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Diseases of Aquatic Organisms

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DAO 110:93-99 (2014)  -  DOI:

Contribution to the DAO Special: 'Microcell parasites of molluscs'

Interlaboratory variability in screening for Bonamia ostreae, a protistan parasite of the European flat oyster Ostrea edulis

Grace Flannery1,*, Sharon A. Lynch1, Matt Longshaw2, David Stone2, Paul Martin2, Andrea Ramilo3, Antonio Villalba3, Sarah C. Culloty

1Aquaculture and Fisheries Development Centre, School of Biological, Earth and Environmental Sciences, University College Cork (UCC), Ireland
2Centre for Environment, Fisheries and Aquaculture Science (Cefas), Weymouth Laboratory, The Nothe, Dorset DT4 8UB, UK
3Centro de Investigaciones Marinas (CIMA), Conseller?a do Medio Rural e do Mar, Xunta de Galicia, Aptdo. 13, 36620 Vilanova de Arousa, Spain
*Corresponding author:

ABSTRACT: The spread of the protozoan parasite Bonamia ostreae is of major concern to the European flat oyster Ostrea edulis industry. Many studies have looked at the sensitivity of individual methods available to screen for B. ostreae, but in this study, 3 separate laboratories examined 4 methods of diagnosis currently used routinely in laboratories: heart imprints, histology, polymerase chain reaction (PCR) and in situ hybridisation (ISH). The results were compared to estimate interlaboratory variability. Heart imprints and histology had the highest reproducibility amongst the 3 laboratories, with greatest agreement between detection of infected and uninfected individuals. PCR had the highest detection level in every laboratory. These positives were related to the presence of confirmed infections but also in unconfirmed infections, possibly due to the presence of traces of B. ostreae DNA in oysters where clinical infections were not observed. PCR, in combination with histology or ISH, provided the most reliable detection levels in every laboratory. Variation in results for PCR and ISH observed between laboratories may be due to the different protocols used by each laboratory for both methods. Overall, the findings from the 3 laboratories indicated that at least 2 methods, with fixed protocols, should be used for the accurate detection and determination of infection prevalence within a sample. This combination of methods would allow for a clearer and more precise diagnosis of B. ostreae, preventing further spread of the disease and providing more accurate detection levels and epidemiological information.

KEY WORDS: Bivalve · Protozoa · Diagnostics · Methodology · PCR · In situ hybridisation · Heart imprints · Histology

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Cite this article as: Flannery G, Lynch SA, Longshaw M, Stone D and others (2014) Interlaboratory variability in screening for Bonamia ostreae, a protistan parasite of the European flat oyster Ostrea edulis. Dis Aquat Org 110:93-99.

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