild and aquacultured oyster populations, but B. ostreae is small and difficult to detect by traditional microscopic methods. We designed a fluorescent in situ hybridization (FISH) assay to sensitively detect B. ostreae in standard histopathological sections of B. ostreae-infected oysters using fluorescently labeled DNA oligonucleotide probes. Hybridization using a cocktail of 3 presumptively B. ostreae-specific, fluorescein iso(thio)cyanate (FITC)-labeled oligonucleotides produced an unambiguous staining pattern of small green rings inside infected oyster hemocytes that was easily distinguished from host tissue background. This pattern is diagnostic for B. ostreae. A negative control cocktail of oligonucleotides containing 2 mismatches relative to target sequences, on the other hand, failed to hybridize at all. B. ostreae-specific probes did not cross-react with a related protist, Haplosporidium nelsoni.
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“Neutral-buffered formalin, Z-fix®, and SafeFix® II are fixatives of choice for molecular applications, such as in situ hybridizations (e.g., Bruce et al. , 1993; Carnegie et al. , 2003; Li et al. , 2006; Small et al. , 2007)”
From Collection techniques for the analyses of pathogens in crustaceans (2017) · cites this paper as Carnegie et al. , 2003
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Research works printed but not linked: 12
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- Farley CA, Wolf PH, Elston RA (1988) A long-term study of 'microcell' disease in oysters with description of a new genus, Mikrocytos (g. n.), and two new species, Mikro- cytos mackini (sp. n.) and Mikrocytos roughleyi (sp. n.). Fish Bull 86(3):581-593
- Friedman CS, McDowell T, Groff JM, Hollibaugh JT, Manzer D, Hedrick RP (1989) Presence of Bonamia ostreae among populations of the European flat oyster, Ostrea edulis Linné, in California, USA. J Shellfish Res 8(1):133-137
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Collection techniques for the analyses of pathogens in crustaceans
Shields, J. D. (2017) · Journal of Crustacean Biology · doi:10.1093/jcbiol/rux077 · Cited by 16
Metadata
- Authors
- Carnegie, R., Barber, B., & Distel, D.
- Venue
- Diseases of Aquatic Organisms
- Publisher
- Inter-Research Science Center
- Published
- 2003
- Volume
- 55
- Issue
- 3
- Pages
- 247-252
- ISSN
- 0177-5103 (ISSN-L)
- 1616-1580
- OpenAlex
- W1977869924
- Cited by
- 36
- References
- 0
- Citations by year
- 2025: 1 · 2024: 2 · 2022: 1 · 2021: 1 · 2019: 1 · 2017: 1 · 2016: 2 · 2015: 1 · 2014: 2 · 2013: 2
- FWCI
- 2.84
- Type
- journal article
- Language
- en
- Open access
- bronze
- doi.org
- Locations
- Diseases of Aquatic Organisms (publishedVersion)
- PubMed (publishedVersion)
- W&M Publish (College of William & Mary) (submittedVersion)
- Funders
- U.S. Department of Agriculture
- Dartmouth College
- Maine Agricultural and Forest Experiment Station
- University of Massachusetts Dartmouth
- New Hampshire Sea Grant, University of New Hampshire
- Cooperative State Research, Education, and Extension Service
- Topics
- Life Sciences › Agricultural and Biological Sciences › Aquatic Science › Marine Bivalve and Aquaculture Studies
- Life Sciences › Immunology and Microbiology › Immunology › Aquaculture disease management and microbiota
- Life Sciences › Immunology and Microbiology › Parasitology › Parasitic Infections and Diagnostics
- Keywords
- Bonamia ostreae, Ostrea edulis, oyster parasites, fluorescence in situ hybridization, histopathology, hemocytes, parasite detection, Haplosporidium nelsoni, cross-reactivity, protist parasites
- MeSH terms
- Animals, Hemocytes, Host-Parasite Interactions, Ostreidae, DNA Probes, In Situ Hybridization, Fluorescence, Haplosporida
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