1887

Microbiology Society logo

Volume 128, Issue 10

Interrelationships Between Physical State, Phenotypsic Stability and Transferability of β-Lactamase Genes in Free

Abstract

We have evaluated 66 -lactamase-producing (-Lac) clinical isolates of for the presence of plasmid DNA, ability to conjugate with an type b recipient, and stability of the -Lac phenotype. Of the 66 strains, 23 (35%) contained a plasmid demonstrable by either of two methods of cleared cell lysate preparation. The molecular weight of 17 of the plasmids was approximately 30 × 10. Four strains carried a plasmid of mol.wt 3 × 10 and single strains carried plasmids of mol. wt 36 × 10 or 50 × 10. Of 23 plasmids that were unstable , 10 (43%) had molecular weights of 30 × 10 and coded only for -lactamase. Stability of the -Lac phenotype was observed in all 43 strains in which plasmid DNA was not demonstrated by physical methods. When -Lac,‘plasmid-free’ strains were conjugated with strain Eagan ( type b), ampicillin-resistant transconjugants were isolated in 21 out of 43 matings (the lower limit of detection was 10 per donor cell per 30 min mating). With few exceptions, homologous (type b × type b) crosses with -Lac, plasmid-containing donors were more efficient (frequency 10–10) than heterologous (untypable × type b) matings (frequency 10–10). -Lac,‘plasmid-free’ strains were inefficient donors (frequency 10–10) irrespective of serotype, but all transconjugants from these crosses were efficient donors (frequency 10–10) and contained a plasmid of mol. wt 30 × 10 that was unstable in 9 cases out of 26 (35%). The increase in transferability and phenotypic instability of plasmid-bearing -Lac strains reinforces the concept of a chromosomal locus for -lactamase genes in‘plasmid-free’ -Lac strains. Carriage of -lactamase genes on 30 × 10 mol. wt R plasmids aids transferability at the expense of genetic stability.

  • Received:
  • Revised:
  • Published Online:
© Society for General Microbiology 1982

Erratum

An erratum has been published for this content:
CORRIGENDA
Loading

Article metrics loading...

/content/journal/micro/10.1099/00221287-128-10-2353
1982-10-01
2024-05-05
Loading full text...

Full text loading...

/deliver/fulltext/micro/128/10/mic-128-10-2353.html?itemId=/content/journal/micro/10.1099/00221287-128-10-2353&mimeType=html&fmt=ahah

References

  1. Alexander H. E., Leidy G. 1953; Induction of streptomycin resistance in sensitive Hemophilus influenzae by extracts containing desoxyribonucleic acid from resistant Hemophilus influenzae. Journal of Experimental Medicine 97:17–31
     [Google Scholar]
  2. Anderson P., Johnston R. B. 1972; Human serum activities against Hemophilus influenzae type b. Journal of Clinical Investigation 51:31–38
     [Google Scholar]
  3. Bendler J. W. III 1976; Physical size of the donor locus and transmission of Haemophilus influenzae ampicillin resistance genes by deoxyribonucleic acid-mediated transformation. Journal of Bacteriology 125:197–204
     [Google Scholar]
  4. Broda P. 1979 Plasmids pp. 12–22 Oxford & San Francisco: W. H. Freeman;
     [Google Scholar]
  5. Daum R. S., Syriopoulou V. PH., Smith A. L., Scheifele D. W., Willard J. E. 1981; Loss of plasmid DNA coding for β-lactamase during experimental Haemophilus influenzae infection. Journal of Infectious Diseases 143:548–553
     [Google Scholar]
  6. Daum R. S., Zutter M., Johnson E. J., Willard J. E. 1982; Pathogenic efficiency of plasmid containing ampicillin resistant H. influenzae type b. Abstract 961 Pediatric Research 16:239A
     [Google Scholar]
  7. Degraaff J., Elwell L. P., Falkow S. 1976; Molecular nature of two beta-lactamase-specifying plasmids isolated from Haemophilus influenzae type b. Journal of Bacteriology 126:439–446
     [Google Scholar]
  8. Elwell L. P., DeGraaff J., Seibert D., Falkow S. 1975; Plasmid-linked ampicillin resistance in Haemophilus influenzae type b. Infection and Immunity 12:404–410
     [Google Scholar]
  9. Elwell L. P., Saunders J. R., Richmond M. H., Falkow S. 1977; Relationships among some R plasmids found in Haemophilus influenzae. Journal of Bacteriology 131:356–362
     [Google Scholar]
  10. Farrar W. E., O’Dell N. 1974; Beta-lactamase activity in ampicillin-resistant Haemophilus influenzae. Antimicrobial Agents and Chemotherapy 6:625–629
     [Google Scholar]
  11. Gunn B. A., Woodall J. B., Jones J. F., Thorns-Berry C. 1974; Ampicillin-resistant Haemophilus influenzae. Lancet 2:845
     [Google Scholar]
  12. Hansen J. B., Olsen R. H. 1978; Isolation of large bacterial plasmids and characterization of the P2 incompatibility group plasmids pMGl and pMG5. Journal of Bacteriology 135:227–238
     [Google Scholar]
  13. Harkess N. K., Murray M. L. 1978; Restriction enzyme analysis of plasmids from Haemophilus influenzae. Antimicrobial Agents and Chemotherapy 13:802–808
     [Google Scholar]
  14. Hedges R. W., Jacob A. 1974; Transposition of ampicillin resistance from RP4 to other replicons. Molecular and General Genetics 132:31–40
     [Google Scholar]
  15. Heffron F., Rubens C., Falkow S. 1975a; Translocation of a plasmid DNA sequence which mediates ampicillin resistance: molecular nature and specificity of insertion. Proceedings of the National Academy of Sciences of the United States of America 72:3623–3627
     [Google Scholar]
  16. Heffron F., sublett R., Hedges R. W., Jacob A., Falkow S. 1975b; Origin of the TEM beta- lactamase gene found on plasmids. Journal of Bacteriology 122:250–256
     [Google Scholar]
  17. Jahn G., Laufs R., Kaulfers P. M., Kolenda H. 1979; Molecular nature of two Haemophilus influenzae R-factors containing resistances and the multiple integration of drug resistance transposons. Journal of Bacteriology 138:584–597
     [Google Scholar]
  18. Laufs R., Fock R. 1979; Characterization of genes specifying ampicillin resistance in bacterial isolates using a single-strand specific nuclease for analysis of plasmid DNA-DNA duplexes. Journal of General Microbiology 111:233–237
     [Google Scholar]
  19. Laufs R., Kaulfers P. M. 1977; Molecular characterization of a plasmid specifying ampicillin resistance and its relationship to other R-factors from Haemophilus influenzae. Journal of General Microbiology 103:277–286
     [Google Scholar]
  20. Laufs R., Kaulfers P. M., Jahn G., Teschner U. 1979; Molecular characterization of a small Haemophilus influenzae plasmid specifying β-lacta- mase and its relationship to R factors from Neisseria gonorrhoeae. Journal of General Microbiology 111:223–231
     [Google Scholar]
  21. Medeiros A. A., O’Brien T. F. 1975; Ampicillin resistant Haemophilus influenzae type b possessing a TEM-type beta-lactamase but little permeability barrier to ampicillin. Lancet 1:716–718
     [Google Scholar]
  22. Meyers J. A., Sanchez D., Elwell L. P., Falkow S. 1976; Simple agarose gel electrophoretic method for the identification and characterization of plasmid deoxyribonucleic acid. Journal of Bacteriology 127:1529–1537
     [Google Scholar]
  23. Morse S. I. 1980; The Hemophilus-Bordetella group. In Microbiology, 3rd edn. pp. 793–794 Davis B. D. others Edited by Hagarstown, Maryland: Harper & Row;
     [Google Scholar]
  24. Moxon E. R., Smith A. L., Averill D. R., Smith D. H. 1974; Haemophilus influenzae meningitis in infant rats after intranasal inoculation. Journal of Infectious Diseases 129:154–162
     [Google Scholar]
  25. O’Callaghan C. H., Morris A., Kirby S. M., Shingler A. H. 1972; Novel method for detection of β-lactamase by using a chromogenic cephalosporin substrate. Antimicrobial Agents and Chemotherapy 1:283–288
     [Google Scholar]
  26. Roberts M. C., Smith A. L. 1980; Molecular characterization of ‘plasmid-free’ antibiotic resistant Haemophilus influenzae. Journal of Bacteriology 144:476–479
     [Google Scholar]
  27. Saunders J. R., Sykes R. B. 1977; Transfer of a plasmid specified beta-lactamase gene from Haemophilus influenzae. Antimicrobial Agents and Chemotherapy 11:339–344
     [Google Scholar]
  28. Saunders J. R., Elwell L. P., Falkow S., Sykes R. B., Richmond M. H. 1978; β-Lactamases and R-plasmids of Haemophilus influenzae. Scandinavian Journal of Infectious Diseases 13s:16–22
     [Google Scholar]
  29. Slaney L., Albritton W. L. 1979; Transferable antibiotic resistance in Haemophilus influenzae. Abstract H79 79th Annual Meeting of the American Society for Microbiology, p. 132
     [Google Scholar]
  30. Stuy J. H. 1979; Plasmid transfer in Haemophilus influenzae. Journal of Bacteriology 139:520–529
     [Google Scholar]
  31. Stuy J. H. 1980; Chromosomally integrated conju- gative plasmids are common in antibiotic-resistant Haemophilus influenzae. Journal of Bacteriology 142:925–930
     [Google Scholar]
  32. Sykes R. B., Matthew M., O’Callaghan C. H. 1975; R-factor mediated β-lactamase production by Haemophilus influenzae. Journal of Medical Microbiology 8:437–441
     [Google Scholar]
  33. Vega R., Sadoff H. L., Patterson M. J. 1976; Mechanisms of ampicillin resistance in Haemophilus influenzae type b. Antimicrobial Agents and Chemotherapy 9:164–168
     [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/micro/10.1099/00221287-128-10-2353
Loading
Interrelationships Between Physical State, Phenotypsic Stability and Transferability of β-Lactamase Genes in Haemophilus influenzae
Microbiology 128, 2353 (1982); https://doi.org/10.1099/00221287-128-10-2353
/content/journal/micro/10.1099/00221287-128-10-2353
/content/journal/micro/10.1099/00221287-128-10-2353
Loading

Data & Media loading...

Most cited Most Cited RSS feed

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error