J. Fd Hyg. Safety 17(1), 31 35 (2002) ½ y s ³ s Á Á xá wá Á *Á½ t t t w Prevalence f Staphylcccus aureus in Kimbap Yunsk Kang, Sunkyung Yn, Seunghyeup Jwa, Dngha Lee, Gun-J W, Yngsik Park* and Changmin Kim 'PPE.JDPCJPMPHZ%JWJTJPO,PSFB'PPE %SVH"ENJOJTUSBUJPO4FPVM,PSFB *OEVTUSZ*OGPSNBUJPO%JWJTJPO,PSFB)FBMUI*OEVTUSZ%FWFMPQNFOU*OTUJUVUF4FPVM,PSFB ABSTRACT Accrding t fd pisning statistic data f 2000, the fd pisning utbreaks have ccurred mainly by meat (27.9%), shellfish and its prcessed prducts (26.0%), and ready-t eat meals (24.0%) such as Kimbap and packed lunch bxes. The majr causative fd pisning bacteria were Salmnella spp. (35.6%), Staphylcccus aureus (11.3%) and Vibri parahaemlyticus (3.2%). In this study, we cnducted the islatin and enumeratin fr S. aureus in Kimbap. This mnitring data will be applied t the fllwing study, risk assessment. The Kimbap samples were cllected frm department stres, cnvenient stres and snack bars lcated in Seul, Busan, Daejen, and Gwangju. The verall islatin rate f S. aureus frm 214 Kimbap samples was 34.1% and the average cunt was 623 cells. Entertxin typing test fr islates shwed 42.5%, 4.1% and 2.7% fr type A, B and C, respectively. There was n significant seasnal difference in S. aureus islatin, but the average cunt in summer(793 cfu/g) was 1.8 times higher than that f winter(446 cfu/g). Key wrds ý Staphylcccus aureus, Risk assessment, Kimbap ù, y s ³ w, ƒ. 2000 ü m, t q ƒœt w t(½, ) 24.0% wš, y Salmnella spp. w Staphylcccus aureusƒ 11.3% 2 wš. S. aureus y y ³ 1,2,3,4,5), t w š ³. ³ w w œ», m Ÿ w s wš š w v wš 6,7,8,9), t 10). S. aureus w Clstridium btulinum ƒ ³ tü w w x 11,12). s ³ ù entertxin w ³ S. aureus w. Entertxin 26,000~35,000 Da s rp 5,13), w 9ƒ, entertxin Authr t whm crrespndence shuld be addressed. A, B, C, D, E, F, G, H, I, J(SEA, SEB, SEC, SED, SEE, SEF, SEG, SEH, SEI, SEJ)x 14,15,25). Entertxin w S. aureus x wƒ ƒ x w š16,17,18), x jù, Ax ù D w ƒ š š 12,16,19). entertxin trypsin, chymtrypsin, renin papain wz w w 10), ü w x 100 C 30 ƒ w q 5,12,14). Entertxin w w ƒ w q w ù k entertxin 1µg 14,20,21), w w ³ ww ew. wr, y ƒ t w S. aureus entertxin s w š š, 25~50%ƒ y s ³ ³, 15~20% entertxin ³wš 21). S. aureus t S. aureus, ³ entertxin 31
32 :VOTPPL,BOHFUBM w 5), t w w w. S. aureus s t w, t ³»zƒ» y s ³ ƒ w, w w w 5)., ü t wù ½ y s ³ q w š, ³ w x w, ƒ w y s ³ w. t ³» Á³ w ù, ü ¾ w ƒ, x w m ü w sƒ wš, ½ y s ³ w» Á³ w» y wš w. x ½ y s ³ q w 2000 1 l 2000 9 ¾,,, Ÿ m ½, y, r máq xk ù 214 w w. t ³ S. aureus ATCC 13565, S. aureus ATCC 19095, S. aureus ATCC 27664 w. x 25 g ³ w 10% NaCl ƒ tryptic sy brth (Difc, USA) 225 ml š stmacher 1 ³ y k z 37 C 24 ³ w. ³ egg ylkƒ ƒ mannitl salt agar (Difc, USA) baird parker agar(difc, USA) ƒ ƒ w 37 C 16~24 z x ù kü w. Brain heart infusin agar(difc, USA) sq w z, gram, cagulase x, API Staph(Bi-Meriux, France) w w 22,23). x (0.85% NaCl) 10 w ³ y w -3¾ z 9ml ³ 1ml w -1 10 10 wš ƒ k baird parker agar egg ylkƒ ƒ mannitl salt agar 100 µl 3 w 37 C, 48 z x (Baird parker agar: clny y, mannitl salt egg ylk agar: clny y) w 22). y x Á x k x 10% NaCl ƒ tryptic sy agar(difc, USA) 37 C 18~24 w z, w s ³ y w, s ƒ ³ y ³ API Staph w w 24). ³ w EDTAƒ ƒ cagulase plasma(bbl, USA) w cagulase x w, q ³ S. aureus w. Entertxin y x Entertxin y x reversed passive latex agglutinatin kit(set-rpla, Denka seiken, Japan) w 15). ³ Brain heart infusin(bhi) brth 18 wš 8,000 g 20 w w z 10 w w. 25 µl micrplate 5 ƒwš ƒ latex A, B, C, D latex 25 µl ƒwš 10 w 18~20 ew z, xk w x y w. m 214 ½ x (Table 1, Table 2), 73 y s ³ s³ 34.1% ³, 214 w s³³ 623 cfu/g.,, Ÿ ƒ ƒ 40%, 28%, 37%, 31 % ƒ ù, w. máq xk 42.8% y (25.7%) r (12.2%) (Fig 1). ƒ œ (53.1%) š, ƒ û œ r (0%). 1» 1~2 (s³» 0.45 C) 7~9 (s³» 26.3 C) 34% ù, 1~2 (s³» 0.45 C) s³ 7~9 (s³» 26.3 C) s³ ƒƒ 446 cfu/g,
1SFWBMFODFPG4UBQIZMPDPDDVTBVSFVTJO,JNCBQ 33 Table 1. Prevalence f Staphylcccus aureus frm Kimbap purchased at different regins in Krea N. f islates/samples (%) Regin N. f islates/samples (%) Restaurant C.V.S. a Department Stre Seul 21/52(40.4) 17/32(53.1) 1/9(11.1) 3/11(27.3) Busan 15/54(27.8) 13/36(36.1) 0/10(0.0) 2/8(25.0) Daejen 21/57(36.8) 16/37(43.2) 2/11(18.2) 3/9(33.3) Gwangju 16/51(31.4) 13/33(39.4) 2/11(18.2) 1/7(14.3) Islatin rate(%) 73/214(34.1) 59/138(42.8) 5/41(12.2) 9/35(25.7) a C.V.S., Cnvenience stre Table 2. Enumeratin f Staphylcccus aureus in Kimbap Ppulatin (cfu/g) N. f islates N. f islates Department Restaurant C.V.S. a Stre less than 100 41 31 4 6 100~1,000 17 14 1 2 1,000~10,000 12 11 - b 1 10,000~50,000 3 3 - - Ttal 73 59 5 9 a C.V.S., Cnvenience stre. b -, Nt detected. Fig. 3. Detectin rate f Staphylcccal entertxins Fig. 1. Islatin and enumeratin f S. aureus in Kimbab (Islatin rate: 34.1%, Ave. ppulatin: 623 cfu/g) Fig. 2. Islatin rate by seasn 793 cfu/g 1.8 ùkû (Fig 2). ³ 73 w entertxin x A type 42.5 %(31 /73 ), B type 4.1%(3 /73 ), D type 2.7%(2 / 73 ) s (Fig. 3). š m ½ w, q xk, y s ³,,,, Ÿ ƒ 28~40% w j ùkû. q xk ³ y (25.7%) r (12.2%) (42.8%) q š ƒw, w (s³ 903 cfu/g) y (67 cfu/g) m q ƒ ù, r (16 cfu/g) þ máq y ³ r ³ 4~50 ƒ û ùkû., ½ w ù, r þ máq ½ w k q.
34 :VOTPPL,BOHFUBM ³ w entertxin x, A type³ 42.5%(31 /73 ) ƒ ùkû, t w y s ³ entertxin type Aƒ š ew 16,19). w w 34% ùkû, ³ 1.8 ƒ ùkù w, p ƒ y s ³ w ƒw m ew 14). wz m ½ y s ³ w sƒ w sƒ ww, ½ w y s ³ w z» w v ƒ.» ( y: HMP-99- F-06-0001, t ƒ w w sƒ w ) w w w. 2000 ü m, t 27.9%, q ƒœt 26.0%, w t(½, ) 24.0% wš, ³ Salmnella spp.ƒ 28.8% 1, Vibri parahaemlyticusƒ 13.5% 2, Staphylcccus aureusƒ 8.7% 3 wš. ½ y s ³ w s w w sƒ» y wš w.,,, Ÿ y, r, w ½ 214 w y s ³ x, ³ 34.1% š, s³³ 623 cfu/g. ³ w entertxin x A type 42.5%(31 /73 ), B type 4.1%(3 /73 ), D type 2.7%(2 /73 ) s. w máq x k 42.8% y (25.8%) r (12.3%) ùkû, s³³ w ù, (793 cfu/g) (446 cfu/g) ³ 1.8 ùkû. š x 1. Kim, D.H., Kwn, K.R., Lee, K.H., Ju, Y.R., Oh, K.S. and Kark, H.S.: Studies n Staphylcccal fd-pisning and entertxin prductivity, Reprt f Natinal Institute f Health, 26, 111-121 (1989). 2. Ryu, P.Y., Kim, Y.I., Rhee, J.H., Chung, S.S., Ahn, T.H., Shin, J.H., Ryang, D.W., Kim, Y.H.: Antimicrbial resistance and plasmid prfile f methicillin-resistant S. aureus, J. Infectin, 27(1), 15-29 (1995). 3. Yi, D.H. and Hng, H.S.: Islatin and cultural cnditins f Actinmycetes strain prducing effective antibitic fr MRSA,», 24 (1999). 4. Jung, C.H. and Kang, H.J.: Antimicrbial susceptibilities and -lactamase prductin f S. aureus islated frm bulk milk and dmestic animals, Kr. J. Vet. Publ. Hlth., 17(3) (1993) 5. ½ š, ½ y, ½k,, : w, š, pp.251-256 (2000). 6. Suk, S.U. and Park, S.C.: Staphylcccal infectins, J. Infectin, 17(2), 115-122 (1985). 7. Kim, D.H., Kim, B.S., Lee, K.H., Ju, Y.R., Oh, K.S., Kark, H.S. and Chi. Y.K.: A study n diagnsis f Staphylcccal fd pisning and entertxin, The Reprt f Natinal Institute f Health, 27(1), 56-63 (1990). 8. Lee, K.U., Kim, Y.J., Lee, S.M., Lee, M.S. and Kim, B.H.: Nnstaining methd fr determinatin f gram reactins and cmparisin f the API Staph(DMS) and Cwan and Steel prcedures fr the identificatin f Staphylcccus and Micrcccus, Reprt f NIH Krea, 25, 277-288 (1988). 9. Ch, D.T.: Significance f methicillin-resistant S. aureus as a nscmial pathgen, J. Infectin, 27(1), 11-13 (1995). 10. : S. aureus entertxin e ph, š ª }Œ «. û w w (1992). 11. Kim, D.H., Kwn, K.R., Lee, K.H., Ju, Y.R., Oh, K.S. and Kark, H.S.: Study n Staphylcccal entertxin, Reprt f NIH Krea, 25, 297-307 (1988). 12. Atanassva, V., Meindl, A., Ring, C.: Prevalence f Staphylcccus aureus and staphylcccal entertxins in raw prk and uncked smked ham-a cmparisn f classical culturing detectin and RFLP-PCR, Int. J. Fd Micrbil., 68, 105-113 (2001). 13. Shin, H.K., Kim, J.B., Lee, J.H., Kim, Y.K. and Kim, C.K.: Develpment f the simple immunassays fr the measurement f Staphylcccal entertxins, J. Anim. Sci. Tech., (1993).
1SFWBMFODFPG4UBQIZMPDPDDVTBVSFVTJO,JNCBQ 35 14., y, y, ½, : t w, ƒ, pp.45-133 (1999). 15. Chen, T.R., Hsia, M.H., Chiu, C.S., Tsen, H.Y.: Develpment and use f PCR primers fr the investigatin f C1, C2 and C3 entertxin types f Staphylcccus aureus strains islated frm fd-brne utbreaks, Int. J. Fd Micrbil., 71, 63-70 (2001). 16. Hepner, E.: Fd pisning and Salmnella infectins in England and Wales, 1976-1978, Publ. Hlth. Lnd., 94, 337-349 (1980). 17. Nterman, S. and Heuvelman, C.J. : Cmbined effect f water activity, ph and sub-ptimal temperature n grwth and entertxin prductin f S. aureus, J. Fd Sci., 48, 1832-1840 (1983). 18. Vernn, E. and Tillett, H.E.: Fd pisning and Salmnella infectins in England and Wales, 1969-1972, Publ. H1th. Lnd., 88, 225-235 (1974). 19. Casman, E.P., Bennett, R.W., Drsey, A.E., and Issa, J.A.: Identificatin f a. furth Staphylcccal entertxin, entertxin D, J. Bacteril., 94, 1875-1882 (1967). 20. : The Effect f temperature and time n the multiplicatin f Staphylcccus in fds, Krean Jurnal f Public Health, 9(2), 381-387 (1972). 21. Brwn, M.H.: Meat micrbilgy, Applied Science Publishers. Lndn and New Yrk, pp.269-486 (1982). 22. Bacterilgical Analytical Manual, Chap. 12, Fd and Drug Administratin, 8th Ed. (1995). 23. tœ ( ), t t, pp. 107 (2000). 24. Nrinaga, M., Asak, K., Takashi, M., Masat, A.: An utbreak f fd pisning due t egg ylk reactin-nagative Staphylcccus aureus. Int. J. Fd Micrbil, 64, 361-366 (2001). 25. Mnday, S. R. and Bhach, G.A. : Use f multiplex PCR t detect classical newly described pyrgenic txin genes in staphylcccal islates, J. Clin. Micrbil., 37, 3411-3414 (1999).