Comparative Evaluation of molecular diagnostic tests for Nucleospora salmonis and prevalence in migrating juvenile salmonids from the Snake River, USA

Samantha Badil, Diane G. Elliott, Tomofumi Kurobe, Ronald Hedrick, Kathy Clemens, Marilyn Blair, Maureen K. Purcell

Research output: Contribution to journalArticle

2 Citations (Scopus)

Abstract

Nucleospora salmonis is an intranuclear microsporidian that primarily infects lymphoblast cells and contributes to chronic lymphoblastosis and a leukemia-like condition in a range of salmonid species. The primary goal of this study was to evaluate the prevalence of N. salmonis in out-migrating juvenile hatchery and wild Chinook salmon Oncorhynchus tshawytscha and steelhead O. mykiss from the Snake River in the U.S. Pacific Northwest. To achieve this goal, we first addressed the following concerns about current molecular diagnostic tests for N. salmonis: (1) nonspecific amplification patterns by the published nested polymerase chain reaction (nPCR) test, (2) incomplete validation of the published quantitative PCR (qPCR) test, and (3) whether N. salmonis can be detected reliably from nonlethal samples. Here, we present an optimized nPCR protocol that eliminates nonspecific amplification. During validation of the published qPCR test, our laboratory developed a second qPCR test that targeted a different gene sequence and used different probe chemistry for comparison purposes. We simultaneously evaluated the two different qPCR tests for N. salmonis and found that both assays were highly specific, sensitive, and repeatable. The nPCR and qPCR tests had good overall concordance when DNA samples derived from both apparently healthy and clinically diseased hatchery rainbow trout were tested. Finally, we demonstrated that gill snips were a suitable tissue for nonlethal detection of N. salmonis DNA in juvenile salmonids. Monitoring of juvenile salmonid fish in the Snake River over a 3-year period revealed low prevalence of N. salmonis in hatchery and wild Chinook salmon and wild steelhead but significantly higher prevalence in hatchery-derived steelhead. Routine monitoring of N. salmonis is not performed for all hatchery steelhead populations. At present, the possible contribution of this pathogen to delayed mortality of steelhead has not been determined.

Original languageEnglish (US)
Pages (from-to)19-29
Number of pages11
JournalJournal of Aquatic Animal Health
Volume23
Issue number1
DOIs
StatePublished - Apr 13 2011

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Snake River
Salmonidae
diagnostic techniques
hatcheries
quantitative polymerase chain reaction
hatchery
Oncorhynchus tshawytscha
river
polymerase chain reaction
testing
salmonid
Oncorhynchus mykiss
amplification
monitoring
Microsporidia
DNA
leukemia
probes (equipment)
gills
chemistry

ASJC Scopus subject areas

  • Aquatic Science

Cite this

Comparative Evaluation of molecular diagnostic tests for Nucleospora salmonis and prevalence in migrating juvenile salmonids from the Snake River, USA. / Badil, Samantha; Elliott, Diane G.; Kurobe, Tomofumi; Hedrick, Ronald; Clemens, Kathy; Blair, Marilyn; Purcell, Maureen K.

In: Journal of Aquatic Animal Health, Vol. 23, No. 1, 13.04.2011, p. 19-29.

Research output: Contribution to journalArticle

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