Comparison of Antibiotic Resistance of Blood Culture Strains and Saprophytic Isolates in the Presence of Biofilms, Formed by the Intercellular Adhesion (ica) Gene Cluster in Staphylococcus epidermidis

  • CHO BONG-GUM (Laboratory of Enteric Infections, Department of Microbiology, National Institute of Health, Department of Applied Biochemistry, College of Natural Sciences, Konkuk University) ;
  • KIM CHEORL-HO (National Research Laboratory for Glycobiology, Ministry of Science and Technology of Korean Government and Department of Biochemistry and Molecular Biology, Dongguk University COM) ;
  • LEE BOK KWON (Laboratory of Enteric Infections, Department of Microbiology, National Institute of Health) ;
  • CHO SEUNG-HAK (Laboratory of Enteric Infections, Department of Microbiology, National Institute of Health)
  • Published : 2005.08.01

Abstract

To elucidate the question of whether biofilm formed by the intercellular adhesion (ica) gene cluster has influences on antibiotic resistance in Staphylococcus epidermidis, we compared 124 skin strains with strains isolated from 50 blood cultures that cause septicemic diseases. The results revealed that the blood culture isolates were more resistant to the antibiotics tested than the saprophytic isolates. Moreover, antibiotic multiresistance was more prevalent in the clinical isolates. In the blood culture isolates, $46\%$ of the strains were resistant to three or more antibiotics, whereas only $12\%$ of the saprophytic isolates were resistant to three or more antibiotics. Interestingly, these characteristics were highly correlated with the biofilm formed by the ica gene cluster. In biofilm-producing strains, $84\%$ of the blood culture isolates and $44\%$ of the saprophytic isolates were antibiotic multiresistant, whereas only $22\%=;and\;9\%$, respectively, were antibiotic multiresistant in biofilm-nonproducing strains. Additionally, in the biofilm-producing ica-positive strains, $89\%$ of the blood culture isolates and $57\%$ of the saprophytic isolates were antibiotic multiresistant. However, the rate of the antibiotic multiresistance in the ica-negative strains was very low, thus indicating that the biofim formed by the lea gene cluster in S. epidermidis is an important pathogenic factor in association with the antibiotic multiresistance.

Keywords

References

  1. Anderl, J. N., M. J. Franklin, and P. S. Stewart. 2000. Role of antibiotic penetration limitation in Klebsiella pneumoniae biofilm resistance to ampicillin and ciprofloxacin. Antimicrob. Agents Chemother. 44: 1818-1824 https://doi.org/10.1128/AAC.44.7.1818-1824.2000
  2. Arciola, C. R., L. Baldassarri, and L. Montanaro. 2001. Presence of icaA and icaD genes and slime production in a collection of staphylococcal strains from catheter-associated infections. J. Clin. Microbiol. 39: 2151-2156 https://doi.org/10.1128/JCM.39.6.2151-2156.2001
  3. Baselga, R., J. Albizu, M. De la Cruz, E. Del Cacho, M Barberan, and B. Amorena. 1993. Phase variation of slime production in Staphylococcus aureus: Implications in colonization and virulence. Infect. Immun. 61: 4857-4862
  4. Bezek, D. M. 1998. Genus identification and antibiotic susceptibility patterns of bacterial isolates from cows with acute mastitis in a practice population. J. Am. Vet. Med. Assoc. 212: 404-406
  5. Chambers, H. F. 1988. Methicillin-resistant staphylococci. Clin. Microbiol. Rev. 1: 173-186 https://doi.org/10.1128/CMR.1.2.173
  6. Chang, M. M. and K. Merritt. 1992. Microbial adherence on poly(methyl methacrylate) (PMMA) surfaces. J. Biomed. Mater. Res. 26: 197-207 https://doi.org/10.1002/jbm.820260206
  7. Cho, S. H., K. Naber, J. Hacker, and W. Ziebuhr. 2002. Detection of the icaADBC gene cluster and biofilm formation in Staphylococcus epidermidis isolates from catheter-related urinary tract infections. Int. J. Antimicrob. Agents 19: 570-575 https://doi.org/10.1016/S0924-8579(02)00101-2
  8. Christensen, G. D., W. A. Simpson, A. L. Bisno, and E. H. Beachey. 1982. Adherence of slime producing strains of Staphylococcus epidermidis to smooth surfaces. Infect. Immun. 37: 318-326
  9. Christensen, G. D., W. A. Simpson, J. J. Younger, L. M. Baddour, F. F. Barrett, D. M. Melton, and E. H. Beachey. 1985. Adherence of coagulase-negative staphylococci to plastic tissue culture plates: A quantitative model for the adherence of staphylococci to medical devices. J. Clin. Microbiol. 22: 996-1006
  10. Christensen, G. D., L. Baldassarri, and W. A. Simpson. 1994. Colonization of medical devices by coagulase-negative staphylococci, pp. 45-78: In A. L. Bisno and F. A. Waldvogel (eds.), Infections Associated with Indwelling Medical Devices, 2nd Ed. ASM Press, Washington, D.C., U.S.A
  11. Chung, T. w., U. H. Jin, and C. H. Kim. 2003. Salmonella typhimurium LPS confers its resistance to antibacterial agents of baicalin of Scutellaria baicalensis george and novobiocin: Complementation of the rfaE gene required for ADP-L-glycero-D-manno-heptose biosynthesis of lipopolysaccharide. J. Microbiol. Biotechnol. 13: 564-570
  12. Emori, T. G. and R. P. Gaines. 1993. An overview of nosocomial infections, including the role of the microbiology laboratory. Clin. Microbiol. Rev. 6: 428-442 https://doi.org/10.1128/CMR.6.4.428
  13. Frebourg, N. B., S. Lefebvre, S. Baert, and J. F. Lemeland. 2000. PCR-based assay for discrimination between invasive and contaminating Staphylococcus epidermidis strains. J. Clin. Microbiol. 38: 877-880
  14. Galdbart, J. O., J. Allignet, H. S. Tung, C. Ryden, and N. El Solh. 2000. Screening for Staphylococcus epidermidis markers discriminating skin-flora strains and those responsible for infections of joint prostheses. J. Infect. Dis. 182: 351-355 https://doi.org/10.1086/315660
  15. Gerke, C., A. Kraft, R. SliBmuth, O. Schweitzer, and F. G6tz. 1998. Characterization of the N-acetylglucosaminyltransferase activity involved in the biosynthesis of the Staphylococcus epidermidis polysaccharide intercellular adhesin (PIA). J. Biol. Chem. 273: 18586-18593 https://doi.org/10.1074/jbc.273.29.18586
  16. Gristina, A. G, P. Naylor, and Q. Myrvik. 1988. Infections from biomaterials and implants: A race for the surface. Med. Prog. Technol. 14: 205-224
  17. Heilmann, C., C. Gerke, F. Perdreau-Remington, and F. Gotz. 1996. Characterization of Tn 917 insertion mutants of Staphylococcus epidermidis affected in biofilm formation. Infect. Immun. 64: 277-282
  18. Heilmann, C., O. Schweitzer, C. Gerke, N. Vanittanakom, D. Mack, and F. Gotz. 1996. Molecular basis of intercellular adhesion in the biofilm-forming Staphylococcus epidermidis. Mol. Microbiol. 20: 1083-1091 https://doi.org/10.1111/j.1365-2958.1996.tb02548.x
  19. Heilmann, C., M. Hussain, G Peters, and F. Gotz. 1997. Evidence for autolysin-mediated primary attachment of Staphylococcus epidermidis to a polystyrene surface. Mol. Microbiol. 24: 1013-1024 https://doi.org/10.1046/j.1365-2958.1997.4101774.x
  20. Henry, S. L. and K. P. Galloway. 1995. Local antibacterial theraphy for the management of orthopaedic infections. Pharmacokinetic considerations. Clin. Pharmacokinet. 29: 36-45 https://doi.org/10.2165/00003088-199529010-00005
  21. Hussain, M., M. Heilmann, C. von Eiff, F. Pedreau-Remington, and G. Peters. 1997. A 140-kilodalton extracellular protein is essential for the accumulation of Staphylococcus epidermidis strains on surfaces. Infect. Immun. 65: 519-524
  22. Jirku, V., J. Masak, and A. Cejkova. 2001. Reduced susceptibility of a model Saccharomyces cerevisiae biofilm to osmotic upshifts. J. Microbiol. Biotechnol. 11: 17-20
  23. Kloos, W. E. and T. L. Bannerman. 1994. Update on clinical significance of coagulase-negative staphylococci. Clin. Microbiol. Rev. 7: 117-140 https://doi.org/10.1128/CMR.7.1.117
  24. Lewis, K. 2001 Riddle of biofilm resistance. Antimicrob. Agents Chemother. 45: 999-1007 https://doi.org/10.1128/AAC.45.4.999-1007.2001
  25. MaCK, D., W. Fischer, A. Krokotsch, K. Leopold, R. Hartmann, H. Egge, and R. Laufs. 1996. The intercellular adhesion involved in biofilm accumulation of Staphylococcus epidermidis is a linear ${\beta}$-1,6-linked glucosaminoglycan: Purification and structural analysis. J. Bacteriol. 178: 175-183 https://doi.org/10.1128/jb.178.1.175-183.1996
  26. Nilsson, M., L. Frykberg, J. J. Flock, L. Pei, M. Lindberg, and B. Gruss. 1998. A fibrinogen-binding protein of Staphylococcus epidermidis. Infect. Immun. 66: 2666-2673
  27. Potera, C. 1999. Forging a link between biofilms and disease. Science 283: 1837-1838 https://doi.org/10.1126/science.283.5409.1837
  28. Rupp, M. E. and G. L. Archer. 1994. Coagulase-negative staphylococci: Pathogens associated with medical progress. Clin. Infect. Dis. 19: 231-245 https://doi.org/10.1093/clinids/19.2.231
  29. Sambrook, J., E. F. Fritsch, and T. Maniatis. 1989. Molecular Cloning: A Laboratory Manual, 2nd Ed. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N. Y., U.S.A
  30. Schumacher-Perdreau, P., C. Heilmann, G Peters, F. Gotz, and G. Pulverer. 1994. Comparative analysis of a biofilmforming Staphylococcus epidermidis strain and its adhesionpositive, accumulation-negative mutant M7. FEMS Microbial. Lett. 117: 71-78 https://doi.org/10.1111/j.1574-6968.1994.tb06744.x
  31. Shin, J. W, J. K. Kang, K. J. Jang, and K. Y. Kim. 2002. Intestinal colonization characteristics of Lactobacillus spp. isolated from chicken cecum and competitive inhibition against Salmonella typhimurium. J. Microbiol. Biotechnol. 12: 576-582
  32. Stewart, P. S. 2002. Mechanisms of antibiotic resistance in bacterial biofilms. Int. J. Med. Microbiol. 292: 107-113 https://doi.org/10.1078/1438-4221-00196
  33. Stone, G, P. Wood, L. Dixon, M. Keyhan, and A. Matin. 2002. Tetracycline rapidly reaches all the constituent cells of uropathogenic Escherichia coli biofilms. Antimicrob. Agents Chemother. 46: 2458-2461 https://doi.org/10.1128/AAC.46.8.2458-2461.2002
  34. Tack, K. J. and L. D. Sabath. 1985. Increased minimum inhibitory concentrations with anaerobiasis for tobramycin, gentamicin, and amikacin, compared to latamoxef, piperacillin, chloramphenicol, and clindamycin. Chemotherapy 31: 204-210 https://doi.org/10.1159/000238337
  35. Tanaka, G, M. Shigeta, H. Komatsuzawa, M. Sugai, H. Suginaka, and T. Usui. 1999. Effect of the growth rate of Pseudomonas aeruginasa biofilms on the susceptibility to antimicrobial agents: Beta-Iactams and tluoroquinolones. Chemotherapy 45: 28-36 https://doi.org/10.1159/000007162
  36. Van de Belt, H., D. Neut, W Schenk, J. R. van Horn, H. C. van Der Mei, and H. C. Busscher. 2001. Staphylococcus aureus biofilm formation on different gentamicin-loaded polymethylmethracrylate bone cements. Biomaterials 22: 1607-1611 https://doi.org/10.1016/S0142-9612(00)00313-6
  37. Walters III, M. C., F. Roe, A. Bugnicourt, M. J. Franklin, and P. S. Stewart. 2003. Contributions of antibiotic penetration, oxygen limitation, and low metabolic activity to tolerance of Pseudomonas aeruginosa biofilms to ciprotloxacin and tobramycin. Antimicrob. Agents Chemother. 47: 317-323 https://doi.org/10.1128/AAC.47.1.317-323.2003
  38. Zabinski, R. A., K. J. Walker, A. J. Larsson, J. A. Moody, G W. Kaatz, and J. C. Rotschafer. 1995. Effect of aerobic and anaerobic environments on anti staphylococcal activities of five fluoroquinolones. Antimicrob. Agents Chemother. 39: 507-512 https://doi.org/10.1128/AAC.39.2.507
  39. Ziebuhr, W, C. Heilmann, F. G6tz, P. Meyer, K. Wilms, E. Straube, and J. Hacker. 1997. Detection of the intercellular adhesion gene cluster (ica) and phase variation in Staphylococcus epidermidis blood culture strains and mucosal isolates. Infect. Immun. 65: 890-896
  40. Ziebuhr, W, V. Krimmer, S. Rachid, J. Loessner, F. Gotz, and J. Hacker. 1999. A novel mechanism of phase variation of virulence in Staphylococcus epidermidis: Evidence for control of the polysaccharide intercellular adhesin synthesis by alternating insertion and excision of the insertion sequence element IS256. Mol. Microbial. 32: 345-356 https://doi.org/10.1046/j.1365-2958.1999.01353.x