Bacillus spp. & B. cereus Isolated in Dried Marine Products

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1 J. Fd Hyg. Safety 21(3), (2006) q s B. cereus ³ w w Á½ y Bacillus spp. & B. cereus Islated in Dried Marine Prducts Hee-Jin Ham and M-Sang Kim 4FPVM)FBMUI&OWJSPONFOUBM3FTFBSDI*OTUJUVUF4FPVM,PSFB 3FDFJWFE"VHVTU"DDFQUFE4FQUFNCFS ABSTRACT 37 Bacillus spp strains(43.0%=37/86), and 18 B. cereus(20.9%=18/86) were islated n 86 dried marine prducts. Each islates were B. cereus 48.6%(18/37), B. mycides 13.5%(5/37), B. cagulus 5.4%(2/37), B. firmus 5.4%(2/37), B. circulus 2.7%(1/37), B. stearthermphilus 2.7%(1/37), B. pumilus 2.7%(1/37), B. spp. 8.1%(3/37), and Brebacillus brevis 10.8%(4/37) etc. Key wrds : Dried marine prducts, Bacillus cereus ³ ³ Bacillus anthracis ƒ ù» w y ³x,,, s,,, jš. w 1) ³ p B. cereus, m, œ» m t y»zƒ ƒ t, t, w, ƒ œt,, ƒœt, š t w. 2) B. cereus w x mx w mx ü» ƒ š, m, ùkü,, q p š t š, x 8-16» ƒ š m,, É, w, v, t w t ù kù. 2) Bacillus spp. t, m t q jš š w ³. 3) B. cereus w t, 2) t, 3), 4) t, 5) š t, 6) z t, 7) w, 8), 9), 10) g 11) x, 2), 3) ƒ wz, 4) y, 6), 7) w p sü wz, 8) ³ ü 12) w y w š., g Authr t whm crrespndence shuld be addressed. w w k s w ³ B. cereus Bacillus spp. s q w q ƒœ t w z w» œg ww. x 2006 q ƒ ew û w w s 86 xw š,, (,, ) 24, k ( k, ) 20, es 11, e 7 š»k 24 ( s 5, Õ» 4, 4, s 3, yw 2, q 2, 1, ½ 1, w es 1, s 1). x ³ x t t tœ 7. x, 8. x w 13) xw. 2ml 18 ml Tryptic Sy Brth(Difc, USA) 37Û0.5 C 48 w ³w š, ³ 1 w Egg Ylk 5-10% Bacillus cereus selective supplement ƒw MYP Agar (Oxid, England) w 37Û0.5 C 18~24 w, Bacillus ³ w Tryptic Sy Agar wš w 159

2 160 AppCtyAfxfyiF RfyrDtx ³ y w z yw x e z API CHB kit(bimerieux, France) w ³ w. w x Disc diffusin methd w BBL t w š Natinal cmmittee fr Clinical Labratry Standards(NCCLS)(Vilanva, 1988) w q w. w x w B. cereus ³ 18 w w š w antimicrbial drugs discs Streptmycin(S), Kanamycin(K), Gentamycin(GM), Amxicillin/clavulanic acid(amc), Chlramphenicl(C), Cephalxin(CF), Ampicillin/sulbactam(SAM), Sulfamethxazle/trimethprim(SXT), Ciprflxacin(CIP), Ticarcillin (TIC), Tetracycline(Te), Amikacin(AN), Sulfnamides(S3), Ceftriaxne(CRO), Nalidixic acid(na), š Cefxitin (FOX) 16 w. š s 86 w x, 37 (43.0%=37/86) Bacillus sppƒ š B. cereus 18 (20.9%=18/86). w 37 Bacillus spp, B. cereusƒ 48.6%(18/37) ƒ š, B. mycides 13.5%(5/37), B. firmus 5.4%(2/37), B. cagulus 5.4%(2/ 37) š B. circulans 2.7%(1/37) ƒƒ ùkû (Table 1). 6) š Bacillus ³ w 81%(68/84) B. cereus, 17.9%(15/84) B. thuringiensis, š 1.2%(1/84) B. mycides w. wr 6) B. cereus B. thuringiensis, B. mycides w š šw. ½ 7) t B. cereus 5 C 50 C 35 C 40 C 6 C w w w üþ B. cereus ³ ƒ B. circulans 2 C w š šw x ew š, ½ 7) B. cereus ³ ƒ Ÿ w y š š,,, yw w y w ü s y» w š š w x ewš. wr, B. cereus y» s x w ƒ yw š ³ w w š 10 C w w üþ B. cereus ³ ƒ š š, þ. t 3) bacteria/g 4 k š, Bacillus spp. s w y y š z, w t m»w k. 3) w, ½ ³ 7) B. cereus yw ³ w š B. cereus x wš» š šw. 37 Bacillus spp w, 58.3%(14/24), k 40.0%(12/20), s 36.4% (4/11), e 14.3%(1/7), s 20.0%(1/5), Õ» 100%(4/4), š 25%(1/4) ƒƒ ù kûš, 18 B. cereus w, 25%(6/24), k 10%(2/20), s 27.3%(3/ 11), e 14.3%(1/7), s 20.0%(1/5), Õ» 50%(2/4), š 25%(1/4) ƒƒ ùkû (Table 2). Giffel 14) 1996 B. cereus w w, 35.6%(56/157), Table 1. Bacillus spp. strains islated and identified frm dried marine salted prducts in Garak whlesale market Identified Strains Dried Dried Dried Dried Dried Dried Dried Percentages Dried Dried Dried Dried Dried Dried Dried Alaska file red beka white green brwn squid pllack shells shrimp anchvy bnit saury Ttal pllack fishes algae squid bait algae algae (%) N. f samples Bacillus cereus B. mycides B. firmus B. cagulus B. circulans B. stearthermphilus B. pumilus Bacillus spp Brebacillus brevis Ttal

3 5fhtww 5hpƒp B wf pity7ƒtpiffƒtypiƒ i h %(3/5), ƒ 55.6%(5/9), 90%(9/10), 90% (9/10), w 100%(6/6), q k(pasta) 50%(4/8) ùkûš, Meena w 12), 18%(18/100), š» 10%(10/100), 8%(8/100), š» 7.1%(5/70), w 16.7%(5/30), sl m e 26.7%(4/15), g 20.0% (4/20) ùkû, Jang 9) ½ 38.6%(17/ 44), 42.9%(15/35), 29.0%(9/31), x 50.0%(13/26), ½ z 2) 13.6%(16 ), 20.7%(17 ) ƒƒ ùkû šw B. cereusƒ t ³. w, 9%ƒ, k 10.3%, 27%, ³ 2%, z 17%, š j 52% B. cereusƒ š. 3) 40%, 80%, 30% B. cereusƒ š š. 7) Table 2. Bacillus spp and B. cereus islates distributin n 32 dried marine prducts Prduct Name N. f samples N. f Bacillus spp N. f B. cereus Dried squids ( ) Dried alaska pllacks ( k) Dried file fishes ( es) Dried anchvy ( e) Dried whitebait ( s) Dried beka squid ( Õ») Dried shrimp ( ) Dried cd meat ( ) Dried green algae ( q ) Dried sea mussel ( yw) Dried red algae ( ½) Dried bnit ( s) Dried brwn algae ( ) Dried saury (w es) Ttal w 18 B. cereus ³ yw r, Hemlysisƒ 94.4%(17/18) ùkù Nur 15) Market samples 89.74%, Piyawan 16) x 95.5%(19/20) ew š, Mtility 22.2%(4/18)ƒ ùkù Nur 15) Market samples 74.35% ùkü. 18 ³, ribse 80%, glucse 100%, fructse 100%, N-acetylglucsamine 93.3%, arbutine 60%, esculine 100%, salicine 53.3%, maltse 100%, š trehalse 93.3% ƒƒ ùkù Giffel ) ribse 99%, glucse 100%, fructse 100%, N-acetylglucsamine 100%, arbutine 99%, esculine 100%, salicine 99%, maltse 100%, š trehalse 100% Giffel 1996 ribse 18) 97%, glucse 100%, fructse 100%, N-acetylglucsamine 97%, arbutine 85%, esculine 82%, salicine 65%, maltse 100%, š trehalse 100% ew š, ½ 4) glucse 100%, N- acetylglucsamine 100%, esculine 100%, maltse 100%, š trehalse 66.7% ew, Kim 11) esculine 100%, š trehalse 100% ew ù, ½ 4) salicine 0% ùkü (Table 3). B. cereus w, ùk ü w Amikacin(100%), Kanamycin(94.4%), Chlramphenicl(94.4%), Streptmycin(77.8%), š Ciprflxacin(77.8%) š, ü ùkü w Amxicillin/clavulanic acid(100%), Ticarcillin(100%), Ceftriaxne(100%), Cephalxin(94.4%), Ampicillin/sulbactam(94.4%), cefxitin(88.8%) š Sulfamethxazle/trimethprim (72.2%) (Table 4). ½ 2) t w B. cereus w Bacitracin (86.1%), Chlramphenicl(97.2%), Gentamicin(100%), Kanamycin(86.1%), Streptmycin(97.2%), š Vancmycin Table 3. Bichemical characteristics f Bacillus spp. islated frm dried salted marine prducts in Garak whlesale market *Numbers is % Strains Bacillus cereus B. mycides B. firmus B. cagulus B. circulans B. stearthermphilus B. pumilus Bacillus spp Breb.acillus brevis *1=glycerl, 2=erythritl, 3=D-arabinse, 4=L-arabinse, 5=D-ribse, 6=D-xylse, 7=L-xylse, 8=D-ardnitl, 9=methyl-βD-xylpyranside, 10=D-galactse, 11=D-glucse, 12=D-fructse, 13=D-mannse, 14=L-srbse, 15=L-rhamnse, 16=dulcitl, 17=insitl, 18=D-mannitl, 19=D-srbitl, 20=Methyl-aD-mannpyranside, 21=Methyl-aD-glucpyranside, 22=N-acetylglucsamine, 23=amygdaline, 24=arbutine, 25=esculine, 26=salicine, 27=D-cellbise, 28=D-maltse, 29=D-lactse, 30=D-mellibise, 31=D-saccharse, 32=D-trehalse, 33=inuline, 34=D-melezitse, 35=D-raffinse, 36=amidn, 37=glycgene, 38=xylitl, 39=gentibise. 40=D-turanse, 41=D-xylse, 42=D-tagatse, 43=D-fucse, 44=L-fucse, 45=D-arabitl, 46=L-arabitl, 47=ptassium glucnate, 48=ptassium 2-cetglucnate, 49=ptassium 5-cetglucnate.

4 162 AppCtyAfxfyiF RfyrDtx Table 4. Antimicrbial discs used fr diffusin susceptibility test f 18 strains B. cereus islated frm dried marine prducts Antimicrbial drugs Cncentratin Susceptible (%) Intermediate (%) Resistant (%) Streptmycin(S) 10mcg Kanamycin(K) 30mcg Gentamycin(GM) 10mcg Amxicillin/clavulanic acid(amc) 30ug Chlramphenicl(C) 30ug Cephalxin(CF) 30ug Ampicillin/sulbactam(SAM) 20ug Sulfamethxazle/trimethprim(SXT) 23.75mcg/1.25mcg Ciprflxacin(CIP) 5ug Ticarcillin(TIC) 75ug Tetracycline(Te) 30mcg Amikacin(AN) 30ug Sulfnamides(S 3 ) 300ug Ceftriaxne(CRO) 30ug Nalidixic acid(na) 30ug Cefxitin(FOX) 30ug (100%), Ampicillin(100%), Penicillin G(100%), š Rifampicin(69.5%) ü ùkü w, Ampicillin wš ew š, Meena 12), š»,, w, š g w, Ampicillin(100%), Bacitracin(100%), Clistin (100%), Ctrimxazl(100%), Cephtaxime(100%), š Carbenicillin(100%) ùkü ew, Kiymet w 10), Ciprflxacin ùkü ew ù, Amxicillin 15 9, Gentamicin ƒ ü ùkü ew. wr, ½ w w 8) B. cereus w, Tetracycline, Streptmycin, Kanamycin, Rifampicin, Chlramphenicl, Bacitracin, š Erythrmycin, Penicillin G, Cephalsprin, Ampicillin, Plymyxin B, Bacitracin ü ùkþ š ew. B. cereus ³ ü, AMC, CF, SAM, SXT, TIC, CRO, FOX ü ³ ƒ 4, AMC, CF, SAM, SXT, TIC, TE, S3, CRO, FOX, Table 5. The multiple drugs resistance pattern f B. cereus n 18 strains islated frm dried marine prducts The multiple drugs resistance pattern N. f strains AMC+CF+SAM+SXT+TIC+CRO+FOX 4 AMC+CF+SAM+SXT+TIC+TE+S 3 +CRO+FOX 2 AMC+CF+SAM+SXT+TIC+S 3 +CRO+FOX 2 AMC+CF+SAM+TIC+CRO+FOX 2 AMC+CF+SAM+SXT+TIC+TE+S 3 +CRO+FOX 1 AMC+CF+SAM+SXT+TIC+CRO+NA+FOX 1 AMC+CF+SAM+TIC+S 3 +CRO+FOX 1 AMC+CF+SAM+SXT+TIC+CRO 1 AMC+SXT+TIC+CRO 1 S+AMC+CF+SAM+SXT+TIC+CRO+NA+FOX 1 S+AMC+CF+SAM+TIC+CRO+NA+FOX 1 S+AMC+CF+SAM+TIC+CRO+FOX 1 Ttal 18 AMC, CF, SAM, SXT, TIC, S3, CRO, FOX š AMC, CF, SAM, TIC, CRO, FOX ü ³ ƒƒ 2 ùkû. AMC, TIC, š CRO 18 w 100% ü ùkþ (Table 5). s 86 w x, 37 (43.0%=37/86) Bacillus sppƒ 18 (20.9%=18/86) B. cereusƒ. 37 B. cereus 48.6%(18/37), B. mycides 13.5%(5/37), B. cagulus 5.4%(2/37), B.

5 5fhtww 5hpƒp B wf pity7ƒtpiffƒtypiƒ i h 163 firmus 5.4%(2/37), B. circulus 2.7%(1/37), B. stearthermphilus 2.7%(1/37), B. pumilus 2.7%(1/37), B. spp. 8.1%(3/37), š Brebacillus brevis 10.8%(4/37) ùkù ³ B. cereusƒ ƒ ùkù w. š x 1. p y,, 4.2. ³ (B. cereus), pp (1999). 2. ½ w, ½,,,, ½,, : t B. cereus k pattern, J. f Life Science, 14(4), (2004). 3. x, ½, k, ½x : ü t B. cereus s w, J. Anim. Sci. & Technl. (Kr.) 42(2), (2000). 4. ½,,, ½w : w B. cereus ³ ƒ w ƒ wz p w, Kr. J. f Micrbilgy, 33(2), (1997). 5., w yw w : t B. cereus w s, pp 1-32, (2004). 6. x, w t œw w : š t Txigenic B. cereus grup s y, pp 1-98, (2003). 7. ½, w w w, z t B. cereus w, pp 1-55, (2000). 8. ½, w yw w : w l w B. cereus w p s wz, pp 1-121, (2000). 9. Ji-Hyun Jang, N-A Lee, Gun-J W, and Jng-Hyun Park : Prevalence f B. cereus Grup in Rice and Distributin f Entertxin Genes, Fd Sci. Bitechnl., 15(2), (2006). 10. Kiymet guven, Mehmet burcin mutlu, and Ozgur avci : Incidence and characterizatin f B. cereus in meat prducts cnsumed in Turkey, J. f Fd Safety, 26, (2006). 11. Hye-jung Kim, Dng-sun Lee, and Hyun-dng Paik : Characterizatin f B. cereus islates frm Raw Sybean Spruts, J. f Fd Prtectin, 67(5), (2004). 12. B.S, Meena, K.N. Kapr and P.K. Agarwal : Occurence f Multi-drug Resistant B. cereus in Fds, J. Fd Sci. Technl., 37(3), (2000). 13. t t : tœ,, pp (2005). 14. M.C. Giffel, R.R. Beumer, S. Leijendekkers and F.M. Rmbuts : Incidence f B. cereus and B. subtilis in fds in the Netherlands, Fd Micrbilgy, 13, (1996). 15. M.A. Nur, M. Thami, and A.A.K. Khater : Identificatin and Characterizatin f B. cereus grup islated frm different surces, Egyptian J. Dairy Sci., 30, 1-14 (2002). 16. Piyawan Gasaluck, Keik Ykyama, Tshi Kimura, and Isa sugahara; The ccurrence f B. cereus in Lcal Thai Traditinal Fds, J. Antibac. Antifung. Agents, 24(5), (1996). 17. M.C. Giffel, R.R. Beumer, B.A. Slaghuis and F.M. Rmbuts : Occurrence and characterizatin f (psychrtrphic) B. cereus n farms in th e Netherlands., Netherlands Milk & Dairy J., 49, (1995). 18. M.C. Giffel, R.R. Beumer, M.H. Bnestr and F.M. Rmbuts : Incidence and characterizatin f B. cereus in tw dairy prcessing plants, Netherlands Milk & Dairy J., 50, (1996).