조피볼락(Sebastes schlegeli) 선충(Nematode: Philometridae)에 대한 분자생물학적 동정 및 PCR 검출법 개발
Molecular Identification and Development of a PCR Assay for the Detection of a Philometrid Nematode in Rockfish Sebastes schlegeli
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초록
Nematode infection in the epithelial tissue of cultured rockfish Sebastes schlegeli was first reported in 2012. Since then, nematode infections have caused serious economic losses in rockfish aquaculture on the west coast of Korea. Taxonomic and life cycle information for this parasite are currently unknown. In this study, 18S rRNA and cytochrome c oxidase subunit I (COI) genes were used for molecular identification and polymerase chain reaction (PCR) to detect the invisible stages of this parasite. Nucleotide sequences of the 18S rRNA of the rockfish nematode showed 98% identity with that of Philometra morii. Therefore, this rockfish nematode was classified to the Philometridae family. However, we could not identify it to genus level using 18S rRNA. Its COI nucleotide sequences shared 85% and 82% identities with those of Bursaphelenchus sinensis and Philometra overstreeti, respectively. In addition, two gene-specific primer sets were designed based on the 18S rRNA gene to detect the intermediate host and nematode larvae. These primers were specific to this rockfish nematode without cross-reacting to other pathogens. The detection limit of the PCR assay using these primers was 1,000 copies of nematoda plasmid DNA. Therefore, the PCR assay described here is suitable for the detection of nematode DNA within rockfish. In addition, this PCR assay could be used to detect nematode larvae and the intermediate host.
Abstract
Nematode infection in the epithelial tissue of cultured rockfish Sebastes schlegeli was first reported in 2012. Since then, nematode infections have caused serious economic losses in rockfish aquaculture on the west coast of Korea. Taxonomic and life cycle information for this parasite are currently unknown. In this study, 18S rRNA and cytochrome c oxidase subunit I (COI) genes were used for molecular identification and polymerase chain reaction (PCR) to detect the invisible stages of this parasite. Nucleotide sequences of the 18S rRNA of the rockfish nematode showed 98% identity with that of Philometra morii. Therefore, this rockfish nematode was classified to the Philometridae family. However, we could not identify it to genus level using 18S rRNA. Its COI nucleotide sequences shared 85% and 82% identities with those of Bursaphelenchus sinensis and Philometra overstreeti, respectively. In addition, two gene-specific primer sets were designed based on the 18S rRNA gene to detect the intermediate host and nematode larvae. These primers were specific to this rockfish nematode without cross-reacting to other pathogens. The detection limit of the PCR assay using these primers was 1,000 copies of nematoda plasmid DNA. Therefore, the PCR assay described here is suitable for the detection of nematode DNA within rockfish. In addition, this PCR assay could be used to detect nematode larvae and the intermediate host.
참고문헌 (15)
- Edmands S, "Allozyme and mitochondrial DNA evidence of population subdivision in the purple sea urchin Strongylocentrotus purpuratus", Mar Biol, 126, pp.443~450 (1996)
- Edward JN, "Diagnosis and Treatment, Fish Disease", pp.166~170 (1996), Blackwell, Iowa State University Press
- González-Solís D, "A new species of Dentiphilometra (Nematoda: Philometridae)from the musculature of the gray snapper Lutjanus griseus (osteichthyes) off the Caribbean coast of Mexico", J Parasitol, 93, pp.1132~1135 (2007)[DOI]
- 한상현, "미토콘드리아 COI 유전자 서열의 다형성과 반수체형에 근거한 한국산 붉바리(Epinephelus akaara)의 유전적 구조와 계통 유연관계", 생명과학회지, 24(6), pp.626~632 (2014), 한국생명과학회
- Herbert PDN, "Biological identifications through DNA barcodes", Proc Biol Sci, 270, pp.313~321 (2003)[DOI]
- 강주희, "고래회충 검출을 위한 육안검사법과 중합효소연쇄반응-제한효소절편길이다형성의 비교", 한국식품위생안전성학회지, 23(4), pp.314~318 (2008), 한국식품위생안전성학회
- "Korean Statistical Information Service"[URL]
- Moravec F, "Paraphilometroides nemipteri gen. et sp. N. (nematoda : philometridae)from the marine fish nemipterus peronei(valenciennes)from Malaysia", Folia Parasitol, 36, pp.345~350 (1989)
- Moravec F, "Dentiphilometra monopteri n. gen., n. sp. (nematoda: philometridae) from the abdominal cavity of the ricefield eel monopterus albus in china", J Parasitol, 88, pp.961~966 (2002)[DOI]
- Nyaku ST, "Characterization of the two intra-individual sequence variants in the 18S rRNA gene in the plant parasitic nematode, Rotylenchulus reniformis", PLoS ONE, 8, pp.e60891~ (2013)[DOI]
- Quiazon KMA., "Studies on philometrid and anisakid nematodes infecting marine fishes in Japanese waters", 박사, pp.2009
- 서한길, "서해안 천수만 일대 양식 조피볼락(Sebastes schlegeli)의 상피 선충 감염현황", 한국수산과학회지, 47(5), pp.603~610 (2014), 한국수산과학회
- Stephanie P, "Use of molecular tools in identification of philometrid larvae in fishes: technical limitations parallel our poor assessment of their biodiversity", Parasitol Res, 109, pp.1725~1730 (2011)[DOI]
- Wang GT, "Seasonal dynamics and distribution of Philometroides fulvidraconi (Philometridae) in the bullhead catfish, Pseudobagrus fulvidraco (Richardson)", J Fish Dis, 25, pp.621~625 (2002)[DOI]
- Wijová M, "Evolutionary relationships of Spirurina (Nematoda: Chromadorea: Rhabditida) with special emphasis on dracunculoid nematodes inferred from SSU rRNA gene sequences", Int J Parasitol, 36, pp.1067~1075 (2006)[DOI]