Quantification of Enterocytozoon hepatopenaei (EHP) in Penaeid Shrimps from Southeast Asia and Latin America Using TaqMan Probe-Based Quantitative PCR

Patharapol Piamsomboon; Seong-Kyoon Choi; Bambang Hanggono; Yani Lestari Nuraini; Fatma Wati; Kathy F J Tang; Song Park; Dongmi Kwak; Man Hee Rhee; Jee Eun Han; Ji Hyung Kim

doi:10.3390/pathogens8040233

Quantification of Enterocytozoon hepatopenaei (EHP) in Penaeid Shrimps from Southeast Asia and Latin America Using TaqMan Probe-Based Quantitative PCR

Pathogens. 2019 Nov 12;8(4):233. doi: 10.3390/pathogens8040233.

Authors

Patharapol Piamsomboon¹, Seong-Kyoon Choi^{2

3}, Bambang Hanggono⁴, Yani Lestari Nuraini⁴, Fatma Wati⁴, Kathy F J Tang⁵, Song Park², Dongmi Kwak⁶, Man Hee Rhee⁶, Jee Eun Han⁶, Ji Hyung Kim⁷

Affiliations

¹ Department of Veterinary Medicine, Faculty of Veterinary Science, Chulalongkorn University, Bangkok 10330, Thailand.
² Core Protein Resources Center, DGIST, Daegu 42988, Korea.
³ Division of Biotechnology, DGIST, Daegu 42988, Korea.
⁴ Fish Health and Environmental Laboratory, Brackishwater Aquaculture Development Center, Situbondo 68351, Indonesia.
⁵ Yellow Sea Fisheries Research Institute, Chinese Academy of Fishery Sciences, Qingdao 266071, China.
⁶ College of Veterinary Medicine, Kyungpook National University, Daegu 41566, Korea.
⁷ Infectious Disease Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 34141, Korea.

Abstract

We developed a qPCR assay based on the β-tubulin gene sequence for the shrimp microsporidian parasite Enterocytozoon hepatopenaei (EHP). This assay reacted with the hepatopancreas (HP) of EHP-infected shrimps, and the highest copy numbers were found in HP and feces samples from Southeast Asian countries (10⁶-10⁸ copies mg^-1), while HP samples from Latin America, Artemia, and EHP-contaminated water showed lower amounts (10¹-10³ copies mg^-1 or mL^-1 of water). No false positive was found with the normal shrimp genome, live feeds, or other parasitic diseases. This tool will facilitate the management of EHP infection in shrimp farms.

Keywords: Enterocytozoon hepatopenaei; microsporidian; qPCR; shrimp parasite.

Abstract

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