534 강경태ㆍ김민주ㆍ박선영ㆍ최종덕ㆍ허민수ㆍ김진수., Choi et al. (1998), Jeong et al. (2004), Ha et al. (2005), Lee (2006), Baker (2016), Vugia et al. (2013) 2003.,,,,. HACC

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1 한수지 49(5), , 2016 Original Article Korean J Fish Aquat Sci 49(5), ,2016 HACCP 구축을위한굴 (Crassostrea gigas) 가공공장의위해평가 강경태 김민주 1 박선영 1 최종덕 1 허민수 2 김진수 1 * 한국식품안전관리인증원, 1 경상대학교해양식품생명의학과 / 해양산업연구소, 2 경상대학교식품영양학과 Risk Assessment of Oyster Crassostrea gigas Processing Site For an HACCP System Model Kyung Tae Kang, Min Joo Kim 1, Sun Young Park 1, Jong-Duck Choi 1, Min Soo Heu 2 and Jin-Soo Kim 1 * Korea Institute for Food Safety Management Accreditation, Daejeon 34917, Korea 1 Department of Seafood and Aquaculture Science, Gyeongsang National University, Tongyeong 53064, Korea 2 Department of Food Science and Nutrition, Gyeongsang National University, Jinju 52828, Korea This study assessed the risk of an oyster-shucking site to establish the hazard analysis critical control point (HACCP) system model by measuring viable cell counts, coliform group Staphylococcus aureus foreign material on oysters, oyster-producing equipment, and washing water. The viable cell count and coliform group levels of the harvested raw oysters were 4.00 log CFU/g and MPN/100 g, while those of washed oysters were 2.99 log CFU/g and ( ) 10 MPN/100 g, respectively. After washing the oysters, no Escherichia coli or pathogenic bacteria (E. coli O157:H7, Listeria monocytogenes, S. aureus, Salmonella spp., Vibrio parahaemolyticus, and Clostridium perfringens) were detected. Regardless of the location of foreign matter, up to 100% more metallic and non-metallic foreign matter was detected at 1.5 mmφ than at 3.5 mmφ, using a metal detector with increased sensitivity. According to the results, the critical control points (CCP) are the washing and metal-detection processes. These results can be used as basic data to improve sanitation at oyster-shucking sites in factories with an HACCP system. Key words: Oyster, HACCP, Risk assessment, Critical control point, Critical limit 서론 30, 60% (Ministry of Oceans and Fisheries, 2016).,,,,,, (Linehan et al., 1999; Hosoi et al., 2003; Kim et al., 1981; Sakaguchi and Murat, 1989). 11 4, 5, (Kim et al., 2002)., (Park et al., 2006).,., (Lee et al., 2010; Oh et al., 2012; Iritani et al., 2014)., FDA EU. HACCP (hazard analysis and critical control point) system Korean J Fish Aquat Sci 49(5) , October 2016 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. Received 28 July 2016; Accepted 3 October 2016 *Corresponding author: Tel: Fax: address: jinsukim@gnu.ac.kr Copyright 2016 The Korean Society of Fisheries and Aquatic Science 533 pissn: , eissn:

2 534 강경태ㆍ김민주ㆍ박선영ㆍ최종덕ㆍ허민수ㆍ김진수., Choi et al. (1998), Jeong et al. (2004), Ha et al. (2005), Lee (2006), Baker (2016), Vugia et al. (2013) 2003.,,,,. HACCP. 재료 재료및방법 D (Fig. 1) cm 2 swab (APHA, 2001). ( ) ( 1 L), (121, 15 min) ( 300 g). (4 ) 24. 생식용탈각굴의제조공정과이의코드 (A) (B), (C) (D), (E), (F), (G), (H), (I).. Fig. 1. ph ph 5 g, 9 (v/w) ph meter (691, Metrohm, Switzerland). Process Fig. 1. Manufacturing process and sample code of oyster Crassostrea gigas shucked for eating raw. 일반세균수, 대장균군및대장균,.,,., ((Ministry of Food and Drug Safety, 2016a). (25 g), 0.85% 225 ml stomacher (Bag Mizer 400VW, Interscience, France) 30. PCA (plate count agar), (35 1, 48 ),. 5, (lactose broth), brilliant green lactose bile (2%) broth (35 1, ), (most probable number, MPN)/100 g. EC broth (44.5 1, 24 ) durham tube /100 g. 식중독균 Post-harvest oyster Washing Cold storing Code A B C Weighing/Inner packing Cooling Washing Shucking G F E D Metal detecting Outer packing Oyster shucked for eating raw H I J E. coli O157:H7, Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Vibrio parahaemolyticus, Clostridium perfringens 6. (Ministry of Food and Drug Safety, 2015),. E. coli O-157:H7 (25 g) modified EC broth (mec broth; Difco, USA) 225 ml, (35-

3 굴가공공장의위해평가 , 24 ) MacConkey sorbitol (cefixime, potassium tellurite ) ( ). sorbitol EMB ( ). Listeria monocytogenes (25 g) Listeria enrichment broth (Difco, USA) 225 ml Oxford agar (Difco, USA), (30 24 ). PALCAM agar (Difco, USA) (30 24 ), API listeria kit (Biomerieux, France). Staphylococcus aureus (25 g) 10% NaCl tryptic soy broth (TSB; Difco, USA) 225 ml ( ) Baird-Parker agar (Difco, USA), ( )., ( ),. Salmonella spp. (25 g) tryptic soy broth (TSB; Difco, USA) 225 ml (37 24 ) 0.1 ml Rappaport-Vassiliadis (Difco, USA) 10 ml 2 (42 24 ). 2 MacConkey agar (Difco, USA) (35 24 ) (nutrient agar, Difco, USA) (35 24 ) API 20E (Biomerieux, France). Vibrio parahaemolyticus 2% NaCl (Alkaline pepton water, ph 8.6) TCBS TSI API 20E (Biomerieux, France). Clostridium perfringens 1 ml cooked meat (Difco, USA) TSC (Difco, USA) TSC. 노로바이러스오염도측정 (Ministry of Food and Drug Safety, 2015). Log CFU/g Viable cell count Coliform group Fig. 2. Change of viable cell counts and coliform group during manufacture of oyster Crassostrea gigas shucked for eating raw. 1 Process codes (A-J) are the same as shown in Fig. 1. 수세및금속검출한계기준설정 (SAMOO, Korea) (60-90 s), 20 kg (20-40 L/ min 10 L/min ), (E. coli O-157:H7, Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Vibrio parahaemolyticus Clostridium perfringens). (QTM-WA25hh-M210, QUALITEC, Korea),, (90-110%, 10% ), (Fe STS), (Fe mm, STS mm ) (, ). 통계처리 44 A B C D E F G H I J SAS system (Cary, NC, USA) (ANOVA test), Duncan (P<0.05). 결과및고찰 생식용탈각굴의선도및생산원 부자재에대한생물학적위해요소분석 ph 4.00 log CFU/g (Fig. 2) 6.49 (Fig. 3) CFU/g, CFU/g, CFU/g (Kim et al., 2007)., ph 6.4, , 5.8, 5.2 (Kim 45 Process codes MPN/100 g

4 536 강경태ㆍ김민주ㆍ박선영ㆍ최종덕ㆍ허민수ㆍ김진수 ph a2 6.51a 6.45 ab 6.40 bc 6.40 bc 6.39 bc 6.37 bc 6.34 cd 6.33 cd 6.28 d A B C D E F G H I J Process codes 1 Fig. 3. Change of ph during manufacture of oyster Crassostrea gigas shucked for eating raw. 1 Process codes (A-J) are the same as shown in Fig Different letters on the data indicate a significant difference at P<0.05. et al., 2007). ph., [, (E. coli O157:H7, Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Vibrio parahaemolyticus, Clostridium perfringens )] Table log CFU/g MPN/100 g,. Ministry of Food and Drug Safety (2016b) 230 MPN/100 g,. Table 1. Biological hazard analysis for manufacturing oyster Crassostrea gigas shucked for eating raw in polyethylene bag Biological hazard Viable cells Unit Postharvest water for Drinking Polyethylene bag oyster processing ND ND E. coli O157:H7 ND Listeria monocytogenes ND - - Staphylococcus ND - - aureus CFU/g Salmonella spp. ND - - Vibrio parahaemolyticus Clostridium perfringens ND - - ND - - Norovirus ND - - MPN/100 g E. coli or ml 2 ND ND 1 ND: Not detected. 2 -: Not determined.., (Ministry of Environment, 2015) 100 CFU/mL /100 ml.,. 제조공정별생물학적위해요소및선도의변화 Fig. 2,. (A), (B), (C), (D), (E), (F), (G), (H), (I) (J). (A) 4.00 log CFU/g, (B) 3.74 log CFU/g, (C) 3.36 log CFU/g, (D) 3.48 log CFU/g, (E) (F) 2.99 log CFU/g 3.01 log CFU/g., cold shock (Kim, 2000),,.,, 1. (G) 3.10 log CFU/g, (H) 2.98 log CFU/g, (I) 2.97 log CFU/g, 2.94 log CFU/g, log CFU/g.,,,,.

5 굴가공공장의위해평가 537 Table 2. Microbiological evaluation in working place for manufacturing oyster Crassostrea gigas shucked for eating raw (log CFU/100 cm 2 ) Workings room Equipment Viable cell count Coliform group Staphylococcus aureus Worktable 3.40 ND 1 ND Shucking room Basket 3.26 ND ND Knife 3.00 ND ND Washing room Washing machine 2.51 ND ND Worktable 3.00 ND ND Packing room Packing machine 1.18 ND ND Worktable 1.65 ND ND 1 ND: Not detected. (A) MPN/100 g, (B) MPN/100 g, (C) MPN/100 g, (D) MPN/100 g, (E) (F) MPN/100 g MPN/100 g.,,. (G) MPN/100 g, (H) MPN/100 g, (I) MPN/100 g, MPN/100 g, ( ) 10 MPN/100 g.,,,,.,. (, E. coli O157:H7, Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Vibrio parahaemolyticus Clostridium perfringens) ( ).,,. ph Fig. 3. ph (P<0.05),., ph,. ph,, TCA cycle lactic acid (David and Michael, 2014). 작업환경위생.,,, 100 cm 2, Table log CFU/100 cm 2, 3.26 log CFU/100 cm 2, 3.00 log CFU/100 cm 2, 2.51 log CFU/100 cm 2, 3.00 log CFU/100 cm 2, 1.18 log CFU/100 cm 2, 1.65 log CFU/100 cm 2., Harrigan and McCance (1976) 2.70 log CFU/100 cm 2, log CFU/100 cm 2, 3.40 log CFU/100 cm 2

6 538 강경태ㆍ김민주ㆍ박선영ㆍ최종덕ㆍ허민수ㆍ김진수, 1.00 log CFU/100 cm 2. Harrigan and McCance (1976) (2.51 log CFU/100 cm 2 ), (1.18 log CFU/100 cm 2 ), (1.65 log CFU/100 cm 2 ), (3.26 log CFU/100 cm 2 ), (3.00 log CFU/100 cm 2 ) (3.00 log CFU/100 cm 2 ), (3.40 log CFU/100 cm 2 ).,..,.,,,, 100 cm 2., Harrigan and McCance (1976). 중요관리점 (CCP) 결정및한계기준 (CL) 설정 CCP CP CCP (Lee, 2011) CCP 2. (60-90 ), 20 kg (20-40 L/min 10 L/min ), Fig log CFU/g, 20 L/ min 3.02 log CFU/g, 30 L/min 2.85 log CFU/g, 40 L/min 2.91 log CFU/g, 30 L/min,., Log CFU/g Before washing L/min 30 L/min 40 L/min Washing water quantity 1 Fig. 4. Viable cell counts of washed oyster Crassostrea gigas as affected by washing water quantity. 1 Oyster weight used in this experiment is 20 kg. 30 L/min E. coli O157:H7, Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Vibrio parahaemolyticus Clostridium perfringens ( )., 20 kg, L/ min., (90-110%, 10% ), (Fe STS), (Fe mm, STS mm ) (, ) Table 3. (Fe STS) Fe 1.0 mm, STS 3.0 mm ( )., 2 mm, Kvenberg and Schwalm (2000) 8 mm. (Fe STS) 90%, 30-70%,,., 90%. (Fe STS) 100% Fe 1.5 mm, STS 3.5 mm 100%, 60-80%

7 굴가공공장의위해평가 539 Table 3. Detection rate of a metal detector as affected by position, sensitivity, diameter and kind of standard sample Position Right Middle Sensitivity Diameter (mmφ) Fe Detectionrate Diameter (mmφ) STS Detection rate Position Sensitivity Diameter (mmφ) Fe Detection rate Diameter (mmφ) STS Detection rate Middle Left , 100%, 1.5 mm, 3.5 mm. (Fe STS) 110% Fe 1.5 mm, STS 3.5 mm 100%, %., 100%, 1.5 mm, 3.5 mm. 사사 2016 ( ) References American public health association (APHA) Compendium of methods for the microbiological examination of foods (4th Ed), American public health association, Washington, DC, U.S.A., Baker LL Public perceptions and pesticides A case study of the Willapa Bay oyster bed spraying permit of Master Thesis, University of Washington, Washington, DC, U.S.A. Choi JD, Jeong WG and Kim PH Bacteriological study of sea water and oyster in Charan bay, Koraea. J Korean Fish Soc 31, David LN and Michael MC Lehninger Principles of Biochemistry (6th Ed). Word Science Publishing Co., Seoul, Korea, Chapter 9. Ha SH, Woo GJ, Kwak HS, Hwang IG and Choi WS Simplified procedure for detection of poliovirus and norovirus in oyster. Korean J Food Sci Technol 37, Harrigan WF and McCance ME The examination of food processing plant. In Laboratory Method in Food and Microbiology. Academic Press, New York, U.S.A. Hosoi M, Kubota S, Toyohara M, Toyohara H and Hayashi I Effect of salinity change on free amino acid content in Pacific oyster. Fish Sci 69, Iritani N, Kaida A, Abe N, Kubo H, Sekiguchi JI, Yamamoto SP, Goto K, Tanaka T and Noda M Detection and genetic characterization of human enteric viruses in oysterassociated gastroenteritis outbreaks between 2001 and 2012 in Osaka City, Japan. Jeong IG, Park JH, Yoo HD and Choi JD Sanitary survey of the sea water and oyster in the south coastal designated area. J Ins Marine Industry 17,

8 540 강경태ㆍ김민주ㆍ박선영ㆍ최종덕ㆍ허민수ㆍ김진수 Kim CY, Pyeun JH and Nam JN Decomposition of glycogen and protein in pickled oyster during fermentation with salt. J Korean Fish Soc 14, Kim JS Cold Storage of Foods. Hyoil Publishing Co., Seoul, Korea, Kim JS, Heu MS, Kim HS and Ha JH Fundamental and Application of Seafood Processing. Hyoil Publishing Co., Seoul, Korea, , Kim JS, Yeum DM., Kang HG, Kim IS, Kong CS, Lee TG and Heu MS Fundamentals and Application for Canned Foods. Hyoil Publishing Co., Seoul, Korea, 59-62, Kvenberg JE and Schwalm DJ Use of microbial data for hazard analysis and critical control point verification - Food and drug administration perspective. J Food Protection 63, Lee H Microbiological population of Vibrio parahaemolyticus in oyster wholesale seafood markets. J Fd Hyg Safety 21, Lee JS Development of HACCP Plan for Packaged Tofu Manufactured by Automated Process System. Master Thesis, University of Chungang, Seoul, Korea. Lee TS, Oh EG, Yoo HD, Ha KS, Yu HS, Byun HS and Kim JH Impact of rainfall events on the bacteriological water quality of the shellfish growing area in Korea. Korean J Fish Aquat Sci 43, KFAS Linehan LG, O'Connor TP and Burnell G Seasonal variation in chemical composition and fatty acid profile of Pacific oyster (Crassostrea gigas). Food Chem 64, Ministry of Environment Environmental Enforcement Ordinance of the Act No Quality standard for drinking water. Retrieved from eq=176558&efyd= #0000 on Jun 15. Ministry of Food and Drug Safety Method of Investigation of cause in Food poisoning. Retrieved from mfds.go.kr/index.do?searchkey=title:contents&mid=695& searchword=foodpoisoning&cd=&pageno=1&seq=20737 &cmd=v on Apr 3. Ministry of Food and Drug Safety. 2016a. Korean Food. Code. Chapter 9. General analytical method ~ Retrieved from portal/ safefoodlife/food/foodrvlv/foodrvlv.do on Apr 3. Ministry of Food and Drug Safety. 2016b. Korean Food. Code. Chapter 6. General analytical method Retrieved from foodrvlv/foodrvlv.do on Apr 3. Ministry of Oceans and Fisheries Statistical Yearbook of Fisheries Production. Retrieved from jsp/sf/ss/ss_kind_law_list.jsp?menudepth= on Jun 15. Oh EG, Yoo HD, Yu HS, Ha KS, Shin SB, Lee TS, Lee HJ, Kim JH and Son KT Removal of fecal indicator bacteria from bivalves under natural and electrolyzed water. Korean J Fish Aquat Sci 45, KFAS Park WJ, Jwa MK, Hyun SH, Lim SB and Song DJ Microbial and quality changes during storage of raw oyster treated with high hydrostatic pressure. J Korean Soc Food Sci Nutr 35, Sakaguchi M and Murat M Seasonal variations of free amino acids on oyster whole body and adductor muscle. Nippon Suisan Gakkaishi 55, Vugia DJ, Farzaneh T, Anna EN, Michael H and Patricia MG Impact of 2003 State regulation on raw oyster-associated Vibrio vulnificus illnesses and deaths, California, U.S.A. Emerging Infectious Diseases 19, dx.doi.org/ /eid

생식용굴가공공장의위생안전성 2,,., Jeong et al. (2004), Yu et al. (206) Bang et al. (2008) norovirus norovirus, Manatawee et al. (20) ice-bath, Xiao et al. (20) mod

생식용굴가공공장의위생안전성 2,,., Jeong et al. (2004), Yu et al. (206) Bang et al. (2008) norovirus norovirus, Manatawee et al. (20) ice-bath, Xiao et al. (20) mod 한수지 50(2), 20-29, 207 Original Article Korean J Fish Aquat Sci 50(2),20-29,207 생식용굴 (Crassostrea gigas) 작업장의위생안전성에대한모니터링 강경태 박선영,3 최종덕 김민주,3 허민수 2,3 김진수,3 * 한국식품안전관리인증원, 경상대학교해양식품생명의학과 / 해양산업연구소, 2 경상대학교식품영양학과,