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Resistance to dicloxacillin, methicillin and oxacillin in methicillin-susceptible and methicillin-resistant Staphylococcus aureus detected by dilution and diffusion methods.

A total of 54 Staphylococcus aureus strains of varying methicillin resistance were investigated for their resistance to methicillin, oxacillin and dicloxacillin by different diffusion tests. Inhibition zones were measured around locally prepared paperdiscs with 10 micrograms methicillin, 5 and 10 micrograms oxacillin, 5 and 10 micrograms dicloxacillin, PDM paperdiscs with 10 micrograms methicillin or oxacillin and Neo-sensitabs tablets with methicillin or oxacillin. All diffusion tests were performed both with Mueller-Hinton agar and Danish Blood agar as well as at 37 degrees C and 30 degrees C and read after overnight incubation. Differences in zone diameter under different conditions were found to be independent of the susceptibility level of the strains. Seventeen of the strains were detected as methicillin-resistant (MRSA) by two methods including high inoculum and prolonged incubation at 30 degrees C. The minimum inhibitory concentration (MIC) of the 54 strains was determined by a plate dilution method at 30 degrees C and 37 degrees C. A 10 micrograms locally prepared methicillin disc detected all MRSA strains with no false reactions either at 37 degrees C or 30 degrees C on Mueller-Hinton agar. Investigations with oxacillin discs had to be performed at 30 degrees C or with a 5 microgram disc in order to detect correctly. PDM paperdiscs gave reactions identical to the corresponding locally prepared discs. Methicillin Neo-sensitabs detected all MRSA strains but also included a few susceptible strains among the resistant ones. Addition of blood increased the number of not-detected MRSA strains. All 17 MRSA strains were susceptible to dicloxacillin by the dilution method, and the disc diffusion test showed similar results. Dicloxacillin discs therefore did not detect the presence of MRSA strains. The implications of replacement of the methicillin/oxacillin disc by a dicloxacillin disc are discussed.

Denmark↗

femA, which encodes a factor essential for expression of methicillin resistance, affects glycine content of peptidoglycan in methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains.

femA is a chromosomally encoded factor, occurring naturally in Staphylococcus aureus, which is essential for the expression of high-level methicillin resistance in this organism. The production of a low-affinity penicillin-binding protein, PBP2a or PBP2', which is intimately involved with methicillin resistance in S. aureus, is not influenced by femA. To elucidate a possible physiological function of the 48-kDa protein encoded by femA, several related methicillin-resistant, methicillin-susceptible, and Tn551 insertionally inactivated femA mutants were analyzed for possible changes in cell wall structure and metabolism. Independent of the presence of mec, the methicillin resistance determinant, all femA mutants had a reduced peptidoglycan (PG) glycine content (up to 60% in the molar ratio of glycine/glutamic acid) compared to that of related femA+ parent strains. Additional effects of femA inactivation and the subsequent decrease in PG-associated glycine were (i) reduced digestion of PG by recombinant lysostaphin, (ii) unaltered digestion of PG by Chalaropsis B-muramidase, (iii) reduced cell wall turnover, (iv) reduced whole-cell autolysis, and (v) increased sensitivity towards beta-lactam antibiotics. Also, the PG-associated glycine content of a femA::Tn551 methicillin-susceptible strain was restored concomitantly with the methicillin resistance to a level almost equal to that of its femA+ methicillin-resistant parent strain by introduction of plasmid pBBB31, encoding femA.

Acetylglucosamine↗

Does nasal cocolonization by methicillin-resistant coagulase-negative staphylococci and methicillin-susceptible Staphylococcus aureus strains occur frequently enough to represent a risk of false-positive methicillin-resistant S. aureus determinations by molecular methods?

By analyzing the colonization of the anterior nares in cardiothoracic surgery patients on admission, nasal cocolonization by methicillin-susceptible Staphylococcus aureus and methicillin-resistant coagulase-negative staphylococci was detected in 8/235 (3.4%) specimens. Consequently, in a low-methicillin-resistant S. aureus (MRSA) setting, a molecular MRSA screening test targeting the mecA gene and an S. aureus-specific gene in parallel and applied directly to clinical specimens would be associated with an unacceptable positive predictive value of about 40%.

Bacterial Proteins↗

[The beta-lactamase activity in methicillin-resistant Staphylococcus aureus--a comparison of the beta-lactamase from a methicillin-resistant and from a methicillin-sensitive strain of Staphylococcus aureus].

Strains of Methicillin-resistant Staphylococcus aureus (MRSA) are more frequently beta-lactamase-negative than those of Methicillin-susceptible S. aureus (MSSA). In the strains of MRSA, the beta-lactamase activity tends to be inversely related to the MIC's of Methicillin. Such a marked tendency has been observed in recent years.

Methicillin↗

Effects of temperature, NaCl, and methicillin on penicillin-binding proteins, growth, peptidoglycan synthesis, and autolysis in methicillin-resistant Staphylococcus aureus.

Methicillin-resistant Staphylococcus aureus strains produce a fifth penicillin-binding protein (PBP), PBP 2', with low affinity for beta-lactam antibiotics that is believed to represent a beta-lactam-insensitive peptidoglycan transpeptidase. In an effort to evaluate the adequacy of PBP 2' as an explanation of methicillin resistance, PBP 2' production and the responses of growth and peptidoglycan synthesis to methicillin under different environmental conditions have been compared. In the heterogeneous methicillin-resistant strain DU4916-K7, less PBP 2' was produced at 40 degrees C than at 30 degrees C, but inclusion of 5% (wt/vol) NaCl in the medium at 40 degrees C boosted PBP 2' production and allowed growth of the organism in the presence of 10 micrograms of methicillin per ml. When exponential-phase cultures were challenged with methicillin, growth and peptidoglycan synthesis were much more resistant at 30 degrees C than at 40 degrees C. Inclusion of NaCl in medium rendered growth and peptidoglycan synthesis more methicillin resistant at 40 degrees C. Hence, there was a good correlation between PBP 2' production and methicillin-resistant peptidoglycan synthesis under these conditions. However, PBP 2' production was increased by NaCl at 30 degrees C without markedly affecting the susceptibilities of growth and peptidoglycan synthesis to methicillin. Pregrowth of cells with methicillin, which was expected to boost PBP 2' production, seemed to increase the susceptibilities of growth and peptidoglycan synthesis to methicillin. Patterns of growth and peptidoglycan synthesis susceptibilities to methicillin which were similar to those described above were found in chloramphenicol-inhibited cultures, in which presumably no induction of PBP 2' could occur during the methicillin challenge period. Complex effects were noted in the combination of subinhibitory methicillin and NaCl. Growth of cells in the presence of NaCl stimulated their autolytic activity, which was further increased by growth with subinhibitory methicillin in addition to NaCl. It appears that NaCl enhances methicillin resistance by stimulating PBP 2' production and providing osmotic support but opposes it by stimulating autolytic activity which is exacerbated by the very low cross-linking of peptidoglycan in methicillin-resistant strains grown in the presence of methicillin.

Bacterial Proteins↗

Differential methicillin susceptibilities of peptidoglycan syntheses in methicillin-resistant Staphylococcus aureus.

The mechanism of staphylococcal resistance to methicillin is unknown. Peptidoglycan synthesis was studied in a methicillin-resistant and a derived methicillin-sensitive Staphylococcus aureus strain. Although the methicillin minimum inhibitory concentration for growth of the methicillin-resistant strain was 1,600 micrograms/ml, peptidoglycan synthesis by the organism incubated in a wall synthesis solution was inhibited about 90% by 5 micrograms of methicillin per ml. In contrast, high concentrations of methicillin added to actively growing cultures of the methicillin-resistant strain had little effect on growth or peptidoglycan synthesis. Peptidoglycan synthesis in chloramphenicol-treated cultures was more susceptible to methicillin than it was in actively growing cultures of the methicillin-resistant strain. It is proposed that in this strain cell wall thickening peptidoglycan synthesis which predominates in cell wall synthesis solution and chloramphenicol-treated cultures is methicillin sensitive, whereas peptidoglycan synthesis involved in cell division, primarily in the region of the septum, which predominates in actively growing cultures is methicillin resistant. Both cell wall thickening and septal peptidoglycan syntheses are methicillin sensitive in the methicillin-sensitive strain.

Chloramphenicol↗

Eagle-type methicillin resistance: new phenotype of high methicillin resistance under mec regulator gene control.

We report a novel phenotype of methicillin resistance, designated "Eagle-type" resistance, which is characteristic in its resistance to high concentrations of methicillin (64 to 512 microg/ml) and susceptibility to low concentrations of methicillin (2 to 16 microg/ml). The type of resistance was expressed in mutant strains selected with high concentrations (e.g., 128 to 512 microg/ml) of methicillin from the pre-methicillin-resistant Staphylococcus aureus strain N315, whose mecA gene transcription is strongly repressed by the mecI gene-encoded repressor protein MecI. The Eagle-type mutant strains harbored no mutation in the mecI gene or in the operator region of mecA gene to which MecI repressor is supposed to bind. In the representative Eagle-type strain h4, repression of mecA gene transcription and penicillin-binding protein 2' production were found to be released by exposing the cells to a high concentration (128 microg/ml) of methicillin but not to lower concentrations (1 and 8 microg/ml) of methicillin. The strain h4 expressed paradoxical susceptibility (Eagle effect) to the cytokilling activity of methicillin. Experimental deletion of mecI gene from the chromosome of h4 by mecI-specific gene substitution converted its Eagle-type resistance to homogeneously high methicillin resistance. We cloned two novel genes, designated hmrA and hmrB, from genomic library of h4, which conferred Eagle-type resistance to N315 when introduced into the cell in multiple copies. The genes were shown to confer homogeneous methicillin resistance to the heterogeneously methicillin-resistant strain LR5 when they were introduced into on multicopy plasmids. This result strongly indicated that the genetic alteration responsible for the expression of the Eagle phenotype is identical, or equivalent in its effect, to the genetic alteration underlying heterogeneous-to-homogeneous conversion of methicillin resistance in S. aureus.

Acyl Carrier Protein↗

Comparison of minimum inhibitory concentrations to methicillin in heterogeneous and homogeneous methicillin resistant Staphylococcus aureus.

OBJECTIVE: To determine the minimum inhibitory concentration (MIC) to methicillin and compare it with the type of methicillin resistance in methicillin resistant Staphylococcus aureus (MRSA) isolated from clinical samples, and to evaluate the usefulness of beta-lactamase resistant penicillins such as cloxacillin for treatment of minor MRSA infections. DESIGN: Sixty-five strains of MRSA isolated from clinical samples were studied. Homogeneous and heterogeneous resistance to methicillin of these strains was determined by an efficiency of plating technique. The MICs to methicillin of all homogeneously resistant strains and an equal number of heterogeneously resistant strains were determined by the 'E' test. SETTING: General Hospital, Colombo; De Soysa Maternity Hospital for Women; Lady Ridgeway Hospital for Children. RESULTS: 171 strains of S aureus were isolated from specimens processed in the laboratory of the Department of Microbiology, Faculty of Medicine Colombo, during a 14-month period. Seventy (40.9%) were methicillin resistant. Fifty-three MRSA strains (81.5%) were heterogeneously resistant to methicillin. Only 12 of 65 strains were homogeneously resistant. Nine of 12 of the homogeneously resistant strains were from surgical wards. MICs for methicillin in the two groups were compared. Six of 12 (50%) homogeneously resistant strains had very high levels of resistance to methicillin (MIC > 200 micrograms/ml). In contrast the MICs of all the heterogeneously resistant strains ranged from 16 micrograms/ml to 96 micrograms/ml. CONCLUSIONS: The majority of clinical isolates of MRSA from hospitals in Colombo, Sri Lanka were heterogeneously resistant to methicillin with lower MICs for methicillin than the homogeneously resistant strains. Thus, beta-lactamase resistance penicillins such as cloxacillin may have a place in the treatment of minor MRSA infections. Surveillance studies to determine if there is an increase of homogeneously methicillin resistant MRSA are required.

Adult↗

Prevalence of methicillin-sensitive and methicillin-resistant Staphylococcus aureus in pregnant women.

OBJECTIVE: To estimate the extent of Staphylococcus aureus vaginal-rectal colonization among pregnant women as severe S aureus infections have emerged in pregnant and postpartum women and infants. METHODS: We conducted a prospective surveillance study for methicillin-sensitive S aureus and methicillin-resistant S aureus on all routine de-identified vaginal-rectal prenatal group B streptococcus (GBS) screening cultures submitted to the microbiology laboratory of a tertiary-care facility from January to July 2005. Standard microbiologic techniques and molecular analyses were used to detect community-associated methicillin-resistant S aureus strains. As opposed to health care-associated methicillin-resistant S aureus isolates, community-associated methicillin-resistant S aureus isolates were defined as those possessing the type IV or type V staphylococcal chromosomal cassette mec element and usually lacking a multidrug-resistant phenotype. RESULTS: A total of 2,963 GBS screening cultures were analyzed, from which 743 (25.1%, 95% confidence interval [CI] 23.5-26.7%) GBS isolates and 507 (17.1%, 95% CI 15.7-18.5%) S aureus isolates were identified. Group B streptococcus colonization was significantly associated with S aureus colonization (prevalence odds ratio 2.1, 95% CI 1.7-2.5, P < .001). Of the S aureus isolates, 14 (2.8%, 95% CI 1.4-4.2%) were methicillin-resistant, and 13 of these were determined to be community-associated methicillin-resistant S aureus. CONCLUSION: The prevalence of S aureus colonization identified in GBS screening cultures from pregnant women was substantial and associated with GBS co-colonization. Although we do not advocate routine screening of pregnant women for methicillin-sensitive S aureus and methicillin-resistant S aureus colonization, we recommend continued monitoring of both methicillin-sensitive S aureus and methicillin-resistant S aureus infections in this population and their infants.

Anti-Bacterial Agents↗

The effect of methicillin on the fatty acid composition of total polar lipid in methicillin resistant Staphylococcus aureus.

The effect of two concentrations of methicillin on the fatty acid (FA) distribution in intracellular total polar lipid (TPL) of the log-phase cultures of a methicillin resistant Staphylococcus aureus strain No. 5814R was studied during a period of 2 h. Half the MIC of methicillin (= 1000 micrograms/ml) caused 18.6% increase in branched-FAs and a same decrease in straight-FAs, while one MIC (= 2000 micrograms/ml) of the drug induced a moderate change in those of TPL. The ratio of branched-FAs to straight-FAs increased from 1.24 to 1.56 in the presence of 1/2 x MIC of methicillin and reduced from 1.24 to 0.87 in the presence of 1 X MIC of the antibiotic. In TPL of the control cultures it gradually decreased from 1.24 to 0.77. It is concluded that under the effect of methicillin, FA composition of TPL in methicillin resistant cocci does not change as dramatically as in methicillin sensitive ones indicating lipid synthesis in methicillin resistant S. aureus to be less sensitive to the action of methicillin than in methicillin susceptible strains. This may contribute to the resistance against the lytic effect of the drug. Membrane lipid properties seem to be involved in the mechanisms of methicillin resistance.

Fatty Acids↗

Clinical comparative study on the activity of cefamandole in the treatment of serious staphylococcal infections caused by methicillin-susceptible and methicillin-resistant strains.

Ninety-two microbiologically documented staphylococcal infections were treated with cefamandole in an open comparative study on the clinical efficacy of this cephalosporin in the therapy of infections caused by both methicillin-susceptible and methicillin-resistant Staphylococcus aureus and coagulase-negative Staphylococcus spp. The majority of the episodes (86 of 92) were treated with cefamandole alone, and six were treated with cefamandole in association with other antibiotics. In the evaluable S. aureus infections, 34 of 46 (73.9%) due to methicillin-susceptible strains and 12 of 16 (75%) due to methicillin-resistant strains responded to therapy. In particular, among the patients infected by methicillin-susceptible S. aureus 6 of 9 cases of septicemia, 0 of 2 cases of endocarditis, 2 of 2 cases of pneumonia, 2 of 3 osteoarticular infections, 8 of 12 cases of peritonitis in patients with chronic renal failure in continuous ambulatory peritoneal dialysis (CAPD), 13 of 15 skin-soft tissue infections, and 3 of 3 urinary tract infections responded to therapy. Among those due to methicillin-resistant strains, cure was achieved in 2 of 4 cases of septicemia, 0 of 1 case of endocarditis, 9 of 10 skin-soft tissue infections, and 1 of 1 urinary tract infection. In the evaluable infections caused by coagulase-negative staphylococci, 9 of 11 (81.8%) due to methicillin-susceptible and 15 of 17 (88.2%) due to methicillin-resistant strains responded to therapy. In particular, among patients infected by methicillin-susceptible, coagulase-negative staphylococci, 4 of 4 cases of septicemia, 0 of 1 case of endocarditis, 1 of 1 case of pneumonia, 1 of 1 case of peritonitis in CAPD, 2 of 3 infections of skin-soft tissue, and 1 of 1 urinary tract infection responded to therapy. Among patients infected by methicillin-resistant, coagulase-negative staphylococci were cured 5 of 6 cases os septicemia, 6 of 6 cases of peritonitis (in CAPD), 4 of 4 infections of skin-soft tissue, and 0 of 1 urinary tract infection.

Adult↗

Detection of methicillin resistance in coagulase-negative staphylococci initially reported as methicillin susceptible using automated methods.

Reliable detection of methicillin resistance in coagulase-negative staphylococci (CNS) is required for appropriate therapy of serious infections from these pathogens. To determine the most accurate method of measuring methicillin resistance in CNS initially reported as methicillin susceptible by automated methods, we compared mecA detection by polymerase chain reaction (PCR) with phenotypic methods. One hundred eighty-eight blood culture isolates of CNS that were initially reported as susceptible to methicillin using commercial methods (Vitek or MicroScan) were tested by agar dilution, disk diffusion, oxacillin salt agar screen plate, and a multiplex PCR assay using primer sets for mecA and 16S rRNA. Sixteen isolates (8.5%) previously reported as methicillin susceptible by automated methods contained the mecA gene. MICs of these isolates ranged from 0.5 microgram/mL to > or = 128 micrograms/mL. Ten of these isolates had MICs equal to or below the NCCLS breakpoint of 2 micrograms/mL. Six of the 10 isolates (4 with MICs of 0.5 microgram/mL and 2 with MICs of 2 micrograms/mL) did not grow on any of the oxacillin screen plates after 48 h of incubation at 30 degrees C or 35 degrees C. All six isolates were induced to grow in the presence of oxacillin at 128 micrograms/mL by serial passaging on plates containing increasing concentrations of antibiotic. Retesting with MicroScan and Vitek detected methicillin resistance in 7 and 10 isolates, respectively. Disk diffusion testing with incubation for 48 h proved to be the next best method after PCR for detection of methicillin resistance (15 of 16 isolates). Commercial automated methods and some methods recommended by National Committee for Clinical Laboratory Standards may not detect methicillin resistance in CNS that carry the mecA gene and have MICs just below breakpoint.

Bacterial Proteins↗

Induction of beta-lactamase and methicillin resistance in unusual strains of methicillin-resistant Staphylococcus aureus.

Two unusual, heterogeneously-resistant, strains of Staphylococcus aureus appeared resistant to oxacillin, but susceptible to methicillin by disc diffusion methods. In agar dilution tests, both strains were oxacillin-resistant. One was susceptible to methicillin, and the other gave a paradoxical reaction, with growth only on plates containing low (0.5, 1 and 2 mg/l) and high (32 and 64 mg/l) concentrations of antibiotic. Induction of methicillin resistance was tested by inoculating each strain on to agar plates containing an inhibitory concentration of methicillin (8 mg/l), and then placing discs containing inducers (oxacillin, nafcillin, methicillin and CBAP [2-(2'-carboxyphenyl) benzoyl-6-aminopenicillanic acid]) on the agar surface. Colonies grew only around discs containing effective inducers. Oxacillin and CBAP were much more potent inducers of methicillin resistance and beta-lactamase than was nafcillin or methicillin. These data suggest that the mechanism that regulates induction of the low-affinity penicillin binding protein (PBP-2') may be altered in these strains. Similar mechanisms appear to induce both beta-lactamase and methicillin resistance.

Bacterial Proteins↗

In vitro susceptibility of methicillin-resistant and methicillin-susceptible Staphylococcus aureus strains to N-formimidoyl thienamycin.

A total of 82 clinical isolates of methicillin-resistant Staphylococcus aureus and 21 isolates of methicillin-susceptible S. aureus were studied for in vitro susceptibility to N-forminidoyl thienamycin at incubation temperatures of 30 and 35 degrees C. The disk diffusion test results were correlated with the macrobroth dilution test by means of the error rate-bounded method of analysis. Both methicillin-susceptible and (to a lesser degree) methicillin-resistant strains were generally susceptible to the antibiotic as judged from their minimum inhibitory concentrations. The discrepancy between in vitro results obtained at 30 and at 35 degrees C was not very remarkable. However, tolerance of N-formimidoyl thienamycin was observed in 37% of methicillin-resistant strains and 24% of methicillin-susceptible strains at an incubation temperature of 30 degrees C; at 35 degrees C, the values were 54% (methicillin-resistant strains) and 14% (methicillin-susceptible strains).

Anti-Bacterial Agents↗

The in vitro activity of ramoplanin (A-16686/MDL 62,198), vancomycin and teicoplanin against methicillin-susceptible and methicillin-resistant Staphylococcus spp.

Ramoplanin (A-16686/MDL 62,198) is a novel lipoglycopeptide antimicrobial, comprised of three closely related polypeptides containing chlorinated phenyl moieties and D-mannose, isolated from the fermentation products of Actinoplanes sp. ATCC 33076. The antimicrobial activity of ramoplanin is limited to Gram-positive bacteria and its reportedly unacceptable administration side-effects suggest that any potential clinical role will be limited to the topical therapy of superficial skin infections and the eradication of bacteria, representing a possible nosocomial cross-infection source, from carriage sites. In this study the MICs of ramoplanin have been determined for methicillin-susceptible and methicillin-resistant isolates of Staphylococcus aureus, S. epidermidis and S. haemolyticus and compared with those of two glycopeptide antimicrobials, vancomycin and teicoplanin. MICs were determined using an agar incorporation technique in Mueller-Hinton medium with an inoculum of 10(5) cfu. Ramoplanin was 2-8 times more active than either vancomycin or teicoplanin against methicillin-susceptible and methicillin-resistant isolates of S. aureus and methicillin-susceptible isolates of S. epidermidis. Isolates of methicillin-resistant S. epidermidis and both methicillin-susceptible and -resistant isolates of S. haemolyticus were generally less susceptible to teicoplanin than to vancomycin. Ramoplanin was significantly more active than either vancomycin or teicoplanin against these isolates. These results suggest that the clinical evaluation of ramoplanin as a topical antibacterial agent for the control of superficial infections caused by Staphylococcus spp. and for the eradication of methicillin-resistant S. aureus from carriage sites, is justified.

Anti-Bacterial Agents↗

[Susceptibility of methicillin-sensitive and methicillin-resistant staphylococci to ciprofloxacin].

Investigations were carried on 352 strains of Staphylococcus aureus and 105 strains of coagulase-negative staphylococci, isolated from various clinical materials and derived from some regions of the country. Methicillin-resistance was tested by an antibiotic dilution method in solid Mueller-Hinton medium with addition of 2% NaCl. Staphylococci with MIC higher than 4 micrograms/ml were considered as resistant. The same method for testing resistance to ciprofloxacin was used. Only one strain (S. aureus) was resistant to both ciprofloxacin and methicillin. All remaining strains of staphylococci were ciprofloxacin-susceptible. The MIC for all of them was not higher than 2 micrograms/ml, regardless of resistance to methicillin. Some slight differences in MIC50 and MIC90 values were found between MRSA and MSSA and they were, respectively, 1 microgram/ml and 0.25 microgram/ml, and 2 micrograms/ml and 1 microgram/ml. Mean MIC of ciprofloxacin for MRSA was 1.1 microgram/ml and for MSSA it amounted to 0.4 microgram/ml. Range of MIC was following: MRSA--0.12 microgram/ml--16 micrograms/ml and MSSA--0.12-2 micrograms/ml. Coagulase-negative staphylococci, both methicillin-resistant and methicillin-sensitive, exhibited same value for MIC50, MIC90 and MIC range and they were following: 0.5 microgram/ml, 1 microgram/ml and 0.12-1 micrograms/ml. Both groups differed slightly in mean MIC values which was 0.6 microgram/ml for methicillin-resistant strains and 0.47 microgram/ml for methicillin-sensitive staphylococci. It seems that within staphylococci isolated in Poland there is no correlation between resistance to methicillin and ciprofloxacin, which is frequently pointed out by other authors.

Ciprofloxacin↗

Genetic characterisation of resistance to metal ions in methicillin-resistant Staphylococcus aureus: elimination of resistance to cadmium, mercury and tetracycline with loss of methicillin resistance.

Susceptibility to six metal ions--cadmium (Cd), mercury (Hg), arsenate (Asa), arsenite (Asi), antimony (Sb) and zinc (Zn)--was tested in 23 independent isolates of methicillin-resistant Staphylococcus aureus (MRSA) obtained from Guy's Hospital (GH) during 1984-1986, which included 10 isolates of the UK epidemic EMRSA-1 strain. Strains were also tested for resistance to antibiotics and the nucleic-acid-binding compounds propamidine isethionate and ethidium bromide. A further 19 methicillin-resistant isolates, including 10 EMRSA-1 were obtained from other sources. Ten methicillin-sensitive, antibiotic sensitive isolates were from Guy's Hospital. Resistance to Hg was associated with methicillin resistance in 19 of the 20 EMRSA-1 isolates, all of which were resistant to Cd. Resistance to Cd and Hg was found in 13 out of 22 other MRSA isolates. Hg resistance was not present in the methicillin-sensitive isolates which were often (13 out of 19) moderately resistant to Cd. Multiple resistance to metal ions, including resistance to Hg, Asa, Asi and Sb, was uncommon. Resistance to Cd (MIC greater than 32 mg/L or 8-16 mg/L) was associated with increased resistance to Zn. In 11 of the consecutive MRSA isolates from Guy's Hospital seven distinct strains were recognised by phage type. Methicillin resistance in these strains varied from 16 to 1024 mg/L at 30 degrees C with a 2-8-fold lower minimum inhibitory concentration at 37 degrees C indicating some degree of heterogeneity. Representatives of the EMRSA-1 strain had the higher levels of resistance. Loss of methicillin resistance occurred in 0.2-5.0% of colonies tested after storage at room temperature in 10 of these isolates.(ABSTRACT TRUNCATED AT 250 WORDS)

Bacteriophages↗