Molecular detection of Hematodinium spp. in Norway lobster Nephrops norvegicus and other crustaceans.
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.
Small
H J
HJ
Division of Environmental and Evolutionary Biology, Institute of Biomedical and Life Sciences, University of Glasgow, UK. hamish@vims.edu
Neil
D M
DM
Taylor
A C
AC
Atkinson
R J A
RJ
Coombs
G H
GH
D033304:Amphipoda / Q000469:parasitology; D000818:Animals; D001483:Base Sequence; D003386:Brachyura / Q000469:parasitology; D003445:Crustacea / Q000469:parasitology*; D017931:DNA Primers / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry; D004275:DNA, Ribosomal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D004141:Dinoflagellida / Q000235:genetics* / Q000302:isolation & purification*; D005880:Gills / Q000469:parasitology; D006321:Heart / Q000469:parasitology; D043143:Hepatopancreas / Q000469:parasitology; D017403:In Situ Hybridization / Q000662:veterinary; D008969:Molecular Sequence Data; D008121:Nephropidae / Q000469:parasitology*; D016133:Polymerase Chain Reaction / Q000379:methods / Q000662:veterinary*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D016415:Sequence Alignment / Q000662:veterinary
D017931:DNA Primers; D015342:DNA Probes; D004275:DNA, Ribosomal; D021903:DNA, Ribosomal Spacer