한수지 50(4), , 2017 Original Article Korean J Fish Aquat Sci 50(4), ,2017 우리나라주요김생산해역의해수및물김에대한위생학적평가 정상현 신순범 오은경 조미라 1 윤민철 1 이희정 1 손광태 2 *

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한수지 50(4), 343-35, 207 Original Article Korean J Fish Aquat Sci 50(4),343-35,207 우리나라주요김생산해역의해수및물김에대한위생학적평가 정상현 신순범 오은경 조미라 윤민철 이희정 손광태 2 * 국립수산과학원남해수산연구소양식산업과, 국립수산과학원식품위생가공과, 2 국립수산과학원남동해수산연구소 Sanitary Evaluation for Seawater and Laver Pyropia sp. in the Major Laver Growing Areas, Korea Sang Hyeon Jeong, Soon Bum Shin, Eun Gyoung Oh, Mi-Ra Jo, Min Chul Yoon, Hee Jung Lee and Kwang-Tae Son 2 * Aquaculture Industry Division, South Sea Fisheries Research Institute, National Institute Fisheries Science, Yeosu 59780, Korea Food Safety and Processing Research Division, National Institute Fisheries Science, Busan 46083, Korea 2 Southeast Sea Fisheries Research Institute, National Institute Fisheries Science, Tongyoung 53085, Korea Sanitary evaluation of seawater and Pyropia sp. laver collected from the five major laver growing areas in Korea was performed four times over the course of a year. The seawater quality in four of these five areas was regarded as the clean area according to Korean criteria, but the seawater at one investigation site in Seoheon area was found to exceed the standard for fecal coliform. In the bacteriological safety analysis of laver (raw source), the percentages of samples not conforming to Chinese criteria at the five sites were 55.6% (Seocheon), 70.0% (Shinan), 8.8% [Jindo (Haenam)], 63.6% (Wando), and 28.6% [Goheung (Jangheung)]. Pathogenic bacteria were not detected in all laver samples. The food safety of laver (raw source) based on heavy metal concentration was confirmed using Korean criteria; the concentrations of heavy metals in laver samples collected from the major laver growing areas were 0.008-0.632 mg/kg wet weight (ww) lead, 0.024-0.37 mg/kg ww cadmium, 0.908-2.892 mg/kg ww total arsenic, and 0.003-0.03 mg/kg ww total mercury. Therefore, pollution source management and periodic monitoring of heavy metals may be required to improve the food safety of laver produced in these laver growing areas. Key words: Laver, Coliform group, E. coli, Heavy metal, Food safety 서론,, (Cardozo et al., 2007; Kim et al., 203), (Choi et al., 204; Bae et al., 205; Kim et al., 205; Ra et al., 206). 40% 30-40%, (Jimenez-Escrig and Sanchez-Muniz, 2000; Lee and Lee, 204).,,,, 0% (Kim et al., 204). 20 40, 32% (FAO, 205). 206 8, 3 5 (MOF, 206).. https://doi.org/0.5657/kfas.207.0343 Korean J Fish Aquat Sci 50(4) 343-35, August 207 This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial Licens (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Received August 207; Revised 6 August 207; Accepted 2 August 207 *Corresponding author: Tel: +82. 55. 640. 4760 Fax: +82. 55. 64. 2036 E-mail address: ktson@korea.kr Copyright 207 The Korean Society of Fisheries and Aquatic Science 343 pissn:0374-8, eissn:2287-885

344 정상현ㆍ신순범ㆍ오은경ㆍ조미라ㆍ윤민철ㆍ이희정ㆍ손광태.,,,,, (Ock, 200)., (Mallin et al., 2000; Glasoe et al., 2004; Walters et al., 20; Qiao et al., 203; Delbecque and Verdoodt, 206)., (Ok et al., 2007; Son et al., 202; Kim and Yoon, 203; Kim et al., 206; Yang et al., 206),., 75% 4 [,, ( ), ( )] 8% (KMI, 205). 재료및방법 조사지점및시료채취 5 [,,, ( ), ( )].,. 0 3, 206, 2, 3 ( 2) 4.(Fig. ). 250 ml (Duran, Germany), 0-30 cm,, Whirl-pak (Nasco, USA), 0, 24. 위생지표세균및병원성세균분석, Laboratory procedures for the examination of seawater and shellfish (APHA, 985), Escherichia coli Bacteriological Analytic Manual (FDA, 998) ( ) (KFDA, 206)., 3. 일반세균수 ml, /0 ( :Sterile Phosphate Buffer 9) ml /00 ml 2 Petri dish 44-46 Plate Count Agar (PCA) (Difco, USA) Petri dish 5-20 ml, 35 0.5 24-48. 30-300, ml g (Colony Forming Unit, CFU). 대장균군, 분변계대장균및 E. coli 0 ml ( :Sterile Phosphate Buffer 9) 0 ml 2 Lauryl tryptose broth (Difco, USA) 5.0 0. ml 5 35 0.5 24-48. Brilliant Green Bile (2%) broth (Difco, USA) EC broth (Difco, USA) Loop (Kartell, Italy). Brilliant Green Bile (2%) broth EC broth 35 0.5 24-48 44.5 0.2 24. E. coli EC broth EMB agar (Merk, Germany) 35 24, PCA, VITEK system (biomerieux, France)., E. coli 00 ml 00 g (Most Probable Number, MPN). 병원성세균분석 (,,

우리나라주요김생산해역에대한위생학적평가 345 Fig. The sampling stations for seawater and laver Pyropia sp. in the major laver growing areas [Seocheon, Shinan, Wando, Jindo (Haenam) and Goheung (Jangheung)]. ) Bacteriological Analytic Manual (FDA, 998) ( ) (KFDA, 206), MPN (FDA, 998). VITEK system. (2 ). 중금속분석 (Freeze dryer, EYELA, FDU-20, Dokyo, Japan) 3,, (CRM; Certified Reference Materials) 7405-a (Hijiki; AIST, Tsukuba, Japan) DORM- 4 (Fish protein; NRC-CNRC, Ottawa, Ontario, Canada)

346 정상현ㆍ신순범ㆍ오은경ㆍ조미라ㆍ윤민철ㆍ이희정ㆍ손광태 566b (Oyster tissue; National Institute of Standard Technology, Gaithersburg, Maryland, USA), (AOAC international, 2002). 0% (v/v) (Nitric acid, Merk, Germany) 8 M cm. 납 (Pb), 카드뮴 (Cd), 총비소 (Total-As) 분석 0.5 g 65% 5 ml 0 ml (Pyrex, USA), 37. 2 20 20 bar 2, 230, 50 bar 5, 230, 50 bar 0. 2% (v/v) 50 ml, 0.45 m, (ICP-MS, Perkin Elmer, Waltham, USA) 3.,000 mg/kg (Merck, Darmstadt, Germany). 총수은 (Total-Hg) 분석 0. g (Combustion gold amalgamation method) (Automatic Mercury analyzer, DMA-80, Milestone, S&T, Italy) 3. CRM 7405-a. 650 90, 650 80, (Amalgamation) 850 2. Easy-DOC3 (Easy-DOC3 for DMA, Ver. 3.30, Milestone, USA). 통계처리 P<0.05 (Duncan s multiple test), SAS Version 9.2/Enterprise Guide 4.2 (SAS Institute Inc., Cary, NC, USA). 해수의수질및물김의안전성평가 (MOF, 203), 43 MPN/00 ml 0%., (KFDA, 206) ( ) 00,000 CFU/g, 30 MPN/00 g, 30,000 CFU/g (,,, ). (EU) Regulation (EC) No 854 E. coli 3 (Class A, B, C), A E. coli 230 MPN/00 g (European Commission, 2004). EU., 0.3 mg/kg,.0 mg/kg, 5.0 mg/kg, 0. mg/kg. Table. Bacteriological quality of the seawater harvested from the major laver growing areas Sea area MPN/00 ml CFU/mL No. of sampling Coliform group Fecal coliform Plate count, at 35 stations No. of samples Seocheon <.8-240 <.8-79 <30-540 0( 2 ) 40 Shinan <.8-7.8 <.8-4.5 <30-30 (0) 44 Jindo (Haenam) <.8-7 <.8-4.5 <30-36 6(0) 64 Wando <.8- <.8-4.5 <30-<30 2(0) 48 Goheung (Jangheung) <.8- <.8-2.0 <30-<30 4(0) 56 Total <.8-240 <.8-79 <30-540 63() 252 The number in brackets is that the percentage of samples which exceed fecal coliform concentration, 43 MPN/00 ml for a five-tube test exceeds 0% of total samples at each sampling station according to Korea Criteria for the clean area (MOF, 203). 2 The station No. 0 of seawater sampling stations in laver growing area, Seochen was exceeded by the standard of Korea Criteria for the clean area.

우리나라주요김생산해역에대한위생학적평가 347 결과및고찰 미생물학적위생성상확인및안전성평가 (MOF, 203) 4 (,, )., 4 MPN/00 ml, 43 MPN/00 ml 0%. 4, 43 MPN/00 ml 0%. 5, 77 252 <.8-240 <.8-79 MPN/00 ml, <30-540 CFU/mL (Table ). 0 40 <.8-240 <.8-79 MPN/00 ml, <30-540 CFU/mL, 0, 79 MPN/00 ml. 0 ( 50 ) ( ) (Todd and Campbell, 2002)., 4 67 ( ) 22 <.8-7 <.8-4.5 MPN/00 ml, <30-36 CFU/ ml,. 4 6 48, E. coli <8-490, <8-0 <8-20 MPN/00 g, <30-5,700 CFU/g (Table 2)., EU, (55.6%, 5/9), (70.0%, 7/0), ( ) (8.8%, 9/), (63.6%, 7/), ( ) (28.6%, 2/7) Table 2. Bacteriological quality of the laver Pyropia sp. (raw source) harvested from the major laver growing areas Sea area Range MPN/00 g CFU/g Coliform group Escherichia coli Plate count, at 35 >30 >230 2 >30,000 Range Range No. % No. % No. % Seocheon <8-490 5 55.6 <8-<8 0 0.0 70-3,000 0 0.0 9 Shinan <8-490 7 70.0 <8-<8 0 0.0 <30-2,400 0 0.0 0 Jindo (Haenam) <8-230 9 8.8 <8-8 0 0.0 50-5,700 0 0.0 Wando <8-230 7 63.6 <8-<8 0 0.0 <30-,300 0 0.0 Goheung (Jangheung) <8-230 2 28.8 <8-<8 0 0.0 80-,300 0 0.0 7 No. of samples Total <8-490 30 62.5 <8-8 0 0.0 <30-5,700 0 0.0 48 China Criteria for the bacteriological safety of seaweeds. 2 EU Criteria for the classification of sea area (European Commission, 2004). Table 3. Monitoring for pathogenic bacteria in the laver Pyropia sp. (raw source) harvested from the major laver growing areas in February, 207 Sea area Pathogenic bacteria Salmonella sp. Shigella spp. Staphylococcus aureus Vibrio parahaemolyticus Seocheon ND ND ND(<30) 2 ND(<30) 2 Shinan ND ND ND(<30) ND(<30) 3 Jindo (Haenam) ND ND ND(<30) ND(<30) 3 Wando ND ND ND(<30) ND(<30) 3 Goheung (Jangheung) ND ND ND(<30) ND(<30) 3 Not detected. 2 The number in brackets is the result for quantitative analysis (Unit : MPN/00 g). No. of samples

348 정상현ㆍ신순범ㆍ오은경ㆍ조미라ㆍ윤민철ㆍ이희정ㆍ손광태 (30 MPN/00 g ). ( ) (Glasoe et al., 2004),,, biofilm (Singh and Reddy, 204)., 5,,, (Table 3).,,, (5 ) 3. (,, ), 2-3. Table 4. The concentrations of four heavy metals in the laver Pyropia sp. (raw source) harvested from the major laver growing areas Sea area Seocheon Shinan Jindo (Haenam) Wando Goheung (Jangheung) Sampling station Heavy metals (mg/kg, wet wt) Pb Cd Total-As Total-Hg No. of samples A 0.03-0.20 0.08-0.37.469-2.33 0.004-0.008 4 B 0.009-0.39 0.089-0.20.97-.66 0.003-0.0 3 C 0.00-0.024 0.095-0.03 0.908-.698 0.004-0.007 2 Total 0.050±0.073ab 0.07±0.08c.529±0.385a 0.006±0.003ab 9 A 0.00 0.34.36 0.006 B 0.039-0.632 0.024-0.062.395-2.892 0.004-0.03 3 C 0.08-0.26 0.04-0.07.470-.926 0.004-0.009 4 Total 0.25±0.208b 0.06±0.033a.796±0.496a 0.007±0.003b 8 A 0.05-0.025 0.027-0.0.099-2.26 0.005-0.0 3 B 0.008-0.09 0.054-0.096 0.90-2.087 0.005-0.006 3 C 0.008-0.06 0.042-0.086 0.920-.809 0.004-0.005 4 Total 0.09±0.06a 0.068±0.024ab.480±0.464a 0.006±0.002ab 0 A 0.02-0.022 0.0-0.37.08-.578 0.003-0.006 3 B 0.03-0.023 0.058-0.0.060-2.023 0.004-0.005 4 C 0.009-0.025 0.03-0.094.279-2.065 0.004-0.006 3 Total 0.06±0.006a 0.092±0.03bc.462 ± 0.353a 0.005±0.00a 0 A 0.036-0.043 0.029-0.4.027-.882 0.003-0.006 3 2 B 0.09-0.085 0.097-0.29.334-.874 0.003-0.004 2 C 0.028-0.058 0.053-0..58-.753 0.005-0.007 2 Total 0.044±0.022ab 0.092±0.037bc.54±0.337a 0.004±0.002a 7 Total 0.048±0.099 0.084±0.032.549±0.4 0.005±0.002 44 (Range) (0.008-0.632) (0.024-0.37) (0.908-2.892) (0.003-0.03) The same column with different small letters indicate significant difference (P<0.05). 2 The one of three laver samples was harvested from the laver growing area in Jangheung.

우리나라주요김생산해역에대한위생학적평가 349. 중금속함량분석및안전성평가 4 (Cd, Pb, Hg As) 5 [,,, ( ), ( )] 4 6 45,, (Pb) 0.008-0.632 (0.048 0.099) mg/kg, (Cd) 0.024-0.37 (0.084 0.032) mg/kg, (Total- As) 0.908-2.892 (.549 0.4) mg/kg (Total-Hg) 0.003-0.03 (0.005 0.002) mg/kg (Table 4).,, ( ) 0.09 0.06 mg/kg, 0.25 0.208 mg/kg,, 0.06 0.033 mg/kg, 0.07 0.08 mg/kg., 5, ( ) 0.004 0.002 mg/kg, 0.007 0.003 mg/kg. (P<0.05).,.462 0.353 mg/kg,.796 0.496 mg/kg, (P<0.05)., 4 ( ),.,, ( ) (Ra et al., 202; El-Said, 203; Delbecque and Verdoodt, 206),, ( )., 0.3 mg/kg 0.5 mg/kg,,.0 mg/kg 5.0 mg/kg 5.0 mg/ kg, 0.5 mg/kg 0. mg/kg. 4. Yang et al. (206),,,. 사사 207 (R207057). References AOAC International. 2002. AOAC guidelines for single laboratory validation of chemical methods for dietary supplements and botanicals. Gaithersburg, MD, U.S.A. APHA (American Public Health Association). 970. Recommended procedures for the examination of seawater and shellfish. 4th Ed. American Public Health Association, Washington D.C., U.S.A, -47. Bae JH, Jung H and Kim MJ. 205. Economic feasibility and impact analysis on energy transformation project of macro-algae biomass. New and Renewable Energy 6, 29-38. https://doi.org/0.7849/ksnre.205.06..2.29. Cardozo KHM, Guaratini T, Barros MP, Falcao VR, Tonon, AP, Lopes NP, Campos S, Torres MA, Souza AO, Colepicolo P and Pinto E. 2007. Metabolites from algae with economical impact. Comp Biochem Physiol C Toxicol Pharmacol 46, 60-78. https://doi.org/0.06/j.cbpc.2006.05.007. Choi M, Yoo DI and Shin Y. 204. Preparation of lip balm utilizing functionalities of colorants extracted from marine algae. Text Color and Finish 26, 24-30. https://doi.org/0.5764/ tcf.204.26.2.24. Delbecque N and Verdoodt A. 206. Spatial patterns of heavy

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Walters SP, Thebo AL and Boehm AB. 20. Impact of urbanization and agriculture on the occurrence of bacterial pathogens and stx genes in coastal waterbodies of central California. Water Res 45, 752-762. https://doi.org/0.06/j. watres.200..032. Yang, WH, Lee HJ, Lee SY, Kim SG and Kim GB. 206. Heavy metal contents and food safety assessment of processed seaweeds and cultured lavers. J Korean Soc Mar Environ Energy 9, 203-20. https://doi.org/0.7846/jkosmee.206.9.3.203. 우리나라주요김생산해역에대한위생학적평가 35