Trypto casein soy broth

Iron(III) chloride hexahydrate (FeCl₃.6H₂O, 97.0−102.0%), iron(II) chloride hydrate (FeCl₂.H₂O, 99%), ammonium nitrate (NH₄NO₃, 99.999%), N-hydroxysuccinimide (NHS, 98+%), ethylenediaminetetraacetic acid tetrasodium salt hydrate (EDTA, 98%) and tetraethyl orthosilicate (TEOS, ≥99.999% metals basis) were bought at AlfaAesar. The n-Cetyltrimethylammonium bromide (CTAB), (3-aminopropyl) triethoxysilane (APTES, ≥98%), thioglycolic acid (≥99.9%), succinic anhydride (≥99% GC), (+)-sodium L-ascorbate (crystalline, ≥98%), N-3-dimethylaminopropyl-n-ethylcarbodiimide hydrochloride (EDC, ≥97%), acetone (≥99.5% GC), 4-morpholineethanesulfonic acid (MES, ≥99%), TRIS (hydroxymethyl)aminomethane hydrochloride (TRIS HCl) and phosphate-buffered saline (PBS, tablets) were purchased from Sigma Aldrich. Ethanol absolute (EtOH, extra pure), hydrochloric acid (HCl, 37%) and sodium hydroxide (NaOH) were bought at Scharlab, SL. Oleic acid (OA, 65.0–88.0 %) was acquired from Honeywell Fluka and sodium chloride (NaCl, 99.0–100.5%) at PanReac AppliChem. Bacto^TM Glycerol was bought from Becton Dickinson&Co (Sparks, MD, USA). Muller Hinton Broth (MHB) and Trypto Casein-Soy Broth (TSB) were obtained from Biokar Diagnostics. All reagents were used as acquired, without any further purification, and all solutions, unless otherwise indicated, were prepared with deionized Millipore miliQ water.

Wild birds (n = 200) were purchased alive on the Sousse market, Tunisia, between January and February 2022 during the legal hunting period. Birds were caught in Bizerte, Zaghouan, or Gabès (Fig. S1), and kept in aviaries for as short a time as possible, without being fed. The intestine of each bird was collected immediately after the death of the bird using a sterile scalpel and stored at −20°C. Intestinal content was emptied in 10 mL of Trypto-casein soy broth (Biokar), homogenized and incubated for 18–24 h at 37°C. Overnight cultures were inoculated on selective MacConkey agar plates supplemented with cefotaxime or imipenem (2 mg/L), for the detection of ESC- or CP-resistant Enterobacterales. One colony per morphology and per plate was picked up. Identification was performed using API20E galleries (bioMérieux).

Wild birds (n = 200) were purchased alive on the Sousse market, Tunisia, between January and February 2022 during the legal hunting period. Birds were caught in Bizerte, Zaghouan, or Gabès (Fig. S1), and kept in aviaries for as short a time as possible, without being fed. The intestine of each bird was collected immediately after the death of the bird using a sterile scalpel and stored at −20°C. Intestinal content was emptied in 10 mL of Trypto-casein soy broth (Biokar), homogenized and incubated for 18–24 h at 37°C. Overnight cultures were inoculated on selective MacConkey agar plates supplemented with cefotaxime or imipenem (2 mg/L), for the detection of ESC- or CP-resistant Enterobacterales. One colony per morphology and per plate was picked up. Identification was performed using API20E galleries (bioMérieux).