Narrow host range of some streptococcal R plasmids.
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Biomedical subjects
Publications and source records attributed to T Horodniceanu.
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Of 20 clinical isolates of group A, B, G, D (Streptococcus bovis), and viridans streptococci, 5 transferred their antibiotic resistance markers into streptococcal recipients at a low frequency (10(-4) to 10(-8)) in the apparent absence of extrachromosomal elements. All strains carried genetic markers for high-level resistance to streptomycin, kanamycin, neomycin, lividomycin A, and ribostamycin, as well as resistance to macrolides and related drugs, tetracycline, and chloramphenicol.
Ten isolates of Streptococcus faecium were found to be resistant to penicillin, tetracycline, macrolides and related drugs, streptomycin, and kanamycin, and four strains were resistant to chloramphenicol. Six of these 10 strains transferred all their resistance markers (except penicillin) by conjugation at a low frequency (10(-7) to 10(-9)). Several plasmids of different molecular weights were found in each of the wild-type strains. In 5 of 11 transconjugant strains, R plasmids were detected which had molecular weights identical to those of the plasmids found in the corresponding donor strain. Each of the six other transconjugants harbored one plasmid with a size different from those found in the corresponding donor strain, suggesting the occurrence of molecular events during or after conjugative transfer. None of the five tetracycline-resistant transconjugants contained detectable satellite DNA, HindIII restriction enzyme fingerprints of S. faecium resistance plasmids were different from the HindIII patterns of macrolide, aminoglycoside, and tetracycline resistance plasmids from other strains of streptococci.
One hundred strains of group A, B, C, D (S faecalis, S. faecium, S. bovis) F, G, S. pneumoniae and viridans streptococci were studied. All these strains were clinical isolates from infective endocarditis and fron upper respiratory, skin, genital and urinary tract infections. These stains were resistant to one or several antibiotics : tetracycline, macrolide and related drugs, chloramphenicol, aminoglycosides (high-level resistance to streptomycin, kanamycin, gentamicin), and penicillin. Conjugative transfer of antibiotic resistance markers (except penicillin) into streptococcal recipients was obtained at a high frequency (10(-1) to 10(-4)) for 12 strains and at a low frequency (10(-5) to 10(-8)) for 29 strains. R plasmids carrying various groups of resistance markers were isolated with different molecular weights. Enzyme restriction analysis showed the existence of different molecular species of streptococcal plasmids. All attempts to detect extrachromosomal DNA in 17 wild-type strains and in the corresponding transconjugants were unsuccessful.
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Two streptococcal isolates of groups C and G harbored conjugative R plasmids with molecular weights of 17 X 10(6) (pIP646) and 20 X 10(6) (pIP920). These plasmids carried genetic markers for resistance to macrolides and related drugs, as well as to chloramphenicol (pIP920), and have very similar HindIII restriction enzyme patterns.
Two antibiotic-resistant isolates of Streptococcus pneumoniae were investigated for conjugative transfer of their drug resistance markers into streptococcal (groups B and D) and pneumococcal (encapsulated and non-encapsulataed) recipients. Of these, 7 wild-type donor pneumococci transferred all their resistance markers (except Pc [penicillin], Su [sulfonamide], and Tp [trimethoprim]) into group D Streptococcus and non-encapsulated S. pneumoniae recipients at a low frequency (10(-5) to 10(-6)). The resistance markers transferred were Tc (tetracycline); Tc and Cm (chloramphenicol); Tc and MLS (macrolides, lincosamides, and streptogramin B); Tc, MLS, Km (kanamycin), and Cm. The transconjugants obtained retransferred their resistance markers into appropriate streptococcal or pneumococcal recipients or both. The resistance markers of streptococcal transconjugants could not be cured by chemical agents. All attempts to detect extra-chromosomal deoxyribonucleic acid from pneumococcal or streptococcal transconjugants were unsuccessful. The molecular weight of a streptococcal conjugative R plasmid (pIP501) was investigated after transfer into the non-encapsulated S. pneumoniae recipient and was found to be similar to that of the wild-type group B Streptococcus host (20 x 10(6)).
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Each of three isolates of Streptococcus faecalis subsp. zymogenes harbored three R plasmids and a hemolysin-bacteriocin plasmid. The plasmids carried by one of these strains were physically characterized after their conjugative transfer. In each strain one of the plasmids carried genetic markers for resistance to gentamicin, kanamycin, sisomicin, netilmicin, and tobramycin.
Resistance plasmids isolated from Streptococcus agalactiae (group B) and S. faecalis (group D) have been compared in regard to resistance markers, molecular weight, and DNA-DNA homology. Three of them (pIP501, pIP612, and pIP613) have been found to confer identical (or very similar) resistance patterns (erythromycin, lincomycin, and streptogramin B, respectively) and to have similar molecular weights (19.8 x 10(6), 22.7 x 10(6), and 17.6 x 10(6), respectively) and a high level of DNA-DNA homology in hybridization experiments (90 to 100%). These results are compatible with the view that these plasmids may derive from one common ancestor, and/or that they can be transferred between unrelated Streptococcus strains belonging to the same or different groups.
Sixteen metabolically deficient dwarf-colony mutants of Escherichia coli were isolated from urine culture and represented about 2% of all E. coli isolated during the same period. In 14 cases, mutants were isolated from debilitated patients: elderly persons or patients in the terminal stages of a chronic disease. In 15 of these subjects, deficient dwarf-colony mutants appeared to be the true cause of urinary tract infection, since there was leukocyturia and important bacteriuria, and organisms were obtained in pure culture. Study of metabolic deficiencies on Davis synthetic medium and nutritive agar resulted in the identification of eleven deficiencies in cysteine, two in thiamine, two in thymidine, and one in glutamine. Study of resistance to antibiotics revealed that nine were susceptible to all antibiotics, three were resistant to tetracycline alone, two were resistant to two antibiotics (chloramphenicol-tetracycline, streptomycin-tetracycline), and two were resistant to three antibiotics (ampicillin-chloramphenicol-tetracycline, ampicillin-streptomycin-tetracycline). Resistance was coded for by conjugative plasmids in five strains.
Deficient dwarf colony (DDC) mutants of E. coli K 12, harboring or no resistance plasmids, were obtained in vitro. The R plasmids of parental strains and to DDC mutants were transfered by conjugation to normal colony, and to DDC mutants of E. coli K 12; the frequencies of transfer were similar for all strains studied.
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Three recombinant plasmids, resulting from recombination between an R plasmid of the FI incompatibility group and the F of HfrH, were introduced in a temperature-sensitive dnaA mutant to isolate Hfr-type-donors. All of the temperature-insensitive clones isolated from two of the three recombinant plasmids had the same origin and transfer pattern as the parental HfrH strain.
Two plasmids determining resistance to tetracycline (RIP500) and to chloramphenicol, erythromycin, lincomycin, and pristinamycin I (RIP501) were isolated from a strain of Streptococcus agalactiae. The frequency-of-resistance loss is very low for RIP500 (<3 x 10(4)) but higher for RIP501 (the efficiency was dependent upon the curing agents and incubation temperature and varied between 0.5 and 96%). Derivatives susceptible to all drugs were also obtained. RIP500 and RIP501 have similar molecular weights (17.9 x 10(6) and 20 x 10(6), respectively) and represent different percentages of total deoxyribonucleic acid (0.4 and 4%, respectively). The number of copies of RIP500 and RIP501 per cell is different, and these plasmids are likely replicated under different kinds of control (stringent and/or relaxed). No plasmid deoxyribonucleic acid was found in a derivative of strain B96 susceptible to all drugs.
Recombinant plasmids between an R plasmid of the FI group (R162/3) and the sex factor F or HfrH were produced after the conjugal transfer of this R plasmid into HfrH. Three types of recombinant plasmids were identified after the mating of HfrH (R162/3) with recA and rec+ recipients. One specimen of each type (pIP218, pIP222, pIP226) was studied in this report. All three recombinant plasmids carry the same genetic information for resistance to antibiotics (CSSuT) retained from R162/3. pIP218 retained all the other properties from F of HfrH: derepression for pilus synthesis, mobilization of the chromosome for the proximally transferred HfrH genes (thr, leu, proA), interference with T7 propagation, and ability to be cured by acridine orange. pIP222 retained from F of HfrH the derepression for pilus synthesis and the same polarity of chromosome transfer (thr, leu, proA), while pIP226 retained the interference with T7 propagation and acridine orange curing. Physical studies revealed that replication control and/or recovery of F and pIP218 as covalent circles of deoxyribonucleic acid are similar, and are different from R162/3. The new plasmids are more likely the result of a substitutive recombination event than a fusion. We propose genetic maps of these recombinant plasmids, showing the unequal participation of the parental plasmids in their formation.
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