Antimicrobial resistance was evaluated in one isolate per sample. The minimum inhibitory concentration (MIC ) was determined using the EUCAMP2 plates (Thermo Scientific, West Palm Beach, FL). The tests were performed according to the manufacturer instructions. The following antibiotics were evaluated: gentamicin, ciprofloxacin, nalidixic acid, tetracycline, streptomycin, and erythromycin. Campylobacter jejuni ATCC 33560 was used as the quality control strain. Epidemiological breakpoint values from the European Committee on Antimicrobial Susceptibility Testing were considered to determine bacterial antibiotic resistance (EUCAST, 2015 ).
Eucamp2 plates
Manufactured by Thermo Fisher Scientific
Sourced in Spain, United States
The EUCAMP2 plates are multi-well culture plates designed for cell culture applications. They feature a standard 96-well format and are made of tissue culture-treated polystyrene. The plates are intended for in vitro use.
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Lab products found in correlation
4 protocols using eucamp2 plates
1
Antimicrobial Resistance Evaluation in Campylobacter
2
Antimicrobial Susceptibility of Campylobacter
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Campylobacter strains were subjected to an antimicrobial susceptibility test (AST) using the broth microdilution method. Sensititre Campylobacter EUCAMP2® plates (Thermo Fisher Scientific®, Madrid, Spain) were used according to the manufacturer’s instructions. Each strain was tested against six different antimicrobials: nalidixic acid (NAL), ciprofloxacin (CIP), erythromycin (ERY), tetracycline (TET), gentamycin (GEN), and streptomycin (STR). The susceptibility or resistance of C. jejuni and C. coli isolates to antimicrobials was determined using the epidemiological cut-off values (ECOFF) established by the European Committee on Antimicrobial Susceptibility Testing (EUCAST, 2023) (Table 1 ) [35 ]. For other Campylobacter species, the minimum inhibitory concentration (MIC) cut-off values for C. jejuni were applied as they were the most restrictive ones. MDR was defined as resistance to at least three different antimicrobial families [36 (link)].
3
Antimicrobial Resistance Profiling of Isolates
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The minimum inhibitory concentrations (MIC) for six antimicrobials (ciprofloxacin, erythromycin, gentamicin, nalidixic acid, streptomycin, and tetracycline) were determined by broth microdilution using Thermofisher Sensititre EUCAMP2 plates according to the manufacturer’s protocol. Plates were incubated microaerobically at 41 °C for 24 h. MICs were interpreted using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) epidemiological cut-off (ECOFF) values as wild-type or non-wild-type [27 (link)]. ECOFFs distinguish microorganisms without (wild-type) and with phenotypically detectable acquired resistance mechanisms (non-wild-type) to the antimicrobial in question (https://mic.eucast.org/ ). Isolates with non-wild-type susceptibilities have been termed microbiologically ‘resistant’, although it is recognised this is not necessarily synonymous with clinical resistance. Multidrug resistance was defined as resistance to three or more antimicrobial classes [27 (link)]. In the panel of antimicrobials tested the quinolone class is represented by ciprofloxacin and nalidixic acid.
4
Antimicrobial Resistance Profiling via WGS
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For isolates characterized by WGS, antimicrobial resistance was evaluated using minimum inhibitory concentration (MIC) (EUCAMP2 plates; Thermo Scientific, USA). The following antibiotics (and dilution ranges) were evaluated in the panel: gentamicin (GEN; 0.12–16 mg/L), ciprofloxacin (CIP; 0.12–16 mg/L), nalidixic acid (NAL; 1–64 mg/L), tetracycline (TET; 0.5–64 mg/L), streptomycin (STR; 0.25–16 mg/L), and erythromycin (ERY; 1–128 mg/L). MIC classification was interpreted as wild-type (susceptible) or non-wild-type (resistant) using the epidemiological cutoff (ECOFF) defined by European Committee on Antimicrobial Susceptibility (EUCAST) (EFSA et al., 2022 ). The relatedness between AMR genotypes and phenotypes was calculated by dividing the count of resistant isolates based on their genotype by the count of isolates that showed phenotypic resistance (Dahl et al., 2021 (link)).
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