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Diseases of Aquatic Organisms

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DAO 63:197-204 (2005)  -  doi:10.3354/dao063197

Parvicapsula pseudobranchicola (Myxosporea) in farmed Atlantic salmon Salmo salar: tissue distribution, diagnosis and phylogeny

A. Nylund1,*, E. Karlsbakk1, P. A. Sæther2, C. Koren3, T. Larsen4, B. D. Nielsen5,A. E. Brøderud6, C. Høstlund2, K. R. Fjellsøy2, K. Lervik7, L. Rosnes1

1Department of Biology, University of Bergen, PO Box 7800, 5020 Bergen, Norway
2 MarinHelse AS, Parkveien 5, 9060 Lyngseidet, Norway
3Fiskehelsetjenesten i Tromsø, PO Box 2047, 9265 Tromsø, Norway
4Norut NIBR Finnmark, Follums vei 33, 9510 Alta, Norway
5Flatvoll, 9151 Storslett, Norway
6HeMiTec, Hovedveien 2, 9151 Storslett, Norway
7Grindav., 7970 Kolvereid, Norway

ABSTRACT: Parvicapsula pseudobranchicola infections in farmed Atlantic salmon in Norway are associated with low-grade to significant mortalities. The parasite is found as mature spores in pseudobranchs, but has also been detected in the gills, liver and kidney. Diagnosis has relied on the detection of Parvicapsula spores, with the pseudobranch being the preferred organ. A better understanding of the epizootiology of this myxosporean is a prerequisite for appropriate management and control. Hence, early detection of infections and life cycle studies are needed. We sequenced the small subunit (ssu) rDNA (18S) from P. pseudobranchicola and developed a sensitive diagnostic PCR protocol. This allowed us to (1) identify appropriate tissues for diagnostic assays, (2) examine the intraspecific variation in ssu rDNA in the parasite’s Norwegian range, (3) examine annelid potential primary hosts and (4) obtain additional ssu rDNA sequences of marine Parvicapsula species to perform a phylogenetic study. Primers were constructed targeting the ssu rDNA from P. minibicornis. With these we obtained a partial ssu sequence of the P. pseudobranchicola type isolate. A new set of primers (PCF3/PCR3) was constructed for diagnostic purposes. These were tested against DNA from the host and several myxozoan species infecting Norwegian salmon. The primers give a positive product of 203 bp and pick out P. pseudobranchicola in salmonids. They also amplify the congeners P. unicornis and P. asymmetrica infecting unrelated fish. The PCR protocol developed showed a greater sensitivity than light microscopy. The pseudobranchs were always positive and are the recommended organ for PCR diagnostics. There was no sequence variation between geographic isolates from farmed salmon. Preliminary examinations of marine polychaetes and oligochaetes collected from farm sites with parvicapsulose problems were negative. A comparison of the sequence of the ssu rDNA from P. pseudobranchicola with that of other myxozoans shows that it groups closely together with P. unicornis and P. asymmetrica. The closest relative to this group is P. minibicornis.

KEY WORDS: Parvicapsula pseudobranchicola · P. unicornis · P. asymmetrica · ssu rDNA · Molecular phylogeny · Diagnostic PCR

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