Microbiological Evaluation of Raw Vegetables

Transcription

1 J. Fd Hyg. Safety 21(4), (2006) ƒ x Áx Á yá½ Á Áš Á½ Á 1 y ƒ w w w Micrbilgical Evaluatin f Raw Vegetables SeungHye Jung, Myung Je Hur, Jeng Hwa Ju, Kyung-Ae Kim, Sung Suck Oh, Jng Myung G, Yng Hee Kim, Jengs Im 1 *ODIFPO.FUSPQPMJUBO*OTUJUVUFPG1VCMJD)FBMUIBOE&OWJSPONFOU3FTFBSDI (BDIPO6OJWFSTJUZPG.FEJDJOFBOE4DJFODF(SBEVBUFTDIPPMPGQVCMJDIFBMUI 3FDFJWFE/PWFNCFS"DDFQUFE%FDFNCFS ABSTRACT The purpse f this study is t evaluate micrbilgical cntaminatin f leafy vegetables. Ttal aerbic bacteria and clifrms were mnitred t get the cntaminatin levels and Staphylcccus aureus, Bacillus cereus, Clstridium perfringens, Escherichia cli, Escherichia cli O157:H7, Salmnella spp., Vibri parahaemlyticus, Listeria mncytgenes, Yersinia enterclitica, Campylbacter jejuni t detect pathgens with risk f fdpisning frm fresh vegetables. The clny cunt f ttal aerbes and clifrms was als perfrmed t determine the efficacy f washing with tab water by cmmn cnsumers. 124 samples which are divided int 8 kinds f vegetables - Sesame leaf, Drpwrt, Chinese cabbage, Krean leek, Lettuce, Crwn daisy, Pimpinella brachycarpa, Chicry were sampled in 2 whlesale markets in Inchen. Mean cunts f ttal aerbic bacteria fr individual vegetables ranged frm CFU/g t CFU/g and ttal clifrms were frm CFU/ g t CFU/g. Bth shw the peaks in summer n this study frm March t September. Decrease rates after washing with tab water averaged 81.0% and 82.5% in ttal aerbic bacteria and clifrm cunts respectively. Staphylcccus aureus was islated 8.1%, Bacillus cereus 14.5%, Clstridium perfringens 5.6%, Escherichia cli 18.5%. 11 samples shwed verlapped bacterial cntaminatin. Fr respective vegetables Staphylcccus aureus islated frm 0.0% t 22.2%, Bacillus cereus frm 0.0% t 29.4%, Clstridium perfringens frm 0.0% t 23.1%, Escherichia. cli frm 0.0% t 35.0%. Escherichia cli O157:H7, Salmnella spp., Vibri parahaemlyticus, Listeria mncytgenes, Yersinia enterclitica, Campylbacter jejuni were nt islated. This study is expected t be available as the reference fr the basal data f pathgens in fresh vegetables. Key wrds: leafy vegetables, ttal aerbes, clifrms, fdpisning pathgens, the efficacy f washing ù ù ù y, t w ƒ. ù 1) w š š ƒ w eš., y, y k w ƒ Á w w. w š ù, ¾» ù eš. 2-6) w, ƒ w ƒ Authr t whm crrespndence shuld be addressed.» ù w, n ƒ w, š, y 7) w ƒ l ƒ ù w»¾ ƒ. ù y m m w. w š, ù y œ w ww. y» ³ ³ w» š, ƒ z w. w, ³,» w w 250

2 .JDSPCJPMPHJDBM&WBMVBUJPOPG3BX7FHFUBCMFT 251 w t w ƒ š w, ³ w» y wš w. x l 9 ¾ x w. ƒƒ q w z x w, z 7 ü x w. Á ³ ƒ 4,8) š w w j» kwš, m š w» w. ³ tœ x w x w. 8) z y» ³ sƒ y» ³ d tœ ³ sq w. z ù 10 g 90 ml ³ ƒw z m f(bagmixer, Interscience, U.S.A.) w 1 ³ yw. z 10 g ƒ 3 5z w. ³ y -5¾ 1 ml ³ 9 ml 10-1 ~10 w. ƒ 1 ml 43~45 C SPCA(Standard Plate Cunt Agar, Oxid, England) 15 ml ³ rp w z 35Û1 C w» 48 w z w. z ³ sƒ ³ d tœ g p w w w. ³ w w š, ƒ 1 ml 43~45 C DLA(Desxychlate lactse agar, Oxid, England) 15ml ³ rp w z 35Û1 C w» 24 w x x w. Salmnella spp. 25 g w 225 ml peptne water(oxid, England) ƒwš m f w 1 ³ yw z 35 C 18Û2 ³ w. 0.1 ml w 10 ml Rappaprt-Vassiliadis(Oxid, England) w 42 C 24Û2 w. ³ XLDw (Oxid, England) w 35 C 24 w z, y x w. sq mw 35 C 18~24 w z, TSI w 35 C 24 w z Á w( ), ƒ (³ y ) H 2 S, urease, indle ³ w API 20E kit(bimerieux, France) y w. š, Salmnella O wx (Denka Seiken, Japan) Hwx (Difc, U.S.A.) w. Staphylcccus aureus 25 g w 225 ml 10% NaCl ƒ Tryptic Sy (Oxid, England) ƒw m f w 1 ³ yw z 35~37 C 16 ³ w. ³ Baird-Parker agar w 37 C 24~48 w. ykw y z y x w. z Bld agar(, w ) 37 C 18~24 w β-hemlysis y w z s ³ y wš, cagulase(bimerieux, France) x w. š API Staph kit(bimerieux, France) y z y s ³ q w. Vibri parahaemlyticus 25 g w 225 ml 3% APW ƒw z m f w 1 ³ yw z 35 C 18~24 ³ w. ³ TCBSw (Oxid, England) w 35 C 18~24 w. sq 2~4mm w w y x w. 0, 3, 8 10% NaCl ƒw Nutrient agar(oxid, England) 35 C 18~24 w ü xw 3, 8, 10% y w. TSI (Difc, U.S.A.) 35 C 18~24 w ƒ, šd y, ƒ, yy y wš Oxidase, ONPG y API 20E kit y w. Chlstridium perfrigenes 25 g 225 ml š m f w 1 ³ yw. ³ y 1 ml Cked Meat Medium(Oxid,England) w 37 C 18~24 x» w. TSC(Oxid, England)

3 252 4FVOH)ZF+VOHFUBM ³ w 37 C 18~24 x» w ykw y y x w. sq mw 37 C 18~24 y» w ³ y w. Bld agar 37 C 18~24 x» w z β-hemlysis y wš ³ y wš, rapid ID 32A(Bimerieux, France) w. Listeria mncytgenes 25 g Listeria enrichment brth(oxid, England) 225 ml ƒwš m f w 1 ³ yw z 30 C 24 ³ w. Oxfrd agar (Oxid, England) w 30 C 24 w z Bld agar w β-hemlysis,, catalase y w. mtility, mannitl w, rhanmnse w, xylse w y w. z ³ y š, CAMP test Staphlycccus aureus, Rhdcccus equi ùkù Listeria mncytgenes q w. E. cli E.cli O157:H7 25 g ³ EC(Oxid, England) 225 ml ƒwš m f w 1 ³ yw z 35 C 24 ³ w. EMB w w 35 C 24 wš, Ÿ k y TSA w 35 C 24 w. ³ y wš KIA A/A, gas, yy y w z Indle, Citrate, Urease y wš API 20E kit y w. ³ y ³ O157 wx w x x wš, O157 y ³ H7 x x x w. Yersinia enterclitica 25 g w 225ml PSBB ƒwš m f w 1 ³ yw z 10 C 10 w. ³ CIN w w 30 C 24 w. CIN w ƒ ƒƒ TSI w, 35 C 24 z šd š ƒ yy ƒ w ³ kw 25 C, 37 C y w. w, urea x, citrate ³ API 20E y w q w. Bacillus cereus 25 g w 225 ml ƒwš m f w 1 ³ yw z MYPw w 30 C 24 w. z ykw y y w. yw 24 w w. MYP w x w m w wš 30 C 24 z w s ¼ xk ³ y ³, nitratey, VP, β-hemlysis, x» s y w q w. Campylbacter jejuni 25 g Hunt (Oxid, England) 100 ml š m f w 1 ³ yw z 35 C y» 24~48 w. Campylbacter agar with bld(oxid, England) 24 z y», y» k bld agar 24 w. y» š y» catalase, xidase y wš, API Campy(Bimerieux, France) y w. š,» y» ³ ³ y» ³ ³ s³ Table 1.. y» ³ s³ ef CFU/g > ù Á CFU/g > CFU/g > CFU/g > CFU/g > à CFU/ Table 1. Cunts f aerbic bacteria and clifrms in fresh vegetables Vegetables Mean cunts f ttal aerbic bacteria 1) (CFU/g) Mean cunts f clifrms 2) (CFU/g) Sesame leaf Drpwrt Chinese cabbage Krean leek Lettuce Crwn daisy Pimpinella brachycarpa Chicry ) ttal 124 samples, 2) ttal 122 samples

4 .JDSPCJPMPHJDBM&WBMVBUJPOPG3BX7FHFUBCMFT 253 g> ù CFU/g. ³ s³ CFU/g > ef CFU/g > ù CFU/g > CFU/g > CFU/g > à CFU/g > ù CFU/g > CFU/g. ù» > > > > >ƒ > y» ³ d š2), y» ³, ³ >à 3) > ùkù w parsley, dill 9) ³ ƒ ƒ ƒ m w t ƒ q w. w t ƒ. x y» ³ ³ Õ ew. w, w w k š w, m ƒ dw. 2) ù» ³ 82.1% w MPN z y w w w MPN 10) 3) 100% š Vilet red bile Agar w 100%. y» ³ s³ r ƒ û ƒ» à 3 ( CFU/g) 7 ( CFU/g), ù 9 ( CFU/g) 7 ( CFU/g), 9 ( CFU/g) 6 ( CFU/g), 9 ( CFU/g) 7 ( CFU/g), 3 ( CFU/g) 6 ( CFU/g), 4 ( CFU/ g) 7 ( CFU/g), ù 3 ( CFU/g) 7 ( CFU/g), ef 9 ( CFU/g) 7 ( CFU/g) ùkû. y» ³ ƒ 3, 4 9 ƒ û š 6, 7 ƒ (Fig. 1). ³ s³ r ƒ û ƒ» à 3 ( CFU/g) 6 ( CFU/g), ù 3 ( CFU/g) 7 ( CFU/g), 3 ( CFU/g) 7 ( CFU/g), 9 ( CFU/ Fig. 1. Mnthly variatins f ttal aerbic bacterial cunts in individual fresh vegetables. Fig. 2. Mnthly variatins f clifrm bacterial cunts in individual fresh vegetables.

5 254 4FVOH)ZF+VOHFUBM g) 7 ( CFU/g), 3 ( CFU/g) 6 ( CFU/g), 4 ( CFU/g) 5 ( CFU/g), ù 3 ( CFU/g) 4 ( CFU/g), ef 9 ( CFU/g) 7 ( CFU/g) ùkù y» ³ w 7 e» y ew (Fig. 2). Ueda 2, 6, 9 y» ³ ³ j y š wš11), 3, 4, 5, 6 j y ù 7 j s š 9 û. w,,, ƒ, ƒ j ûš ³ w w š w. 10) y» ³ ³ z w Table 2 w. 5z z y» ³ ef 92.6%, 87.2%, 82.3%, à 79.0%, ù 77.5%, 77.3%, 76.6%, ù 71.7% ùkù, ƒ p, s³ 81.0%., 20z 5z w 6~7 z 2) š 4 4 cm 6 L š v p r w % ³ 12) û. w x ƒ, ƒ ƒ ww 80% w Table 2. Reductin in micrbial cunts after 5 times washing fresh vegetables with tab water Vegetables Reductin in ttal aerbic bacterial cunts (%) Reductin in clifrm bacterial cunts (%) sesame leaf Drpwrt Chinese cabbage Krean leek Lettuce Crwn daisy Pimpinella brachycarpa Chicry mean w. w, z ƒ 12)». š ƒ ƒ ƒw 13) w ù þ w š, ƒ w 3 e w ³ ƒ 95.5% 4) w ƒ ü, ee z w w. ³ 92.3%, ef 91.8%, 88.4%, 85.0%, à 82.3%, ù 79.2%, 66.4%, ù 63.8% ùkù p, s³ y» ³ w 82.5%. ³ w t w Á k ƒ ƒw j š.,» ³ 124 w w w š, Staphylcccus aureus ef 22.2%, 17.6%, 12.0%, à 10.0% š, ù,, ù 8.1%. 890 Staphylcccus spp.ƒ» 22.4%, 15.0% ù Staphylcccus aureus 4 š, x, q, dill, Nrway 14) w. ù,, q,, 15) w ü š w ƒœ, ) Bacillus cereus ef 22.2%, 29.4%, ù 20.0%, 15.4%, 16.0%, à 10.0% š, ù 14.5%. 36% w 11) Ueda û ù w. wr Bacillus cereusƒ w. 13) Clstridium perfringens ù 13.3%, 23.1%, 8.0% š ef,,, ù, à 5.6%. ù» 10.0%,

6 .JDSPCJPMPHJDBM&WBMVBUJPOPG3BX7FHFUBCMFT 255 Table 3. Bacterial species islated frm fresh vegetables N. f Islates(%) Vegetables Staphylcccus aureus Bacillus cereus Clstridium perfringens Escherichia cli Chicry Chinese cabbage Crwn daisy Drpwrt Krean leek Lettuce Pimpinella brachycarpa sesame leaf Ttal %, 8.0%, 8.0%, ù 28.0% 6) û š, ù wš k w». Escherichia cli ef 11.1%, 5.9%, 17.6%, ù 13.3%, 7.7%, 32.0%, Ã 35.0% š ù 18.5%. w r MacCnkey brth MPN Teague Levine agar(bbl) 86.1% 10) w MacCnkey brth MPN 65.6% 16) û. ù 2) w ù» 23.2%, w w. ù x 3M Petrifilm E. cli cunt xw 30% ù m yw blue clny w. w, 11% 11) ù ù 10,16) w w e. Escherichia cli t ³ x w ³ ƒ ù, ù 7) ƒ š, m y k š. w Bacillus cereus + Escherichia cliƒ 7, Clstridium perfringens + Bacillus cereusƒ 2, Staphylcccus aureus + Escherichia cliƒ 1, Staphylcccus aureus + Bacillus cereusƒ 1 11, w / (%) ef 66.7%, 0.0%, 33.3%, ù 40%, 20.0%, 13.3%, Ã 22.2% 0.0% ~ 66.7% w w ƒ ƒ š ƒ. Staphylcccus aureus, Bacillus cereus, Clstridium perfringens, Escherichia cli 4ƒ ³ w w. / ƒ 6 8/ %, 5 23/ %, 4 8/ %, 7 6/ %, 9 2/ %, 3 0/7 0.0%. 3 š 9 ûš 5,6 Table 4. Sample Numbers f verlapped islatin in fresh vegetables vegetables B. cereus + E. cli C. perfringens + B.cereus S. aureus + E. cli S. aureus + B. cereus Ttal sesame leaf Drpwrt Chinese cabbage Krean leek Lettuce Crwn daisy Pimpinella brachycarpa Chicry Ttal

7 256 4FVOH)ZF+VOHFUBM Table 5. Islatin rates f Staphylcccus aureus, Bacillus cereus, Clstridium perfringens, Escherichia cli mnthly Mnth N. f islates Ttal N. f islates(%) Ttal r y» ³ ³ ew,»» š» û» û w w. Escherichia cli O157:H7, Salmnella spp., Vibri parahaemlyticus, Listeria mncytgenes, Yersinia enterclitica, Campylbacter jejuni Escherichia cli O157:H7 g ù 17)ƒ 870 w š14) ù» w x 2). Salmnella spp. r, š10) w» 3.0% 2)ƒ,, 11,14) w. v w Vibri parahaemlyticus,, yás d w ƒ š q sww ù. Listeria mncytgenes, Yersinia enterclitica ³ š. 0.5% 14) w» 2,3) š, Yersinia enterclitica. y»³» ù y ù w ƒ ³ Campylbacter jejuni. ƒ,, y w,, w,,, w. 7), y, m, ƒœ,, y ƒ j 15) ƒ y z, y 5 z, qt. 13),» w, ƒœ,, fp w ƒ v w ƒ œ, w w v». ƒ 7) y ( d, t l,»y y, y)ƒ 100%, t y w w w x yw. 18) w y» ³ ³ q w» œwš w, ³ ƒ ³ w. w w t w ƒ ƒ w ³» y w» w. w, mw ƒ w š w. 7) ³ ƒ w w ³ x wš, ³ š w w xw ƒ ƒ w w w y w w q. y w. sƒw. w» w y» ³ ³ Staphylcccus aureus, Bacillus cereus, Clstridium perfringens, Escherichia

8 .JDSPCJPMPHJDBM&WBMVBUJPOPG3BX7FHFUBCMFT 257 cli, Escherichia cli O157:H7, Salmnella spp., Vibri parahaemlyticus, Listeria mncytgenes, Yersinia enterclitica, Campylbacter jejuni ³ w. w y» ³ ³ w ww ù z» w z w. Ã, ù,,,,, ù, ef w. ƒƒ w y» ³ s³ CFU/g l CFU/g š, ³ CFU/g l CFU/g ùkû. ƒ 3 l 9» š e. w z y» ³ 81.0% š ³ 82.5%. ³ Staphylcccus aureus 8.1%, Bacillus cereus 14.5%, Clstridium perfringens 5.6%, Escherichia cli 18.5%. 11ƒ ƒ ³. ƒƒ w Staphylcccus aureusƒ š 0.0% 22.2%, Bacillus cereusƒ 0.0% 29.4%, Clstridium perfringens 0.0% 23.1%, Escherichia cli 0.0% 35.0%. Escherichia cli O157:H7, Salmnella spp., Vibri parahaemlyticus, Listeria mncytgenes, Yersinia enterclitica, Campylbacter jejuni. w w» y ƒ w». š x 1., ½,,, w,,»y: w w l, J. Fd Hyg. Safety, 16(4): (2001), 2.,,, ûx,,, w, ½, :» ³, Ÿ t t, 21(2002), 3.,, x,, y, ³y, ½ š, w, ½, w : m sƒ, J. Fd Hyg. Safety, 20(1): (2005), 4. ½, ³, w :, w t wz, 19(2): (2004). 5., w û, «v,, ½, ¼, ³: m ³ k, The Annual Reprt f KFDA, 7: 364~ 365 (2003). 6. š: y š,» ( ) Clstridium perfringens, KFDA, 14 (2002). 7. Brackett,R.E.: Incidence, cntributing factrs, and cntrl f bacterial pathgens in prduce. Pstharvest Bilgy and Technlgy, 15(3): (1999). 8. t t : tœ, t t, (2005). 9. Aycicek,H., Oguz,U.,Karci,K.: Determinatin f ttal aerbic and indicatr bacteria n sme raw eaten vegetables frm whlesalers in Ankara, Turkey. Internatinal Jurnal f Hygiene and Envirnmental Health, 209(2): (2006). 10. Garcia-Villanva,R.B. and Galvez,V.R.: Cntaminatin n fresh vegetables during cultivatin and marketing. Internatinal Jurnal f Fd Micrbilgy, 4(4): (1987). 11. Ueda, S. and Kuwabara, Y.: Bacterilgical study n fresh vegetables. Jurnal f Antibacterial and Antifungal Agents, Japan, 26(12): (1998). 12. Adams,M.R., Hartley,A.D. and Cx,L.J.: Factrs affecting the efficacy f washing prcedures used in the prductin f prepared salads. Fd Micrbilgy, 6(2): (1989), 13. ž:, ª}, 23(2): (2003). 14. Jhannessen,G.S., Lncarevic,S. and Kruse,H.: Bacterilgical analysis f fresh prduce in Nrway. Internatinal Jurnal f Fd Micrbilgy, 77(3): (2002). 15. Burnett,S.L. and Beuchat,L.R.: Human pathgens assciated with raw prduce and unpasteurized juices and difficulties in decntaminatin. Jurnal f Industrial Micrbilgy & Bitechnlgy, 25(6): (2000). 16. Kiran,P.A., Kamat,D.B.: Micrbilgical quality f fresh leafy vegetables, salad cmpnents and ready-t-eat salads: an evidence f inhibitin f Listeria mncytgenes in tmates. Internatinal Jurnal f Fd Sciences and Nutritin, 52(1): (2001). 17. Hammnd, R.M., Bdager, D., Ward, K.V. and Rwan, A.: Case studies in fdbrne illness in Flrida frm fresh prduce. HrtScience, 36(1): (2001). 18. ž «: Á w š, ŽƒŽ, 31(1): (2003).