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Plasmid-mediated production of a new colonization factor (F42) in enterotoxigenic Escherichia coli

Wanderley Dias da Silveira^I; Tomomasa Yano^II; Joao L. de Azevedo^III; Antonio F.P. de Castro^II

^IDepartamento de Biologia Celular, Universidade Estadual de Campinas, Caixa Postal619, 13081 Campinas, SP, Brasil. Send correspondence to W.D.S.
^IIDepartamento de Microbiologia e Imunologia, Universidade Estadual de Campinas, 13081 Campinas, SP, Brasil
^IIIInstituto de Genética, Escola Superior de Agricultura Luiz de Queiroz, Universidade de São Paulo, Caixa Postal 82, 13400 Piracicaba, SP, Brasil

ABSTRACT

Strain 567/7 of enterotoxigenic Escherichia coli (ETEC) isolated in São Paulo, SP, Brazil, from a pig with neonatal diarrhea was shown to produce thermostable enterotoxin (STa) and a new colonization factor, provisionally designated F42 (Yano et al., 1986). In this study, the presence of plasmids in strain 567/7 was correlated with resistance to several antibiotics, and with presence of a col factor, as well as with F42 adhesin and STa enterotoxin production. Using agarose gel electrophoresis techniques, six plasmids were found in this strain. A plasmid of 65.1 megadaltons (Md) codes for tetracyclin resistance. Resistance to streptomycin, kanamycin and colicin V production was coded by a 48.8 Md conjugative plasmid, whereas resistance to kanamycin only was also related to another small 5.8 Md plasmid. A nonconjugative 21.1 Md plasmid was shown by transformation to be responsible for coding both the F42 and STa virulence factors produced by strain 567/7. Two other plasmids (2.3 Md, and 2.6 Md) were considered to be cryptic.

Keywords: plasmid-mediated; colonization factor; enterotoxigenic; Escherichia coli.

REFERENCES

Azevedo, J.L. and da Costa, S.O.P. (1973). Exercfcios Práticos de Genética. EDUSP, 288 p.

Bastarrachea, F. and Willetts, W.S. (1967). The elimination by acridine orange of F30 from recombination-deficient strains of Escherichia coli K12. Genetics 59: 155-166.

Birnboin, H.C. and Doly, J. (1979). A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acid Res. 7: 1513-1522.

Bounchaud, D.H., Scavizzi, M.R. and Chabbert, Y.A. (1969). Elimination by ethidium bromide of antibiotic resistance in Enterobacteria and Staphylococci. J. Gen. Microbiol. 54: 417-425.

Dean, A.G., Ching, A.C., Williams, R.G. and Harden, L.B. (1972). Test for Escherichia coli enterotoxin using infant mice: Application in a study of diarrhea in children in Honolulu. J. Infect. Dis. 125: 407-411.

Elvell, L.P. and Shipley, P.L. (1980). Plasmid-mediated factors associated with virulence of bacteria to animals. Annu. Rev. Microbiol. 34: 465-496.

Evans, D.G., Silver, R.P., Evans, D.J., Jr., Chase, D.G. and Gorbach, S.L. (1975). Plasmid controlled colonization factor associated with virulence in Escherichia coli enterotoxigenic for humans. Infect. Immun. 12: 656-657.

Fairbrother, J.M., Lariviere, S. and  Lallier, R. (1986). New fimbrial antigen F165 from Escherichia coli serogroup 0115 strain isolated from piglets with diarrhea. Infect. Immun. 51: 10-15.

Gaastra, W. and  de Graaf, F.K. (1982). Host-specific fimbrial adhesins of noninvasive entero-toxigenic Escherichia coli strains. Microbiol. Rev. 46: 129-161.

Gyles, C.L., So, M. and Falkow, S. (1974). The enterotoxin plasmids of Escherichia colt J. Infect. Dis. 130: 40-49.

Hirota, Y. (1960). The effect of acridine dyes of mating factors in Escherichia coli. Genetics 46: 57-64.

Mandel, M. and Higa, A. (1970). Calcium dependent bacteriophage DNA infection. J. Mol. Biol. 53:154.

Maniatis, T. Tritch, E.F. and Sambrook, J. (1983). Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, 545 pp.

Meyers, J.A., Sanches, D., Elwell, L.P. and Falkow, I. (1976). Simple agarose electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. J. Bacteriol. 127: 1529-1537.

Orskov, I. and Orskov, F. (1983). Serology of Escherichia coli fimbriae.Prog. Allergy 33: 80-125.

Reis, M.H.L., Alfonso, M.H.T., Trabulsi, L.R., Mazaitis, A.J., Maas, R. and Maas, W.K. (1980). Transfer of a CFA/I-ST plasmid promoted by a conjugative plasmid in a strain of Escherichia coli of serotype 0128ac: H12. Infec. Immun. 29: 140-143.

Scaletsky, I.C.A., Monteiro da Silva, M.L., Reis, M.H.L., Alfonso, M.H.T. and Trabulsi, L.R. (1983). Ent plasmids, col plasmids and drug resistance in Escherichia coli strains of human origin. Rev. Brasil. Genet. VI: 1-14.

Smith, H.W. and Halls, S. (1968). The transmissible nature of the genetic factor in Escherichia coli that controls enterotoxin production. J. Gen. Microbiol. 52: 319-334.

Smith, H.W. and Linggood, M.A. (1972). Further observations on Escherichia coli enterotoxin with particular regard to those produced by atypical piglet strains and by calf and lamb strains: The transmissible nature of these enterotoxins and of a K antigen possessed by calf and lamb strains. J. Med. Microbiol. 5: 243-250.

Tomoeda, M., Inuzuka, M., Kubo, N. and Nakamura, S. (1968). Effective elimination of drug resistance and sex factors in Escherichia coli by sodium dodecyl sulfate. J. Bacteriol. 95: 1078-1089.

Winther, M.D., Pickard, D. and Dougan, D. (1985). A novel method for increasing bacterial cellular yields of a funbrial subunit polypeptide. FEMS Microbiol. Letters 28: 193-197.

Yano, T., Leite, D.S., Camargo, I.J.B. and Castro, A.F.P. (1986). A probable new adhesive factor (F42) produced by enterotoxigenic Escherichia coli isolated from pigs. Microbiol. Immunol.30:495-508.