Influence of inducible cross-resistance to macrolides, lincosamides, and streptogramin B-type antibiotics in Enterococcus faecium on activity of quinupristin-dalfopristin in vitro and in rabbits with experimental endocarditis.

The influence of inducible cross-resistance to macrolides, lincosamides, and streptogramin B (MLS(B)) type antibiotics (inducible MLS(B) phenotype) on the activity of quinupristin-dalfopristin was investigated against Enterococcus faecium in vitro and in rabbits with experimental endocarditis. In vitro, quinupristin-dalfopristin displayed bacteriostatic and bactericidal activities against a MLS(B)-susceptible strain similar to those against two strains with the inducible MLS(B) phenotype. In addition, induction of the two MLS(B)-resistant strains with quinupristin (0.016 to 1 microg/ml) or quinupristin-dalfopristin (0.08 to 0.25 microg/ml) increased the MICs of quinupristin from 8 microg/ml to 32 to > 128 microg/ml, but did not modify the MIC of dalfopristin (2 microg/ml) or quinupristin-dalfopristin (0.5 microg/ml). In a rabbit endocarditis model, quinupristin-dalfopristin was as active as amoxicillin against the MLS(B)-susceptible E. faecium strain. In contrast, the activity of quinupristin-dalfopristin was significantly decreased in animals infected with either of the two inducible MLS(B)-resistant strains (P < 0.05), although no mutants resistant to quinupristin-dalfopristin were detected. Against the clinical strain with the inducible MLS(B) phenotype, quinupristin-dalfopristin was not effective and was less active than amoxicillin (P < 0.001); however, the activity of the combination of amoxicillin and dalfopristin-quinupristin was superior to that of amoxicillin (P < 0.01). The different impact of the inducible MLS(B) phenotype in E. faecium on the activity of quinupristin-dalfopristin in vitro and in experimental endocarditis may be related to the reduced diffusion of dalfopristin compared with that of quinupristin into cardiac vegetations that we previously reported. This result emphasizes the importance of the constant presence of dalfopristin at the site of infection to ensure synergism with quinupristin.     

Antibiotic resistance patterns of aerobic coryneforms and furazolidone-resistant Gram-positive cocci from the skin surface of the human axilla and fourth toe cleft

Samples of skin surface bacteria from 28 healthy subjects plated directly on to selective and non-selective media revealed that the proportion of aerobic coryneforms and furazolidone-resistant Gram-positive cocci (FURECs) resistant to erythromycin was significantly greater in the fourth toe cleft than in the axilla (P < 0.05). There were more erythromycin-resistant bacteria than tetracycline-resistant bacteria at both sites (P = 0.001 for the toe cleft; P < 0.01 for the axilla). In total, 160 distinct isolates were obtained, of which 42 were FURECs and 118 were aerobic coryneforms. Of these, 153 (96%) were resistant to erythromycin and 66 (41%) to tetracycline. All except seven of the tetracycline-resistant strains were also resistant to erythromycin. The resistant isolates belonged to a variety of species. CDC group ANF corynebacteria were most numerous and composed 31% of all isolates. The majority (76%) of FURECs were identified as Micrococcus luteus. MIC determinations on selected strains revealed that tetracycline-resistant FURECs were sensitive to doxycycline and minocycline, as were most tetracycline-resistant coryneforms. Nine coryneform isolates were cross-resistant to all three tetracyclines. Only a minority of erythromycin-resistant FURECs (21%) demonstrated a macrolide-lincosamide-streptogramin type B (MLS)-resistant phenotype with inducible or constitutive cross-resistance to clindamycin and the type B streptogramin, pristinamycin IA. Twenty-nine erythromycin-resistant FURECs had a novel phenotype distinct from MLS and macrolide-streptogramin type B resistance. In contrast, most coryneforms (79%) were MLS resistant. Among the remainder, two unusual erythromycin resistance phenotypes were apparent, both of which differed from the unusual phenotype in FURECs. This study has revealed that the non-staphylococcal aerobic flora of skin contains a considerable reservoir of tetracycline and erythromycin resistance determinants. The three unusual macrolide resistance phenotypes may be associated with novel resistance mechanisms.     

Antimicrobial susceptibility testing and phenotyping of Neisseria gonorrhoeae isolated from patients with ophthalmia neonatorum in Nairobi, Kenya.

Antimicrobial susceptibility testing, auxotyping-serotyping, and plasmid analysis were performed on 41 ocular isolates, 7 nasopharyngeal isolates, and 18 cervical isolates of Neisseria gonorrhoeae obtained during a recent treatment trial of gonococcal ophthalmia neonatorum in Nairobi, Kenya. Fourteen distinct serovar-auxotype patterns were observed with IB-1/Pro-strains which accounted for 59% of the isolates. Infection with multiple types of gonococci appeared to occur in 22% of the mothers since 4 of 18 paired maternal cervical and neonatal ocular isolates had mismatched serovar-auxotype patterns. Among 10 treatment failure isolates only 1 had a mismatched serovar-auxotype pattern. Six (15%) of the ocular isolates were penicillinase-producing N. gonorrhoeae (PPNG). Five had the 4.4-megadalton (Md) beta-lactamase plasmid and one had the 3.2-Md beta-lactamase plasmid. The 24.5-Md plasmid was found in 5 of 6 PPNG strains and in 8 of 35 non-PPNG strains (P less than 0.02). For most antimicrobial agents, PPNG and non-PPNG strains showed similar patterns of susceptibility. Ceftriaxone was the most active of the antibiotics tested, with all strains having an MIC less than or equal to 0.06 mg/liter. Among non-PPNG strains, 15 (43%) had a penicillin MIC greater than or equal to 2 mg/liter and were considered intrinsically resistant to penicillin. Overall, non-PPNG intrinsically resistant strains had greater resistance to other antibiotics than did non-intrinsically resistant strains (P less than or equal to 0.006). The Mtr phenotype was found in 53% of these strains.     

Emergence of 4'',4"-aminoglycoside nucleotidyltransferase in enterococci.

Enterococcus faecium BM4102 was resistant to macrolide-lincosamide-streptogramin B-type (MLS) antibiotics; tetracycline-minocycline; and high levels of kanamycin, neomycin, tobramycin, and dibekacin but not gentamicin. This aminoglycoside resistance phenotype is new in enterococci. The genes conferring resistance to aminoglycosides and MLS antibiotics in this strain were carried on a plasmid, pIP810, that was self-transferable to to other Enterococcus strains. Resistance to tobramycin and structurally related aminoglycosides, kanamycin, neomycin, and dibekacin, was due to synthesis of a 4',4"-aminoglycoside nucleotidyltransferase. Homology was detected by hybridization between pIP810 DNA and a probe specific for a gene encoding an enzyme with identical site specificity in staphylococci. The bacteriostatic activity of amikacin apparently was not affected by the presence of the enzyme, although it was modified in vitro. However, the bactericidal activity of amikacin and the synergism of this aminoglycoside with penicillin were abolished.     

Antimicrobial susceptibility of non-enterococcal intrinsic glycopeptide-resistant Gram-positive organisms

Non-enterococcal Gram-positive bacteria that are intrinsically vancomycin-resistant have been infrequently isolated in association with serious infections. However, well-documented infections have lately been reported with increasing frequency. Because these organisms may be pathogens, we tested the MICs of 19 antimicrobial agents by the agar dilution method for predicting susceptibility. The activity of these antimicrobial agents was assessed against 28 strains (Lactobacillus rhamnosus, 6; Lactobacillus acidophilus, 1; Lactobacillus casei, 1; Lactobacillus fermentum, 2; Lactobacillus brevis, 1; Lactobacillus plantarum, 1; Weissella confusa, 2; Leuconostoc mesenteroides, 7; Leuconostoc lactis, 4; Pediococcus acidilactici, 2; Pediococcus pentosaceus, 1), isolated from clinical specimens in an Argentinian university hospital from 1997 to 2003. The MICs of penicillin for 67% of the Lactobacillus strains and 100% of the Leuconostoc spp. and Pediococcus spp. strains tested were in the 0.25-2 microg/mL range. Erythromycin was the most active antimicrobial overall. Multiresistance was observed in 2 strains (Lactobacillus rhamnosus, 1; Lactobacillus plantarum, 1).     

Distribution of erythromycin esterase and rRNA methylase genes in members of the family Enterobacteriaceae highly resistant to erythromycin

The distribution of nucleotide sequences related to ereA, ereB, and ermAM was studied by colony hybridization in 112 strains of members of the family Enterobacteriaceae that are highly resistant to erythromycin. The ereA and ereB genes encoding erythromycin esterases type I and II, respectively, were detected in strains inactivating the 14-membered macrolides erythromycin and oleandomycin. Because all 52 strains resisting these antibiotics by inactivation were detected by ereA (n = 23), ereB (n = 23), or both probes (n = 6), only two classes of genes accounted for this resistance phenotype. The ermAM gene encoding a streptococcal rRNA methylase was detected in 21 strains of Escherichia coli and two strains of Klebsiella spp. Determination of the MICs of macrolide, lincosamide, and streptogramin (MLS) antibiotics demonstrated a correlation between hybridization with ermAM and the so-called MLS resistance phenotype. The presence of 11 strains coresistant to MLS antibiotics that did not hybridize to the ermAM probe suggests that, as in gram-positive organisms, MLS resistance in members of the family Enterobacteriaceae involves more than one class of rRNA methylase. Numerous strains (n = 18) were found to produce both an erythromycin esterase type II and an rRNA methylase. Physical linkage between ereB and ermAM may be responsible for the codissemination of the genes. Despite their exogenous origin, ereB and ermAM are already disseminated in various genera of the Enterobacteriaceae.     

Combination of quinupristin-dalfopristin and gentamicin against methicillin-resistant Staphylococcus aureus: experimental rabbit endocarditis study

The combination of quinupristin-dalfopristin (Q-D) and gentamicin was tested against two strains of gentamicin- and dalfopristin-susceptible methicillin-resistant Staphylococcus aureus (MRSA). One strain was susceptible to macrolides, lincosamides, and streptogramin B type antibiotics (MLS(B)), and the other was constitutively resistant to these antibiotics by virtue of the ermA gene. The checkerboard method and time-kill curves showed that the combination of Q-D and gentamicin was indifferent. A rabbit endocarditis model simulated the pharmacokinetics achieved in humans receiving intravenous injections of Q-D (7.5 mg/kg of body weight three times a day) and gentamicin (3 mg/kg once daily). For the MLS(B)-susceptible strain, a 4-day regimen reduced mean bacterial titers (MBT) in vegetations from 8.5 +/- 0.8 log CFU/g (control group) to 4.1 +/- 2.6 (gentamicin), 3.0 +/- 0.9 (Q-D), and 2.6 +/- 0.5 log CFU/g (Q-D plus gentamicin). For the strain constitutively resistant to MLS(B), a 4-day regimen reduced MBT in vegetations from 8.7 +/- 0.9 log CFU/g (control group) to 5.0 +/- 2.2 (gentamicin), 5.2 +/- 2.2 (Q-D), and 5.1 +/- 2.4 log CFU/g (Q-D plus gentamicin). The differences between control and treatment groups were significant for both strains (P < 0.0001), although there was no significant difference between treatment groups. No resistant variant was isolated from vegetations, and no significant difference in MBT in vegetations of treatment groups after 1-day regimens was observed. This experimental study found no additive benefit in combining Q-D and gentamicin against dalfopristin- and gentamicin-susceptible MRSA.     

Transferable vancomycin and teicoplanin resistance in Enterococcus faecium.

Enterococcus faecium BM4165 and BM4178, isolated from immunocompromised patients, one treated with vancomycin, were inducibly resistant to high levels of the glycopeptide antibiotics vancomycin and teicoplanin but susceptible to the new lipopeptide daptomycin (LY146032). Strain BM4165 was also resistant to macrolidelincosamide-streptogramin B-type (MLS) antibiotics. The genes conferring resistance to glycopeptides and to MLS antibiotics in strain BM4165 were carried on plasmids pIP819 and pIP821, respectively; pIP819 also carried genes that encoded resistance to MLS antibiotics. The two plasmids, which were distinct although related, were self-transferable to other E. faecium strains. Plasmid pIP819 could also conjugate to E. faecalis, Streptococcus sanguis, S. pyogenes, S. lactis, and Listeria monocytogenes, in which it conferred inducible glycopeptide resistance, but not to S. aureus. Glycopeptide-inactivating activity was not detected, and the biochemical mechanism of resistance remains unknown. Based on this first report of transferable resistance to glycopeptides, we anticipate dissemination of resistance to these antibiotics in gram-positive cocci and bacilli in which it can be phenotypically expressed.     

High-level resistance to gentamicin in Enterococcus faecium.

During a six-month period in a hospital in Ireland, four patients were infected (isolation from blood cultures) and two were colonized (isolation from rectal swabs) with strains of Enterococcus faecium highly resistant to gentamicin. MICs of gentamicin were greater than 1000 mg/L for all six strains, and each possessed a plasmid of approximately 50 MDa. Resistance to gentamicin was transferable by conjugation from two of the six strains, and was associated with transfer of the 50 MDa plasmid. This plasmid hybridized with a DNA probe specific for the bifunctional AAC(6')-APH(2") aminoglycoside-modifying enzyme. The mechanism of high-level gentamicin resistance in these strains appeared therefore, to be identical to that encountered in Enterococcus faecalis strains worldwide and reported in E. faecium strains in the USA.     

Presence of two unique genes encoding macrolide-lincosamide-streptogramin resistance in members of the Bacteroides fragilis group as determined by DNA-DNA homology

The relationship of a previously described, plasmid-encoded macrolide-lincosamide-streptogramin (MLS) resistance determinant to Bacteroides fragilis group clinical isolates from 11 different patients with bacteroides infections was assessed using in situ hybridization. Most of the strains were found to have DNA sequences homologous with the bacteroides MLS plasmid, pBF4, but this homology was exclusively to chromosomal DNA. Two organisms isolated from different sites in a single patient did not share homology with the plasmid, and probably represent a second type of MLS gene. The MLS resistance plasmids, pBF4 (from Bact. fragilis) and pE194 (from Staphylococcus aureus), did not share homologous sequences; therefore at least one of the bacteroids MLS genes is different from that found in Gram-positive organisms.     

Transition mutations in the 23S rRNA of erythromycin-resistant isolates of Mycoplasma pneumoniae.

Erythromycin is the drug of choice for treatment of Mycoplasma pneumoniae infections due to its susceptibility to low levels of this antibiotic. After exposure of susceptible strains to erythromycin in vitro and in vivo, mutants resistant to erythromycin and other macrolides were isolated. Their phenotypes have been characterized, but the genetic basis for resistance has never been determined. We isolated two resistant mutants (M129-ER1 and M129-ER2) by growing M. pneumoniae M129 on agar containing different amounts of erythromycin. In broth dilution tests both strains displayed resistance to high levels of several macrolide-lincosamide-streptogramin B (MLS) antibiotics. In binding studies, ribosomes isolated from the resistant strains exhibited significantly lower affinity for [14C]erythromycin than did ribosomes from the M129 parent strain. Sequencing of DNA amplified from the region of the 2S rRNA gene encoding domain V revealed an A-to-G transition in the central loop at position 2063 of M129-ER1 and a similar A-to-G transition at position 2064 in M129-ER2. Transitions at homologous locations in the 23S rRNA from other organisms have been shown to result in resistance to MLS antibiotics. Thus, MLS-like resistance can occur in M. pneumoniae as the result of point mutations in the 23S rRNA gene which reduce the affinity of these antibiotics for the ribosome. Since they involve only single-base changes, development of resistance to erythromycin in vivo by these mechanisms could be relatively frequent event.     

Location of antibiotic resistance markers in clinical isolates of Enterococcus faecalis with similar antibiotypes.

Eight wild-type strains of Enterococcus faecalis, resistant to chloramphenicol (Cmr), erythromycin (Emr), tetracycline (Tcr), and minocycline (Mnr), were examined for the genetic basis of their antibiotic resistance, Five of the strains transferred all of their antibiotic resistance markers by conjugation, while the other three strains transferred only Tcr and Mnr. Cmr and Emr determinants were localized by DNA-DNA hybridization experiments, in which the Cmr gene of plasmid pIP501, of group B Streptococcus origin, and the Emr gene of transposon Tn917, of E. faecalis origin, served as probes. A chromosomal location was found for the nonconjugative Cmr and Emr markers of one wild-type strain. In two strains these markers were carried by nonconjugative plasmids, and in the other strains they were carried by plasmids that transferred by conjugation. Plasmids isolated from three transconjugants resistant to tetracycline but susceptible to minocycline bore nucleotide sequences homologous to the tetL gene. Nucleotide sequences homologous to conjugative transposon Tn916, of E. faecalis origin, were detected by hybridization in the tetracycline-minocycline-resistant transconjugants. Three of these transconjugants were plasmid free, while four harbored conjugative cryptic plasmids. Sequences homologous to Tn916 were also found on two conjugative plasmids, one of which appeared to be a conjugative cryptic plasmid that had acquired chromosomal Tcr Mnr markers during transfer.     

Aminocyclitol-modifying enzymes specified by chromosomal genes in Staphylococcus aureus.

A genetic analysis of aminocyclitol resistance in two strains of Staphylococcus aureus was performed. Resistance of strain FK170, isolated in Zurich, was due to the production of a 3'-phosphotransferase [APH(3')]. Strain 5532, isolated in London, produced a 2"-phosphotransferase [APH(2")] and a 6'-N-acetyltransferase [AAC(6')]. Plasmid deoxyribonucleic acid (DNA) was isolated by isopycnic centrifugation from the two parent strains, as well as from susceptible variants and from resistant transductants of both strains. Comparative analysis of plasmid and from resistant transductants of both strains. Comparative analysis of plasmid DNA by centrifugation in sucrose gradients revealed that strain FK170 harbored a 2.7-megadalton tetracycline R-plasmid and a 36-megadalton cryptic plasmid. Strain 5532 contained an 18.5-megadalton penicillinase plasmid. No evidence for plasmid location of the markers for aminocyclitol resistance could be obtained.     

Macrolide-lincosamide-streptogramin B (MLS) resistance in cutaneous propionibacteria: definition of phenotypes

Erythromycin-resistant propionibacteria isolated from the skin of antibiotic-treated acne patients were found to express four different patterns of resistance to a set of eight MLS antibiotics. The majority of isolates (48/89 strains) were constitutively resistant to 14- and 16-membered ring macrolide, lincosamide and streptogramin B-type antibiotics. MICs of josamycin (0.5-16 mg/l) and spiramycin (0.5-128 mg/l) were lower than those of erythromycin, oleandomycin and tylosin (64 to less than 512 mg/l). Two strains of Propionibacterium granulosum exhibited inducible MLS resistance. Both 14- and 16-membered ring macrolides and virginiamycin S induced clindamycin resistance in these strains. Fifteen isolates demonstrated a similar phenotype to the inducible strains but were non-inducible by erythromycin. A fourth group of strains demonstrated high level resistance to all five macrolides tested (MIC greater than or equal to 128 mg/l) but were sensitive to virginiamycin S. The phenotype displayed by these strains is not compatible with the expression of methylase genes as currently known, nor with the action of an erythromycin esterase which hydrolyses only 14-membered ring macrolides. The resistance patterns of 12 isolates could not be classified into any of the above phenotypic classes. Therefore, the majority of erythromycin resistant propionibacteria express MLS resistance which is phenotypically similar to that coded for by several well characterized RNA methylase (erm) genes.     

Comparative evaluation of VITEK 2 for antimicrobial susceptibility testing of group B Streptococcus

OBJECTIVES: Intrapartum antibiotic prophylaxis is recommended to prevent neonatal group B streptococcal (GBS) disease in colonized women, and penicillin or aminopenicillin constitute the first-line antibiotics. Most isolates are resistant to tetracycline, and resistance to macrolide-lincosamide-streptogramin (MLS) antibiotics is increasing. Therefore, laboratory testing for MLS resistance in GBS is now recommended for penicillin-allergic patients. The aim of this study was to compare the antimicrobial susceptibility of GBS as determined by the VITEK 2 system (bioMérieux, Marcy l'Etoile, France), agar diffusion methods and PCR-genotypic detection of resistance genes. METHODS: One hundred and ten unrelated selected GBS clinical isolates were studied. The antibiotics tested (VITEK 2 and agar diffusion method) were benzylpenicillin, ampicillin, erythromycin, clindamycin, co-trimoxazole, tetracycline, kanamycin, streptomycin and vancomycin. A standardized double-disc (DD) diffusion test was performed for MLS antibiotics. Genotypic characterization of tetracycline, MLS and aminoglycoside resistance genes was performed by PCR. RESULTS: All strains were susceptible to benzylpenicillin, ampicillin and vancomycin [category agreement (CA) between VITEK 2 and the diffusion method was 100%]. Ninety-five (86%) strains were resistant to tetracycline (CA was 98.9%). Eighty-one strains (73.6%) harboured an MLS resistance phenotype; 50 (61.8%) an MLS(B)-constitutive phenotype, 25 (30.8%) an MLS(B)-inducible phenotype and 6 (7.4%) an M phenotype. The agreement between data of VITEK 2 and the DD diffusion test for the detection of MLS(B)-constitutive, MLS(B)-inducible and M phenotype isolates was 76%, 36% and 100%, respectively. Almost all discrepancies were due to failure to detect erythromycin resistance by VITEK 2. CONCLUSIONS: VITEK 2 allows accurate determination of GBS susceptibility for the majority of antibiotics, but has to be improved for erythromycin. Thus, the DD diffusion test remains the most simple and reliable method for macrolide resistance detection among this streptococcal species.     

Mobilization of the penicillinase gene in Enterococcus faecalis.

Enterococcus faecalis SF4855 is a beta-lactamase-producing isolate resistant to high levels of gentamicin, with determinants for these resistances on the chromosome. SF4855 transferred both determinants into E. faecalis FA2-2 and UV202 at a frequency of 10(-9) in the presence of the MLS plasmid pYN120. beta-Lactamase and gentamicin resistance probes hybridized to three locations on the chromosome of FA2-2 transconjugants on contour-clamped homogeneous electric field electrophoresis. The study results suggest mobilization of the beta-lactamase determinant.     

Characterization of antibiotic-resistant Corynebacterium striatum strains.

Antibiotic-resistant Corynebacterium strains were isolated from 14 Harborview and one Veterans Administration Hospital patients in Seattle during the period 1987-90. These clindamycin-erythromycin resistant strains were shown to hybridize with the ermCd gene, which was cloned from a Corynebacterium diphtheriae plasmid and encodes for a rRNA methylase. Thirteen of these strains also hybridized with the tetM gene probes, and were tetracycline resistant. The ermCd gene could be transferred, by conjugation, while the tetM gene was not transferable.     

Serious streptococcal infections produced by isolates resistant to streptogramins (quinupristin/dalfopristin): case reports from the SENTRY antimicrobial surveillance program

The emergence and sustained prevalence of Gram-positive organisms resistant to antimicrobials has been of interest for over a decade. Quinupristin/dalfopristin (formerly RP 59500 or Synercid) is a new injectable streptogramin combination that has been reported to have activity against Gram-positive organisms, even those with documented MLS(B) resistance. However, the two case reports presented here illustrate three well-documented Streptococcus spp. strains (S. mitis, S. pneumoniae) to be resistant to quinupristin/dalfopristin (MICs at 3, 8, and 12 microg/ml) following referral as routine isolates in the SENTRY Antimicrobial Surveillance Program. The S. pneumoniae pleural fluid isolate was cross-resistant to erythromycin. Both bacteremic S. mitis strains were resistant to macrolides (erythromycin, azithromycin, clarithromycin), lincosamides (clindamycin), and fluoroquinolones. Patient histories indicated no prior use of MLS class antimicrobials for the S. mitis case, but the patient having the S. pneumoniae isolate did receive prior treatment of erythromycin and clindamycin. All isolates had modestly increased penicillin MICs of 0.12 microg/ml. The mode of resistance to quinupristin/dalfopristin was not evident (sat A-negative by PCR); and these cases illustrate the existence of streptogramin-resistant isolates before the introduction of this antimicrobial class into human clinical practice.     

Plasmid-determined resistance to fosfomycin in Serratia marcescens.

Multiple-antibiotic-resistant strains of Serratia marcescens isolated from hospitalized patients were examined for their ability to transfer antibiotic resistance to Escherichia coli by conjugation. Two different patterns of linked transferable resistance were found among the transconjugants. The first comprised resistance to carbenicillin, streptomycin, and fosfomycin; the second, and more common, pattern included resistance to carbenicillin, streptomycin, kanamycin, gentamicin, tetracycline, chloramphenicol, sulfonamide, and fosfomycin. The two types of transconjugant strains carried a single plasmid of either 57 or 97 megadaltons in size. Both of these plasmids are present in parental S. marcescens strains resistant to fosfomycin. The 57-megadalton plasmid was transformed into E. coli.     

Penicillin-binding components of penicillin-susceptible and -resistant strains of Streptococcus pneumoniae.

Three strains of Streptococcus pneumoniae with low-level resistance to penicillin, one strain with resistance to penicillin, and three strains susceptible to penicillin were compared. The three susceptible strains had very similar patterns of penicillin-binding components (PBCs) as detected by fluorography after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Gels of these strains showed four major bands and a single minor band. All of the resistant strains showed different PBC patterns from those obtained for susceptible strains. The most marked changes were observed with the most resistant strain SPR110, which showed markedly reduced intensity of PBC 3 and a reduction in intensity of PBCs 1 and 2, as well as additional bands in the region of band 2. Low-level resistant strains had band densities different from the susceptible strains principally for PBC 3. No beta-lactamase activity or plasmid was detected in the resistant strains. An apparent difference in affinity for cloxacillin was also demonstrated between some of the PBCs of susceptible strain S. pneumoniae SPS101 and resistant strain S. pneumoniae SPR110. Penicillin resistance in the low-level resistant and the resistant strains was associated with PBCs possessing different properties from those seen in the three susceptible strains. Reduction in affinity for penicillin or a reduction in the amount of a PBC protein, or both, are the probable explanations of penicillin resistance in these strains.