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Journal of Clinical Microbiology logoLink to Journal of Clinical Microbiology
. 1995 Dec;33(12):3146–3149. doi: 10.1128/jcm.33.12.3146-3149.1995

Screening for antimicrobial resistance in fecal samples by the replica plating method.

M Osterblad 1, T Leistevuo 1, P Huovinen 1
PMCID: PMC228661  PMID: 8586690

Abstract

Replica plating can be used for the detection of antibiotic resistance in normal flora. We have evaluated this application of the replica plating method by comparing it with a five-colony method. The replica plating method uses a single plate for each antibiotic, with a concentration just above that for borderline resistance. By the five-colony method, five colonies per sample were picked, chosen to represent all different colony morphologies present, and MICs were determined by a standard agar dilution method. The gram-negative, aerobic floras of 131 fecal samples were screened for resistance to ampicillin, cefuroxime, nalidixic acid, trimethoprim, sulfamethoxazole, and tetracycline by both methods. The rate of resistance detection by the two methods did not differ statistically for any of the antibiotics tested. The breakpoint concentrations used for the replica plates in the study gave results similar to those produced by the agar dilution method and the breakpoint values of the National Committee for Clinical Laboratory Standards and can thus be recommended. As the only currently used resistance detection method, replica plating facilitates an exact determination of the percentage of resistant colonies/total number of colonies (between 1 and 100%) in a sample. This revealed an uneven distribution, with only 23% of the samples having resistance frequencies in the range of 10 to 85%; usually, the resistant flora either was a small minority or was very dominant in samples with resistance. This phenomenon was present for all of the antibiotics.

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Selected References

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  1. Amyes S. G., Tait S., Thomson C. J., Payne D. J., Nandivada L. S., Jesudason M. V., Mukundan U. D., Young H. K. The incidence of antibiotic resistance in aerobic faecal flora in south India. J Antimicrob Chemother. 1992 Apr;29(4):415–425. doi: 10.1093/jac/29.4.415. [DOI] [PubMed] [Google Scholar]
  2. Bonten M., Stobberingh E., Philips J., Houben A. High prevalence of antibiotic resistant Escherichia coli in faecal samples of students in the south-east of The Netherlands. J Antimicrob Chemother. 1990 Oct;26(4):585–592. doi: 10.1093/jac/26.4.585. [DOI] [PubMed] [Google Scholar]
  3. Cooke E. M., Ewins S., Shooter R. A. Changing faecal population of Escherichia coli in hospital medical patients. Br Med J. 1969 Dec 6;4(5683):593–595. doi: 10.1136/bmj.4.5683.593. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cooke E. M. Properties of strains of Escherichia coli isolated from the faeces of patients with ulcerative colitis, patients with acute diarrhoea and normal persons. J Pathol Bacteriol. 1968 Jan;95(1):101–113. doi: 10.1002/path.1700950112. [DOI] [PubMed] [Google Scholar]
  5. Corpet D. E. Antibiotic residues and drug resistance in human intestinal flora. Antimicrob Agents Chemother. 1987 Apr;31(4):587–593. doi: 10.1128/aac.31.4.587. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Datta N. Drug resistance and R factors in the bowel bacteria of London patients before and after admission to hospital. Br Med J. 1969 May 17;2(5654):407–411. doi: 10.1136/bmj.2.5654.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Degener J. E., Smit A. C., Michel M. F., Valkenburg H. A., Muller L. Faecal carriage of aerobic gram-negative bacilli and drug resistance of Escherichia coli in different age-groups in Dutch urban communities. J Med Microbiol. 1983 May;16(2):139–145. doi: 10.1099/00222615-16-2-139. [DOI] [PubMed] [Google Scholar]
  8. Hartley C. L., Richmond M. H. Antibiotic resistance and survival of E coli in the alimentary tract. Br Med J. 1975 Oct 11;4(5988):71–74. doi: 10.1136/bmj.4.5988.71. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Heritage J., Dyke G. W., Johnston D., Lacey R. W. Selection of resistance to gentamicin and netilmicin in the faecal flora following prophylaxis for colo-rectal surgery. J Antimicrob Chemother. 1988 Aug;22(2):249–256. doi: 10.1093/jac/22.2.249. [DOI] [PubMed] [Google Scholar]
  10. Huovinen P., Mattila T., Kiminki O., Pulkkinen L., Huovinen S., Koskela M., Sunila R., Toivanen P. Emergence of trimethoprim resistance in fecal flora. Antimicrob Agents Chemother. 1985 Aug;28(2):354–356. doi: 10.1128/aac.28.2.354. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Katouli M., Lund A., Wallgren P., Kühn I., Söderlind O., Möllby R. Phenotypic characterization of intestinal Escherichia coli of pigs during suckling, postweaning, and fattening periods. Appl Environ Microbiol. 1995 Feb;61(2):778–783. doi: 10.1128/aem.61.2.778-783.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Katsanis G. P., Spargo J., Ferraro M. J., Sutton L., Jacoby G. A. Detection of Klebsiella pneumoniae and Escherichia coli strains producing extended-spectrum beta-lactamases. J Clin Microbiol. 1994 Mar;32(3):691–696. doi: 10.1128/jcm.32.3.691-696.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. LEDERBERG J., LEDERBERG E. M. Replica plating and indirect selection of bacterial mutants. J Bacteriol. 1952 Mar;63(3):399–406. doi: 10.1128/jb.63.3.399-406.1952. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Langlois B. E., Dawson K. A., Leak I., Aaron D. K. Effect of age and housing location on antibiotic resistance of fecal coliforms from pigs in a non-antibiotic-exposed herd. Appl Environ Microbiol. 1988 Jun;54(6):1341–1344. doi: 10.1128/aem.54.6.1341-1344.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Lester S. C., del Pilar Pla M., Wang F., Perez Schael I., Jiang H., O'Brien T. F. The carriage of Escherichia coli resistant to antimicrobial agents by healthy children in Boston, in Caracas, Venezuela, and in Qin Pu, China. N Engl J Med. 1990 Aug 2;323(5):285–289. doi: 10.1056/NEJM199008023230501. [DOI] [PubMed] [Google Scholar]
  16. Levy S. B., Marshall B., Schluederberg S., Rowse D., Davis J. High frequency of antimicrobial resistance in human fecal flora. Antimicrob Agents Chemother. 1988 Dec;32(12):1801–1806. doi: 10.1128/aac.32.12.1801. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Levy S. B. Microbial resistance to antibiotics. An evolving and persistent problem. Lancet. 1982 Jul 10;2(8289):83–88. doi: 10.1016/s0140-6736(82)91701-9. [DOI] [PubMed] [Google Scholar]
  18. Lewis M. J. Transferable drug resistance and other transferable agents in strains of Escherichia coli from two human populations. Lancet. 1968 Jun 29;1(7557):1389–1393. doi: 10.1016/s0140-6736(68)91973-9. [DOI] [PubMed] [Google Scholar]
  19. Linton K. B., Lee P. A., Richmond M. H., Gillespie W. A., Rowland A. J., Baker V. N. Antibiotic resistance and transmissible R-factors in the intestinal coliform flora of healthy adults and children in an urban and a rural community. J Hyg (Lond) 1972 Mar;70(1):99–104. doi: 10.1017/s0022172400022130. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Moorhouse E. C. Transferable drug resistance in enterobacteria isolated from urban infants. Br Med J. 1969 May 17;2(5654):405–407. doi: 10.1136/bmj.2.5654.405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Murray B. E., Mathewson J. J., DuPont H. L., Ericsson C. D., Reves R. R. Emergence of resistant fecal Escherichia coli in travelers not taking prophylactic antimicrobial agents. Antimicrob Agents Chemother. 1990 Apr;34(4):515–518. doi: 10.1128/aac.34.4.515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Rolland R. M., Hausfater G., Marshall B., Levy S. B. Antibiotic-resistant bacteria in wild primates: increased prevalence in baboons feeding on human refuse. Appl Environ Microbiol. 1985 Apr;49(4):791–794. doi: 10.1128/aem.49.4.791-794.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Shaw E. J., Datta N., Jones G., Marr F. M., Froud W. J. Effect of stay in hospital and oral chemotherapy on the antibiotic sensitivity of bowel coliforms. J Hyg (Lond) 1973 Sep;71(3):529–534. doi: 10.1017/s0022172400046519. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Sturtevant A. B., Cassell G. H., Bobo R. A., Feary T. W. Effect of antibiotic treatment on the incidence of infectious drug resistance among intestinal lactose-fermenting bacteria isolated from burn patients. Infect Immun. 1971 Mar;3(3):411–415. doi: 10.1128/iai.3.3.411-415.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Tullus K., Berglund B., Fryklund B., Kühn I., Burman L. G. Influence of antibiotic therapy on faecal carriage of P-fimbriated Escherichia coli and other gram-negative bacteria in neonates. J Antimicrob Chemother. 1988 Oct;22(4):563–568. doi: 10.1093/jac/22.4.563. [DOI] [PubMed] [Google Scholar]
  26. VOSTI K. L., MONTO A. S., RANTZ L. A. The importance of sample size in studies based upon the serologic classification of escherichia coli. Proc Soc Exp Biol Med. 1962 Oct;111:201–204. doi: 10.3181/00379727-111-27744. [DOI] [PubMed] [Google Scholar]

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