Minimum Inhibitory Concentration Of Chlorhexidine Research Articles

Antibacterial effect of resveratrol extract compared to chlorhexidine mouthwash against primary cariogenic pathogen, Streptococcus mutans.

Modern dental caries prevention methods have focused on using herbal products that ideally inhibit the critical cariogenic bacteria (Streptococcus mutans). The present study compared antibacterial efficacy of the resveratrol herbal extract and 0.12% chlorhexidine (CHX) against Streptococcus mutans (S. mutans). In this in vitro study the pure powder of resveratrol (Bulksupplement) was dissolved in dimethyl sulfoxide as its solvent to produce a 1000-µg/mL concentration of resveratrol solution, which was later used at different dilutions. The antibacterial effects of resveratrol solution and 0.12% CHX mouthrinse on the standard strain of S. mutans were determined using a minimum inhibitory concentration (MIC) test in a tube, minimum bactericidal concentration (MBC) test in a solid medium, and a well diffusion test to measure the zone of inhibition. The data were analyzed using Kruskal-Wallis, Bonferroni, and Man-Whitney tests (α=0.05). The MIC and MBC of resveratrol was 250 µg/mL. In addition, this extract exhibited a diameter of 6.67 mm for the inhibition zone at only the 1000-µg/mL concentration. The MIC of CHX was 15.6 µg/mL, and its MBC was 31.25 µg/mL. The highest growth inhibition zone of CHX was 16.67 mm. Resveratrol extract exhibited a dose-dependent antibacterial (bacteriostatic and bactericidal) activity against S. mutans. Although it was not as effective as CHX, it might be a suitable alternative to prevent dental caries. Key words:Antibacterial agents, Chlorhexidine digluconate, mouthwashe, resveratrol, streptococcus mutans.

Synergistic effect of chlorhexidine and azoles on candida biofilm on titanium surface.

Candida infections of orthopedic implants are one of the most detrimental orthopedic implant-related complications with unsuccessful treatment and a poor prognosis. Most orthopedic Candida infections form biofilms and have resistance to the commonly used antifungal agents. This study aimed to develop a novel combination of normally prescribed drugs against Candida biofilm on orthopedic implants. We cultured 26 clinical isolates of Candida strains to form biofilm without titanium sheets or on titanium sheets, which are the most commonly used materials for permanent or orthopedic implants. The checkerboard method was used to evaluate the synergistic effects of chlorhexidine (CHL) and azoles on these Candida biofilms. For the evaluation of synergistic effects, we constructed the cell viability assay by fluorescence staining and CFU reduction hot map of Candida. Twenty-six clinical isolates of Candida strains formed biofilm in 96-well plates without titanium sheets, and we selected 9 of them to form biofilm on titanium sheets in 24-well plates. In Candida biofilm formed in 96-wells, the synergistic rates of CHL with fluconazole, itraconazole, and voriconazole were 61% (16/26), 65% (17/26), and 23% (6/26), respectively. When compared to the blank control group, CHL monotherapy significantly inhibited biofilm formation on titanium sheets (P < 0.05). We demonstrated 100% synergistic rates of the CHL and fluconazole combination against Candida biofilm formation on titanium sheets, and the minimum inhibitory concentration of CHL and FLU decreased four- to eight-fold. We concluded that CHL combined with azoles inhibited the Candida biofilm formation 96-wells or on titanium sheets and has the potential to control the infections of orthopedic implants.

EVALUATION AND COMPARISON OF ANTIBACTERIAL EFFICACY OF ALBIZIA LEBBECK (L.) BENTH., BAUHINIA VARIEGATA L. AND CHLORHEXIDINE MOUTHWASHES: IN-VITRO STUDY

Aim: Herbal medicines have therapeutic benefits, especially in plaque and gingivitis control. Therefore, the study aims to establish the minimum inhibitory concentration (MIC) of Albizia lebbeck (L.) Benth. (Siris tree), Bauhinia variegata Linn. (Mountain Ebony) and Chlorhexidine mouthwashes and to assess and compare the antibacterial effect of Albizia lebbeck (L.) Benth. mouthwash (Group I), Bauhinia variegata L. mouthwash (Group II) and Chlorhexidine (Group III) on plaque. Materials and Methods: The MIC of herbal mouthwashes and Chlorhexidine were determined against standard strains of bacteria involved in gingivitis. The antibacterial action of these mouthwashes on the supragingival plaque was determined and compared. Fifteen plaque samples were collected from fifteen individuals with gingivitis and were sent to the laboratory for processing. Data obtained was tabulated and subjected to statistical analysis using analysis of variance and paired t-test. Bonferroni post hoc test was used to know the difference between the pairs of mouthwashes. Analysis of covariance was also done to adjust for the baseline differences. Results: The MIC of Group I, Group II and Group III mouthwashes ranged 0.8 µg/ml-12.5 µg/ml, 0.4 µg/ml-25 µg/ml and 0.4 µg/ml-25 µg/ml, respectively, against the tested bacterial strains. Group I and Group II mouthwashes did exhibit antibacterial activity. However, the antibacterial efficacy of herbal mouthwashes exhibited significant differences when compared with Chlorhexidine. Conclusion: Group I and Group II mouthwashes showed antibacterial activity. However, these herbal mouthwashes showed less antibacterial action when compared to Group III.

In Vitro Antimicrobial Activity of Piper betle Leaf Extract and Some Topical Agents against Methicillin-Resistant and Methicillin-Susceptible Staphylococcus Strains from Canine Pyoderma.

As multidrug-resistant methicillin-resistant staphylococci (MRS) is becoming more prevalent in canine pyoderma, the discovery of new therapeutic options is required. This study aimed to test the antimicrobial activity of crude Piper betle leaf extract and some topical antimicrobial agents against canine Staphylococcus clinical strains by determining the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC). The results showed that the mean MICs of chlorhexidine, miconazole, crude P. betle leaf extract, azelaic acid, and benzoyl peroxide against Staphylococcus strains were 1.41, 1.62, 252.78, 963.49, and 1342.70 mg/L, respectively. Therefore, betel leaf extract demonstrated a superior efficacy to azelaic acid and benzoyl peroxide. Furthermore, the ratio of MBC/MIC of betel leaf extract was 1.75, indicating its bactericidal action. When applied to methicillin-resistant S. pseudintermedius (MRSP) and methicillin-susceptible S. pseudintermedius (MSSP), betel leaf extract was equally efficient towards both groups. S. pseudintermedius strains were more susceptible to betel leaf extract than S. schleiferi subsp. coagulans. In gas chromatography-mass spectrometry analysis, eugenol and hydroxychavicol appeared to be the major components of betel leaf extract. Given its efficacy, dogs with pyoderma could benefit from the use of betel leaf extract as a topical antimicrobial alternative.

Minimum inhibitory concentrations of chlorhexidine- and lactic acid-based teat disinfectants: An intervention trial assessing bacterial selection and susceptibility

Teat disinfection is a recommended preventive tool to improve udder health and to prevent new intramammary infections. However, side effects are discussed, such as bacterial selection of less-susceptible bacteria with the application of certain teat disinfectants. The objective of this study was to assess the species composition and bacterial in vitro susceptibility by means of an interventive trial. For this purpose, 3 different postmilking teat treatments (disinfection with 0.215% chlorhexidine or 3.5% lactic acid, or control group with no dipping) were applied to 28 cows in a 6-d intervention approach using a split-udder design. Milk samples were taken before and after intervention. Bacteria were cultured and differentiated to species or genus level by MALDI-TOF mass spectrometry. Minimum inhibitory concentrations (MIC) were determined, and MIC changes over time were recorded. Susceptibilities to chlorhexidine and lactic acid were compared between species of the genera Staphylococcus, Streptococcus, Corynebacterium, and others. Species composition changed during the intervention. Under the treatment of chlorhexidine and lactic acid, the proportion of coagulase-negative staphylococci (CNS) decreased. An increased proportion of species belonging to the genus Corynebacterium was observed especially under the application of lactic acid. Although both teat disinfectants were basically effective, isolates differed in their susceptibility to both teat disinfectants. Populations of CNS, Staphylococcus aureus, and Corynebacterium spp. showed significantly lower absolute MIC values for chlorhexidine. Compared with other species, Corynebacterium spp. showed the lowest susceptibility for chlorhexidine as well as for lactic acid. A significant increase in MIC values after 6 d of intervention was observed with the lactic acid treatment in all isolates, as well as in CNS. This increase can be interpreted as either adaptation of isolates or displacement of more-susceptible species by less-susceptible species. Further studies using long-term intervention might reveal more pronounced effects on MIC values and species composition.

Chlorhexidine leads to the evolution of antibiotic-resistant Pseudomonas aeruginosa.

Antimicrobial resistance is a major public-health concern. We evaluate chlorhexidine role in selection of resistant Pseudomonas aeruginosa mutants and their antibiotic cross-resistance. Mutation frequency and mutation rate after short-term exposure to sub-inhibitory concentrations of chlorhexidine were compared to those after spontaneous chlorhexidine-exposure, in P. aeruginosa PAO1 strain. Chlorhexidine-resistant mutants were generated, either by serial passage in increasing chlorhexidine concentrations or by single exposure to lethal chlorhexidine concentration. The generated mutants were tested for cross-resistance to different antibiotics, by determination of minimum inhibitory concentrations (MIC). The accompanied phenotypic changes in membrane permeability, outer membrane proteins (OMP), and efflux function were evaluated. The effect of exposure to chlorhexidine on MexAB-OprM, MexEF-oprN, and MexXY efflux pumps expression was investigated. No significant change was recorded between the mutation frequencies and mutation rates after short-term exposure to sub-inhibitory concentrations of chlorhexidine and after spontaneous chlorhexidine-exposure, in P. aeruginosa PAO1 strain. Twelve stable mutants, with ≥ eight-fold increase in chlorhexidine MIC, were generated. Several mutants showed increase in the MIC of colistin, cefepime, ceftazidime, meropenem, ciprofloxacin, and amikacin; seven mutants expressed meropenem cross-resistance. This was accompanied by decreased outer membrane permeability and changes in OMP. Using efflux pump inhibitor, chlorhexidine resistance was reverted in most isolates. Exposure to sub-inhibitory concentration of chlorhexidine induced the expression of MexXY efflux pump. Some resistant mutants had overexpressed MexXY efflux pump. Chlorhexidine can select P. aeruginosa strains with antibiotic cross-resistance. This necessitates implementing special protocols for chlorhexidine use and re-evaluation of its benefit versus risk in personal-care products.

Nystatin effect on chlorhexidine efficacy against Streptococcus mutans as planktonic cells and mixed biofilm with Candida albicans.

ObjectiveThe aim of this study was to evaluate the effect of nystatin on the efficacy of chlorhexidine against Streptococcus mutans in planktonic cells and mixed biofilm with Candida albicans.Material and methodsS. mutans ATCC 25,175 in suspension and also combined with C. albicans ATCC 18,804 in biofilm were cultured. Minimum inhibitory concentration (MIC), crystal violet colorimetric assay, and colony-forming unit counting (CFUs/mL) were performed.ResultsAn increased MIC of chlorhexidine against S. mutans was observed when the drugs were administered mixed in a single formulation and with time intervals in between, except for the 30-min interval. The biofilm optical density (OD) in treatments using chlorhexidine and nystatin combined did not significantly differ from chlorhexidine alone. Either in biofilm colorimetric assay or determination of CFUs, the combined treatments with nystatin administered before chlorhexidine had less effect on chlorhexidine efficacy.ConclusionsNystatin interferes with the action of chlorhexidine against S. mutans. The antimicrobial effectiveness of the combined drugs depends on their concentration, time interval used, and the planktonic or biofilm behavior of the microorganisms.Clinical relevanceIn view of the great number of patients that can receive a prescription of chlorhexidine and nystatin concomitantly, this study contributes to the knowledge about the effect of the combined drugs. Given the high prevalence of prescriptions of chlorhexidine and nystatin in dentistry, dental professionals should be aware of their possible antagonistic effect.

Chlorhexidine MICs Remain Stable Among Antibiotic-Resistant Bacterial Isolates Collected from 2005 to 2019 at Three US Sites

Background: Chlorhexidine bathing reduces bacterial skin colonization and prevents infections in specific patient populations. As chlorhexidine use becomes more widespread, concerns about bacterial tolerance to chlorhexidine have increased; however, testing for chlorhexidine minimum inhibitory concentrations (MICs) is challenging. We adapted a broth microdilution (BMD) method to determine whether chlorhexidine MICs changed over time among 4 important healthcare-associated pathogens. Methods: Antibiotic-resistant bacterial isolates (Staphylococcus aureus from 2005 to 2019 and Escherichia coli, Klebsiella pneumoniae, and Enterobacter cloacae complex from 2011 to 2019) were collected through Emerging Infections Program surveillance in 2 sites (Georgia and Tennessee) or through public health reporting in 1 site (Orange County, California). A convenience sample of isolates were collected from facilities with varying amounts of chlorhexidine use. We performed BMD testing using laboratory-developed panels with chlorhexidine digluconate concentrations ranging from 0.125 to 64 μg/mL. After successfully establishing reproducibility with quality control organisms, 3 laboratories performed MIC testing. For each organism, epidemiological cutoff values (ECVs) were established using ECOFFinder. Results: Among 538 isolates tested (129 S. aureus, 158 E. coli, 142 K. pneumoniae, and 109 E. cloacae complex), S. aureus, E. coli, K. pneumoniae, and E. cloacae complex ECVs were 8, 4, 64, and 64 µg/mL, respectively (Table 1). Moreover, 14 isolates had an MIC above the ECV (12 E. coli and 2 E. cloacae complex). The MIC50 of each species is reported over time (Table 2). Conclusions: Using an adapted BMD method, we found that chlorhexidine MICs did not increase over time among a limited sample of S. aureus, E. coli, K. pneumoniae, and E. cloacae complex isolates. Although these results are reassuring, continued surveillance for elevated chlorhexidine MICs in isolates from patients with well-characterized chlorhexidine exposure is needed as chlorhexidine use increases.Funding: NoneDisclosures: None

Genetic resistance determinants to fusidic acid and chlorhexidine in variably susceptible staphylococci from dogs

BackgroundConcern exists that frequent use of topically-applied fusidic acid (FA) and chlorhexidine (CHX) for canine pyoderma is driving clinically relevant resistance, despite rare description of FA and CHX genetic resistance determinants in canine-derived staphylococci. This study aimed to determine minimum inhibitory concentrations (MICs) and investigate presence of putative resistance determinants for FA and CHX in canine-derived methicillin-resistant (MR) and -susceptible (MS) staphylococci. Plasmid-mediated resistance genes (fusB, fusC, fusD, qacA/B, smr; PCR) and MICs (agar dilution) of FA and CHX were investigated in 578 staphylococci (50 MR S. aureus [SA], 50 MSSA, 259 MR S. pseudintermedius [SP], 219 MSSP) from Finland, U.S.A., North (NUK) and South-East U.K. (SEUK) and Germany. In all isolates with FA MIC ≥64 mg/L (n = 27) fusA and fusE were amplified and sequenced.ResultsFA resistance determinants (fusA mutations n = 24, fusB n = 2, fusC n = 36) were found in isolates from all countries bar U.S.A. and correlated with higher MICs (≥1 mg/L), although 4 SP isolates had MICs of 0.06 mg/L despite carrying fusC. CHX MICs did not correlate with qacA/B (n = 2) and smr (n = 5), which were found in SEUK SA, and SP from NUK and U.S.A.ConclusionsIncreased FA MICs were frequently associated with fusA mutations and fusC, and this is the first account of fusB in SP. Despite novel description of qacA/B in SP, gene presence did not correlate with CHX MIC. Selection pressure from clinical use might increase prevalence of these genetic determinants, but clinical significance remains uncertain in relation to high skin concentrations achieved by topical therapy.

In vitro Antibacterial Effect of Hydroalcoholic Extract of Lawsonia inermis, Malva sylvestris, and Boswellia serrata on Aggregatibacter actinomycetemcomitans.

Background:Considering the increased rate of microbial resistance to antibiotics and chemical side effects of antibiotics and antiseptics used for the treatment of periodontal disease, there is a need for an alternative antimicrobial agent with fewer complications. Medicinal herbs have recently become popular as novel antimicrobial agents. This study aimed to assess the antibacterial effects of hydroalcoholic extracts of Lawsonia inermis, Malva sylvestris, and Boswellia serrata on Aggregatibacter actinomycetemcomitans.Materials and Methods:Hydroalcoholic extracts of the three medicinal plants were obtained by the maceration technique and A. actinomycetemcomitans was cultured. Antimicrobial efficacy of the three medicinal plants was compared with that of 0.2% chlorhexidine (CHX) according to the Clinical and Laboratory Standards Institute protocol using agar disc diffusion and broth microdilution techniques. All tests were repeated three times.Results:Hydroalcoholic extracts of all three plants had antimicrobial activity against A. actinomycetemcomitans. The minimum inhibitory concentration (MIC) of L. inermis, M. sylvestris, and B. serrata was 78.1, 156.2, and 1666 μg/mL with no significant difference between them. The MIC of CHX was 3.33 μg/mL, which was significantly higher than that of B. serrata extract.Conclusion:Given that further in vivo studies confirm other properties of these extracts and their safety in terms of cytotoxicity and mutagenicity, hydroalcoholic extracts of L. inermis and M. sylvestris may be used in mouthwashes or local delivery systems to affect periodontal biofilm.

Pseudomonas aeruginosa adapts to octenidine in the laboratory and a simulated clinical setting, leading to increased tolerance to chlorhexidine and other biocides

Octenidine is frequently used for infection prevention in neonatal and burn intensive care units, where Pseudomonas aeruginosa has caused nosocomial outbreaks. To investigate the efficacy and impact of using octenidine against P.aeruginosa. Seven clinical isolates of P.aeruginosa were exposed to increasing concentrations of octenidine over several days. Fitness, minimum bactericidal concentrations after 1min, 5min and 24h, and minimum inhibitory concentrations (MICs) of a variety of antimicrobials were measured for the parental and octenidine-adapted P.aeruginosa strains. Octenidine and chlorhexidine MICs of a population of P.aeruginosa isolated from a hospital drain trap, exposed to a diluted octenidine formulation four times daily for three months, were also tested. Some planktonic cultures of P.aeruginosa survived >50% of the working concentration of an in-use octenidine formulation at the recommended exposure time. Seven strains of P.aeruginosa stably adapted following continuous exposure to increasing concentrations of octenidine. Adaptation increased tolerance to octenidine formulations and chlorhexidine up to 32-fold. In one strain, it also led to increased MICs of antipseudomonal drugs. Subsequent to continuous octenidine exposure of a multi-species community in a simulated clinical setting, up to eight-fold increased tolerance to octenidine and chlorhexidine of P.aeruginosa was also found, which was lost upon removal of octenidine. Incorrect use of octenidine formulations may lead to inadequate decontamination, and even increased tolerance of P.aeruginosa to octenidine, with resulting cross-resistance to other biocides.

Stability and efficacy of combined nystatin and chlorhexidine against suspensions and biofilms of Candida albicans

ObjectiveNystatin and chlorhexidine are extensively used in oral medicine; however, there is some controversy about the possibility of these drugs showing antagonism. To clarify this issue, this study investigated the efficacy and stability of nystatin and chlorhexidine in combination. DesignAn in vitro study was conducted to analyze the effect of nystatin and chlorhexidine combined on Candida albicans ATCC 18804, using the drugs mixed as a single formulation and as independent formulations used sequentially with different time intervals between them. The minimum inhibitory concentration (MIC) and effects on C. albicans suspensions and biofilms were evaluated. Also, the stability of nystatin and chlorhexidine in a mixture was tested by high performance liquid chromatography (HPLC). ResultsWhen nystatin and chlorhexidine were mixed in a single formulation, there was no significant difference in MIC compared to that of the drugs used alone (as the only treatment). However, when these drugs were used as independent formulations, sequentially with time intervals in between, their MICs were higher than the respective MIC of the drug used alone, except for the MIC of chlorhexidine with a 10-min interval. Nystatin/chlorhexidine combinations showed lower activity against C. albicans biofilms, except for that with a 30-min interval. The drugs when combined showed high percentages of degradation at all the times evaluated. ConclusionsThe combination of nystatin and chlorhexidine seems to interfere with the efficacy of the drugs and to increase their rate of degradation.

Minimum Inhibitory Concentration of Chlorhexidine and Cetylpyridinium Chloride against a Mixture of Two Species of Oral Streptococci

Minimum Inhibitory Concentration of Chlorhexidine and Cetylpyridinium Chloride against a Mixture of Two Species of Oral Streptococci

Impact of Brief Exposure to Drugs with Antifungal Properties on the Susceptibility of Oral Candida dubliniensis Isolates to Lysozyme and Lactoferrin

Objective: Lysozyme and lactoferrin have anti-candidal activity. Candida dubliniensis is associated with oral candidiasis. Candida infections are managed with nystatin, amphotericin B, caspofungin, ketoconazole, fluconazole, and chlorhexidine. Candida species undergo a brief exposure to therapeutic agents in the mouth. There is no data on the influence of limited exposure to antimycotics on the sensitivity of C. dubliniensis to lactoferrin and lysozyme. Hence, this study observed the changes in the sensitivity of C. dubliniensis to anti-candidal action of lactoferrin and lysozyme after transitory exposure to sub-lethal concentrations of antifungals. Materials and Methods: After determination of the minimum inhibitory concentration (MIC), 20 C. dubliniensis isolates were exposed to twice the concentration of MIC of nystatin, amphotericin B, caspofungin, ketoconazole, fluconazole, and chlorhexidine for 1 h. Drugs were removed by dilution and thereafter the susceptibility of these isolates to lysozyme and lactoferrin was determined by colony-forming unit quantification assay. Results: Exposure of C. dubliniensis to nystatin, amphotericin B, caspofungin, ketoconazole, fluconazole, and chlorhexidine resulted in an increase in susceptibility to lysozyme by 9.45, 30.82, 30.04, 50.64, 55.60, and 50.18%, respectively (p < 0.05 to p < 0.001). Exposure of C. dubliniensis to nystatin, amphotericin B, caspofungin, ketoconazole, fluconazole, and chlorhexidine resulted in an increase in susceptibility to lactoferrin by 13.54, 16.43, 17.58, 19.60, 21.32, and 18.73, respectively (p < 0.05 to p < 0.001). Conclusion: Brief exposure to nystatin, amphotericin B, caspofungin, ketoconazole, fluconazole, and chlorhexidine enhances the antifungal effect of lysozyme and lactoferrin on C. dubliniensis isolates in vitro.

Spread of Klebsiella pneumoniae ST395 non-susceptible to carbapenems and resistant to fluoroquinolones in North-Eastern France

Fluoroquinolones (FQs) are a potential treatment for infections caused by extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae that are susceptible to these agents. Owing to increasing non-susceptibility to carbapenems among Enterobacteriaceae, in this study FQ resistance mechanisms were characterised in 36 ertapenem-non-susceptible Klebsiella pneumoniae isolated from North-Eastern France in 2012. The population structure was described by pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). Among the 36 isolates, 13 (36%) carried a carbapenemase encoding-gene. Decreased expression of the OmpK35-encoding gene might be considered a major resistance determinant that could explain the non-susceptibility to carbapenems. The carbapenemase-producing isolates carried the well-known IncL pOXA-48a plasmid. All 36 K. pneumoniae isolates also harboured a FQ resistance determinant. The aac(6')-Ib-cr gene was the major plasmid-mediated quinolone resistance (PMQR) determinant found in K. pneumoniae (89%; 32/36). MLST identified five sequence types (STs), with the most common being ST395 (69%; 25/36), followed by ST147 (17%; 6/36). ST395 strains showed ertapenem minimum inhibitory concentrations (MICs) ranging from 0.75-32μg/mL. Klebsiella pneumoniae ST395 isolates did not show enhanced biofilm formation or environmental survival but showed higher chlorhexidine MICs compared with ST147 isolates. These findings showed that (i) K. pneumoniae ST395 carrying pOXA-48a has spread in North-Eastern France, (ii) aac(6')-Ib-cr is predominant in carbapenem-non-susceptible K. pneumoniae, (iii) K. pneumoniae ST395 is resistant to chlorhexidine and (iv) FQs as an alternative to β-lactams to treat ertapenem-non-susceptible K. pneumoniae are compromised.

Detection of Antiseptic Resistance Genes among Staphylococcus aureus Colonising Nurses and Coagulase-Negative Staphylococci Isolated from Clinical Specimens at Teaching Hospitals in Southwest of Iran

Background: The wide application of antibiotics and antiseptics for patient therapy and medical equipment and surfaces disinfection has resulted in the emergence of resistant microorganisms. Staphylococcus aureus and coagulase-negative Staphylococci (CoNS) are found as a part of the normal resident flora in human so that up to two-thirds of the healthy populations are permanently or transiently colonized by S. aureus and CoNS. Chlorhexidine is an antiseptic agent particularly effective against Gram-positive bacteria. It is widely used for hygienic hand wash to prevent transmission of Staphylococci nosocomial infections. The plasmid-borne qacA/B, qacC, and smr genes confer resistance to cationic antiseptic agents in S. aureus and CoNS. Objectives: The objective of the current study was to characterize the antibiotic resistance and susceptibility to quaternary ammonium compounds (QACs) in methicillin-sensitive S. aureus (MSSA), methicillin-resistant S. aureus (MRSA), methicillin-sensitive coagulase-negative staphylococci (MSCoNS), and methicillin-resistant coagulase-negative Staphylococci (MRCoNS). Methods: In this study, the antibiotic susceptibility and resistance to Chlorhexidine in 120 Staphylococcal strains were evaluated by disc diffusion and Minimum inhibitory concentration (MIC) of Chlorhexidine gluconate (CHG) methods, respectively. The MICs of CHG were determined in triplicate by broth micro-dilution, and the presence of mecA, qacA/B, qacC, and smr genes was examined by PCR assay. Results: Of total 60 S. aureus isolates, 51 (85%) were MRSA, and of 60 CoNS, 7 (11.66%) were MRCoNS. The results showed that the MIC of Chlorhexidine for all 120 isolates was 1-16 _g/mL. 15 (12.5%) isolates carried qacA/B gene, 26 (21.7%) carried qacC gene, and 38 (31.7%) carried smr gene. Conclusions: Maintenance of MRSA isolates in the attendance of low amounts of antiseptics could result in the decreased susceptibility to antiseptics.

Epidemiology and microbiological characterization of clinical isolates of Staphylococcus aureus in a single healthcare region of the UK, 2015.

We investigated the epidemiology and characterization of isolates of Staphylococcus aureus within the Yorkshire and Humber (YH) region in the UK. In July 2015, each laboratory within YH (n = 14) was assigned two consecutive days during which all clinical isolates of S. aureus were collected. Isolates were tested for antibiotic susceptibilities and the presence of genes encoding methicillin resistance (mecA and mecC), Panton-Valentine leukocidin (PVL) (lukS-PV), and efflux-mediated chlorhexidine resistance (qacA); isolates were also characterized by spa-types. Minimum inhibitory concentrations (MICs) to chlorhexidine were determined by the broth dilution method. Of 520 isolates collected, 6·2% were methicillin-resistant S. aureus (MRSA, all mecA-positive) and mupirocin resistance was low [0·8%, 95% confidence interval (CI) 0·3-2·0] and only found in MRSA. Carriage of the qacA gene was identified in 1·7% (95% CI 0·8-3·3) of isolates and 3·5% (95% CI 2·2-5·4) had a chlorhexidine MIC of 4 mg/l. The PVL gene was infrequent (3·7%, 95% CI 2·4-5·6). Genotyping identified 234 spa-types that mapped to 22 clonal complexes. Comparison of these current data with previous work suggest that the widespread use of staphylococcal decolonization regimens over the past decade or more has not had an adverse impact on resistance rates, PVL carriage or the prevalence of specific S. aureus lineages.

Effect of daily chlorhexidine bathing on the acquisition of methicillin-resistant Staphylococcus aureus in a medical intensive care unit with methicillin-resistant S aureus endemicity.

Universal decolonization is recommended in intensive care units (ICUs) that have unacceptably high rates of methicillin-resistant Staphylococcus aureus (MRSA) despite implementation of basic prevention strategies. An interrupted time series study was performed to evaluate the effect of daily chlorhexidine bathing on the acquisition of MRSA in a medical ICU with MRSA endemicity. There was a 14-month control period and a 16-month chlorhexidine bathing period. Segmented Poisson regression analysis was performed to assess the impact of daily chlorhexidine bathing on the incidence density of MRSA. Also, chlorhexidine susceptibility testing with polymerase chain reaction for the qacA/B gene was performed on MRSA isolates collected during the chlorhexidine bathing period. There was a significant reduction in trend (-0.056; 95% confidence interval, -0.095 to -0.017; P = .005) of incidence density of MRSA despite a significant increase in both level and trend of MRSA prevalence rates during the chlorhexidine bathing period. However, there was no significant reduction in level of incidence density of MRSA during the interventional period. Minimum inhibitory concentration of chlorhexidine and the detection rates of the qacA/B gene for a total of 174 MRSA isolates did not increase during the chlorhexidine bathing period. Daily chlorhexidine bathing resulted in a significantly decreasing trend of MRSA acquisition rates irrespective of increased MRSA prevalence rates in the medical ICU. There was no shift of chlorhexidine-resistant MRSA strains.

Interaction of chlorhexidine with trisEDTA or miconazole in vitro against canine meticillin-resistant and -susceptible Staphylococcus pseudintermedius isolates from two UK regions.

Topical therapy is an important alternative to systemic antibacterial therapy for treatment of canine superficial pyoderma in light of the emergence of multidrug-resistant staphylococci. Chlorhexidine is widely used in shampoo products alone or in combination with miconazole or tromethamine-ethylenediaminetetraacetic acid (trisEDTA). Comparisons of these combinations have not been made. To determine minimum inhibitory concentrations (MICs) of combinations of chlorhexidine/miconazole and chlorhexidine/trisEDTA in vitro in a collection of Staphylococcus pseudintermedius (SP) from northern (NUK) and southeastern (SEUK) United Kingdom (UK) sources. MICs of chlorhexidine, miconazole, trisEDTA and combinations of chlorhexidine/miconazole (1:1) or chlorhexidine/trisEDTA (80:16:1 and 80:5:1) were determined for 196 canine SP isolates from NUK [49 meticillin-resistant (MRSP), 50 meticillin-susceptible (MSSP)] and fom SEUK (48 MRSP, 49 MSSP) using agar dilution. TrisEDTA alone did not inhibit growth. Chlorhexidine/miconazole MICs (median = 0.5 mg/L) were lower than those of either drug alone (P < 0.05) and lower than chlorhexidine/trisEDTA MICs (median = 1 mg/L; P < 0.0005) in each bacterial type and from both regions, except for miconazole in NUK MSSP. An additive interaction was noted between chlorhexidine and miconazole or trisEDTA (80:16:1 ratio) in 79 and 43 isolates, respectively, whereas antagonism between chlorhexidine and trisEDTA was noted for three isolates. NUK isolates were more susceptible than SEUK isolates (P < 0.05), except MRSP exposed to chlorhexidine and the chlorhexidine/trisEDTA (80:16:1) combination. These low MICs are likely to be exceeded by topical therapy. Evaluation of the mechanisms by which chlorhexidine combinations interact to reduce MICs is warranted, in view of increasing concerns of biocide tolerance in staphylococci.

Chlorhexidine and mupirocin susceptibilities in methicillin-resistant Staphylococcus aureus isolates from bacteraemia and nasal colonisation.

Chlorhexidine and mupirocin have been increasingly used in healthcare facilities to eradicate methicillin-resistant Staphylococcus aureus (MRSA) carriage. The aim of this study was to determine the prevalence and mechanisms of chlorhexidine and mupirocin resistance in MRSA from invasive infections and colonisation. MRSA isolates obtained from blood and nasal samples between 2012 and 2014 were analysed. Susceptibility to mupirocin was determined by disk diffusion and Etest and susceptibility to chlorhexidine by broth microdilution. The presence of mupA and qac (A/B and C) genes was investigated by PCR. Molecular typing was performed in high-level mupirocin-resistant (HLMR) isolates. Mupirocin resistance was identified in 15.6% of blood isolates (10.9% HLMR) and 15.1% of nasal isolates (12.0% HLMR). Presence of the mupA gene was confirmed in all HLMR isolates. For blood isolates, chlorhexidine minimum inhibitory concentrations (MICs) ranged from ≤0.125 to 4mg/L and minimum bactericidal concentrations (MBCs) from ≤0.125 to 8mg/L. In nasal isolates, chlorhexidine MICs and MBCs ranged from ≤0.125 to 2mg/L. The qacA/B gene was detected in 2.2% of MRSA isolates (chlorhexidine MIC range 0.25-2mg/L) and the qacC gene in 8.2% (chlorhexidine MIC range ≤0.125-1mg/L). The prevalence of qacC was 18.9% in HLMR isolates and 3.6% in mupirocin-susceptible isolates (P=0.009). Most of the HLMR isolates (97.1%) belonged to ST125 clone. These results suggest that chlorhexidine has a higher potential to prevent infections caused by MRSA. In contrast, mupirocin treatment should be used cautiously to avoid the spread of HLMR MRSA.