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1 J. Fd Hyg. Safety Vol. 22, No. 3, pp. 179~191 (2007),QWTPCN QH (QQF *[IKGPG CPF 5CHGV[ Available online at Î ÜôÚ ò Ù Â 1  2 Âú äâ Ö 3  Ùý* w w w, 1 w tœw, 2 w yw, 3 w yl w Survey of the Level of Microbial Contamination in Fish Farms on the Jeju-Island Young-Gun Moon, Jin-Hwan Ha 1, Chang-Hee Kang 2, Chun-Bok Song, Myung-Cheol Oh 3, and Moon-Soo Heo* Faculty of Marine Science, Cheju National University, Jeju , Korea 1 Dept. of Food Science and Technology, Cheju National University, Jeju , Korea 2 Dept. of Chemistry, Cheju National University, Jeju , Korea 3 Dept. of Tourism hotel Culinary Arts, Jeju college of Technology, Jeju , Korea (Received December 11, 2006/Accepted July 20, 2007) ABSTRACT The purpose of this study was to monitor and compare the contamination levels of heterotrophic and pathogenic (total s, Escherichia coli O157:H7, Salmonella spp., Vibrio parahaemolyticus, Staphylococcus aureus and Bacillus cereus). The samples collected at fish farms located in the Jeju-island were investigated from March to August in The contamination levels of S. aureus, B. cereus and Salmonella spp. ranged CFU/ml, CFU/ml and CFU/ml, respectively, and 85% of moist pellet feeds were contaminated with S. aureus, B. cereus and total s. But total s and E. coli O157:H7 were not detected in oliver flounder (Paralichthys olivaceus). For the production of safe oliver flounder (Paralichthys olivaceus), education of sanitation for employees, control of fish feed, and continuous monitoring for microorganism will be required. Key words: Fish farm in Jeju-island, Microbial contamination, pathogenic, Paralichthys olivaceus, fish farm sea water, feed õ w j» w» t y ƒ ƒ š. w» (WTO) s y t yƒ w x wù š. ü, x ùk ù t w ƒ j š w 1) t t t y». p, y y,, w *Correspondence to: Moon-Soo Heo, Faculty of Marine Science, Cheju National University, Jeju , Korea Tel: , Fax: msheo@cheju.ac.kr Ê ƒ» š w. t wù y š y wd š. ù w ƒw t w ƒw w ùküš 2). p e(paralichyhus olivaceus) ú y zù z q š ƒœt w. t»w ƒ y w ³ t ³ w ƒ 3-7). ù q w w ƒ wš. wš x t ³ ù ³ w 8). w 179
2 180 Young-Gun Moon et al. ³ tvù ƒ wš ³ w ƒ w j ƒ. ¾ ƒ t w j ƒ. w w w w ù w 2 w w., tvù ü w 2 w ú ƒ w. w p swš. s ƒ. š w tœ 9) wš t ³, š ³ s ³, l w w w w. Ì Ý ßá ò l 8 ¾ e 21 w (Fig. 1). ò Ì (,, ) 1L ³ (Y&K, KOREA) w x w š, w (expension or extruded pellet, EP) w (moist pellet, MP) v Polypropylene bag(daihan, KOREA) w w, x n w w w ( e) w. ³ w 4 o C w x w x w. Þ Ý ý w 1 ml w 9 ml ³ 0.85% w w, w e ³ w w z 1 g 9 ml ³ 0.85% ³ p(daihan) w ³ y k z w w. šx ³ y k z w w z 1 ml w w. ƒ w ³ Marine agar(difco, USA), ³ Micriobiology Chromocult Coliformen- Agar(Merck, Germany) w z ³ d w. Escherichia coli O157:H7 æð ³ ³ ƒ EC broth(bd, USA) w w 37 o C 24 ³ e z w š ³ ³ 100 µl w E. coli O157:H7 k Fig. 1. Location of the sampling site (black circle) in Jeju-island, Korea.
3 Survey of the Level of Microbial Contamination in Fish Farms on the Jeju-Island 181 MacConkey Sorbitol agar(difco) w w z 37 o C 24 w z, Sorbitol ww w White colony w x w, y x Eosin Methylene Blue agar(emb agar; BBL, USA) w y w. EMB agar ³ Gram staining, Triple sugar iron(tsi; Difco) test, Indol, Methyl red, Voges proskauer, citrate utilization(imvic) test, Lysine decarboxylase test, Motility test w w. ³ API 20E kit(biomerieux, France) w y x w. Salmonella spp. æð ³ ³ Lee et al. 10) w ƒ 1 ml selenite broth (Difco) w 37 C 24 ³w ³ o Salmonella spp. k MacConkey agar(difco) w 37 C 24 w o w³ y w. k ³ Triple Sugar Iron Agar(Difco) w w y x w, API 20E kit(biomerieux) w. Vibrio parahaemolyticus æð parahaemolyticus Bacteriological Analytical Manual 11) w w. yw x w w ³ Alkaline pepton water 30 o C 24 ³ wš k Thiosulfate, Citrate, Bile Salts, Sucrose(TCBS) agar(difco) w y w z y x k kw TSI agar(difco) w z y x w š, API 20NE(bioMerieux) w w. Staphylococcus aureus æð S. aureus ³ w 1 ml w 10% NaCl ƒ Tryptic Soy Broth(Difco) 9 ml z 37 o C 24 ³ w. ³ ³ Mannitol Salt Agar(BD) z w 37 o C 24 w z mannitol w y n ykw y w S. aureus y x w. š y x sq Gram w s ³ y w. w k ³ coagulase x w z API Staph(bioMerieux) w. Bacillus cereus æð B. cereus ³ w x Mannitol ww w colony x wš w ùy (Lecithinase ) w colony ky x w w 1 ml w Mannitol-Egg yolk-polymixin agar(oxoid, ENGLAND) w 30 o C 24 w y w. ŠŒ Ý ò ý Ñ Table 1, 2, l 8 ¾ 21 w w. v S1 S2 w w Table 1. Cell number of heterotrophic and pathogenic isolated in the fish farm waters in March S1 S2 S4 Sea water parahaemolyticus Salmonella spp. B. cereus S. aureus E. coli (CFU/mL) I* ND ND ND ND ND O*** ND ND ND ND I* ND ND ND ND ND O*** ND ND ND ND I* ND ND ND O*** ND I* ND ND ND ND ND O*** ND ND ND ND ND I* ND ND ND ND ND O*** ND ND ND ND
4 182 Young-Gun Moon et al. Table 1. Continued (CFU/mL) S10 S14 S21 Sea water parahaemolyticus Salmonella spp. B. cereus S. aureus E. coli I* ND ND ND ND O*** ND ND ND ND ND O*** ND ND ND ND I* ND ND ND O*** ND ND ND ND I* ND ND ND ND ND O*** ND ND ND ND ND I* ND ND ND O*** ND ND ND I* ND ND ND ND O*** ND ND I* ND ND ND ND ND ND O*** ND ND ND ND ND I* ND ND ND O*** ND ND ND I* ND ND ND ND O*** ND ND ND ND O*** N N N N N N N I* ND ND ND ND ND O*** ND ND ND ND ND I* ND ND ND 5, ND O*** ND ND ND I* ND ND ND ND ND O*** ND ND ND ND I* ND ND 2, ND ND O*** ND ND ND ND I* ND ND ND ND ND O*** ND ND ND ND I* ND ND ND ND O*** ND ND ND * I; In sea water, ** B; Breeding sea water, *** O; out sea water, N; No sampling, ND; Not detect.
5 Survey of the Level of Microbial Contamination in Fish Farms on the Jeju-Island 183 Table 2. Cell number of heterotrophic and pathogenic isolated in the fish farm waters in July (CFU/mL) S1 S2 S4 S10 S14 Sea water parahaemolyticus Salmonella spp. B. cereus S. aureus E. coli I* ND ND ND ND ND ND B** ND ND ND ND O*** ND ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND O*** ND ND ND ND ND I* ND ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND O*** N N N N N N N I* ND ND ND ND B** ND ND ND ND O*** ND ND ND ND O*** N N N N N N N I* ND ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND ND I* ND ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND ND O*** N N N N N N N I* ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND ND I* ND ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND ND I* ND ND ND ND ND B** ND ND ND ND O*** ND ND ND ND O*** N N N N N N N I* ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND ND
6 184 Young-Gun Moon et al. Table 2. Continued. (CFU/mL) S21 Sea water parahaemolyticus Salmonella spp. B. cereus S. aureus E. coli O*** N N N N N N N I* ND ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND ND ND I* ND ND 2, ND ND O*** ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND I* ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND ND * I; In sea water, ** B; Breeding sea water, *** O; out sea water, N; No sampling, ND; Not detect. Table 3. Cell number of heterotrophic and pathogenic isolated in the fish farm waters in August S1 S2 S4 Sea water parahaemolyticus Salmonella spp. B. cereus S. aureus E. coli (CFU/mL) I* ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND ND ND I* ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND ND I* ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND ND ND
7 Survey of the Level of Microbial Contamination in Fish Farms on the Jeju-Island 185 Table 3. Continued. S10 S14 Sea water parahaemolyticus Salmonella spp. B. cereus S. aureus E. coli (CFU/mL) I* ND ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND ND I* ND ND ND ND ND B** ND ND ND ND O*** ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND O*** ND ND ND ND I* ND ND ND ND ND B** ND ND ND ND O*** ND ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND ND ND I* ND ND ND ND B** ND ND ND ND O*** ND ND ND ND I* ND ND ND ND ND B** ND ND ND ND ND O*** ND ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND ND ND I* ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND ND ND I* ND ND ND ND ND ND B** ND ND ND ND ND ND O*** ND ND ND ND ND ND I* ND ND ND ND ND S21 B** ND ND ND ND ND O*** ND ND ND ND * I; In sea water, ** B; Breeding sea water, *** O; out sea water, N; No sampling, ND; Not detect. w wù wš. w 3 w ³ parahaemolyticusƒ ƒƒ CFU/mL, CFU/mL, ³ CFU/mLƒ, 7 8 w. 7 8 w S. aureusƒ, ù 7 8 ww ƒ š. 3 w ƒ ù 7 8 S. aureus wš 3 v œ w y w. S4 3 ³. š Salmonella spp. B. cereus, S. aureusƒ 8 8 ww
8 186 Young-Gun Moon et al. w w ƒ. 3 7 w,, ³ parahaemolyticusƒ ~ CFU/mL š 3 ³ CFU/ml ù 7 8 w ùkù. l(table 4) 7 w S. aureus B. cereusƒ š» S. aureusƒ w MP ƒ t x ù 2 ¾ w e ƒ. 3 parahaemolyticus, S. aureusƒ w 8 Table 4. Cell number of heterotrophic and pathogenic isolated in the fish farm feed in July (CFU/mL) Sea water E. coli Salmonella spp. B. cereus S. aureus parahaemolyticus S1 MP ND ND ND ND ND ND S2 MP ND ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND S4 MP ND ND ND ND ND MP ND ND ND EP ND ND ND ND ND MP ND ND ND ND EP ND ND ND ND ND MP ND ND ND ND EP ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND S10 MP ND ND ND ND ND MP ND ND ND ND EP ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND ND MP ND ND EP ND ND ND ND S14 MP ND ND ND ND ND MP ND ND ND ND EP ND ND ND ND ND MP ND ND ND ND EP ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND ND MP ND ND ND ND ND S21 EP ND ND ND ND ND ND MP; Moist pellet, EP; Extruded pellet, ND; Not detect
9 Survey of the Level of Microbial Contamination in Fish Farms on the Jeju-Island 187 ³ y Áò w ù» w. 3 v w ww parahaemolyticusƒ ù 7 8. ù 8 S. aureus, ³ w w 8 ü y w overflow x ù. y z w k ƒ w. w S. aureus, ³. 3 wš Table 5. Cell number of heterotrophic and pathogenic isolated in the fish farm feed in August (CFU/ml) Sea water E. coli Salmonella spp. B. cereus S. aureus parahaemolyticus S1 MP ND ND ND ND ND ND S2 MP ND ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND S4 MP ND ND ND ND ND ND MP ND ND ND EP ND ND ND ND ND ND MP ND ND ND ND EP ND ND ND ND ND MP ND ND ND ND EP ND ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND ND MP ND ND ND ND ND ND EP ND ND ND ND ND ND S10 MP ND ND ND ND ND MP ND ND ND ND EP ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND ND MP ND ND ND EP ND ND ND ND ND S14 MP ND ND ND ND ND MP ND ND ND ND EP ND ND ND ND ND MP ND ND ND ND EP ND ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND ND MP ND ND ND ND ND EP ND ND ND ND ND ND MP ND ND ND ND ND S21 EP ND ND ND ND ND ND MP; Moist pellect, EP; Extruded pellet, ND; Not detect.
10 188 Young-Gun Moon et al. Áòw r. overflow x. S10,,,, S14,,,,,,, S21 w S. aureus, B. cereus š ³. S. aureus ³, B. cereus ù,, w ù» w. 3, S4 8, š S w Salmonella spp.ƒ s, m,, ƒ. j ù w ù ƒ w» w. w,,,, ~S21 q w. š w w w ƒ w š. òì Ñ š EP, MP v Polypropylene bag w w x w w. ùkü l w w (Table 4, 5). MP EP S. aureusƒ. ü (š, ¾ù, ) w w, ³, z, k, yww, j» pellet wš, k w MPxk wš. yw w w» É ƒ. ù w MP» w w wš. EP 4 q wš s t» e EP ƒ w Table 6 ùkü. S1, S2, S4, S14 EP wš Table 6 t»w. ü ³ 12) œ ³ e, ƒ w ƒ w w ³ š ƒƒ w (%). (P) w 2.7% w w š w ³. swš wš w ³» ƒ w ù w ƒ l. w MP EP w ƒ w ƒ, s, w. MP w ù w w ù MP ƒ k ƒ w. EP w z û w. Ñ ùkü l e w w (Table 7, 8). ³, E. coli O157: H7, Salmonella spp., parahaemolyticus e. S. aureus, B. cereus e. e y k» w w. S. aureus, B. cereus ù w l qƒ e v w»ƒ 7, 8 Table 6. Nutrient composition(%) of commercial EP diets for olive flounder, Paralichthys olivaceus by marked on packing sheet. Nutrient Fish farm Crude protein Crude lipid Crude fiber Crude ash Ca P A Co , 5 B Co , 7, 8, 9, 11, 12, 20, 21 C Co , 15, 16, 17 D Co , 19 1 ; The foreign company Ë; above addition, ; below addition
11 Survey of the Level of Microbial Contamination in Fish Farms on the Jeju-Island 189 Table 7. Cell number of heterotrophic and pathogenic isolated in the Paralichyhus olivaceus in July (CFU/ml) S1 S2 S4 S10 S14 region parahaemolyticus Salmonella spp. B. cereus S. aureus E. coli Kidney ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND Kidney ND ND ND ND ND ND Kidney ND ND ND ND ND Liver ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND Kidney ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND S21 Liver ND ND ND ND ND; Not detect, w ƒ». p e v w,,,,, S21 k w ³ ƒ w, s kƒ. w q w. p w overflow x œ. w e ü B. cereus. S1,,,,,,, S21 e
12 190 Young-Gun Moon et al. Table 8. Cell number of heterotrophic and pathogenic isolated in the Paralichyhus olivaceus in August S1 S2 S4 S10 S14 region parahaemolyticus Salmonella spp. B. cereus S. aureus E. coli (CFU/ml) Kidney ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND Kidney ND ND ND ND ND ND Kidney ND ND ND ND ND Liver ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND Kidney ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ss 2 ND ND ND Liver ND ND ND ND ND ND Kidney ND ND ND ND ND S21 Liver ND ND ND ND ND ND; Not detect S. aureusƒ MP w w ù š w. l» l 8 ¾ 21 w w w,, e
13 Survey of the Level of Microbial Contamination in Fish Farms on the Jeju-Island 191 w. t ³ ³, š Salmonella spp., Escherichia coli O157:H7, Staphylococcus aureus, Bacillus cereus, Vibrio parahaemolyticus w. ww p w w w ƒ ü. w S. aureus, B. cereus and Salmonella spp. ƒƒ CFU/mL, CFU/mL and CFU/mL. w ü w w MP 85% S. aureus, B. cereus and total s. e w Escherichia coli O157:H7, total s. z w l x w ƒ š t»w w w t ƒe. ò 2005 HACCP w. Ù y : (2002). 2. Ryu, K.: Outbreaks of domestic and foreign food poisoning. Ind. Health. 197, (2004). 3. Ham, H.J., Jin, Y.H and Jung, Y.T.: Distribution of Vibrio parahaemolyticus in fishery products, sold at Garak wholesale market and serological characteristics of isolated strains. J. Fd. Hyg. Safty. 17, (2002). 4. Lee, H.K., Yoon, Y.H, Lee, S.S and Ha, K.H.: Biochemical characteristic of Vibrio isolated from cultured shellfish, Ruditapes philippinarum, and some species of wild shellfish. J. Kor. Soc. Microbiol, 33, (1998). 5. Lee. T.S., Lee, H.J, Oh, E.K, Son, K.T and Park, M.S.: Bacterial flora in oyster shellstock and larva in Koje and Tongyoung area. Bull. Natl Fish. Res. Dev. Inst, 55, (1999). 6. Wi, C.H., Kim, H.O, Song, K.C, Choi, J.I, Kim, J.O, Rho, U and Kim, S.J.: Bacteriological survey of shellfish growing waters on Kamak bay. Bull. Natl. Fish. Res. Dev. Agency, 50, (1995). 7. Yoo, H.D., Oh, E.G, Lee, D.S and Wi, C.H.: Sanitary survey of shellfish growing area in jeollanamdo. Tech. Rep. South Sea Fish. Inst., Korea, (2001). 8. Son, K.T., Oh, E.G, Lee, T.S, Lee, H.J, Kim, P.H and Kim, J.H.: Survey of sanitary indicative and pathogenic in fish farms on the Southern coast of Korea. J. Kor. Fish. Soc. 38, (2005). 9. tœ xz : tœ. t» ³ (1996). 10. Lee, J.M., Park, J.Y., Lee, R.H., Lee, M.S., Yoon, S.Y., Chung, D.H., Lee, J.M. and Oh, S.S.: Microbiological evaluation for HACCP guideline of Korean traditional cookies. J. Fd Hyg. Safety. 20, (2005). 11. Elliot, E.L., Kaysner, C.A. and Tamplin, M.L.: Vibrio cholerae, parahaemolyticus, vulnificus and other Vibrio spp. In: Food and Drug Administration, Bacteriological Analytical Manual, 7th ed., AOAC International, Arlington, pp (1992) y : (2005).
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