Member States shall carry out every two years the sampling, the collection and the antimicrobial susceptibility testing provided for in Article 2 to 4 of each combination of bacterial species and type of sample of animal populations or food categories listed in point 1 of this Part and the specific monitoring of ESBL- or AmpC- or carbapenemase-producing Salmonella spp. and E. coli in accordance with point 4 of this Part in accordance with the following rotation system:

1. (a)

   In the years 2014, 2016, 2018 and 2020 for laying hens, broilers and fresh meat thereof, and fattening turkeys. However, the specific monitoring of ESBL- or AmpC- or carbapenemase-producing indicator commensal E. coli in accordance with point 4.1 shall not be mandatory in the year 2014;

2. (b)

   In the years 2015, 2017 and 2019, for pigs, bovines under one year of age, pig meat and bovine meat.

Member States shall test 170 isolates for antimicrobial susceptibility testing for each combination of bacterial species and type of sample of animal population or food category listed in point 1(a), (b), (c), (e) and (f). However, in Member States with a production of less than 100 000 tonnes of poultry meat slaughtered per year and less than 100 000 tonnes of pig meat slaughtered per year18, they shall test 85 isolates instead of 170 isolates for each corresponding specific combination.

In those Member States where, in any given year, a higher number of isolates for some of the combinations of bacterial species and type of sample of animal population or food category listed in point 1(a), (b), (c), (e) and (f) is available, all isolates or a representative random selection equal to or greater than the number of isolates required in accordance with the first paragraph, shall be included in the antimicrobial susceptibility testing.

In those Member States where, due to a low bacterial prevalence or low number of epidemiological units, in any given year, the number of isolates required in accordance with the first paragraph for some of the combinations of bacterial species and type of sample of animal population or food category listed in point 1(a), (b), (c), (e) and (f), cannot be achieved, all available isolates at the end of the monitoring period shall be included in the antimicrobial susceptibility testing.

For the specific monitoring of ESBL- or AmpC- or carbapenemase-producing indicator commensal E. coli set out in point 4.1, Member States shall analyse 300 samples of each of the animal population and food category, listed in point 1(d). However, in Member States with a production of less than 100 000 tonnes of poultry meat slaughtered per year, less than 100 000 tonnes of pig meat slaughtered per year and less than 50 000 tonnes bovine meat slaughtered per year19 the Member States shall analyse 150 samples instead of 300 samples for each corresponding specific combination.

For the purpose of estimating the proportion of samples containing ESBL- or AmpC- or carbapenemase-producing E. coli amongst the caecal samples collected from broilers, fattening turkeys, fattening pigs, bovines under one year of age, fresh meat of broilers, pig meat and bovine meat in accordance with point 1(d) of this Part, the following method shall apply.

For the detection of ESBL- or AmpC-producing E. coli the method shall start by a pre-enrichment step, followed by inoculation on McConkey agar containing a third generation cephalosporin in a selective concentration according to the most recent version of the detailed protocol for standardisation of the European Union Reference Laboratory for Antimicrobial Resistance31. The microbial species E. coli shall be identified using an appropriated method.

The Member State may decide, based on the epidemiological circumstances, to test in parallel an additional selective plate that inhibits for the growth of AmpC-producing E. coli to facilitate the specific detection of ESBL-producing E. coli. When using this possibility, the results of the additional selective plate that inhibits for growth of AmpC-producing E. coli shall be kept separately when reported in accordance with point 2 of Part B.

Member States may decide to detect for carbapenemase-producing micro-organisms by using selective pre-enrichment and subsequent selective plating on carbapenem-containing media, according to the most recent version of the detailed protocol for standardisation of the European Union Reference Laboratory for AMR32.

One presumptive ESBL- or AmpC- or carbapenemase-producing E. coli isolate obtained from each positive caecal sample and meat sample shall be tested on the first panel of antimicrobials in accordance with Table 1 and further submitted to extended susceptibility testing as set out in point 4.2 if they are resistant to cefotaxime or ceftazidime or meropenem based on the interpretative criteria (epidemiological cut-off values) listed in Table 1.

All presumptive ESBL- or AmpC- or carbapenemase-producing E. coli isolates identified through the selective plating described in point 4.1 as well as all those randomly selected isolates of Salmonella spp. and E. coli that after testing with the first panel of antimicrobials in accordance with Table 1, are resistant to cefotaxime or ceftazidime or meropenem, shall be further tested with a second panel of antimicrobial substances in accordance with Table 4. This panel includes cefoxitin, cefepime and clavulanate synergy test in combination with cefotaxime and ceftazidime for detection of ESBL and AmpC production. In addition the second panel also contains imipenem, meropenem and ertapenem to phenotypically verify the presumptive carbapenemase-producers.

Member States, especially the Member States which have detected a high prevalence of ESBL- or AmpC-producing E. coli by the detection method set out in point 4.1, may characterise the proportion of ESBL- or AmpC-producing E. coli within the whole E. coli population.

That shall be done by enumerating ESBL- or AmpC-producing E. coli and total E. coli present in a sample using dilution methods and subsequent plating onto selective media and non-selective media, according to the most recent version of the detailed protocol of the European Union Reference Laboratory for Antimicrobial Resistance38.

• Description of sampling designs, stratification and randomisation procedures per animal populations and food categories.

• Identifier or code of the isolate

• Bacterial species

• Serovar (for Salmonella spp.)

• Phage type of Salmonella Enteriditis and Salmonella Typhimurium (optional)

• Food-producing animal population or food category

• Stage of sampling

• Type of sample

• Sampler

• The sampling strategy

• Date of sampling

• Date of isolation

• Identifier or code of the isolate given by the laboratory performing the antimicrobial susceptibility testing of the isolate

• Date of susceptibility testing

• Antimicrobial substance

• Minimum Inhibitory Concentration (MIC) value (in mg/L)

• Synergy testing with clavulanic acid for ceftazidime

• Synergy testing with clavulanic acid for cefotaxime