Investigation of Staphylococcus strains with heterogeneous resistance to glycopeptides in a Turkish university hospital

BACKGROUND: The hetero-glycopeptide intermediate staphylococci is considered to be the precursor of glycopeptide intermediate staphylococci especially vancomycin intermediate Staphylococcus aureus (VISA). For this purpose, we aimed to investigate the heterogeneous resistance to glycopeptide and their frequencies in 135 Staphylococcus strains. METHODS: Heterogeneous resistance of Staphylococcus strains was detected by inoculating the strains onto Brain Heart Infusion agar supplemented with 4 mg/L of vancomycin (BHA-V4). Agar dilution method was used for determining MICs of glycopeptides and population analysis profile was performed for detecting frequency of heterogeneous resistance for the parents of selected strains on BHA-4. RESULTS: Eight (6%) out of 135 Staphylococcus strains were exhibited heterogeneous resistance to at least one glycopeptide. One (1.2%) out of 81 S. aureus was found intermediate resistance to teicoplanin (MIC 16 mg/L). Other seven strains were Staphylococcus haemolyticus (13%) out of 54 coagulase negative staphylococci (CoNS). Six of the seven strains were detected heterogeneously reducing susceptibility to vancomycin (MICs ranged between 5–8 mg/L) and teicoplanin (MICs ranged between 32–64 mg/L), and one S. haemolyticus was found heterogeneous resistance to teicoplanin (MIC 32 mg/L). Frequencies of heterogeneous resistance were measured being one in 10(6 )– 10(7 )cfu/ml. MICs of vancomycin and teicoplanin for hetero-staphylococci were determined as 2–6 folds and 3–16 folds higher than their parents, respectively. These strains were isolated from six patients (7%) and two (4%) of health care wokers hands. Hetero-VISA strain was not detected. CONCLUSION: Heterogeneous resistance to glycopeptide in CoNS strains was observed to be significantly more emergent than those of S. aureus strains (vancomycin P 0.001, teicoplanin, P 0.007). The increase MICs of glycopeptide resistance for subpopulations of staphylococci comparing with their parents could be an important clue for recognizing the early steps in the appearance of VISA strains. We suggested to screen clinical S. aureus and CoNS strains, systematically, for the presence of heterogeneously resistance to glycopeptide

Investigation of Chlorhexidine Tolerance and its Association with Antibiotic Resistance in Clinical Gram-Negative Bacilli Isolates

Background: Chlorhexidine (CHX) is one of the most frequently used antiseptic agents in challenging multidrugresistant (MDR) isolates. There is an increasing number of reports on reduced susceptibility (tolerance) to CHX in MDR isolates. We aimed to investigate CHX tolerance in Gram-negative bacilli (GNB) and its possible association with antibiotic resistance. Methods: A total of 84 enteric GNB (ENT) and 40 non-fermentative GNB (NFGNB) isolates were collected from different clinical specimens. Disk diffusion method was performed to differentiate between MDR and non-MDR isolates and tolerance to CHX was determined by a modified agar dilution method. In GNB isolates, CHX tolerance was defined as a minimum inhibitory concentration (MIC) >= 4 mg/L. Results: We detected that 26.2% (22/84) of ENT and 50.0% (20/40) of NFGNB were MDR and the rest were nonMDR isolates. The CHX tolerance rate was detected as 50.0% (10/20) in MDR-NFGNB and 15.0% (3/20) in nonMDR-NFGNB, and this difference was statistically significant (p 0.05). Conclusions: NFGNB isolates had a higher tendency to CHX tolerance than ENT, and antibiotic resistance facilitates the selection of CHX tolerance in NFGNB but not in ENT isolates

Frequency of antiseptic resistance genes in clinical staphycocci and enterococci isolates in Turkey

Abstract Background Disinfectants and antiseptics are biocides widely used in hospitals to prevent spread of pathogens. It has been reported that antiseptic resistance genes, qac’s, caused tolerance to a variety of biocidal agents, such as benzalkonium chloride (BAC) and chlorhexidine digluconate (CHDG) in Staphylococcus spp. isolates. We aimed to search the frequency of antiseptic resistance genes in clinical Staphylococcus spp. and Enterococcus spp. isolates to investigate the possible association with antiseptic tolerance and antibiotic resistance. Methods Antiseptic resistance genes (qacA/B, smr, qacG, qacH, and qacJ) isolated from Gram-positive cocci (69 Staphylococcus spp. and 69 Enterococcus spp.) were analyzed by PCR method. The minimum inhibitory concentrations (MICs) of BAC and CHDG were determined by agar dilution method, whereas antibiotic susceptibility was analyzed by disk diffusion method according to Clinical and Laboratory Standards Institute (CLSI) criteria. Results The frequency of antiseptic resistance genes was found to be high (49/69; 71.0%) in our clinical staphylococci isolates but absent (0/69; 0%) in enterococci isolates. The frequency of qacA/B and smr genes was higher (25/40; 62.5% and 7/40; 17.5%, respectively) in coagulase negative staphylococci (CNS) when compared to Staphylococcus aureus strains (3/29; 10.3%, and 4/29; 13.8%, respectively). In contrast, the frequency of qacG and qacJ genes was higher (11/29; 37.9% and 8/29; 27.5%, respectively) in S. aureus than those of CNS (5/40; 12.5%, 10/40; 25.0%) strains. qacH was not identified in none of the strains. We found an association between presence of antiseptic resistance genes and increased MIC values of BAC (>4 μg/mL) in staphylococci and it was found to be statistically statistically significant (p < 0.01). We also showed that MICs of BAC and CHDG of vancomycin-resistant enterococci (VRE) isolates were significantly higher than those of vancomycin-susceptible enterococci (VSE) isolates (p < 0.01). Conclusions For our knowledge, our study is the first to investigate antiseptic resistance genes in enterococci and also qacG, qacH, and qacJ genes in staphylococci isolates in Turkey. Further studies are needed to revise the biocide policy and to support infection control programs to avoid the development of new resistance mechanisms

IN VITRO SUSCEPTIBILITY OF ENTEROCOCCUS STRAINS TO HIGH LEVEL AMINOGLYCOSIDES AND HEAVY METALS

The widespread use of antimicrobial agents in the hospitals and environmental contamination with heavy metals are increasingly related to resistance progression in microorganisms. The aim of this study was to investigate the resistance of enterococci to high level aminoglycosides and some heavy metals [lead (Pb(+2)), cadmium (Cd(+2)), mercury (Hg(+2)), arsenic (As(+5))]. A total of 39 Enterococcus strains, isolated from stool and rectal swabs of hospitalized patients were included to the study. Twenty of the strains were resistant to glycopeptides (11 were resistant to vancomycin + teicoplanin and 9 were resistant to only vancomycin). Disk diffusion method was performed to determine the high level resistance to aminoglycosides (gentamicin 120 mu g and streptomycin 300 mu g), and agar dilution method was used to detect the sensitivities of the strains against different concentrations (0.005-20 mM begin_of_the_skype_highlighting 005-20 mM &Uuml;CRETSİZ end_of_the_skype_highlighting) of heavy metals. Since there is no specified minimum inhibitory concentration (MIC) breakpoints for heavy metals, resistance criteria described in previous studies were used. Accordingly, enterococci which exhibited MIC &gt;= 1 mM for lead and cadmium, MIC &gt;= 0.1 mM for mercury, and MIC &gt;= 10 mM for arsenic were accepted as resistant. High level aminoglycoside (HLAG) resistance rates were found as 91% (10/11) for vancomycin (V) + teicoplanin (T) resistant and 42% (8/19) for glycopeptide susceptible strains. While all of the isolates were resistant to lead (100%), arsenic (2.6%) and mercury (2.6%) resistance was detected in one isolate for each metal. No cadmium resistance has been detected. In our study, enterococci have exhibited seven different resistance profiles (10 strains were resistant to V + T + HLAG + Pb; 1 was resistant to V + T + Pb; 1 was resistant to V + As + Pb; 1 was resistant to HLAG + Hg + Pb; 8 were resistant to V + Pb; 7 were resistant to HLAG + Pb; 11 were only resistant to Pb). Resistance to antibiotics (aminoglycosides and/or vancomycin and/or teicoplanin) and heavy metals (lead and arsenic and/or mercury) were detected concurrently in 28 (%71.8) of the strains. It was considered remarkable that all of the isolates were resistant to lead and there was no difference between antibiotic-resistant and-susceptible strains in terms of lead resistance. In conclusion, further investigations are needed to reveal the extreme lead resistance and the relations between antibiotic and heavy metal resistances in clinical enterococcus strains. Keywords Author Keywords:Enterococcus; glycopeptide; high level aminoglycoside; heavy metal; susceptibility KeyWords Plus:ANTIBIOTIC-RESISTANCE; STAPHYLOCOCCUS; PLASMID; GENE

Comparison of polymerase chain reaction and conventional methods in detecting methicillin-resistant Staphylococcus aureus

Background: Accurate and rapid detection of methicillin-resistant Staphylococcus aureus is very important in a clinical laboratory setting to avoid treatment failure. Conventional methods were compared against the gold standard polymerase chain reaction (PCR) technique to determine the best combination of the routine procedures. Methodology: Methicillin resistance was investigated in 416 clinical Staphylococcus aureus isolates by PCR, oxacillin agar screening (OAS), oxacillin disk diffusion (ODD) and cefoxitin disk diffusion (CDD) methods. Results: Two hundred and ten (51%) out of 416 S. aureus strains were found to be mecA-positive by PCR. Sensitivity and specificity of the ODD, CDD and OAS methods were detected as follows: 100% and 89%, 99.50% and 100%, and 99.50% and 100%, respectively . Conclusion: Combining the ODD and CDD methods could be a good choice for detecting methicillin resistance in S. aureus strains where mecA PCR cannot be performed. Key Words : Methicillin-resistant Staphylococcus aureus (MRSA), polymerase chain reaction (PCR), oxacillin disk diffusion, cefoxitin disk diffusion, oxacillin agar screening

Prevalence of phenotypic resistance of Staphylococcus aureus isolates to macrolide, lincosamide, streptogramin B, ketolid and linezolid antibiotics in Turkey

The incidence of drug-resistant pathogens differs greatly between countries according to differences in the usage of antibiotics. The purpose of this study was to investigate the phenotypic resistance of 321 methicillin resistance Staphylococcus aureus (MRSA) and 195 methicillin susceptible S. aureus (MSSA) in a total of 516 S. aureus strains to macrolide, lincosamide, streptogramin B (MLS(B)), ketolid, and linezolid. Disk diffusion method was applied to determine MLS(B) phenotype and susceptibility to different antibiotic agents. It was found that 54.6% of the isolates were resistant to erythromycin (ERSA), 48% to clindamycin, 55% to azithromycin, 58.7% to spiramycin, 34.7% to telithromycin, and 0.4% to quinupristin-dalfopristin, respectively. No strain resistant to linezolid was found. The prevalence of constitutive (cMLS(B)), inducible (IMLS(B)), and macrolides and type B streptogramins (M/MS(B)) among ERSA isolates (237 MRSA, 45 MSSA) was 69.6 %, 18.2%, and 12.2 % in MRSA and 28.9%, 40%, and 31.1% in MSSA, respectively. In conclusions, the prevalence of cMLS(B) was predominant in MRSA; while in MSSA strains, iMLS(B) and M/MS(B) phenotype were more higher than cMLS(B) phenotype resistance. The resistance to quinupristin-dalfopristin was very low, and linezolid was considered as the most effective antibiotic against all S. aureus strains. Keywords Author Keywords:Staphylococcus aureus; macrolide; lincosamide; streptogramin B; ketolid; linezolid; ML

Prevalence of phenotypic resistance of Staphylococcus aureus isolates to macrolide, lincosamide, streptogramin B, ketolid and linezolid antibiotics in Turkey

The incidence of drug-resistant pathogens differs greatly between countries according to differences in the usage of antibiotics. The purpose of this study was to investigate the phenotypic resistance of 321 methicillin resistance Staphylococcus aureus (MRSA) and 195 methicillin susceptible S. aureus (MSSA) in a total of 516 S. aureus strains to macrolide, lincosamide, streptogramin B (MLS(B)), ketolid, and linezolid. Disk diffusion method was applied to determine MLS(B) phenotype and susceptibility to different antibiotic agents. It was found that 54.6% of the isolates were resistant to erythromycin (ERSA), 48% to clindamycin, 55% to azithromycin, 58.7% to spiramycin, 34.7% to telithromycin, and 0.4% to quinupristin-dalfopristin, respectively. No strain resistant to linezolid was found. The prevalence of constitutive (cMLS(B)), inducible (IMLS(B)), and macrolides and type B streptogramins (M/MS(B)) among ERSA isolates (237 MRSA, 45 MSSA) was 69.6 %, 18.2%, and 12.2 % in MRSA and 28.9%, 40%, and 31.1% in MSSA, respectively. In conclusions, the prevalence of cMLS(B) was predominant in MRSA; while in MSSA strains, iMLS(B) and M/MS(B) phenotype were more higher than cMLS(B) phenotype resistance. The resistance to quinupristin-dalfopristin was very low, and linezolid was considered as the most effective antibiotic against all S. aureus strains. Keywords Author Keywords:Staphylococcus aureus; macrolide; lincosamide; streptogramin B; ketolid; linezolid; ML

Investigation of in vitro efficacy of meropenem/ polymyxin B and meropenem/fosfomycin combinations against carbapenem resistant Klebsiella pneumoniae strains

The incidence of infections caused by carbapenem-resistant Klebsiella pneumoniae (CRKP) is increasing worldwide, and very limited number of effective antibiotics are available for therapy. In our study, the in vitro efficacy of meropenem/polymyxin B and meropenem/fosfomycin combinations against CRKP strains was investigated. The efficiency of meropenem/polymyxin B and meropenem/fosfomycin combinations was tested by checkerboard microdilution and checkerboard agar dilution methods, respectively, against 21 CRKP strains containing major carbapenem resistant genes (7 blaKPC, 7 blaOXA-48 gene, and 7 blaOXA-48 thorn blaNDM), and seven additional CRKP strains without carbapenemase genes. Among the 28 CRKP strains, the meropenem/polymyxin B combination was synergistic in ten (35.7%), partially synergistic in 12 (42.8%), and indifferent in six (21.4%) isolates. The meropenem/ fosfomycin combination was found to be synergistic in three isolates (10.7%), partially synergistic in 20 (71.4%), and indifferent in five (17.8%). In 21 strains containing carbapenem resistance genes, meropenem/polymyxin B and meropenem/fosfomycin combinations exhibited synergistic/partial synergistic effects in 15 (71.4%) and 16 (76.2%) strains, respectively, compared to 100% synergistic/partial synergistic efficiency in both combinations in seven strains free of carbapenemase genes. No antagonistic effect was detected in either combination.Regardless of presence or absence of carbapenem resistance genes, meropenem/polymyxin B and meropenem/fosfomycin combinations both demonstrated high synergistic and partial synergistic activity against 78.4% and 82.1% of CRKP strains, respectively. Also, they have no antagonistic effects and can be used successfully to prevent therapeutic failure with monotherapy, according to our in vitro studies