Search

Resistance transfer factors are natural conjugative plasmids encoding antibiotic resistance. Some also encode mutagenic DNA repair genes giving resistance to DNA damage and induced mutagenesis. It has been shown that antibiotic resistance has been acquired by recent transposition events; however, we show here that mutagenic repair genes existed much earlier on these types of plasmids. Conjugative plasmids from eight incompatibility groups from the Murray collection of 'pre-antibiotic era' enterobacteria were tested for complementation of mutagenic repair-deficient Escherichia coli umuC36. Although none of these plasmids carry transposon-encoded drug resistance genes, IncI1 and IncB plasmids were identified which restored ultraviolet resistance and induced mutability to umuC36 mutants. Furthermore they increased the UV resistance and induced mutability of wild-type E. coli, Klebsiella aerogenes and Citrobacter intermedius, thus showing that they could confer a general selective advantage to a variety of hosts. Like known mutagenic repair genes, complementation by these plasmid genes required the SOS response of the host cell. Nucleotide hybridisation showed that these plasmids harboured sequences similar to the impCAB locus, the mutagenic repair operon of modern-day IncI1 plasmids. The evolution of mutagenic repair genes is discussed.

Following an outbreak of cross-infection with multiply-resistant Klebsiella aerogenes epidemiological studies demonstrated the association of the use of suction apparatus with the distribution of positive cases. Eighty per cent of 66 cases had been exposed to this equipment. Environmental and clinical isolates were compared with strains of klebsiella recovered from suction apparatus. Colonial morphology, klebecin sensitivity and production, antibiotic resistance pattern and determination of plasmid molecular weight and incompatibility were used to discriminate between strains. A disinfection policy has been introduced to cope with the use of suction apparatus.

SUMMARY: Phage Fo lac is an RNA-containing phage which plates only on strains carrying the plasmid EDP208, a pilus derepressed derivative of the unique incompatibility plasmid Fo lac. A host range mutant, phage Fo lac h, was selected which plated on strains carrying the ungrouped plasmid pPLS:: Tn5 and lysed strains carrying another ungrouped plasmid TP224::Tn10 or the Com9 plasmid R71. An RNA-containing phage, SR, was isolated from sewage on bacteria harbouring plasmid pPLS::Tn5. It was antigenically distinct from the above two phages but had the same host range as phage Fo lac h. Phages Fo lac h and SR adsorbed unevenly to the shafts of the conjugative pili. Another phage, SF, was filamentous and plated or propagated on strains carrying any of the above plasmids as well as on strains harbouring IncD or F-complex plasmids. Plasmids TP224::Tn10 and pPLS::Tn5 were compatible with representative plasmids of all Inc groups also encoding thick flexible pili. The four plasmids EDP208, R71, TP224::Tn10 and pPLS::Tn5 were compatible with one another except for the reaction of TP224::Tn10 in the presence of pPLS::Tn5 which was slightly ambiguous. The host ranges of the bacteriophages, together with the serological relatedness of the thick flexible pili determined by these four compatible plasmids, suggested that they constitute a new complex, here designated S.

The incidence of trimethoprim resistance in enterobacteria causing infection in a London hospital increased from 5.6% in 1970 to 16% in 1979. The proportion of gentamicinresistant aerobic Gram-negative bacilli had risen to 6.5% by 1979. During a 5-month period in 1977, during which no epidemic was recognized, all isolates resistant to either trimethoprim, gentamicin, tobramycin or amikacin were studied. The proportion of enterobacteria resistant to both trimethoprim and gentamicin (3.8% of the total) was significantly higher than expected assuming no correlation between acquisition of resistance characters. The resistance was transferable in 23 % of trimethoprim-resistant and 76% of gentamicin-resistant strains. Trimethoprim resistance was carried by plasmids of seven different incompatibility groups and in at least four instances was part of a transposon. Gentamicin resistance was determined by plasmids of three groups - IncC, IncFII and IncW. Transposition of gentamicin resistance was not shown, though this may have been the means of evolution of the gentamicin R plasmids of IncW, which determined aminoglycoside acetyl-transferase, AAC(3). Some bacterial strains with their plasmids were endemic. There was evidence for these plasmids (i) acquiring new resistance genes by transposition, (ii) losing resistance genes by deletion and (iii) being transferred between bacterial species in the hospital.

In an outbreak of hospital infection caused by Klebsiella aerogenes type K-16 isolates over a 3-month period carried, apparently unaltered, a cryptic 90-Megadalton (Md) plasmid (unclassified) and a multiple-resistance 65-Md plasmid of IncM. The IncM plasmid, identified in environmentally related strains of Citrobacter koseri and Escherichia coli, showed minor variations from that in the klebsiella vector. The IncM plasmids, as well as all wild host strains cured of the IncM plasmids, carried a transposable DNA sequence, encoding trimethoprim and, in every case but one, streptomycin resistance. This transposon appeared identical with Tn7, previously identified in unrelated plasmids in bacteria from different environments.

Colicin factors are thought to fall into two taxonomic groups which differ in, amongst other properties, the molecular weight of the plasmid DNA and the host range of the colicin protein. This hypothesis is supported by the plasmids found in 26 colicinogenic strains. Two small Col factors may have arisen from larger factors, judging from similarities between their colicins.

SUMMARYKlebsiella of capsular type K2 were investigated to find out whether a single epidemic clone was the source of many outbreaks of infection in different hospitals, in different areas over a period of five years. The klebsiellas studied were found to be very similar; they were of the same biotype, had similar klebecin sensitivity patterns and carried multiple drug-resistance plasmids; however, characterization of these plasmids showed that they were heterogeneous. Thus there was not a single epidemic bacterial clone.

Phage pilH alpha was specific for bacterial strains, of various genera, harbouring plasmids of the HI and HII incompatibility groups. Plaque formation was temperature sensitive in that plaques formed at 26 degrees C but not at 37 degrees C. Plaques were fairly clear, irregular in outline and varied from pin point to about 2 mm in diameter on all hosts where plaques were detected. The phage had an isometric hexagonal outline with a diameter of 25 nm. It contained RNA but differed from all but one other plasmid-dependent RNA phage by being sensitive to chloroform. It adsorbed along the length of the shafts of IncHI and HII plasmid-coded pili.

During an outbreak of hospital infection, 71 multiply-resistant klebsiella strains were classified by two extra tests in addition to biotyping, serotyping and sensitivity testing. These were klebecin sensitivity tests (Edmondson & Cooke, 1979) and screening for plasmid DNA molecules by agarose gel electrophoresis of single colony lysates (Eckhardt, 1978). There were 56 examples of an epidemic strain of serotype K21; ten of these differed from 46 identical strains in some minor character. The other 15, although resembling the epidemic strain in biotype and antibiotic resistance differed from it sufficiently to be considered as unrelated.

R plasmids of a new incompatibility group, IncHII, determined the constitutive production of H pili, had high molecular weights, and determined tellurite resistance. They were designated IncHII because, during incompatibility tests, they sometimes eliminated or were eliminated by, previously described IncH plasmids, which they resembled in several respects. Nevertheless, stable and separate coexistence, i.e., compatibility, with plasmids of IncH1, IncH2, and IncH3 was demonstrated. The latter subgroups, members of which are all incompatible with one another, were distinguished on the basis of DNA-DNA hybridization experiments ( N. D. F. Grindley, G. O. Humphreys, and E. S. Anderson, 1973 , J. Bacteriol. 115, 387–398; A. F. Roussel, and Y. A. Chabbert, 1978 , J. Gen. Microbiol. 104, 269–276.); it is proposed that they be called IncHI, the subgroups being HI1, HI2, and HI3.

Conjugative plasmids were common in enterobacteria isolated before the medical use of antibiotics. Plasmid F of Escherichia coli K-12 was one example and we identified others in over 20% of a collection of strains isolated between 1917 and 1954, the Murray collection. In the past 25 years, conjugative plasmids encoding antibiotic resistances have become common in bacteria of the same genera as those of the Murray Collection--Salmonella, Shigella, Klebsiella, Proteus, Escherichia. The present study was made to show whether the 'pre-antibiotic' plasmids belonged to the same groups, as defined by incompatibility tests (Inc groups), as modern R plasmids. Of 84 such plasmids established in E. coli K-12, none with antibiotic resistance determinants, 65 belonged to the same groups as present resistance (R) plasmids. Thus the remarkable way in which medically important bacteria have acquired antibiotic resistance in the past 25 years seems to have been by the insertion of new genes into existing plasmids rather than by the spread of previously rare plasmids.