Water samples (400 mL) were serially diluted in sterile distilled water and filtered through 0.45 µm cellulose nitrate filters (Sartorius) and GN-6 Metricel MCE Membrane Disc Filters (Pall), which were placed on Chromocult® Coliform Agar (Merck Millipore) for 24 h at 37 °C for detection of E. coli (blue colonies), total coliforms (salmon-colored colonies) and other Gram-negative bacteria (transparent colonies). Resistant bacteria were recovered by plating on chromogenic media selective for β-lactamase producing Enterobacteriaceae (CARBA Agar, ChromID, BioMerieux) and extended-spectrum β-lactamases (ESBL Agar, ChromID, BioMerieux), methicillin-resistant S. aureus (MRSA Agar, ChromID, BioMerieux), and vancomycin-resistant enterococci (VRE Agar, ChromID, BioMerieux). Recovered colonies were subjected to previously reported PCR assays for species identification and resistance detection53 –60 (link).
Vre agar
Manufactured by bioMérieux
Sourced in France
VRE Agar is a selective and differential culture medium used for the detection and enumeration of vancomycin-resistant Enterococcus (VRE) in clinical samples. It is designed to inhibit the growth of most other bacteria while allowing VRE to grow and be identified.
Automatically generated - may contain errors
2 protocols using vre agar
1
Comprehensive Microbial Analysis of Water Samples
2
Screening for Antibiotic-Resistant Bacteria
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The collected samples were examined at the Departments of Clinical Microbiology at Aalborg University Hospital, Aarhus University Hospital, Odense University Hospital and Slagelse Hospital. The same method of analysis was applied to all four departments. All analyses followed the procedure described in the protocol article without deviations from the protocol.11 (link)
Briefly outlined, samples were screened with commercially available, selective, chromogenic agar media ((MRSA: CHROMagar MRSA II agar, ESBL: CHROMagar ESBL bi-agar (Becton Dickinson, Heidelberg, Germany)), (CPE: chromID CARBA SMART agar, VRE: chromID VRE agar (bioMérieux, Marcy-l’Etoile, France)). A preceding enhancement broth step was used for both VRE and MRSA. All isolates were identified by mass spectrometry (Matrix-Assisted Laser Desorption-Ionization - time of flight) (MALDI-TOF)) and the presence of resistance genes in MRSA (mecA/mecC) VRE (vanA/vanB) and CPE (blaKPC/blaNDM/blaVIM/blaOXA-48/blaIMP) was confirmed by PCR. ESBL production was confirmed phenotypically (synergism between clavulanic acid and cefotaxime, ceftazidime and/or cefepime).
Briefly outlined, samples were screened with commercially available, selective, chromogenic agar media ((MRSA: CHROMagar MRSA II agar, ESBL: CHROMagar ESBL bi-agar (Becton Dickinson, Heidelberg, Germany)), (CPE: chromID CARBA SMART agar, VRE: chromID VRE agar (bioMérieux, Marcy-l’Etoile, France)). A preceding enhancement broth step was used for both VRE and MRSA. All isolates were identified by mass spectrometry (Matrix-Assisted Laser Desorption-Ionization - time of flight) (MALDI-TOF)) and the presence of resistance genes in MRSA (mecA/mecC) VRE (vanA/vanB) and CPE (blaKPC/blaNDM/blaVIM/blaOXA-48/blaIMP) was confirmed by PCR. ESBL production was confirmed phenotypically (synergism between clavulanic acid and cefotaxime, ceftazidime and/or cefepime).
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