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Diseases of Aquatic Organisms

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DAO 141:91-101 (2020)  -  DOI: https://doi.org/10.3354/dao03525

PCR-RFLP: a targeted method to reveal host specific malacosporean infection profiles (Cnidaria: Myxozoa: Malacosporea)

Paolo Ruggeri1,*, Juliana Naldoni2, Hanna Hartikainen3, Beth Okamura1

1Natural History Museum, Cromwell Rd, Kensington, London SW7 5BD, UK
2Departamento de Ecologia e Biologia Evolutiva, Universidade Federal de São Paulo, Rua Professor Artur Riedel, no. 275, CEP 09972-270, Diadema, SP, Brazil
3School of Life Sciences, University of Nottingham, University Park, Nottingham NG7 2RD, UK
*Corresponding author:

ABSTRACT: Malacosporeans are a group of endoparasitic cnidarians (Myxozoa) that use freshwater bryozoans and fish as final and intermediate hosts, respectively. The malacosporean Tetracapsuloides bryosalmonae causes proliferative kidney disease (PKD), an emerging disease in aquaculture and wild fish populations, including threatened salmonids in Europe and the USA. Mixed infections of malacosporeans are often encountered, and a monitoring tool for screening of multiple malacosporean species in either their fish or bryozoan hosts is therefore desirable. We describe an inexpensive method that combines PCR amplification of the partial 18S rRNA gene (~260 bp) and a single-step restriction fragment length polymorphism (RFLP) method for identification of 10 malacosporean lineages and species. We demonstrate and test this methodology on a set of DNA extracted from malacosporeans infecting fish kidney and tissues sampled from bryozoan colonies and compare the results with Sanger sequencing of the same parasite DNA isolates. The PCR-RFLP and Sanger sequencing methods agreed in 100% of cases. The PCR-RFLP method offers a number of opportunities, including screening large panels of host tissue samples to gain insights into infection patterns, characterizing mixed infections, and confirming highly pathogenic T. bryosalmonae infections. The method can also be further refined as new sequence data become available for malacosporeans.


KEY WORDS: 18S rDNA · Restriction enzymes · Myxozoa · Buddenbrockia spp. · Tetracapsuloides spp. · Mixed infections


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Cite this article as: Ruggeri P, Naldoni J, Hartikainen H, Okamura B (2020) PCR-RFLP: a targeted method to reveal host specific malacosporean infection profiles (Cnidaria: Myxozoa: Malacosporea). Dis Aquat Org 141:91-101. https://doi.org/10.3354/dao03525

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