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1 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, 3 경상대학교수산식품산업화기술지원센터 Safety Monitoring of a Processing Plant for Preparing Raw Oysters Crassostrea gigas for Consumption Kyung Tae Kang, Sun Young Park,3, Jong-Duck Choi, Min Joo Kim,3, Min Soo Heu 2,3 and Jin-Soo Kim,3 * Korea Institute for Food Safety Management Accreditation, Daejeon 3497, Korea Department of Seafood and Aquaculture Science/Institute of Marine Industry, Gyeongsang National University, Tongyeong 53064, Korea 2 Department of Food Science and Nutrition, Gyeongsang National University, Jinju 52828, Korea 3 Research Center for Industrial Development of Seafood, Gyeongsang National University, Tongyeong 53064, Korea This study assessed the safety of raw oysters Crassostrea gigas for consumption during processing in a processing plant. Bacterial contamination (e.g., viable cell counts, coliform groups, Escherichia. coli and pathogenic bacteria) and chemical contamination (e.g., heavy metals and shellfish toxins) were measured on raw oysters, a processing equipment, employees and work areas. No total mercury, lead, paralytic shellfish poison, diarrheic shellfish poison or norovirus was detected in any post-harvested oyster samples. However, the cadmium level ranged from mg/kg. The viable cell count, E. coli and coliform group levels in post-harvested oysters ranged from log CFU/g, ND-20 MPN/00 g and 0-40 MPN/00 g, respectively. The viable contaminating cell counts on employees, equipment and work areas were in the range of log CFU/00 cm 2. Airborne bacteria in the work areas ranged from 0.60 to.8 log CFU/plate/5 min. Thus, no significant health risks were detected in the processing plant. Key word: Oyster, Hazard analysis, Oyster processing workplace, Hygienic safety, Raw oyster for consumption 서론 897, , 60% (Ministry of Oceans and Fisheries, 206).,,,,,, (Linehan et al., 999). 4, 5,.,,.. (Sobsey and Jaykus, 99). FDA EU. Korean J Fish Aquat Sci 50(2) 20-29, April 207 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 3 February 207; Revised March 207; Accepted 4 April 207 *Corresponding author: Tel: Fax: address: jinsukim@gnu.ac.kr Copyright 207 The Korean Society of Fisheries and Aquatic Science 20 pissn:0374-8, eissn:

2 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) modified atmosphere packaging (MAP).,,,,.,,,. 재료및방법 재료 205 S (S), O (O), Process Postharvested oyster First washing Cold storing Code A B C Weighing/ Inner packing Ice cooling Washing Shucking G F E D Metal detecting Outer packing Raw oyster for consumption in PE H I J Fig.. Manufacturing process and sample code of raw oyster Crassostrea gigas for consumption in polyethylene (PE) bag. H (H), D (D) Y (Y) Fig..,. 00 cm 2 (APHA, 200), Glove juice (FDA, 978). ( L), (2, 5 min) ( 300 g). (4 ) 24. 생식용굴 ( 봉지굴 ) 의제조공정도 (A) (B), (C) (D), (E), (F), (G), (H), (I), Fig... 중금속,,,. 0. g, 0. ml (DMA-80, Milestone, Milano, Italy), Easy-DOC3 (Easy-DOC3 for DMA, Ver. 3.30, Milestone, USA). Kim (204),, (MFDS, 206) 70 (4% ) ICP-MS (ELAN DRC II, PerkinElmer, Santa Clara, USA)., (,000 mg/l), (supra-pure grade), 8 MΩ. 일반세균수 25 g ml 0.85% 225 ml stomacher

3 22 강경태ㆍ박선영ㆍ최종덕ㆍ김민주ㆍ허민수ㆍ김진수 (Bag Mizer 400VW, Interscience, France) 30,,, 00 cm % 9 ml. PCA (plate count agar), (35, 48 ),. 대장균군및대장균 25 g ml, 3, 5. (lactose broth) MPN/00 g. EC broth ( , 24 ) durham tube MPN/00 g., 00 cm % 9 ml. ml, ( ),, log CFU/g, log CFU/plate/5 min. 식중독균 Enterohemorrhagic E. coli, Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Vibrio parahaemolyticus Clostridium perfringens, norovirus 7, (MFDS, 206) 9., 3.. Enterohemorrhagic E. coli (25 g) modified EC broth (mec broth; Difco, USA) 225 ml, (35-37, 24 ) Tellurite cefixime-sorbitol macconkey agar (Difco, USA) (35-37, 8 ). sorbitol EMB (35-37, 24 ). Listeria monocytogenes (25 g) Listeria enrichment broth (Difco, USA) 225 ml Oxford agar (Difco, USA), (30, ), 0.6% yeast extract Tryptic soy agar API listeria kit (Biomerieux, France). Staphylococcus aureus (25 g) 0% NaCl Tryptic soy broth (TSB, Difco, USA) 225 ml (35-37, 24 ) Baird- Parker agar (Difco, USA), (35-37, 24 )., (35-37, 24 ),. Salmonella spp. 25 g 225 ml (37, 24 ) 0. ml Rappaport-Vassiliadis (Difco, USA) 0 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) (37, 8-24 ) TCBS (37, 24 ). TSI API 20E (Biomerieux, France). Clostridium perfringens ml cooked meat (Difco, USA) (35-37, 8-24 ) TSC (Difco, USA) (35-37, 8-24 ) TSC.. Norovirus,, RNA, RT-PCR PCR. 패류독소, (MFDS, 206) 9, 6 AOAC (2000). 포장재의화학적검사 (MFDS, 206),,, -hexen, -octene,. 70 (4% ) 70

4 생식용굴가공공장의위생안전성 , 05 2 (L), -hexen -octene GC-MS (GC 6890, Agilent Technologoes, USA). 통계처리 SAS system (Cary, NC, USA) (ANOVA test), Duncan (P<0.05). 결과및고찰 생식용굴생산을위한원부재료분석 5 [ (,, ) (, )] [,, (Enterohemorrhagic E. coli, Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Vibrio parahaemolyticus, Clostridium perfringens norovirus)] Table. 5, mg/kg, log CFU/g, 0-40 MPN/00 g, ND-20 MPN/00 g, D Y 2 7 (Enterohemorrhagic E. coli, Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Vibrio parahaemolyticus, Clostridium perfringens) norovirus. (National Fishery Products Quality Management Service, 206) 0.5 mg/kg, 2.0 mg/kg, 0.8 mg/kg, 50,000 CFU/g, 230 MPN/00 g, (Listeria monocytogenes, Staphylococcus aureus, Salmonella spp., Vibrio parahaemolyticus)., (MFDS, 206) n=5, c=, m=230, M=700 MPN/00 g, (Enterohemorrhagic E. coli, Listeria monocytogenes, Salmonella spp., Vibrio parahaemolyticus, Clostridium perfringens, Staphylococcus aureus) n=5, c=0, m=0/25 g, norovirus. 5. [ Table. Results on the chemical and biological hazards of post-harvested oyster for preparing raw oyster Crassostrea gigas for consumption in polyethylene bag Chemical (mg/kg) Biological (log CFU/g) Hazard Factory S O H D 3 Y Total Hg ND ND ND ND ND Pb ND ND ND ND ND Cd PSP (paralytic shellfish poison) ND ND ND ND ND DSP (diarrhetic shellfish poison) ND ND ND ND ND Viable cell count Enterohemorrhagic E. coli ND ND Listeria monocytogenes ND ND Staphylococcus aureus ND ND Salmonella spp ND ND Vibrio parahaemolyticus ND ND Clostridium perfringens ND ND Norovirus ND ND ND ND ND E. coli (MPN/00 g) ND ND Coliform group (MPN/00 g) ND, Not detected; 2 -, Not determined; 3 Biological data of factory D were quoted from Kang et al. (206).

5 24 강경태ㆍ박선영ㆍ최종덕ㆍ김민주ㆍ허민수ㆍ김진수 (,, )] (,, ) Table 2. 5,, ND-.4 log CFU/g,., 0.00 mg/l, 0.0 mg/l, mg/l, 00 CFU/mL, /00 ml, /00 ml., 5. (,,, -hexene -octene) (,, ) Table 3. (S, O, H, D, Y), -3 mg/l, 2-9 mg/l, -hexene -octene.,,, (D, Y)., (polyetylene, PE) (MFDS, 206), ( ).0 mg/l, 0 mg/l, 30 mg/l, -hexene 3 mg/l -octene 5 mg/l. 5 (S, O, H, D Y),,, -hexene -octene. 생식용굴의생산을위한작업환경위생. (S, O, H, D Y),,, Table 2. Results on the chemical and biological hazards of drinking water for processing raw oyster Crassostrea gigas for consumption in polyethylene bag Chemical (mg/l) Biological Hazard Factory S O H D 2 Y Total Hg ND ND ND ND ND Pb ND ND ND ND ND Cd ND ND ND ND ND Viable cell count (log CFU/mL) ND.08.4 ND ND E. coli (MPN/00 ml) ND ND ND ND ND Coliform group (MPN/00 ml) ND ND ND ND ND ND, Not detected; 2 Biological data of factory D were quoted from Kang et al. (206). Table 3. Results on the chemical and biological hazards in polyethylene (PE) bag for packing raw oyster Crassostrea gigas for consumption Chemical (mg/l) Biological Migrant from PE PE Hazard Factory S O H D 5 Y Pb ND ND ND ND ND PPC ER Hexene ND ND ND ND ND -Octene ND ND ND ND ND Viable cell count (log CFU/mL) ND ND Staphylococcus aureus (log CFU/mL) ND ND E. coli (MPN/00 ml) ND ND Coliform group (MPN/00 ml) ND ND ND, Not detected; 2 PPC, Potassium permanganate consumption; 3 ER, Evaporation residue; 4 -, Not determined; 5 Biological data of factory D were quoted from Kang et al. (206).

6 생식용굴가공공장의위생안전성 25 (00 cm 2 ), Table log CFU/00 cm 2, log CFU/00 cm 2, log CFU/00 cm 2, log CFU/00 cm 2, log CFU/00 cm 2, log CFU/00 cm 2, log CFU/00 cm 2,., Harrigan and McCance (976) 2.70 log CFU/00 cm 2, log CFU/00 cm 2, 3.40 log CFU/00 cm 2. Harrigan and McCance (976) (S, D Y), ( ), ( ), (S, O, D Y), ( ) (H, D Y), (H, D Y).,,., Bryan et al. (982) 6%, 9%, Stauffer (97),,,.. (,,, ) (00 cm 2 ) (O.00 log CFU/00 cm 2 H.40 log CFU/00 cm 2 ), (H.8 log CFU/00 cm 2 ) (H.08 log CFU/00 cm 2 Y 0.60 log CFU/00 cm 2 ),. Harrigan and McCance (976).00 log CFU/00 cm 2. (O H), (H) (H Y). 개인위생 (S, O, H, D Y),, (00 cm 2 ), Table log CFU/hand, log CFU/hand, log CFU/00 cm 2, log CFU/00 cm 2, log CFU/hand, log CFU/hand, log CFU/00 cm 2,.-.44 log CFU/00 cm 2, log CFU/hand, log CFU/hand, log CFU/00 cm 2, log CFU/00 cm 2.,,, Harrigan and McCance (976) Table 4. Microbiological evaluation of equipment used for manufacturing raw oyster Crassostrea gigas for consumption in polyethylene bag (log CFU/00 cm 2 ) Working Shucking Washing Equipment Viable cell count Coliform group Staphylococcus aureus S O H D 2 Y S O H D 2 Y S O H D 2 Y Worktable ND ND ND ND ND ND ND ND Basket ND ND ND ND ND ND ND ND Knife ND ND.08 ND 0.60 ND ND ND ND ND Washing machine ND ND ND ND ND ND ND ND ND ND Worktable ND ND ND ND ND ND ND ND ND ND Inner Packing machine ND ND ND ND ND ND ND ND ND ND packing Worktable ND ND ND ND ND ND ND ND ND ND ND, Not detected; 2 Data of factory D were quoted from Kang et al. (206).

7 26 강경태ㆍ박선영ㆍ최종덕ㆍ김민주ㆍ허민수ㆍ김진수 Table 5. Microbiological evaluation of person worked for manufacturing raw oyster Crassostrea gigas for consumption in polyethylene bag (log CFU 00 cm 2 or hand) Working Shucking Washing Inner packing Employee Viable cell count Coliform group Staphylococcus aureus S O H D 2 Y S O H D 2 Y S O H D 2 Y Hands ND ND ND ND ND ND ND ND ND ND Glove ND ND ND ND ND ND ND ND ND ND Apron ND ND ND ND ND ND ND ND ND ND Boot ND ND ND ND ND ND ND ND ND ND Hands ND ND ND ND ND ND ND ND ND ND Glove ND ND ND ND ND ND ND ND ND ND Apron ND ND ND ND ND ND ND ND ND ND Boot ND ND ND ND ND ND ND ND ND ND Hands ND ND ND ND ND ND ND ND ND ND Glove ND ND ND ND ND ND ND ND ND ND Apron ND ND ND ND ND ND ND ND ND ND Boot ND ND ND ND ND ND ND ND ND ND ND, Not detected; 2 Data of factory D was quoted from Kang et al. (206).,,,,,,., Kim (200) 4.69 log CFU/hand,.00 log CFU/hand,.,,. Martin et al. (2004),., Kjolen and Amdersen (992),.,,,. 공중낙하균 (S, O, H, D, Y) (, ) ( ) Table log CFU/plate/5 min,.8-.4 log CFU/plate/5 min, log CFU/plate/5 min, log CFU/plate/5 min., Seol et al. (2009).7 log CFU/ plate/5 min,.7-.9 log CFU/plate/5 min, log CFU/plate/5 min., Seol et al. (2009) H (.8 log CFU/plate/5 min),,.,.. Kang (990)., Hambraeus et al. (978). 제조공정별생물학적위해요소 Fig. 2.

8 생식용굴가공공장의위생안전성 27 Table 6. Microbiological evaluation of working area for manufacturing raw oyster Crassostrea gigas for consumption in polyethylene bag (log CFU/plate/5 min) Working Area Viable cell count Coliform group S O H D 2 Y S O H D 2 Y Shucking Worktable ND ND ND ND ND Washing Washing machine ND ND ND ND ND Inner packing Packing machine ND ND ND ND ND Outer packing Worktable ND ND ND ND ND ND, Not detected; 2 Data of factory D was quoted from Kang et al. (206). D Y (A) log CFU/g, (B) log CFU/g, (C) log CFU/g, (D) log CFU/g, (E) (F) log CFU/g, log CFU/g.,,.,,. cold shock (Kim, 2000). D Y (G) log CFU/g, (H) log CFU/g, (I) log CFU/g, log CFU/g, log CFU/g.,,,. D Y (A) 0 80 MPN/00 g, (B) MPN/00 g, (C) 75 0 MPN/00 g, (D) MPN/00 g, (E) (F) 44 8 MPN/00 g, MPN/00 g.,. D Viable cell counts (Log CFU/ g) Coliform group (MPN/ 00 g) Fig. 2. Change in viable cell counts and coliform group of raw oyster Crassostrea gigas for consumption in polyethylene bag during processing. Process codes (A-J) are the same as shown in Fig. ; 2 Data of Factory D were quoted from Kang et al. (206). Factory D Factory Y A B C D E F G H I J Process code Y (G) MPN/00 g, (H) MPN/00 g, (I) 40 6 MPN/00 g, MPN/00 g, MPN/00 g., 8 70 Factory D Factory Y A B C D E F G H I J Process code

9 28 강경태ㆍ박선영ㆍ최종덕ㆍ김민주ㆍ허민수ㆍ김진수,,,.,., (National Fishery Products Quality Management Service, 206) 50,000 CFU/g 230 MPN/00 g, (MFDS, 206) n=5, c=, m=230, M=700 MPN/00 g.. D Y,,,,,. 사사 206 ( ) 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., AOAC (Official method of Analysis) Association of Official Analytical Chemists. 7th ed. Maryland, U.S.A., Bang SJ, Park KH, Park MK and Kwon YO The research of pollution of Norovirus in circulating oysters. J Gyeonggi Institute Health Environ 2, Bryan FL, Bartleson CA and Sugi M Hazard analysis of charsiu and roast pork in Chinese restaurant and market. J Food Prot 45, Food and Drug Administration (FDA) Guide lines for effectiveness testing of surgical hands crub (Glove juice test) [Internet]. Fed Rdgist 43, Retrieved from on Feb 5, 207. Hambraeus A, Bengtsson S and Laurell G Bacterial contamination in a modern operating suite. 2. Effect of a zoning system on contamination of floors and other surfaces. J Hyg 80, S Published online. 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. 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 7, Kang KT, Kim MJ, Park SY, Choi JD, Heu MS and Kim JS Risk assessment of oyster Crassostrea gigas processing site for an HACCP system model. Korean J Fish Aquat Sci 49, Kang YJ Analysis and characteristic of airborne bacteria as food contaminants. Korean Dairy Technol 8, 7-4. Kim HK Application of HACCP system for microbiological quality control of sushi production in Japanese restaurants. MS thesis, Chung-Ang University, Seoul, Korea. Kim JS Cold Storage of Foods. Hyoil Publishing Co., Seoul, Korea, Kim KH Concentration and risk assessment of heavy metal in mainly consumed fishes. MS thesis, Gyeongsang National University, Tongyeong, Korea. Kjolen H and Amdersen BM Hand washing and disinfection of heavily contaminated hands-effective or ineffective?. J Hospital Infect 2, 6-7. 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, dx.doi.org/0.06/s (98) Martin MC, Fueyo JM, Gonzalez-Hevia MA and Mendoza MC Generic procedures for identification of enterotoxigenic strains of Staphylococcus aureus from three food poisoning outbreaks. International J Food Microbiol 94, /j.ijfoodmicro Manatawee S, Boonprasop N, Boonprasop S and Sutthirak P. 20. Spoilage bacteria changes during storage of oyster (Crassostrea belcheri) in ice-bath. Thai J Agric Sci 44, Ministry of Environment (ME) Environmental Enforcement Ordinance of the Act No. 28. Quality Standard for Drinking Water [internet]. Retrieved from go.kr/lsinfop.do?lsiseq=76558&efyd =20523#0000 on Feb 5, 207. Ministry of Food and Drug Safety (MFDS) Korean Food Code. Chapter 2. Standard on the common foods, Chapter 6. Standard on the seafoods, Chapter 9. General analytical method [Internet]. Retrieved fromhttps://foodsafetykorea.go.kr/portal/safefoodlife/food/foodrvlv/foodrvlv. do?menu_grp=menu_new0&menu_no=3093 on Feb 5, 207. Ministry of Oceans and Fisheries (MOS) Product Statistic [Internet]. Retrieved from ss_info.jsp?menudepth=0700 on on Feb 8, 207. National Fishery Products Quality Management Service (NFQS) Inspection code of fishery products [Ineternt]. Re-

10 trieved from 3&s=&s2= on Feb 25, 207. Seol HR, Park HS, Park KH, Park AK and Ryu K Microbiological evaluation of foods and kitchen environments in childcare center and kindergarten food service operations. J Korean Soc Food Sci Nutr 38, /jkfn Sobey MD and Jaykus LA. 99. Human enteric viruses and depuration of bivalve molluscs. In: Otwell WS, Rodrick GE and Martin R(Eds), Molluscan shellfish deprivation. CRC Press, Boca Raton, Florida, U.S.A., 7-4. Stauffer LD. 97. Sanitation and the human ingredient. Hospitals 45, Xiao G, Zhang M, Shan L, You Y and Salokhe VM. 20. Extension of the shelf-life of fresh oyster mushs (Pleurotus ostreatus) by modified atmosphere packaging with chemical treatments. Afr J Biotechnol 0, dx.doi.org/0.5897/ajb Yu HS, Park YS, An SR, Park KBWI, Shim KB, Song KC and Lee TS Defecation of norovirus from the oyster Crassostrea gigas by depuration following translocation of the growing area. Korean J Fish Aquat Sci 49, 생식용굴가공공장의위생안전성 29

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

534 강경태ㆍ김민주ㆍ박선영ㆍ최종덕ㆍ허민수ㆍ김진수., Choi et al. (1998), Jeong et al. (2004), Ha et al. (2005), Lee (2006), Baker (2016), Vugia et al. (2013) 2003.,,,,. HACC 한수지 49(5), 533-540, 2016 Original Article Korean J Fish Aquat Sci 49(5),533-540,2016 HACCP 구축을위한굴 (Crassostrea gigas) 가공공장의위해평가 강경태 김민주 1 박선영 1 최종덕 1 허민수 2 김진수 1 * 한국식품안전관리인증원, 1 경상대학교해양식품생명의학과 / 해양산업연구소,