agar and Cross-Infection

The incidence of invasive candidiasis has increased dramatically over the last decades due to a larger number of patients at risk. The diagnosis remains difficult as the clinical presentation is not specific and the biological diagnosis usually takes several days to become positive. We propose in this work through a prospective study to evaluate the contribution of a chromogenic medium CHROMagar(®) (Becton-Dickinson) in the mycological diagnosis of Candida. We selected 680 samples from patients hospitalized in the intensive care unit for epidemiological surveillance over a period of 11 weeks. We treated samples by culture on Sabouraud and on CHROMagar(®). The species identification was performed by chlamydosporulation test and carbohydrate assimilation tests. We found that the CHROMagar(®)Candida evaluated in our work was a valuable tool in the primary culture in differentiating the most frequently isolated yeast species and in better detection of mixed cultures.

Topics: Agar; Axilla; Candida; Candidiasis; Chromogenic Compounds; Cross Infection; Culture Media; Fungal Proteins; Hexosaminidases; Hospitals, Military; Humans; Inpatients; Intensive Care Units; Monophenol Monooxygenase; Mouth; Nasal Cavity; Population Surveillance; Predictive Value of Tests; Prospective Studies; Rectum; Sensitivity and Specificity; Species Specificity; Tunisia

Acinetobacter causes nosocomial infections and its biofilm formation can contribute to the survival on dry surfaces such as hospital environments. Thus, biofilm quantification and visualization are important methods to assess the potential of Acinetobacter strains to cause nosocomial infections. The biofilms forming on the surface of the microplate can be quantified in terms of volume and cell numbers. Biofilm volumes can be quantified by staining using crystal violet, washing, destaining using ethanol, then measuring the solubilized dye using a microplate reader. To quantify the number of cells embedded in the biofilms, the biofilms are scrapped off using cell scrapers, harvested in the saline, vigorously agitated in the presence of glass beads, and spread on Acinetobacter agar. Then, the plates are incubated at 30 °C for 24-42 h. After incubation, the red colonies are enumerated to estimate the number of cells in biofilms. This viable count method can also be useful for counting Acinetobacter cells in mixed-species biofilms. Acinetobacter biofilms can be visualized using fluorescent dyes. A commercially available microplate designed for microscopic analysis is employed to form biofilms. Then, the bottom-surface attached biofilms are stained with SYTO9 and propidium iodide dyes, washed, then visualized with confocal laser scanning microscopy.

Topics: Acinetobacter; Agar; Biofilms; Cross Infection; Fluorescent Dyes; Humans

BACKGROUND Partition curtains are one of the main sources of nosocomial infection in the hospital environment. However, there are no unified standards for monitoring medical textiles across different countries or regions. This study aimed to investigate the accuracy of 2 different sampling methods - swabbing vs RODAC (replicate organism detection and counting) agar plate - in terms of detection of bacterial contamination, and their suitability as monitoring methods for partition curtains and other medical textiles. MATERIAL AND METHODS A total of 24 partition curtains were selected by stratified random sampling. The swabbing technique and RODAC agar plates were the chosen sampling methods. The number of colony-forming units was calculated and colony morphologies and strains on the plates were observed and identified after culturing. RESULTS A total of 192 samples were collected. Of them, 161 pathogenic strains were isolated via the swabbing technique and 309 pathogenic strains were isolated using the RODAC agar plates. The swabbing technique had a higher proportion for gram-positive bacteria (P=0.0004), while RODAC agar plates had a higher proportion for gram-negative bacteria (P=0.72). The detection of bacterial contamination rates using the swabbing technique was superior to that of the RODAC agar plate method (P<0.001). CONCLUSIONS The swabbing technique offers more advantages in terms of detection of bacterial contamination rates and gram-positive bacteria, while the RODAC agar plate is more sensitive for detection of gram-negative bacteria.

Topics: Agar; Bone Plates; Cross Infection; Hospitals; Humans; Research Design

From 1 January to 31 December 2022, fifty-five institutions across Australia participated in the Australian Staphylococcus aureus Surveillance Outcome Program (ASSOP). The aim of ASSOP 2022 was to determine the proportion of Staphylococcus aureus bacteraemia (SAB) isolates in Australia that were antimicrobial resistant, with particular emphasis on susceptibility to methicillin and on characterisation of the molecular epidemiology of the methicillin-resistant isolates. A total of 3,214 SAB episodes were reported, of which 77.5% were community-onset. Overall, 15.0% of S. aureus were methicillin resistant. The 30-day all-cause mortality associated with methicillin-resistant SAB was 21.4%, which was significantly different to the 16.8% all-cause mortality associated with methicillin-susceptible SAB (p = 0.02). With the exception of the β-lactams and erythromycin, antimicrobial resistance in methicillin-susceptible S. aureus was rare. However, in addition to the β-lactams, approximately 31% of methicillin-resistant S. aureus (MRSA) were resistant to ciprofloxacin; 30% to erythromycin; 13% to tetracycline; 11% to gentamicin; and 2% to co-trimoxazole. One MRSA isolate, with a daptomycin MIC of 1.5 mg/L, harboured the A302V mprF and A23V cls2 mutations. When applying the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints, teicoplanin resistance was detected in one MRSA isolate. Resistance to vancomycin or linezolid was not detected. Resistance to non-β-lactam antimicrobials was largely attributable to the healthcare-associated MRSA (HA-MRSA) clone ST22-IV [2B] (EMRSA-15), and to the community-associated MRSA (CA-MRSA) clone ST45-V [5C2&5] which has acquired resistance to multiple antimicrobials including ciprofloxacin, clindamycin, erythromycin, gentamicin, and tetracycline. The ST22-IV [2B] (EMRSA-15) clone is the predominant HA-MRSA clone in Australia. Nonetheless, 86% of methicillin-resistant SAB episodes were due to CA-MRSA clones. Although polyclonal, approximately 72% of CA-MRSA clones were characterised as ST93-IV [2B] (Queensland clone); ST5-IV [2B]; ST45-V [5C2&5]; ST1-IV [2B]; ST30-IV [2B]; ST97-IV [2B]; ST953-IV [2B]; and ST8-IV [2B]. As CA-MRSA is well established in the Australian community, it is important to monitor antimicrobial resistance patterns in community- and healthcare-associated SAB as this information will guide therapeutic practices in treating S. aureus bacteraemia.

Topics: Agar; Anti-Bacterial Agents; Anti-Infective Agents; Australia; Bacteremia; Ciprofloxacin; Cross Infection; Drug Resistance, Bacterial; Erythromycin; Gentamicins; Humans; Methicillin; Methicillin-Resistant Staphylococcus aureus; Staphylococcal Infections; Staphylococcus aureus; Tetracycline

From 1 January to 31 December 2020, forty-nine institutions around Australia participated in the Australian Staphylococcus aureus Sepsis Outcome Programme (ASSOP). The aims of ASSOP 2020 were to determine the proportion of Staphylococcus aureus bacteraemia (SAB) isolates in Australia that were antimicrobial resistant, with particular emphasis on susceptibility to methicillin; and to characterise the molecular epidemiology of the methicillin-resistant isolates. A total of 2,734 SAB episodes were reported, of which 79.7% were community-onset. Of S. aureus isolates, 17.6% were methicillin resistant. The 30-day all-cause mortality associated with methicillin-resistant SAB was 14.2%, which was not significantly different from the 13.3% mortality associated with methicillin-susceptible SAB (p = 0.6). With the exception of the β-lactams and erythromycin, antimicrobial resistance in methicillin-susceptible S. aureus was rare. However, in addition to the β-lactams, approximately 35% of methicillin-resistant S. aureus (MRSA) were resistant to erythromycin, 33% to ciprofloxacin, 13% to tetracycline, 13% to gentamicin and 4% to co-trimoxazole. When applying the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints, teicoplanin resistance was detected in four S. aureus isolates. Resistance was not detected for vancomycin and linezolid. Resistance to non-beta-lactam antimicrobials was largely attributable to two healthcare-associated MRSA (HA-MRSA) clones: ST22-IV [2B] (EMRSA-15) and ST239-III [3A] (Aus-2/3 EMRSA). The ST22-IV [2B] (EMRSA-15) clone is the predominant HA-MRSA clone in Australia. However, 85% percent of methicillin-resistant SAB isolates were community-associated MRSA (CA-MRSA) clones. Although polyclonal, approximately 77% of CA-MRSA clones were characterised as: ST93-IV [2B] (Queensland CA-MRSA); ST5-IV [2B]; ST45-V [5C2&5]; ST1-IV [2B]; ST30-IV [2B]; ST8-IV [2B]; and ST97-IV [2B]. The CA-MRSA clones, in particular ST45-V [5C2&5], have acquired multiple antimicrobial resistance determinants including ciprofloxacin, erythromycin, clindamycin, gentamicin and tetracycline. The multi-resistant ST45-V [5C2&5] clone accounted for 11.0% of CA-MRSA. As CA-MRSA is well established in the Australian community, it is important to monitor antimicrobial resistance patterns in community- and healthcare-associated SAB as this information will guide therapeutic practices in treating S. aureus sepsis.

Topics: Agar; Anti-Bacterial Agents; Anti-Infective Agents; Australia; Bacteremia; Ciprofloxacin; Cross Infection; Drug Resistance, Bacterial; Erythromycin; Gentamicins; Humans; Methicillin; Methicillin-Resistant Staphylococcus aureus; Sepsis; Staphylococcal Infections; Staphylococcus aureus; Tetracyclines

mobile phone plays an essential role in the lives of healthcare professionals in hospitals as far as communication is concerned. However, it can also serve as a source of nosocomial infections. This study aimed at determining the prevalence and antibiotic susceptibility of Methicillin-resistant Staphylococcus aureus (MRSA) and Escherichia coli isolated from mobile phones used by healthcare staff working in three public hospitals in Ghana.. in total, 220 swab samples were collected from 110 mobile phones of healthcare workers at a referral and two public tertiary hospitals in Ghana. Direct spreading of swab samples on agar plates was done. MacConkey agar and Baird Parker agar were used to isolate E. coli and S. aureus, respectively. Clinical Laboratory Standard Institute´s guidelines were followed for susceptibility testing, and S. aureus strains resistant to cefoxitin were considered to be MRSA. All E. coli and MRSA isolates were tested for their susceptibility to antibiotics using European Committee on Antimicrobial Susceptibility Testing (EUCAST) 2018 guidelines with its breakpoints. Obtained qualitative data were analyzed by using Microsoft Excel.. of 110 mobile phones, 78 (70.9%) and 4 (3.6%) were colonized with S. aureus and E. coli, respectively. From the 78 S. aureus isolates, 22 (28%) isolates were MRSA. Fifty percent (50%) (11/22) of the MRSA isolates were multi-drug resistant, of which one isolate was resistant to all antibiotics tested. E. coli isolates had 100 resistances to both ceftriaxone and ceftazidime.. mobile phones used by healthcare workers in hospitals frequently harbor E. coli, S. aureus, MRSA and may be sources of hospital-associated infections.

Topics: Agar; Anti-Bacterial Agents; Cell Phone; Cross Infection; Drug Resistance, Microbial; Escherichia coli; Escherichia coli Infections; Ghana; Health Personnel; Hospitals, Public; Humans; Methicillin-Resistant Staphylococcus aureus; Microbial Sensitivity Tests; Staphylococcal Infections; Staphylococcus aureus

Colonized surfaces, equipment, individuals, and infected patients can be sources for the hospital spread of vancomycin-resistant enterococci (VRE), which is one of the important nosocomial pathogens. The basic epidemiological tools for the prevention and control of hospital-acquired infections are the typing methods. Pulsed-field gel electrophoresis (PFGE) is a highly discriminating method used frequently to detect clonal associations in epidemics and hospital-acquired VRE infections. This study aimed to investigate the presence of cross-contamination, which service or services come to the forefront in case of possible cross-contamination and clonal relationship between VRE strains isolated from rectal swab, clinical and environmental swab samples taken in two different periods in various clinics of Istanbul University Istanbul Medical Faculty Hospital by PFGE method. A total of 125 VREs isolated in two different periods were included in the study. Rectal and environmental swab samples were inoculated on Enterococcosel agar and sodium azide broth, urine samples were inoculated on chromogenic agar, and other clinical samples were inoculated on 5% sheep blood agar and incubated for 18-24 hours. VITEK 2 Compact automation system GP panel (bioMerieux, MarcyL'Etoile, France) was used for the species identification of catalase-negative, gram-positive colonies and disc diffusion and minimum inhibitory concentration (MIC) gradient tests were used to determine vancomycin susceptibility. Multiplex polymerase chain reaction was used to search for vanA and vanB resistance genes in isolates identified as VRE, and PFGE was used to determine clonal association. All isolates were identified as Enterococcus faecium with the vanA resistance gene. It was shown that PFGE clones were divided into six types with 65% similarity (A-F), and in this polyclonal spread, the major type was type A, which contained 108 isolates in 37 subtypes existed in the hospital for years. In patients from whom similar isolates were obtained, the high rate of hospitalizations in the same emergency room or in different emergency services in the same building drew attention. Our results showed that the presence of VRE was established in our hospital, new isolates were added from time to time, and there was a cross-contamination. It was observed that emergency services, where infection control measures were neglected due to working conditions, were among the high-risk areas for VRE contamination. In o

Topics: Agar; Bacterial Proteins; Cross Infection; Electrophoresis, Gel, Pulsed-Field; Emergency Service, Hospital; Enterococcus faecium; Gram-Positive Bacterial Infections; Humans; Vancomycin-Resistant Enterococci

From 1 January to 31 December 2021, forty-eight institutions around Australia participated in the Australian Staphylococcus aureus Surveillance Outcome Programme (ASSOP). The aim of ASSOP 2021 was to determine the proportion of Staphylococcus aureus bacteraemia (SAB) isolates in Australia that were antimicrobial resistant, with particular emphasis on susceptibility to methicillin and on characterisation of the molecular epidemiology of the methicillin-resistant isolates. A total of 2,928 SAB episodes were reported, of which 78.4% were community-onset. Overall, 16.9% of S. aureus isolates were methicillin resistant. The 30-day all-cause mortality associated with methicillin-resistant SAB was 15.0%, which was not significantly different from the 14.4% all-cause mortality associated with methicillin-susceptible SAB (p = 0.7). With the exception of the β-lactams and erythromycin, antimicrobial resistance in methicillin-susceptible S. aureus was rare. However, in addition to the β-lactams, approximately 36% of methicillin-resistant S. aureus (MRSA) were resistant to ciprofloxacin; 30% to erythromycin; 15% to tetracycline; 16% to gentamicin; and 3% to co-trimoxazole. When applying the European Committee on Antimicrobial Susceptibility Testing (EUCAST) breakpoints, teicoplanin resistance was detected in three S. aureus isolates. Resistance to vancomycin or linezolid was not detected. Resistance to non-β-lactam antimicrobials was largely attributable to the healthcare-associated MRSA (HA-MRSA) clone ST22-IV [2B] (EMRSA-15), and the community-associated MRSA (CA-MRSA) clone ST45-V [5C2&5] which has acquired multiple antimicrobial resistance determinants including ciprofloxacin, erythromycin, clindamycin, gentamicin and tetracycline. The ST22-IV [2B] (EMRSA-15) clone is the predominant HA-MRSA clone in Australia. Nonetheless, 85% of methicillin-resistant SAB episodes were due to CA-MRSA clones. Although polyclonal, approximately 68% of CA-MRSA clones were characterised as ST93-IV [2B] (Queensland clone); ST45-V [5C2&5]; ST5-IV [2B]; ST1-IV [2B]; ST30-IV [2B]; and ST97-IV [2B]. As CA-MRSA is well established in the Australian community, it is important to monitor antimicrobial resistance patterns in community- and healthcare-associated SAB as this information will guide therapeutic practices in treating S. aureus bacteraemia.

Topics: Agar; Anti-Bacterial Agents; Anti-Infective Agents; Australia; Bacteremia; Ciprofloxacin; Cross Infection; Drug Resistance, Bacterial; Erythromycin; Gentamicins; Humans; Methicillin; Methicillin-Resistant Staphylococcus aureus; Microbial Sensitivity Tests; Staphylococcal Infections; Staphylococcus aureus; Tetracycline

The increasing prevalence of multidrug resistant bacteria is a problem in the inpatient care setting, and in the emergency care system. The aim of this observational, cross-sectional study was to evaluate the prevalence of pathogens on well-defined surfaces in German ambulances that have been designated as 'ready for service'.. After informed consent was obtained, ambulance surfaces were sampled with agar plates for microbiological examination during an unannounced visit. A standardised questionnaire was used to obtain information regarding the disinfection protocols used at each rescue station.. Methicillin resistant staphylococcus aureus contamination was present in 18 sampling surfaces from 11 out of 150 ambulance vehicles (7%) that were designated as ready for service. Contact surfaces directly surrounding patients or staff were most frequently contaminated with pathogens. However, bacterial contamination was not related to annual missions, methods or frequency of disinfection.. In accordance with previous studies, disinfection and cleaning of areas with direct contact to patients or staff seem to be the most challenging. This should also be reflected in disinfection guidelines and the related continuing education.

Topics: Agar; Ambulances; Bacteria; Cross Infection; Cross-Sectional Studies; Culture Media; Equipment Contamination; Fungi; Humans; Prevalence

This study aimed to correlate the presence of ica genes, biofilm formation and antimicrobial resistance in 107 strains of Staphylococcus epidermidis isolated from blood cultures. The isolates were analysed to determine their methicillin resistance, staphylococcal cassette chromosome mec (SCCmec) type, ica genes and biofilm formation and the vancomycin minimum inhibitory concentration (MIC) was measured for isolates and subpopulations growing on vancomycin screen agar. The mecA gene was detected in 81.3% of the S. epidermidis isolated and 48.2% carried SCCmec type III. The complete icaADBC operon was observed in 38.3% of the isolates; of these, 58.5% produced a biofilm. Furthermore, 47.7% of the isolates grew on vancomycin screen agar, with an increase in the MIC in 75.9% of the isolates. Determination of the MIC of subpopulations revealed that 64.7% had an MIC ≥ 4 μg mL-1, including 15.7% with an MIC of 8 μg mL-1 and 2% with an MIC of 16 μg mL-1. The presence of the icaADBC operon, biofilm production and reduced susceptibility to vancomycin were associated with methicillin resistance. This study reveals a high level of methicillin resistance, biofilm formation and reduced susceptibility to vancomycin in subpopulations of S. epidermidis. These findings may explain the selection of multidrug-resistant isolates in hospital settings and the consequent failure of antimicrobial treatment.

Topics: Adult; Agar; Aged; Biofilms; Cross Infection; Culture Media; Female; Humans; Infant; Infant, Newborn; Male; Methicillin Resistance; Microbial Sensitivity Tests; Middle Aged; Operon; Polymerase Chain Reaction; Staphylococcal Infections; Staphylococcus epidermidis; Tertiary Care Centers; Vancomycin; Vancomycin Resistance

Infections associated with health care services are nowadays widespread and, associated to the progressive emergence of microorganisms resistant to conventional chemical antibiotics, are major causes of morbidity and mortality. One of the most representative microorganisms in this scenario is the bacterium Pseudomonas aeruginosa, which alone is responsible for ca. 13-15% of all nosocomial infections. Bacteriophages have been reported as a potentially useful tool in the diagnosis of bacterial diseases, since they specifically recognize and lyse bacterial isolates thus confirming the presence of viable cells. In the present research effort, immobilization of these biological (although metabolically inert) entities was achieved via entrapment within (optimized) porous (bio)polymeric matrices of alginate and agar, aiming at their full structural and functional stabilization. Such phage-impregnated polymeric matrices are intended for future use as chromogenic hydrogels sensitive to color changes evolving from reaction with (released) intracytoplasmatic moieties, as a detection kit for P. aeruginosa cells.

Topics: Agar; Alginates; Bacteriological Techniques; Biopolymers; Biosensing Techniques; Cells, Immobilized; Cross Infection; Glucuronic Acid; Hexuronic Acids; Humans; Hydrogels; Pseudomonas aeruginosa; Pseudomonas Infections; Pseudomonas Phages

Stenotrophomonas maltophilia (Smalto) is a prominent nosocomial pathogen, commonly isolated in the hospital environment. Multiple Smalto nosocomial outbreaks have been linked to contaminated water sources. This study aimed to develop a medium able to ease healthcare environment Smalto isolation.. Financed, from March 2007 to June 2008, by a university hospital of Amiens' clinical research program, this study allowed Stenotrophomonas maltophilia selective medium with coloured indicator (SM2i) development. SM2i is constituted of Mueller Hinton agar (MH), maltose, DL-methionine, bromothymol blue. The mixture sterilized is refreshed at 50 degrees C, its pH adjusted to 7.1, and render selective by addition of vancomycin, imipenem and amphotericin B. Then, SM2i agar is sunk into 90 cm diameter Petri dish dated and stored at 4 degrees C for 4 weeks. SM2i is developed using Pasteur Institute culture type collection (CIP) strains of Smalto, Burkholderia cepacia, Pseudomonas aeruginosa (Psa) and a Smalto strain of our hygiene laboratory collection. It was validate on Psa imipenem-resistant and Enterococcus faecium vancomycin-resistant strains, then, tested on cold water first jet and faucet cotton-swabs samples. SM2i tests were made in comparison with the MH agar, MH agar plus four paper disks loaded 10 microg of imipenem and Cetrimed agar. Its sensitivity, specificity, positive (PPV) and negative (NPV) predictive values, accuracy, likehood-ratio (LR) and Youden index have been determined.. SM2i agar is better in culturing Smalto test-strains. On SM2i, Smalto colonies are smooth, round, greeny, olive or lime green, have a green olive centre with a peripheral lighter or a dark green centre with an olive green suburb surrounded by a blue halo. SM2i is a selective, specific, predictive, accurate medium to search for Smalto in healthcare environment. In 122 pairs of cold water first jet and taps cotton-swabs samples, Smalto was isolated from 14.8% of water samples, 10.7% of cotton-swabs samples. It was isolated alone in 6.6% of water samples and 2.5% of swab samples. Thus, smalto has biocontaminated 17.2% of cold water taps. Compared to MH agar, SM2i sensitivity, specificity, PPV, NPV, accuracy, LR were 100, 100, 100, 100, 100% and infinity, and 87.5, 100, 100, 98.1, 98.4% and infinity for water and cotton-swabs samples respectively.. SM2i is a selective, specific, predictive medium which can allow easily isolating and identifying accurately Smalto in environmental samples. Its evaluation on clinical samples is on going.

Topics: Agar; Bacteriological Techniques; Cross Infection; Culture Media; Equipment Contamination; Gram-Negative Bacterial Infections; Humans; Indicators and Reagents; Predictive Value of Tests; Sensitivity and Specificity; Stenotrophomonas maltophilia; Water Microbiology

Pre-emptive isolation of suspected methicillin-resistant Staphylococcus aureus (MRSA) carriers is considered essential for controlling the spread of MRSA, but noncolonized patients will be isolated unnecessarily as a result of a delay in diagnosis of 3-5 days with conventional cultures. We determined costs per isolation day avoided, and incremental costs of rapid MRSA screening tests when added to conventional screening, but with decisions on isolation measures based on PCR results. A prospective multicentre study evaluating BD GeneOhm MRSA PCR (`IDI') (BD Diagnostics, San Diego, CA, USA), Xpert MRSA (`GeneXpert') (Cepheid, Sunnyvale, CA, USA) and chromogenic agar (MRSA-ID) (bioMérieux, Marcy-l'Etoile, France) was performed in 14 Dutch hospitals. Among 1764 patients at risk, MRSA prevalence was 3.3% (n=59). Duration of isolation was 19.7 and 16.1 h with IDI and GeneXpert, respectively, and would have been 30.0 and 76.2 h when based on chromogenic agar and conventional cultures, respectively. Negative predictive values (at a patient level) were 99.5%, 99.1% and 99.5% for IDI, GeneXpert and chromogenic agar, respectively. Numbers of isolation days were reduced by 60% and 47% with PCR-based and chromogenic agar-based screening, respectively. The cost per test was €56.22 for IDI, €69.62 for GeneXpert and €2.08 for chromogenic agar, and additional costs per extra isolation day were €26.34. Costs per isolation day avoided were €95.77 (IDI) and €125.43 (GeneXpert). PCR-based decision-making added €153.64 (IDI) and €193.84 (GeneXpert) per patient to overall costs and chromogenic testing would have saved €30.79 per patient. Rapid diagnostic testing safely reduces the number of unnecessary isolation days, but only chromogenic screening, and not PCR-based screening, can be considered as cost saving.

Topics: Agar; Carrier State; Chromogenic Compounds; Cost-Benefit Analysis; Cross Infection; Diagnostic Tests, Routine; Health Care Costs; Humans; Methicillin-Resistant Staphylococcus aureus; Patient Isolation; Polymerase Chain Reaction; Predictive Value of Tests; Prospective Studies; Staphylococcal Infections

Methicillin-resistant Staphylococcus aureus (MRSA) is a leading cause of nosocomial infections that result in extended hospital stays and increased mortality. Therefore, rapid, cost-effective techniques for surveillance and detection of MRSA are critical to the containment and prevention of the spread of MRSA within the health care environment. We examined the ability of 3 chromogenic media (Spectra MRSA, Remel, Lenexa, KS; MRSA Select, Bio-Rad, Redmond, WA; and ChromID MRSA, bioMerieux, Marcy l'Etoile, France) to detect MRSA from routine surveillance specimens following 18, 24, and 48 hours of incubation. Our results indicate that detection of MRSA using all 3 chromogenic media is optimal following 24 hours of incubation. Early examination reduced sensitivity, while extended incubation reduced specificity. In addition, Spectra MRSA identified 2 borderline oxacillin-resistant strains of S aureus that were not detected by the other 2 chromogenic agars evaluated. These strains demonstrate increased basal and inducible resistance to β-lactam antibiotics.

Topics: Agar; Anti-Bacterial Agents; Chromogenic Compounds; Cross Infection; Culture Media; Humans; Methicillin Resistance; Methicillin-Resistant Staphylococcus aureus; Nasal Mucosa; Oxacillin; Predictive Value of Tests; Staphylococcal Infections; Staphylococcus aureus

Topics: Agar; Bacteria; Bacteriological Techniques; Colony Count, Microbial; Cross Infection; Culture Media; Disinfection; Equipment Contamination; Hospital Units; Humans; Pediatrics; Stethoscopes

Increasing antibiotic resistance in gram-negative bacteria has recently renewed interest in colistin as a therapeutic option. The increasing use of colistin necessitates the availability of rapid and reliable methods for colistin susceptibility testing. We compared seven methods of colistin susceptibility testing (disk diffusion, agar dilution on Mueller-Hinton [MH] and Isosensitest agar, Etest on MH and Isosensitest agar, broth microdilution, and VITEK 2) on 102 clinical isolates collected from patient materials during a selective digestive decontamination or selective oral decontamination trial in an intensive-care unit. Disk diffusion is an unreliable method to measure susceptibility to colistin. High error rates and low levels of reproducibility were observed in the disk diffusion test. The colistin Etest, agar dilution, and the VITEK 2 showed a high level of agreement with the broth microdilution reference method. Heteroresistance for colistin was observed in six Enterobacter cloacae isolates and in one Acinetobacter baumannii isolate. This is the first report of heteroresistance to colistin in E. cloacae isolates. Resistance to colistin in these isolates seemed to be induced upon exposure to colistin rather than being caused by stable mutations. Heteroresistant isolates could be detected in the broth microdilution, agar dilution, Etest, or disk diffusion test. The VITEK 2 displayed low sensitivity in the detection of heteroresistant subpopulations of E. cloacae. The VITEK 2 colistin susceptibility test can therefore be considered to be a reliable tool to determine susceptibility to colistin in isolates of genera that are known not to exhibit resistant subpopulations. In isolates of genera known to (occasionally) exhibit heteroresistance, an alternative susceptibility testing method capable of detecting heteroresistance should be used.

Topics: Acinetobacter baumannii; Acinetobacter Infections; Agar; Anti-Bacterial Agents; Bacterial Infections; Colistin; Cross Infection; Culture Media; Drug Resistance, Multiple, Bacterial; Enterobacter cloacae; Enterobacteriaceae Infections; Humans; Intensive Care Units; Microbial Sensitivity Tests; Polymyxin B

With the growing frequency of extended-spectrum beta-lactamases (ESBL) among Enterobacteriaceae, treatment of Gram-negative nosocomial infections requires rapid and reliable detection of this enzyme. Quicolor agar (QC agar) (Salubris Inc., Massachusetts, USA) is a novel chromogenic agar medium changing colour within 4 to 6 h due to the metabolic activity of growing bacteria. This study investigated the use of QC agar compared to Mueller Hinton agar (MH) for the detection of ESBL using disk diffusion and E-test. 100 Enterobacteriaceae isolated at Hacettepe University Hospital, of which 50 were predetermined to be ESBL positive and 50 as negative using the CLSI disk diffusion ESBL (phenotypic confirmatory test) criteria. For disk diffusion and E-test, cefotaxime+/-clavulanate (CT/CTL) and ceftazidime+/-clavulanate (TZ/TZL) were used, and for E-test, cefepime+/-clavulanate (PM/PML) was also used. QC agar rapid ESBL results for all strains were in agreement with the standard overnight procedure. All 50 ESBL positives were detected by both methods. For the 50 ESBL negatives, QC agar rapid results from E-test and disk diffusion were in complete accordance with the overnight MH results. Moreover, E-test detected 8 additional ESBL positive strains that disk diffusion missed. For disk diffusion, CT/CTL alone detected all 50 ESBL positives while TZ/TZL alone missed 5 ESBL positives. E-test CT/CTL alone confirmed all 50 ESBL positives and identified 4 additional ESBL-positive strains. When used together, E-test CT/CTL, TZ/TZL and PM/PML identified a total of 58 ESBL positives among the 100 strains tested. QC agar can be used for rapid and reliable ESBL detection within 4 to 6 h, using disk diffusion and E-test ESBL reagents. This rapid method should be further validated using genotype characterized ESBL and other beta-lactamase positive strains.

Topics: Agar; beta-Lactam Resistance; beta-Lactamases; Cross Infection; Disk Diffusion Antimicrobial Tests; Enterobacteriaceae; Gram-Negative Bacterial Infections; Humans

Active surveillance for methicillin-resistant Staphylococcus aureus (MRSA) is among the strategies recommended by the Society for Healthcare Epidemiology of America for control of nosocomial MRSA infections. Infection control and laboratory personnel desire rapid, sensitive, and inexpensive methods to enhance surveillance activities. A multicenter study was performed to evaluate a new selective and differential chromogenic medium, BBL CHROMagar MRSA (C-MRSA) medium (BD Diagnostics, Sparks, MD), which enables recovery and concomitant identification of MRSA strains directly from nasal swab specimens taken from the anterior nares. Specimens were inoculated to C-MRSA and Trypticase soy agar with 5% sheep blood agar (TSA II, BD Diagnostics). Mauve colonies on C-MRSA at 24 h and 48 h and suspicious colonies on TSA II were confirmed as Staphylococcus aureus by Gram stain morphology and a coagulase test. In addition, the results of C-MRSA were compared to results of susceptibility testing (five different methods) of S. aureus strains isolated on TSA II. A total of 2,015 specimens were inoculated to C-MRSA and TSA II. Three hundred fifty-four S. aureus isolates were recovered; 208 (59%) were oxacillin (methicillin) susceptible and 146 (41%) were oxacillin resistant (MRSA). On C-MRSA, 139/146 or 95.2% of MRSA isolates were recovered, whereas recovery on TSA II was 86.9% (127/146) (P = 0.0027). The overall specificity of C-MRSA was 99.7%. When C-MRSA was compared to each susceptibility testing method, the sensitivity and specificity, respectively, were as follows: oxacillin MIC by broth microdilution, 94.4% and 96.7%; oxacillin screen agar, 94.3% and 96.7%; PBP2' latex agglutination, 93.7% and 98.5%; cefoxitin disk diffusion, 95.0% and 98.1%; and mecA PCR, 95.1% and 98.1%. In this study, C-MRSA was superior to TSA II for recovery of MRSA from surveillance specimens obtained from the anterior nares and was comparable to conventional, rapid, and molecular susceptibility methods for the identification of MRSA isolates.

Topics: Agar; Anti-Bacterial Agents; Chromogenic Compounds; Cross Infection; Culture Media; Humans; Methicillin; Methicillin Resistance; Nasal Mucosa; Oxacillin; Sensitivity and Specificity; Staphylococcal Infections; Staphylococcus aureus; United States

The effect of endotracheal tubes (ETTs) impregnated with chlorhexidine (CHX) and silver carbonate (antiseptic ETTs) against Staphylococcus aureus, methicillin-resistant S. aureus (MRSA), Pseudomonas aeruginosa, Acinetobacter baumannii, and Enterobacter aerogenes [organisms associated with ventilator-associated pneumonia (VAP)], was evaluated in a laboratory airway model. Antiseptic ETTs and control ETTs (unimpregnated) were inserted in culture tubes half-filled with agar media (airway model) previously contaminated at the surface with 10(8) cfu/mL of the selected test organism. After five days of incubation, bacterial colony counts on all ETT segments were determined. Swabs of proximal and distal ends of the agar tract in antiseptic and control models were subcultured. The initial and residual CHX levels, (five days post-implantation in the model) were determined. Cultures of antiseptic ETTs revealed colonization by the tested pathogens ranging from 1-100 cfu/tube, compared with approximately 10(6) cfu/tube for the control ETTs (P < 0.001). Subcultures from proximal and distal ends of the agar tract showed minimal or no growth in the antiseptic ETTs compared with the control ETTs (P < 0.001). The amount of CHX retained in the antiseptic ETTs after five days of implantation was an average of 45% of the initial level. Antiseptic ETTs prevented bacterial colonization in the airway model and also retained significant amounts of the antiseptic. These results indicate that the effectiveness of antiseptic-impregnated ETTs in preventing the growth of bacterial pathogens associated with VAP may vary with different organisms.

Topics: Acinetobacter baumannii; Agar; Carbonates; Chlorhexidine; Colony Count, Microbial; Cross Infection; Culture Media; Disinfectants; Enterobacter aerogenes; Equipment Contamination; Humans; In Vitro Techniques; Intubation, Intratracheal; Methicillin Resistance; Pneumonia, Bacterial; Pseudomonas aeruginosa; Silver Compounds; Staphylococcus aureus; Ventilators, Mechanical

To establish an efficient and sensitive technique for recovering vancomycin-resistant enterococci (VRE) from perianal and environmental samples collected during implementation of control measures for an outbreak of VRE.. Perianal and environmental samples were collected in triplicate on sterile swabs. One swab was used to inoculate a selective broth medium containing 6 pg of vancomycin and 8 pg of ciprofloxacin per mL, one to inoculate Campylobacter agar containing 10 microg/mL of vancomycin, and one to inoculate Enterococcosel agar containing 8 microg/mL of vancomycin.. Samples were collected in the intensive care units of a 600-bed university hospital over a period of 2 months. SAMPLE SELECTION: Patients and their immediate environment were sampled if they resided in a ward with a patient known to be colonized or infected with VRE.. Of the 88 perianal samples obtained from 63 patients, 37 were positive for VRE by broth culture, with 36 also recovered on both types of solid media (sensitivity, 97.3%; negative predictive value, 98.1%). Of the initial samples collected from each of the 63 patients, 20 were positive for VRE by all methods. Of the 500 environmental samples cultured, 139 were positive for VRE in broth, with only 33 recovered on Campylobacter agar (sensitivity, 23.7%; negative predictive value, 77.2%) and 22 on Enterococcosel agar (sensitivity, 15.8%; negative predictive value, 75.2%).. Our data indicate that, when performing surveillance cultures during an outbreak of VRE, use of an enrichment broth medium is required to recover VRE contaminating environmental surfaces; however, direct inoculation to selective solid medium is adequate to recover VRE in patient perianal specimens.

Topics: Agar; Anal Canal; Colony Count, Microbial; Cross Infection; Disease Outbreaks; Enterococcus; Gram-Positive Bacterial Infections; Humans; Infection Control; Sensitivity and Specificity; Specimen Handling; Vancomycin Resistance

This study evaluated a polymerase chain reaction (PCR) method for detection of methicillin-resistant Staphylococcus aureus (MRSA) in specimens referred for nosocomial surveillance. PCR was used to detect the mecA and nuc gene targets using yellow growth on mannitol salt agar containing 6 mg/liter oxacillin (MSO-6) as a source of DNA (N = 645). The diagnostic values for PCR compared with culture methods were 97% specificity, 100% sensitivity, 96% positive predictive value, and 100% negative predictive value. Total cost for PCR per test is $3.62 compared to $4.77 for culture. However, the total cost per specimen is significantly lower due to only 20% of all surveillance specimens producing yellow colonies on MSO-6. The average turnaround time for the PCR method is 48 h compared with 82 h for culture. PCR amplification of mecA and nuc genes using yellow colonies on MSO-6 is a simple, fast, accurate and cost-effective method for routine use in clinical laboratories for detecting MRSA in surveillance specimens.

Topics: Agar; Bacterial Proteins; Bacteriological Techniques; Carrier Proteins; Cross Infection; Culture Media; Endonucleases; Hexosyltransferases; Humans; Mannitol; Methicillin Resistance; Micrococcal Nuclease; Muramoylpentapeptide Carboxypeptidase; Oxacillin; Penicillin-Binding Proteins; Penicillins; Peptidyl Transferases; Polymerase Chain Reaction; Sensitivity and Specificity; Staphylococcal Infections; Staphylococcus aureus

Propionibacterium acnes has been identified as a significant agent of nosocomial infections, including endophthalmitis. Data concerning susceptibility of P. acnes to newer beta-lactam antibiotics and fluoroquinolones are limited. Recent reports suggest that quinolones have activity against these organisms sufficient to warrant further study. We undertook a study to select appropriate antimicrobial agents for use in a rabbit model of P. acnes endophthalmitis. We compared the antibiotic susceptibilities of P. acnes by using the National Committee for Clinical Laboratory Standards method of agar dilution with the E test. Thirteen clinical isolates obtained from eye specimens and three American Type Culture Collection control strains were tested against 14 antibiotics. All the clinical isolates were susceptible by both methods to piperacillin, piperacillin-tazobactam, ampicillin-sulbactam, ticarcillin-clavulanate, cefotaxime, cefotetan, ceftriaxone, cefoxitin, and imipenem in addition to clindamycin but were resistant to metronidazole. The clinical P. acnes isolates also displayed high-level susceptibility to ciprofloxacin, sparfloxacin, and ofloxacin. Almost all the P. acnes strains demonstrated E-test MICs within 2 dilutions of the MICs observed by the agar dilution method. Those few strains for which discrepancies were noted exhibited E-test susceptibilities three- to fivefold dilutions lower than the agar dilution method susceptibilities but only with ampicillin-sulbactam, ticarcillin-clavulanate, and/or clindamycin. On the basis of our study, all of clinical eye isolates were susceptible to these newer antimicrobial agents and the two methods demonstrated similar susceptibility patterns.

Topics: Agar; Animals; Anti-Bacterial Agents; Anti-Infective Agents; Cross Infection; Disease Models, Animal; Endophthalmitis; Fluoroquinolones; Gram-Positive Bacterial Infections; Humans; Lactams; Microbial Sensitivity Tests; Propionibacterium acnes; Rabbits

The E test and the reference agar dilution methods were compared for detecting high-level aminoglycoside resistance (HLAR) among 71 selected clinical isolates, including 62 Enterococcus faecalis and 9 Enterococcus faecium isolates. High-level gentamicin resistance alone was found in 11% (5 E. faecalis and 3 E. faecium strains) and high-level streptomycin resistance was found in 42% (28 E. faecalis, 2 E. faecium strains) of the strains tested, and 31% of the strains demonstrated high-level resistance to both antimicrobial agents (21 E. faecalis and 1 E. faecium strains). The E test detected all HLAR populations, but the streptomycin strip may require recalibration to achieve absolute MIC comparisons with the reference value (twofold less) or the use of an alternative interpretive resistance breakpoint, e.g., > 1,000 micrograms/ml. By the E test, MIC results indicate that ampicillin, imipenem, penicillin, piperacillin, and vancomycin remain active against the HLAR E. faecalis isolates; however, these tested drugs were less effective on the HLAR E. faecium isolates (< 50%).

Topics: Agar; Aminoglycosides; Anti-Bacterial Agents; Cross Infection; Drug Resistance, Microbial; Enterococcus faecalis; Enterococcus faecium; Evaluation Studies as Topic; Gram-Positive Bacterial Infections; Humans; Microbial Sensitivity Tests

In 1946, DIENES observed that non-identical Proteus-strains, when swarming towards each other, were froming distinctly demarcated lines ("DIENES' phenomenon", demarcation (=AGL) phenomenon). Strains of the same origin were amalgamating without demarcation. In 1970, STURDZA studied this phenomenon and commented on its relevance for the epidemiology of nosocomial Proteus-infections. An increased number of Proteus-infections in Berlin was the reason that this procedure was tested for epidemiological purposes. Approx. 300 Proteus mirabilis-strains, tested on normal meat fluid agar, could be divided into 52 AGL-groups. When tested by several investigators, the coordination to any AGL-group seemed to be very subjective. Consequently, a DNase-agar with o-toluidinblue as an indicator, as had been mentioned by CHAMBERS in 1975, was used in order to give a more exact demonstration of AGL. This resulted in a reduction from 52 to 42 AGL-groups. Another 140 newly isolated Proteus-strains belonged to the 42 known as well as to another 42 new AGL-groups. Whether these 84 AGL-groups possess the constancy which is imperative for practical purposes, is however, still rather doubtful.

Topics: Agar; Bacteriological Techniques; Cross Infection; Humans; Proteus Infections; Proteus mirabilis; Tolonium Chloride

A defined agar medium (hereinafter designated caprylate-thallous [CT5 agar) containing 0.01% yeast extract, 0.1% caprylic (n-octanoic) acid, and 0.025% thallous sulfate is highly selective for all Serratia species and effectively discriminates against most non-Serratia strains likely to be in the same habitats. The selectivity of CT agar is demonstrated by the very high efficiency of colony formation (mean, 80.7% of that on a nonselective complex medium) on CT agar by known Serratia strains and the very low efficiency of colony formation (close to zero) on CT agar by bacterial strains known not to be Serratia. The utility of this medium in actual clinical laboratory practice is demonstrated by the more rapid and higher recovery of Serratia on this selective medium as compared to conventional procedures of in-tandem runs of 513 consecutive urine, feces, and sputum specimens. Pigmented and nonpigmented Serratia strains deliberately added to fecal specimens can be selectively and quantitatively recovered on CT agar. CT agar compares favorably with, or in some cases is an improvement over, other selective media which have been recommended for isolating Serratia. This selective CT agar medium could be quite useful in ecological surveys, especially those related to hospital-acquired infections.

Topics: Agar; Bacterial Infections; Caprylates; Cross Infection; Evaluation Studies as Topic; Feces; Humans; Serratia; Species Specificity; Sputum; Thallium; Urine

Topics: Agar; Air Microbiology; Bacteria; Bacteriological Techniques; Biguanides; Cross Infection; Disinfectants; Housekeeping, Hospital; Humans; Locomotion; Quaternary Ammonium Compounds; Water

Topics: Agar; Bacteriocins; Bacteriological Techniques; Bacteriuria; Blood; Bone and Bones; Cross Infection; Humans; Mitomycins; Pseudomonas aeruginosa; Pseudomonas Infections; Sputum; Tracheotomy; Ulcer; Wound Infection

Following the demonstration of massive spread of bacterial contamination throughout the hospital by the wet-mopping techniques in use, quantitative studies were undertaken to determine the source of contamination and to institute measures of control. It was found that mops, stored wet, supported bacterial growth to very high levels and could not be adequately decontaminated by chemical disinfection. Laundering and adequate drying provided effective decontamination, but build-up of bacterial counts occurred if mops were not changed daily or if disinfectant was omitted from the wash-water. Recommendations were based upon the experimental findings.

Topics: Agar; Anti-Bacterial Agents; Bacteria; Bacteriological Techniques; Blood; Cross Infection; Detergents; Equipment and Supplies, Hospital; Gossypium; Humans; Laundering; Phenols; Sterilization; Time Factors

Topics: Agar; Bacteriological Techniques; Cross Infection; Erwinia; Hospitals; Humans

Topics: Agar; Bacteriocins; Bacteriological Techniques; Cross Infection; Culture Media; Humans; Mitomycins; Pseudomonas aeruginosa; Pseudomonas Infections

Topics: Agar; Burns; Cross Infection; Drug Resistance; Enterobacteriaceae; Gentamicins; Humans; Pseudomonas aeruginosa; Pseudomonas Infections

Topics: Agar; Bacteriological Techniques; Cross Infection; Humans; Proteus; Proteus Infections; Species Specificity

Topics: Agar; Air Microbiology; Cross Infection; Culture Media; Disease Reservoirs; Enterobacteriaceae; Escherichia coli; Evaluation Studies as Topic; Feces; Hospital Departments; Humans; Methods; Pediatrics

Topics: Agar; Bacteria; Bacteriological Techniques; Blood; Cross Infection; Humans; Methods; Polymers; Sterilization

Davis, Nour A. (University of Lagos Medical School, Lagos, Nigeria), and G. H. G. Davis. Ecology of nasal staphylococci. J. Bacteriol. 89:1163-1168. 1965.-The rate of nasal carriage of Staphylococcus aureus in Nigerian adults (46%) approximates that found in other countries. The rate in infants under 12 months was ca. 70%, which exceeds that found elsewhere, e.g., England. The incidence of penicillin resistance in nasal staphylococci (50 to 60%) is about the same as has been found in strains isolated from infections in outpatients in urban centers in this country. Mannitol-polymyxin agar was used for the selection and differentiation of coagulase-positive staphylococci and proved to be valuable in such studies. Our results clearly show that the degree of colonization by S. aureus significantly influences, or is influenced by, the rate of incidence of other bacteria in the vestibular flora, particularly in the case of diphtheroids and coagulase-negative cocci. The relationship between the degree of nasal microbial colonization and social and other factors is discussed.

Topics: Adult; Agar; Anti-Bacterial Agents; Carrier State; Child; Coagulase; Cross Infection; Culture Media; Drug Resistance; Drug Resistance, Microbial; Humans; Infant; Mannitol; Medical Staff, Hospital; Nigeria; Nose; Penicillins; Polymyxins; Research; Staphylococcal Infections; Staphylococcus; Staphylococcus aureus