Isolation of Tetracycline-resistant Lactic Acid Bacteria from Kimchi

김치에서 tetracycline 내성 유산균의 분리

  • Kang, Hyo-Jin (Department of Microbiology, College of Natural Sciences, Kyungpook National University) ;
  • Kim, Byung-Chun (Department of Microbiology, College of Natural Sciences, Kyungpook National University) ;
  • Park, Wan (Department of Microbiology, College of Natural Sciences, Kyungpook National University)
  • 강효진 (경북대학교 자연과학대학 미생물학과) ;
  • 김병천 (경북대학교 자연과학대학 미생물학) ;
  • 박완 (경북대학교 자연과학대학 미생물학과)
  • Published : 2004.03.01

Abstract

Tetracycline resistant bacterial strains were isolated from 10 batches of Kimchi among 50 batches collected in Taegu restrict. The MIC of tetracycline ranged between 25 and> 100 ㎖/l. Total genomic DNA preparation from all 10 tetracycline resistant lactic acid bacterial isolates were subjected to PCR amplification with class-specific primers for tet(M) and tet(O). In only one isolate, HJ9, tet(M) was detected. By Southern blotting and hybridization with a tet(M)-specific probe, the tet(M) gene of HJ9 isolate could be localized on a plasmid. The partial nucleotide sequence and deduced amino acid sequence of tet(M) of HJ9 showed 90-99% and 94-100% homology to those of Gram positive bacteria, respectively. With sequencing of 16S rRNA, HJ9 isolate from Kimchi was identified as Lactobacillus sakei. From these results, Kimchi can be considered potential vehicle for the spread of antibiotic-resistant lactic acid bacteria along the food chain to the consumer.

대구지역에서 수집한 50점의 김치 중에서 10점의 김치로부터 tetracycline내성 세균을 분리하였다. 이 균주들의 tetracycline에 대한 MIC는 25-100 mg/l 이상의 범위로 분포하였으며 다른 항생제에 대한 내성도 다양하였다. tet(M), tet(O)특이적 primer를 이용한 PCR에서 HJ9 한 균주에서만 tet(M)유전자가 검출되었으며, tet(M)은 플라스미드상에 존재하는 것으로 나타났다. HJ9 균주의 tet(M)부분 염기서열을 분석한 결과 기존에 보고된 Gram양성균의 tet(M)의 DNA 염기서열 및 아미노산 서열과 각각 90-99%, 94-100%의 높은 상동성을 보였다. 165 rRNA 염기서열을 분석한 결과 HJ9 균주는 Lactobacillus sakei로 동정되었다. 김치를 통하여서도 항생제 내성 유전자의 전파 확산이 가능할 것으로 생각된다.

Keywords

References

  1. Kor. J. Anim. Nutr. Feed. v.12 항생제 및 약품에 대한 내성세균의 문제 마점술
  2. Kor. J. Appl. Microbiol. Bioeng. v.13 유산균 plasmid DNA의 신속 간편한 분리방법 백영진
  3. 제 83차 대한미생물학회 학술대회 초록집 항생제 내성세균의 문제점 이연희
  4. Kor. J. Microbiol. Biotechnol. v.31 벼과식물로부터 질소고정세균의 분리와 nitrogenase 활성 측정 최은화;이상은;윤기순;권덕기;손재근;박승환;한명숙;김사열
  5. Diag. Microbiol. Inf. Dis. v.37 Comparison of antimicrobial resistance phenotypes and resistance genes in Enterococcus faecalis and Enterococcus faecium from humans in the community, broilers, and pigs in Denmark Aarestrup,F.M.;Y.Agerso;P.Gerner-Smidt;M.Madsen;L.B.Jensen https://doi.org/10.1016/S0732-8893(00)00130-9
  6. FEMS Microbiol Lett. v.214 The identification of a tetracycline resistance gene tet(M), on a Tn916-like tansposon, in the Bacillus cereus group Agerso,Y;L.B.Jensen;M.Givskov;M.C.Roberts https://doi.org/10.1111/j.1574-6968.2002.tb11355.x
  7. Proc. KSAM Fall Meeting Health benefits of yogurt cultures Baek,Y.J.
  8. Microbiol. Mol. Biol. Rev. v.65 Health benefits of yogurt cultures Baek,Y.J. https://doi.org/10.1128/MMBR.65.2.232-260.2001
  9. Antimicrob. Agents Chemother v.41 New tetracycline resistance determinants coding for ribosomal protection in Streptococci and nucleotide sequence of tet(T) isolated from Streptococcus pyogenes A498 Clermont,D.;O.Chesneau;G.D.Cespedes;T.Horaud
  10. Trends Microbiol. v.3 Unconstrained bacterial promiscuity: the Tn916-Tn1545 family of conjugative transposons Clewell,D.B.;S.E,Rlannagan;D.D.Jaworski https://doi.org/10.1016/S0966-842X(00)88930-1
  11. EMBo J. v.22 Mehanism of TET(O)-mediatied tetra-cycline resistance Connell,S.R.;C.A.Trieber;G.P.Dinos;E.Einfeldt;D.E.Taylor;K.H.Nierhaus https://doi.org/10.1093/emboj/cdg093
  12. Plasmid v.48 Characterization of the tetracycline rsistance plasmid pMD5057 from Lactobacillus plantarum 5057 reveals a composite structure Danielsen,M. https://doi.org/10.1016/S0147-619X(02)00118-X
  13. Int. J. Food Microbiol v.82 Susceptibility of Lactobacillus sp. to antimicrobial agents Danielsen,M.;A.Wind https://doi.org/10.1016/S0168-1605(02)00254-4
  14. Int. J. Food Microbiol v.64 Lactic acid bacterial diversity in the traditinal Mexican fernented dough pozol as determined by 16S rRNA sequence analysis Escalante,A.;C.Wacher;A.Farres https://doi.org/10.1016/S0168-1605(00)00428-1
  15. Plasmid v.37 Isolation and characterization fo a plasmid from Lactobacillus fermentum conferring erythromycin resistance Fons,M.;T.Hege;M.Ladire;P.Raibaud;R.Ducluzeau;E.Maguin https://doi.org/10.1006/plas.1997.1290
  16. Syst. Appl. Microbiol v.24 Antimicrobial susceptibility of intestinal bacteria from Swiss poultry flocks before the ban of antimicrobial growth promoters Frei,A.;D.Goldenberger;M.Teuber https://doi.org/10.1078/0723-2020-00004
  17. Syst. Appl. Microbiol. v.23 Isolation and identification of tet-racycline resistant lactic acid bacteria from pre-packed sliced meat products Gervers,D.;G.Huys https://doi.org/10.1016/S0723-2020(00)80015-6
  18. FEMS Microbiol. Lett In vitro conjugal transfer of tetracycline resistance form Lactobacillus isolates to other gram-positvie bacteria Gervers,D.;G.Huys;J.Swings
  19. Appl. Environ. Microbiol. v.69 Molecular characterization of tet(M) genes in Lactobacillus isolates from different types of fermented dry sausage Gervers,D.;M.Danielsen;G.Huys;J.Swings https://doi.org/10.1128/AEM.69.2.1270-1275.2003
  20. J. Gen. Appl. Microbiol v.26 Drug resistance plasmids in Lacto-bacillus fermentum Ishiwa,H.;S.Iwata https://doi.org/10.2323/jgam.26.71
  21. J. Dairy Sci. v.75 Characterization of Leu-conostoc isolates from commercial mixed strain mesophilic starter cultures Johanson,E.;A.Kibenich https://doi.org/10.3168/jds.S0022-0302(92)77865-5
  22. Int. J. Food Microbiol v.67 Anti-microbial susceptibility of starter culture bacteria used in Norwegian dairy products Katla,A.K.;H.Kruse;G.Johnsen;H.Herikstad https://doi.org/10.1016/S0168-1605(00)00522-5
  23. Appl. Environ. Microbiol v.64 Antibiotic resistance patterns of enterococci and occurrence of vancomycin-resistant entero-cocci in raw minced beef and pork in Germany Klein,G.;A.Pack;G.Reuter
  24. FEMS Microbiol. Lett. v.139 Intergeneric and intrageneric conjugal transfer of plasmids pAMal;pIL205 and pIP501 in Lactobacillus sake Lanella,P.;M.Zagorec;S.D.Ehrlich;F.Morel-Deville
  25. National committee for Clinical Laboratory Standards Performance standards for antimicobial disk susceptibility tests(6th ed.) National committee for Clinical laboratory Standards
  26. Plasmid v.35 The tetracycline resistance gene tet(M) exhibits mosaic structure Oggioni,M.R.;C.G.Dowson;J.M.Smith;R.Provvedi;G.Pozzi https://doi.org/10.1006/plas.1996.0018
  27. J. Food Prot. v.63 Vancomycin resistance and antibiotic susceptibility of enterococci in raw meat Pavia,M.;C.G.A.Nobile;L.Salpietro;I.F.Angelillo https://doi.org/10.4315/0362-028X-63.7.912
  28. J. Appl. Microbiol v.84 Antibiotic-resistant strains of Enterococcus isolated from Swedish and Danish retailed chicken and pork Quednau,M;S.Ahme;A.C.Petersson;G.Molin https://doi.org/10.1046/j.1365-2672.1998.00463.x
  29. Plasmid v.23 Characterization of plasmids and plasmid-borne macrolide resistance from Lactobacillus sp. strain 100-33 Rinckel,L.A.;D.C.Savage https://doi.org/10.1016/0147-619X(90)90030-G
  30. FEMS Microbiol. Rev. v.19 Tetracycline resistance determinants; mechanisms of action, regulation of expression, genetic mobility, and distribution Roberts,M.C. https://doi.org/10.1111/j.1574-6976.1996.tb00251.x
  31. Antimicrob. Agents Chemother v.35 Tetracycline resistance in Pediococcus species Roberts,M.C.
  32. Molecular cloning: a laboratory manual 2nd ed. Sambrook,J.;E.F.Fritsch;T.Maniatis
  33. Antimicrob. Agents Chemother v.40 Tetracycline resistance mediated by ribosomal protection Taylor,D.E.;A.Chau
  34. Plasmid v.31 Molecular characterization of a plasmid-borne (pGT633) erythromycin resistance determinant (ermGT) from Lactobacillus reuteri 100-163 Tannock,G.W.;J.B.Luchansky;L.Miller;H.Connell;S.Thodeandersen;A.A.Mercer;T.R.Kalenhammer https://doi.org/10.1006/plas.1994.1007
  35. Appl. Environ. Microbiol. v.43 Drug resistance plasmids in Lactobacillus acidophilus and Lactobacillus reuteri Vescovo,M.;L.Morelli;V.Bottazzi
  36. J. Food Prot. v.50 Resistance and sensitivity of meat lactic acid bacteria to antibiotics Vidal,C.A.;D.Collins-Thompson https://doi.org/10.4315/0362-028X-50.9.737
  37. Science v.279 Medical comsequences of antibiotic use in agriculture Witte,W. https://doi.org/10.1126/science.279.5353.996