한수지 53(5), , 2020 Original Article Korean J Fish Aquat Sci 53(5), , 년서해연안패류및양식새우 (Litopenaeus vannamei) 에서분리한 Vibrio parahaemoly

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1 한수지 53(5), , 2020 Original Article Korean J Fish Aquat Sci 53(5), , 년서해연안패류및양식새우 (Litopenaeus vannamei) 에서분리한 Vibrio parahaemolyticus 균주들과새우급성간췌장괴사병 (AHPND) 과의연관성 장광일 박진일 오은경 * 김수미 1 * 국립수산과학원서해수산연구소, 1 국립수산과학원수산방역과 The Relationship Between Acute Hepatopancreatic Necrosis Disease (AHPND) in Shrimp Litopenaeus vannamei and Vibrio parahaemolyticus Strains Isolated from Shellfish and Shrimp of the West Coast of Korea in 2019 Gwang Il Jang, Jin Il Park, Eun-Gyoung Oh* and Sumi Kim 1 * West Sea Fisheries Research Institute, National Institute of Fisheries Science, Incheon 22383, Korea 1 Aquatic Disease Control Division, National Institute of Fisheries Science, Busan 46083, Korea Acute hepatopancreatic necrosis disease (AHPND), previously known as early mortality syndrome (EMS), is an emerging disease in shrimp caused by Vibrio parahaemolyticus. Some V. parahaemolyticus strains are associated with foodborne diseases in humans. To date, studies on the relationship between AHPND and pathogenic V. parahaemolyticus are very limited. In this study, we monitored the thermostable direct hemolysin-related hemolysin (trh) gene and AHPND related genes, such as Photorhabdus insect-related (pir) genes, in 892 strains of V. parahaemolyticus isolated and identified in 24 areas of the West Coast of Korea from May to October The trh gene was detected in 9.6% of the isolates from short neck clam samples. However, the pira and pirb genes related to AHPND were not found in any of the isolates despite using both duplex and nested PCR assays, suggesting that AHPND-related genes were nonexistent in the V. parahaemolyticus strains isolated. This study contributes to the current understanding of the relationship between AHPND and V. parahaemolyticus in Korea, as well as provides data on spatial and seasonal distributions of V. parahaemolyticus. Keywords: Acute hepatopancreatic necrosis disease (AHPND), Vibrio parahaemolyticus, Shrimp, West Coast, Thermostable direct hemolysin-related hemolysin (trh) gene 서론 Vibrio (genus),, 140 (Parte, 2018; Urakawa et al., 2000). (Kongrueng et al., 2015) 12 Vibrio, Vibrio parahaemolyticus, Vibrio vulnificus, Vibrio cholera, Vibrio mimicus, Vibrio hollisae, Vibrio alginolyticus, Vibrio cincinnatiensis, Vibrio damsela, Vibrio fluvialis, Vibrio furnissii, Vibrio harveyi, Vibrio metschnikovii (Farmer et al., 2015; CDC, 2019). (V. cholera), (V. parahaemolyticus), (V. vulnificus) (Thompson et al., 2006)., (V. para- *Corresponding author: Tel: Fax: address: ohdagu@korea.kr; sumikim@korea.kr This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Licens ( which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Korean J Fish Aquat Sci 53(5), , October 2020 Received 9 July 2020; Revised 19 August 2020; Accepted 10 September 2020 저자직위 : 장광일 ( 연구사 ), 박진일 ( 연구사 ), 오은경 ( 연구관 ), 김수미 ( 주무관 ) Copyright 2020 The Korean Society of Fisheries and Aquatic Science 752 pissn: , eissn:

Vibrio parahaemolyticus 균주와새우급성간췌장괴사병 (AHPND) 과의연관성 753 haemolyticus) (Kang et al., 2016; Yang et al., 2017; Park et al., 2018) (thermostable direct hemolysin, TDH) (TDH-related hemolysin, TRH)., V.

2 Vibrio parahaemolyticus 균주와새우급성간췌장괴사병 (AHPND) 과의연관성 753 haemolyticus) (Kang et al., 2016; Yang et al., 2017; Park et al., 2018) (thermostable direct hemolysin, TDH) (TDH-related hemolysin, TRH)., V. parahaemolyticus,, (Sudheeshand and Xu, 2001; Marudhupandi et al., 2017). (acute hepatopancreatic necrosis disease, AHPND) V. parahaemolyticus (Lightner et al., 2012; Zhang et al., 2012) AHPND. V. parahaemolyticus AHPND, AHPND. (Photorhabdus insect-related, Pir) plasmid AHPND (Joshi et al., 2014; Han et al., 2015; Kim and Kim, 2015). V. parahaemolyticus. AHPND 2009 (2010 ), (2011 ), (2012 ), (2013 ), 2017 (Zorriehzahra and Banaederakhshan, 2015; Dhar et al., 2019) Tran et al. (2013) AHPND V. parahaemolyticus, V. parahaemolyticus AHPND V. parahaemolyticus type IV pilus/type IV plasmid (pir toxin) (Kondo et al., 2014; Yang et al., 2014)., Han et al. (2015) AHPND V. parahaemolyticus plasmid (pira/pirb) AHPND (Hwang et al., 2018). V. parahaemolyticus AHPND (Tran et al., 2013; Kondo et al., 2014; Han et al., 2015; Lee et al., 2015), AHPND V. parahaemolyticus V. parahaemolyticus (thermostable direct hemolysin, TDH) (TDH-related hemolysin, TRH) (tdh, trh) (V. parahaemolyticus) (AHPND). 재료및방법 시료채취 V. parahaemolyticus 18, (Muuido), (Yeongheungdo), (Soijakdo), (Seungbongdo), (Cheonsu Bay), (Wonsando), (Iwon-myeon), (Garorim Bay), (Sowon-myeon), (Byeonsan), (Gochang-gun), (Dolipo), (Aphaedo), (Jangsando), (Hampyeong Bay), (Tando Bay), (Maehwado), (Chilsando) 4, (Ganghwa-gun), (Pyeongtaek-si), 13, , Incheon 3 Gyeonggi-do Chungcheongnam-do Jeollabuk-do Jeollanam-do Fig. 1. Sampling sites of shellfish and shrimp Litopenaeus vannamei on the West Coast of Korea during the period May to October in The description of sampling sites is the same as in Table 2.

3 754 장광일ㆍ박진일ㆍ오은경ㆍ김수미 (Taean-gun) (Boryeong-si), (Oyster Crassostrea gigas), (Surf clam Mactra veneriformis), (Short neck clam Venerupis philippinarum), (Ocean quahog Arctica islandica), (Conch Lobatus gigas), (Abalone Haliotis discus hannai) (Shrimp Litopenaeus vannamei) (pond water) (Fig. 1; Table 1)., 10 C. V. parahaemolyticus 균주분리및동정 V. parahaemolyticus (food and drug administration, FDA) bacteriological analytical manual (BAM) (Elliot et al., 1995). ( ) phosphate buffered saline (PBS: 140 mm NaCl, 5 mm anhydrous Na 2 HPO 4, 1.5 mm KH 2 PO 4, ph 7.4), alkaline peptone water (APW; peptone 10 g/l, NaCl 20 g/l, ph ) C (Feng et al., 2017; Yang et al., 2019)., CHROMagar (Paris, France) 35 C 24. CHROMagar V. parahaemolyticus (colony). V. parahaemolyticus, 42 C, ONPG (O-nitrophenyl- -D-galactopyranoside), VITEK system (BioMérieux Vitek, Marcy-l'Étoile, France) V. parahaemolyticus (Park et al., 2019; Song et al., 2020). DNA 추출및병원성관련유전자탐색 tdh trh PCR (polymerase chain reaction). tdh trh VPD- 1/VPD-2 VPR-1/VPR-2 primer sets (Takara, Bio Inc., Otsu, Japan). Tryptic Soy Broth (Difco, Detroit, MU, USA) 10 ml 37 C 18-24, DNA Isolation kit (MO- BIO, Carlsbad, CA, USA) DNA. tdh trh PCR Master mix (Takara Bio Inc., Otsu, Japan) 12.5 μl, primer (10 pmol/μl) 0.2 μl, DNA 2 μl, 25 μl. thermal cycle (Takara Bio Inc., Otsu, Japan) 98 C 10, 55 C 30, 72 C PCR. PCR gel documentation system G BOX (Syngene, Cambridge, UK) amplicon (tdh, 251 bp; trh, 250 bp). (AHPND) (world organization for animal health, OIE) (duplex PCR AP4 nested PCR) (Dangtip et al., 2015; Han et al., 2015). V. parahaemolyticus plasmid Pir toxin AccuPower HotStart PCR premix kit (Bioneer, Korea) VpPirA-284F/R VpPirB-392F/R primer sets duplex PCR (Table 1; Han et al., 2015). Dangtip et al. (2015) AP4-F1/AP4-R1 primers PCR, PCR AP4-F2/AP4-R2 primers AP4 nested PCR (Table 1). PCR QIAxcel Advanced (Qiagen, Hilden, Germany). 결과및고찰 패류및새우양식장의 V. parahaemolyticus 분포 V. parahaemolyticus,, V. parahaemolyticus Table 2. Table 1. PCR primers and sequences of Photorhabdus insect-related (pir) genes used in this study Genes Primers Product (bp) References Photorhabdus insect-related (pir) gene VpPirA-284F: 5'-TGA-CTA-TTC-TCA-CGA-TTG-GAC-TG-3' VpPirA-284R: 5'-CAC-GAC-TAG-CGC-CAT-TGT-TA-3' VpPirB-392F: 5'-TGA-TGA-AGT-GAT-GGG-TGC-TC-3' VpPirB-392R: 5'-TGT-AAG-CGC-CGT-TTA-ACT-CA-3' AP4-F1: 5'-ATG-AGT-AAC-AAT-ATA-AAA-CAT-GAA-AC-3' AP4-R1: 5'-ACG-ATT-TCG-ACG-TTC-CCC-AA-3' AP4-F2: 5'-TTG-AGA-ATA-CGG-GAC-GTG-GG-3' AP4-F2: 5'- GTT-AGT-CAT-GTG-AGC-ACC-TTC-3' Han et al., 2015 Dangtip et al., 2015

4 Vibrio parahaemolyticus 균주와새우급성간췌장괴사병 (AHPND) 과의연관성 755 V. parahaemolyticus (MPN/100 g) 1.E+05 1.E+04 1.E+03 1.E+02 1.E+01 1.E+00 Site 13 Site 14 Site 19 Site 20 May June July August September October November Fig. 2. Monthly variation of V. parahaemolyticus isolated from Oyster Crassostrea gigas farm on the West Coast of Korea. The description of sampling sites is the same as in Table , 20, 204, 4, 125, 147, , , , , , (Table 2). 5 10, V. parahaemolyticus 8 10 MPN (most probable number) (Fig. 2). Park et al. (2019) V. parahaemolyticus %. Lee et al. (2019) 3 (,, ) V. parahaemolyticus, 8 [ colony form unit (CFU)/g], 10 ( CFU/g), 14.6 C V. parahaemolyticus (Lee et al., 2008; Yu et al., 2014; Kim et al., 2016), 5 10 V. parahaemolyticus V. parahaemolyticus 23.1% 26.7% (Table2). (26- M NC PC Fig. 3. Representative results of PCR of the AHPND related genes (pira/pirb) using the duplex PCR for V. parahaemolyticus isolated from shellfish and shrimp Litopenaeus vannamei on the West Coast of Korea. M, size marker; 1, Shrimp (1906 DS-W1); 2, Pond water (1909 DS-W13); 3, Oyster (JN 16-11); 4, Short neck clam (IC 2-1); 5, Conch (CN 4-13); 6, Abalone (CN 3-19); 7, Surf clam (JB 1-7); 8, Ocean quahog (JN 19-20); NC, negative control; PC, positive control. 28 C) (10-15 psu) V. parahaemolyticus, (20-30 C) (4-80 psu) V. parahaemolyticus (Farmer et al., 2015; NIFS, 2018). 병원성관련유전자검출 내열성용혈독소 (tdh) 및내열성용혈독소유사독소 (trh) 유전자검출 V. parahaemolyticus 892

5 756 장광일ㆍ박진일ㆍ오은경ㆍ김수미 (tdh) (trh) tdh, trh 10 5 (9.6%) (Table 3) V. parahaemolyticus tdh trh, tdh %, trh % tdh, V. parahaemolyticus 37 tdh trh (Park et al., 2016). Kang et al. (2017) toxr V. parahaemolyticus 31 tdh trh 4 (9.1%) trh, tdh. V. parahaemolyticus. V. parahaemolyticus (Thompson et al., 1976; Honda and Iida,., 1993; Yamamoto et al., 2008; Malcolm et al., 2015). Table 2. Number of isolates from shellfish and shrimp Litopenaeus vannamei on the West Coast of Korea during the period May to October in 2019 Sites Sampling sites Samples May June July August September October 1 Ganghwa-gun, Incheon #1 - - (2*) 4 (4*) 9 (1*) 5 (21*) 8 2 Ganghwa-gun, Incheon #2-8 (6*) 10 (6*) 9 (4*) 19 (17*) 8 (5*) 3 Pyeongtaek-si, Gyeonggi-do Shrimp Litopenaeus vannamei (Pond water*) (10*) 3 (20*) 12 (19*) 6 (14*) 4 Taean-gun, Chungcheongnam-do (3*) - (7*) Boryeong-si, Chungcheongnam-do - 4 (8*) Jung-gu, Incheon (Muuido) 7 Seocheon-gun, Chungcheongnam-do (Cheonsu Bay) Boryeong-si, Chungcheongnam-do (Wonsando) Short neck clam Buan-gun, Jeollabuk-do (Byeonsan) Venerupis philippinarum Muan-gun, Jeollanam-do (Dolipo) Sinan-gun, Jeollanam-do (Aphaedo) Sinan-gun, Jeollanam-do (Jangsando) Taean-gun, Chungcheongnam-do (Iwon-myeon#1) Taean-gun, Chungcheongnam-do (Iwon-myeon #4) Hampyeong-gun, Jeollanam-do (Hampyeong Bay) Oyster Crassostrea gigas Muan-gun, Jeollanam-do (Tando Bay) Sinan-gun, Jeollanam-do (Maehwado) Ongjin-gun, Incheon (Yeongheungdo) Short neck clam (4 ) 19 Ongjin-gun, Incheon (Soijakdo) Venerupis philippinarum - 8 (1 ) - (21 ) 14 (17 ) 20 (36 ) 31 (19 ) 20 Ongjin-gun, Incheon (Seungbongdo) (Oyster Crassostrea gigas ) (5 ) 12 (29 ) 12 (9 ) 7 (11 ) Taean-gun, Chungcheongnam-do (Garorim Bay) Taean-gun, Chungcheongnam-do (Sowon-myeon) Conch Lobatus gigas Abalone Haliotis discus hannai Gochang-gun, Jeollabuk-do Surf clam Mactra veneriformis Yeonggwang-gun, Jeollanam-do 24 Ocean quahog Arctica islandica (Chilsando) * and in parentheses were isolated from pond water and Oyster, respectively. -, no data.

Vibrio parahaemolyticus 균주와새우급성간췌장괴사병 (AHPND) 과의연관성 757 (a) M 1 2 3 4 5 6 7 8 NC PC (b) M 1 2 3 4 5 6 7 8 NC PC Fig. 4. Representative results of PCR of the AHPND related genes (pira/pirb) using the (a) first PCR and (b) nested PCR for V.

M, size marker; 1, Shrimp (1906 DS-W1); 2, Pond water (1909 DS-W13); 3, Oyster (JN 16-11); 4, Short neck clam (IC 2-1); 5, Conch (CN 4-13); 6, Abalone (CN 3-19); 7, Surf clam (JB 1-7); 8, Ocean

6 Vibrio parahaemolyticus 균주와새우급성간췌장괴사병 (AHPND) 과의연관성 757 (a) M NC PC (b) M NC PC Fig. 4. Representative results of PCR of the AHPND related genes (pira/pirb) using the (a) first PCR and (b) nested PCR for V. parahaemolyticus isolated from shellfish and shrimp on the West Coast of Korea. M, size marker; 1, Shrimp (1906 DS-W1); 2, Pond water (1909 DS-W13); 3, Oyster (JN 16-11); 4, Short neck clam (IC 2-1); 5, Conch (CN 4-13); 6, Abalone (CN 3-19); 7, Surf clam (JB 1-7); 8, Ocean quahog (JN 19-20); NC, negative control; PC, positive control. AHPND 관련유전자검출 V. parahaemolyticus 892 duplex PCR nested PCR AHPND pira pirb (Fig. 3 and Fig. 4, Table 3) AHPND, AHPND AHPND V. parahaemolyticus, 2013 V. parahaemolyticus AHPND (Zhang et al., 2012; Tran et al., 2013). Han et al. (2015) Vibrio Photobacterium 77 AHPND pira/pirb AHPND V. parahaemolyticus pira/pirb., AHPND V. parahaemolyticus pira/pirb. AHPND V. parahaemolyticus pira/pirb. Han et al. (2015) AHPND V. parahaemolyticus 2 1 AHPND. AHPND V. parahaemolyticus 69 kb plasmid (Pir toxins). V. parahaemolyticus AHPND / biofilm (transposition), (conjugation) plasmid pira/pirb Table 3. Detection rates (%) of thermostable direct hemolysin-related hemolysin (trh) gene and AHPND related genes (pira/pirb) in V. parahaemolyticus isolated from shellfish and shrimp Litopenaeus vannamei on the West Coast of Korea during the period May to October in 2019 Areas Number of isolates trh gene Detection rates (%) Detection rates(%) Source of pira/pirb genes Incheon 428 0% - 0% Gyeonggi-do 94 0% - 0% Chungcheongnam-do 284 0% - 0% Jeollabuk-do 34 0% - 0% Jeollanam-do % Short neck clam (October) 0%

7 758 장광일ㆍ박진일ㆍ오은경ㆍ김수미 V. parahaemolyticus. AHPND AHPND V. parahaemolyticus V. parahaemolyticus (Kondo et al., 2014; Yang et al., 2014; Lee et al., 2015; Fu et al., 2017). Fu et al. (2017) 3 V. parahaemolyticus 9 Genbank V. parahaemolyticus 93 V. parahaemolyticus. Fu et al. (2017) V. parahaemolyticus 9 2 pira/pirb AHPND, Genbank V. parahaemolyticus 16 pira/pirb. V. parahaemolyticus FDA_R31 (single nucleotide polymorphisms, SNPs) 35,001 35,889 SNPs. V. parahaemolyticus. V. parahaemolyticus 102. V. parahaemolyticus AHPND. AHPND V. parahaemolyticus V. parahaemolyticus AHPND, V. parahaemolytic AHPND PirA/ PirB. V. parahaemolyticus V. parahaemolyticus AHPND V. parahaemolyticus. AHPND.,. AHPND. 사사 (R ) (R ). References CDC (Centers for Disease Control and Prevention) Vibrio species causing vibriosis. Retrieved from cdc.gov/vibrio/faq.html on May 1, Dangtip S, Sirikharin R, Sanguanrut P, Thitamadee S, Sritunyalucksana K, Taengchaiyaphum S, Mavichak R, Proespraiwong P and Flegel TW AP4 method for two-tube nested PCR detection of AHPND isolates of Vibrio parahaemolyticus. Aquaculture Rep 2, org/ /j.aqrep Dhar AK, Piamsomboon P, Caro LFA, Kanrar S, Adami R Jr and Juan YS First report of acute hepatopancreatic necrosis disease (AHPND) occurring in the USA. Dis Aquat Org 132, Elliot EL, Kaysner CA, Jackson L and Tamplin ML Vibrio cholera, V. parahaemolyticus, V. vulnificus and other Vibrio spp. In: Bacteriological analytical manual. Association of Offical Analytical Chemists ed. FDA, Arlington, U.S.A., Farmer JJ III, Brenner FW, Cameron DN, Birkhead KM and Janda JM Genus Vibrio Pacini 1854, 411 AL In: Bergey s manual of systematics of archaea and bacteria. John Wiley Sons, Hoboken, NJ, U.S.A. org/ / gbm Feng B, Liu H, Wang M, Sun X, Pan Y and Zhao Y Diversity analysis of acute hepatopancreatic necrosis diseasepositive Vibrio parahaemolyticus strains. Aquacul Fish 2, Fu S, Tian H, Wei D, Zhang X and Liu Y Delineating the origins of Vibrio parahaemolyticus isolated from outbreaks of acute hepatopancreatic necrosis disease in asia by the use of whole genome sequencing. Front Microbiol 28, Han JE, Tang KFJ, Tran LH and Lightner DV Photorhabdus insect-related (Pir) toxinlike genes in a plasmid of Vibrio parahaemolyticus, the causative agent of acute hepatopancreatic necrosis disease (AHPND) of shrimp. Dis Aquat Organ 113, Honda T and Iida T The pathogenicity of Vibrio parahaemolyticus and the role of the thermostable direct haemolysin and related haemolysins. Rev Med Microbiol 4, Hwang SD, Hwang JY, Kang BC, Jeon HR, Chun WJ, Lee DW, Sohn SB, Kim SM, Kim SR, Lee KY, Jung MH, Seo JS, Kwon M and Jee BY Whole genome sequence of acute hepatopancreatic necrosis (AHPND) isolated from Korea. In: Proceedings of the Korean Federation of Fisheries Science and Technology Societies (KOFFST) International Conference 2018, Busan, Korea, 492.

8 Vibrio parahaemolyticus 균주와새우급성간췌장괴사병 (AHPND) 과의연관성 759 Joshi J, Srisala J, Truong VH, Chen IT, Nuangsaeng B, Suthienkul O, Lo CF, Flegel TW, Sritunyalucksana K and Thitamadee S Variation in Vibrio parahaemolyticus isolates from a single Thai shrimp farm experiencing an outbreak of acute hepatopancreatic necrosis disease (AHPND). Aquaculture , aquaculture Kang CH, Shin Y, Jang S, Yu H, Kim S, An S, Park K and So JS Characterization of Vibrio parahaemolyticus isolated from oysters in Korea: Resistance to various antibiotics and prevalence of virulence genes. Mar Pollut Bull 118, Kang CH, Shin YJ, Kim WR, Kim YG, Song KC, Oh EG, Kim SK, Yu HS and So JS Prevalence and antimicrobial susceptibility of Vibrio parahaemolyticus isolated from oysters in Korea. Environ Sci Pollut Res 23, doi.org/ /s Kim NE and Kim DH Acute hepatopancreatic necrosis of shrimp and import health measures. J fish Pathol 28, Kim SK, An SR, Park BM, Oh EG, Song KC, Kim JW and Yu HS Virulence factors and antimicrobial susceptibility of Vibrio parahaemolyticus isolated from oyster Crassostrea gigas. Korean J Fish Aquat Sci 49, Kondo H, Tinwongger S, Proespraiwong P, Mavichak R, Unajak S, Nozaki R and Hirono I Draft genome sequences of six strains of Vibrio parahaemolyticus isolated from early mortality syndrome/acute hepatopancreatic necrosis disease shrimp in Thailand. Genome Announc 2, e doi.org/ /genomea Kongrueng J, Yingkajorn M, Bunpa S, Sermwitaya-wong N, Singkhamanan K and Vudhakul V Characterization of Vibrio parahaemolyticus causing acute hepatopancreatic necrosis disease in southern Thailand. J Fish Dis 38, Lee CT, Chen IT, Yang YT, Ko TP, Huang YT, Huang JY, Huang MF, Lin SJ, Chen CY, Lin SS, Lightner DV, Wang HC, Wang AH, Wang HC, Hor LI and Lo CF The opportunistic marine pathogen Vibrio parahaemolyticus becomes virulent by acquiring a plasmid that expresses a deadly toxin. Proc Natl Acad Sci U S A 112, org/ /pnas Lee DS, Shin HY, Park KS and Shin IS Distribution of Vibrio parahaemolyticus and Vibrio vulnificus in seawater and shellfish at Gomso Bay. Korean J Fish Aquat Sci 52, Lee JK, Jung DW, Eom SY, Oh SW, Kim Y, Kwak HS and Kim YH Occurrence of Vibrio parahaemolyticus in oysters from Korean retail outlets. Food Control 19, Lightner, DV, Redman, RM, Pantoja, CR, Noble, BL and Tran L Early mortality syndrome affects shrimp in Asia. Retrieved from GAA-Lightner-Jan12.pdf on May 1, Malcolm TTH, Cheah YK, Radzi CWJWM, Kasim FA, Kantilal HK, John TYH, Martinez-Urtazaf J, Nakaguchig Y, Nishibuchig M and Sona R Detection and quantification of pathogenic Vibrio parahaemolyticus in shellfish by using multiplex PCR and loop-mediated isothermal amplification assay. Food Control 47, foodcont Marudhupandi T, Kumar TTA, Prakash S, Balamurugan J, Dhayanithia NB Vibrio parahaemolyticus a causative bacterium for tail rot disease in ornamental fish, Amphiprion sebae. Aquaculture Reports 8, org/ /j.aqrep NIFS (National Institute of Fisheries Science) Standard manual of whiteleg shrimp farming. NIFS, Busan, Korea. Park K, Mok JS, Kwon JY, Ryu AR and Shim KB Seasonal and Spatial Variation of Pathogenic Vibrio Species Isolated from Seawater and Shellfish off the Gyeongnam Coast of Korea in Korean J Fish Aquat Sci 52, Park K, Mok JS, Ryu AR, Kwon JY, Ham IT and Shim KB Occurrence and virulence of Vibrio parahaemolyticus isolated from seawater and bivalve shellfish of the Gyeongnam coast, Korea, in Mar Poll Bull 137, Park YS, Park K, Kwon JY, Yu HS, Lee HJ, Kim JH, Lee TS and Kim PH Antimicrobial reststance and distribution of virulence factors of Vibrio parahaemolyticus isolated from shellfish farms on the southern coast of Korea. Korean J Fish Aquat Sci 49, KFAS Parte AC LPSN-list of prokaryotic names with standing in nomenclature (bacterio.net), 20 years on. Int J Syst Evol Microbiol 68, ijsem Song X, Zang J, Yu W, Shi X and Wu Y Occurrence and Identification of Pathogenic Vibrio Contaminants in Common Seafood Available in a Chinese Traditional Market in Qingdao, Shandong Province. Front Microbiol 11, Sudheesh PS and Xu HS Pathogenicity of Vibrio parahaemolyticus in tiger prawn Penaeus monodon Fabricius: possible role of extracellular proteases. Aquaculture 196, Thompson CA, Vanderzant C and Ray SM Serological and hemolytic characteristics of Vibrio parahaemolyticus from marine sources. J Food Sci 41, Thompson FL, Austin B and Swings J The biology of vibrios. ASM Press copyright, Washington DC, U.S.A.,

9 760 장광일ㆍ박진일ㆍ오은경ㆍ김수미 Tran L, Nunan L, Redman RM, Mohney LL, Pantoja CR, Fitzsimmons K and Lightner DV Determination of the infectious nature of the agent of acute hepatopancreatic necrosis syndrome affecting penaeid shrimp. Dis Aquat Organ 105, Urakawa H, Yoshida T, Nishimura M and Ohwada K Characterization of depth-related population variation in microbial communities of a coastal marine sediment using 16S rdna-based approaches and quinone profiling. Environ Microbiol 2, Yamamoto A, Iwahori J, Vuddhakul V, Charernjiratrag W, Vose D, Osaka K, Shigematsu M, Toyoufuku H, Yamamoto S, Nishibuchi M and Kasuga F Quantitative modelling for risk assessment of Vibrio parahaemolyticus in bloody clams in southern Thailand. Int J Food Microbiol 124, Yang JH, Mok JS, Jung YJ, Lee KJ, Kwon JY, Park K, Moon SY, Kwon SJ, Ryu AR and Lee TS Distribution and antimicrobial susceptibility of Vibrio species associated with zooplankton in coastal area of Korea. Mar Poll Bull 125, Yang Q, Dong X, Xie G, Fu S, Zou P, Sun J, Wang Y and Huang J Comparative genomic analysis unravels the transmission pattern and intra-species divergence of acute hepatopancreatic necrosis disease (AHPND)-causing Vibrio parahaemolyticus strains. Mol Genet Genomics 294, Yang YT, Chen IT, Lee CT, Chen CY, Lin SS, Hor LI, Tseng TC, Huang YT, Sritunyalucksana K, Thitamadee S, Wang HC and Lo CF Draft genome sequences of four strains of Vibrio parahaemolyticus, three of which cause early mortality syndrome/acute hepatopancreatic necrosis disease in shrimp in China and Thailand. Genome Announc 2, e Yu HS, Oh EG, Shin SB, Park YS, Lee HJ, Kim JH and Song KC Distribution and antimicrobial resistance of Vibrio parahaemolyticus isolated from Korean shellfish. Korean J Fish Aquat Sci 47, KFAS Zhang BC, Liu F, Bian HH, Liu J, Pan LQ and Huang J Isolation, identification, and pathogenicity analysis of a Vibrio parahaemolyticus strain from Litopenaeus vannamei. Prog Fish Sci 33, pone Zorriehzahra MJ and Banaederakhshan R Early mortality syndrome (EMS) as new emerging threat in shrimp industry. Adv Anim Vet Sci 3,

Technical/Specs Sequencing 서비스안내 Standard Sequencing Plasmid/PCR product 를 primer 를이용하여고객이원하는 region 을분석하는서비스이며, 대부분의 normal 샘플에대해 최적화되어있습니다. Full Len

Technical/Specs Sequencing 서비스안내 Standard Sequencing Plasmid/PCR product 를 primer 를이용하여고객이원하는 region 을분석하는서비스이며, 대부분의 normal 샘플에대해 최적화되어있습니다. Full Length Sequencing (Primer walking) 길이가긴 insert 를포함한 plasmid