Isolation and Identification of Aliarcobacter from Fecal Samples

The Aliarcobacter isolates from fecal samples were cultivated in Arcobacter broth (Oxoid GmbH, Wesel, Germany) which was supplemented with three different antibiotics (cefoperazone, amphotericin, and teicoplanin (CAT), Oxoid GmbH). The broth was then spread on plates (Mueller–Hinton agar/CAT/5% defibrinated bovine blood, Sifin GmbH, Berlin, Germany). The incubation criteria for each step were: 48–72 h, 30 °C and microaerophilic atmosphere (5% O₂, 10% CO₂, and 85% N₂). Suspicious colonies were further cultivated and then identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as described before [32 (link),33 (link)]. IVD Bacterial Test Standard and Biotyper 3.1 software were used (Bruker Daltonik GmbH, Bremen, Germany). DNA was purified using the High Pure PCR Template Preparation Kit (Roche Diagnostics, Mannheim, Germany) following the manufacturer’s instructions, and the species identification was confirmed with a multiplex PCR assay [34 (link)].