The first step of the PCR thermo cycling conditions was ramping the temperature to 95 °C and held for 10 min. It was then lowered to 60 °C for 2 min before increasing to 98 °C for 30 s. To allow for efficient gene amplification, this cycle between 60 °C and 98 °C was repeated 40 times. Temperature was then lowered to 60 °C and held for 2 min, before cooling to room temperature. After allowing 30 mins for chips to equilibrate to room temperature, each chip was processed and read utilising the QuantStudio 3D Digital PCR system chip reader. For ARG analysis, the AnalysisSuite™ software was used for quantification of the target gene and any DNA dilutions were considered here. The DNA extracted from each biological replicate was run in triplicate for each gene investigated.