Detection of Minchinia sp., in rock oysters Saccostrea cuccullata (Born, 1778) using DNA probes.

Detection of Minchinia sp., in rock oysters Saccostrea cuccullata (Born, 1778) using DNA probes. Research Abstract Details 

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Detection of Minchinia sp., in rock oysters Saccostrea cuccullata (Born, 1778) using DNA probes. Abstract Text:

Douglas Bearham,Zoe Spiers,Shane Raidal,J Brian Jones,Philip K Nicholls,Douglas Bearham,Zoe Spiers,Shane Raidal,J Brian Jones,Philip K Nicholls,Douglas Bearham,Zoe Spiers,Shane Raidal,J Brian Jones,Philip K Nicholls,

Haplosporidian parasites infect various invertebrate hosts including some commercially important shellfish. Haplosporidium nelsoni (along with Perkinsus marinus) has severely affected Eastern oyster production on the eastern seaboard of the United States and flat oyster production in Europe has been severely impacted by Bonamia ostreae. These parasites are also often present at a very low prevalence and there are a variety of morphologically similar species that can be difficult to differentiate during cytological or histological diagnosis hence the need to develop specific tests. Recently, a Minchinia sp. was described affecting rock oysters (Saccostrea cuccullata) in north Western Australia. In this study, two in situ hybridisation (ISH) assays and a PCR assay have been developed and optimised for use in investigating these parasites. The first ISH assay used a 166bp polynucleotide probe while the second used a 30bp oligonucleotide probe. The specificity of each ISH assay was assessed by applying each probe to a variety of haplosporidian (5), a paramyxian (1) or ciliophora (1) parasites. The polynucleotide probe produced strong hybridisation signals against all of the haplosporidian parasites tested (Minchinia sp., Minchinia teredinis, Bonamia roughleyi, H. nelsoni and Haplosporidium costale) while the oligonucleotide probe recognised only the Minchinia sp. Both probes failed to detect the paramyxian (Marteilia sp.) or the Rhynchodid-like ciliate. The PCR assay amplifies a 220bp region and detected Minchinia sp. DNA from 50ng of genomic DNA extracted from the tissues of infected oysters and 10fg of amplified Minchinia sp. DNA. The assay did not react to oysters infected with H. nelsoni or H. costale. The ability of the PCR and oligonucleotide ISH assay to diagnose Minchinia sp. infected oysters was compared to histological examination from a sample of 56 oysters. The PCR assay revealed 26 infections while histological examination detected 14 infections. The oligonucleotide ISH assay detected 29 infections. The oligonucleotide ISH and PCR assays were found to be significantly more sensitive than histology for detecting the parasite.

Detection of Minchinia sp., in rock oysters Saccostrea cuccullata (Born, 1778) using DNA probes. Publishing Authors By Initials

D Bearham,Z Spiers,S Raidal,JB Jones,PK Nicholls,D Bearham,Z Spiers,S Raidal,JB Jones,PK Nicholls,D Bearham,Z Spiers,S Raidal,JB Jones,PK Nicholls,

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Detection of Minchinia sp., in rock oysters Saccostrea cuccullata (Born, 1778) using DNA probes. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Journal of invertebrate pathology

VOLUME: 97

Page Numbers: 50-60

Journal Abbreviation: J. Invertebr. Pathol.

ISSN: 0022-2011

DAY: 18

MONTH: 07

YEAR: 2007

Detection of Minchinia sp., in rock oysters Saccostrea cuccullata (Born, 1778) using DNA probes. Information

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LANGUAGE: eng

NlmUniqueID: 14067

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Grant and Affiliation Information for Detection of Minchinia sp., in rock oysters Saccostrea cuccullata (Born, 1778) using DNA probes.

AFFILIATION: Murdoch University, School of Veterinary and Biomedical Sciences, Murdoch Drive, Murdoch, WA 6150, Australia.

Country: United States

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MEDLINETA: J Invertebr Pathol

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