PCR detection of Tetramicra brevifilum (Microspora) infection in turbot ( Scophthalmus maximus L.) musculature

This study investigated the spatial distribution of Tetramicra brevifilum spores in the musculature of infected turbot Scophthalmus maximus , with the aim of identifying the most appropriate body locations for diagnostic assays. A PCR protocol optimized for the detection of T. brevifilum spores in t...

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Bibliographic Details
Published in:Parasitology
Main Authors: LEIRO, J., IGLESIAS, R., PARAMÁ, A., ARAGORT, W., SANMARTÍN, M. L.
Format: Article in Journal/Newspaper
Language:English
Published: Cambridge University Press (CUP) 2002
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Online Access:http://dx.doi.org/10.1017/s0031182001001020
https://www.cambridge.org/core/services/aop-cambridge-core/content/view/S0031182001001020
Description
Summary:This study investigated the spatial distribution of Tetramicra brevifilum spores in the musculature of infected turbot Scophthalmus maximus , with the aim of identifying the most appropriate body locations for diagnostic assays. A PCR protocol optimized for the detection of T. brevifilum spores in turbot muscle is also described. In fish showing low- and moderate-intensity infection, the spatial distribution of spores was best fitted by a negative binomial distribution, indicating a clumped spatial pattern; the negative binomial coefficient k was lower for fish with low-intensity infection, indicating a more markedly clumped pattern in these fish. In fish with high-intensity infection, the spatial distribution of spores was best fitted by the Poisson distribution, indicating a random pattern. In both low- and moderate-intensity infection, spores were present at highest density in the musculature adjoining the dorsal fins. Samples for PCR were therefore obtained from this location. PCR amplification was of the small subunit ribosomal DNA (SSUrDNA), using a pair of species-specific primers that amplify the 1250 bp product. The PCR protocol developed showed better sensitivity than microscopical techniques (detection rate by microscopy 25%, versus 42% by PCR), suggesting that it may be useful for routine screening for Tetramicra brevifilum infection in cultured turbot.