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Diseases of Aquatic Organisms

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DAO 69:185-195 (2006)  -  doi:10.3354/dao069185

Molecular detection of Hematodinium spp. in Norway lobster Nephrops norvegicus and other crustaceans

H. J. Small1,4,*, D. M. Neil1, A. C. Taylor1, R. J. A. Atkinson2, G. H. Coombs3

1Division of Environmental and Evolutionary Biology, and 3Division of Infection & Immunity, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK
2University Marine Biological Association, Millport, Isle of Cumbrae KA28 0EG, UK
4Present address: Virginia Institute of Marine Science, College of William and Mary, Gloucester Point, Virginia 23062, USA

ABSTRACT: The Norway lobster Nephrops norvegicus (L.) from the coastal waters of Scotland is seasonally infected by a parasitic dinoflagellate of the genus Hematodinium. Methods used to detect infection include a morphological index (pleopod diagnosis) and several immunoassays. The present study describes the development and application of a set of Hematodinium-specific polymerase chain reaction (PCR) primers and DNA probes based on Hematodinium ribosomal DNA (rDNA). In the PCR assay, a diagnostic band of 380 bp was consistently amplified from total genomic DNA isolated from Hematodinium-infected N. norvegicus. The sensitivity of the assay was 1 ng DNA, which is equivalent to 0.6 parasites. The primer pair also detected Hematodinium DNA in preparations of the amphipod Orchomene nanus, indicating that the amphipod may be infected with the same Hematodinium sp. infecting N. norvegicus. DNA probes detected Hematodinium parasites in heart, hepatopancreas and gill tissues from N. norvegicus, and hepatopancreas and gill tissues from Carcinus maenas, confirming Hematodinium infection in the latter.

KEY WORDS: Hematodinium · Parasite · Norway lobster · Molecular detection

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