title: Development of PCR method for detecting Kudoa iwatai (Myxozoa: Multivalvulida) from rock bream Oplegnathus fasciatus.
abstract: We developed a PCR assay targeting the 28S rDNA of Kudoa iwatai (Multivalvulida: Myxozoa) and investigated the prevalence of infection in rock bream Oplegnathus fasciatus, which is commercially an important aquaculture species in Korea, with this assay. Detection limit of the PCR assay was 2.5 fg/μl with plasmid DNA and 8.6 × 103spores/ml with purified spores, respectively. This PCR assay did not amplify DNA of other Kudoa species (Kudoa septempunctata, Kudoa lateolabracis, Kudoa thyrsites) tested. Sliced muscles of whole body from 318 rock bream (wild and cultured) were examined by this PCR assay and also with the naked eyes. All of the wild fish did not produce amplicons nor did harbor visible Kudoa cysts (0/70). Three of the cultured fish were PCR-positive and also harbored visible Kudoa cysts (3/248, 1.2%). The sequences of amplicons (574 bp) were 100% identical with those of the K. iwatai already registered in Genbank. When the visceral organs of these three fish were examined, visible cysts were not found, but one stomach sample was found to be PCR-positive. There was no difference in the prevalence of infection estimated by PCR assay and the presence of visible Kudoa cysts in our samples. This is thought to be because the development of K. iwatai is already completed and only mature Kudoa cysts existed in our samples.
authors.affiliation: East Coast Life Science Institute, Gangneung-Wonju National University, Gangneung, 25457, South Korea.|Department of Marine Bioscience, Gangneung-Wonju National University, Gangneung, 25457, South Korea.
authors.forename: Chan-Hyeok
authors.initials: CH
authors.lastname: Jeon
authors.affiliation: Pathology Division, National Institute of Fisheries Science, Busan, 46083, South Korea.
authors.forename: Jeung-Wan
authors.initials: JW
authors.lastname: Do
authors.affiliation: Department of Marine Bioscience, Gangneung-Wonju National University, Gangneung, 25457, South Korea.
authors.forename: U-Hwa
authors.initials: UH
authors.lastname: Nam
authors.affiliation: Department of Aqualife Medicine, College of Fisheries and Ocean Science, Chonnam National University, Yeosu, 59626, South Korea.
authors.forename: Wi-Sik
authors.initials: WS
authors.lastname: Kim
authors.affiliation: Department of Marine Bioscience, Gangneung-Wonju National University, Gangneung, 25457, South Korea. jhkim70@gwnu.ac.kr.
authors.forename: Jeong-Ho
authors.identifier: http://orcid.org/0000-0001-7920-8258
authors.initials: JH
authors.lastname: Kim
mesh_terms: D000818:Animals; D017756:Aquaculture; D004275:DNA, Ribosomal / Q000737:chemistry / Q000235:genetics; D005393:Fish Diseases / Q000453:epidemiology / Q000469:parasitology*; D009132:Muscles / Q000469:parasitology; D055764:Myxozoa / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification; D010273:Parasitic Diseases, Animal / Q000453:epidemiology / Q000469:parasitology*; D010473:Perciformes / Q000469:parasitology*; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000662:veterinary; D056910:Republic of Korea / Q000453:epidemiology; D017422:Sequence Analysis, DNA / Q000662:veterinary; D033761:Spores, Protozoan
chemical_list: D004275:DNA, Ribosomal
keywords: Kudoa iwatai; Myxosporean; Myxozoa; Oplegnathus fasciatus; Rock bream