The Mueller–Hinton agar (MHA) (NEOGEN) plates were inoculated by streaking using the cotton swab three times over the entire surface and rotating the MHA plates approximately 60° each time to ensure an even distribution of the inoculums. Then, the MHA plates were left open for three to five minutes to allow for any excess surface moisture to be absorbed [16 ]. The impregnated discs were dispensed onto the surface of the inoculated agar plates using sterile forceps. Discs of commercial ciprofloxacin (1 µg/disc) and fluconazole (1 µg/disc) were used as positive controls for bacterial and fungal pathogens, respectively, and the pure solvent (petroleum) impregnated discs were used as negative controls. Then, the MHA plates were sealed with parafilm and incubated at 37°C for 24 hrs and 27°C for 72 hrs for bacterial and fungal pathogens, respectively. Then, the diameters of the zone of inhibition around each disc were measured to the nearest millimeter along two axes (i.e., 90° to each other) and the means of the two readings were recorded.
Mueller hinton agar (mha)
Manufactured by Neogen
Sourced in United States
The MHA is a laboratory equipment used for the detection and enumeration of microorganisms. It provides a standardized method for the assessment of antimicrobial susceptibility of bacterial isolates.
Automatically generated - may contain errors
Lab products found in correlation
Meropenem, by Liofilchem (1 mentions)
Piperacillin tazobactam, by Liofilchem (1 mentions)
Cefepime, by Liofilchem (1 mentions)
Ciprofloxacin, by Liofilchem (1 mentions)
Imipenem, by Liofilchem (1 mentions)
Anaerobic jar 2.5l, by Thermo Fisher Scientific (1 mentions)
Atcc 29213, by Thermo Fisher Scientific (1 mentions)
4 protocols using mueller hinton agar (mha)
1
Antimicrobial Susceptibility Testing Protocol
2
Antimicrobial Susceptibility Testing: Disc Diffusion and Automated Systems
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Various antibiotic discs (Bioanalyses, Turkey) were used for antimicrobial susceptibility tests (Supplementary Tables S4, S5 ). The tests were performed manually (CLSI, 2017 ) from 1 January 2018, to 19 May 2019. Mueller–Hinton agar (MHA) (Neogen, UK) was used for the disc diffusion test. The diameters of the inhibition zones around each disc were measured and compared with the standard inhibition zone, as recommended by the Clinical Laboratory Standard Institution (CLSI, 2017 ). The results were interpreted as sensitive or resistant; intermediate resistance was also considered as resistance. On the other hand, from 20 May 2019, onward, the antimicrobial susceptibility test was carried out using an automated system with a Vitek®2 AST-P640 card for Gram-positive bacteria (G+) and AST-N327 cards for Gram-negative bacteria (G−) (BioMerieux, France).
3
Antibiotic Sensitivity and MIC Determination
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Antibiotic sensitivity to cefiderocol (MedChemExpress, Monmouth Junction, NJ, USA) and minimum inhibitory concentrations (MIC) for piperacillin/tazobactam, ciprofloxacin, cefepime, meropenem, and imipenem (Liofilchem, Waltham, MA, USA) were obtained by recording the lowest concentration of antibiotic that inhibited growth of bacteria on Mueller Hinton agar (Neogen, Lansing, MI, USA), as described by the manufacturer’s protocol.
4
Antibacterial Susceptibility Screening of Campylobacter
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Phenotyping antibacterial susceptibility screening to ciprofloxacin (5 μg), nalidixic acid (30 μg), erythromycin (15 μg), and tetracycline (30 μg) was conducted as recommended by the World Health Organization (WHO) Advisory Group on Integrated Surveillance of Antimicrobial Resistance guidelines on foodborne bacteria. The test was performed using the Kirby–Bauer disk diffusion method and the results were interpreted using the Clinical and Laboratory Standards Institute (CLSI) guidelines [24 ]. Antibacterial susceptibility test was performed on Mueller-Hinton agar (Neogen Corporation, Lansing, MI, United States) that was supplemented with 10% sheep blood, according to CLSI guidelines. The plates were then sealed in an anaerobic jar 2.5L (Oxoid, UK) each containing gas generating sachet without catalyst in a microaerophilic incubator at 41°C for 24 h. The suspect colonies were subjected to Gram staining and examination under a light microscope for the identification of any potential Campylobacter spp. Antibiotic susceptibility was calculated by the zones of inhibition observed around each antibiotic disc in millimeters. Standard reference strains of Staphylococcus aureus (ATCC® 29213, Thermo Fischer, USA) and C. jejuni ATCC (33560) were used as quality controls [22 ,25 (link)].
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