Emb agar

The bacterial specimens were cultured using eosine methylene blue (EMB) agar (bioMérieux, Marcy-l’Étoile, France) plates. To record colony morphology of the bacterial isolates, EMB plates were inoculated and incubated at 37 °C for 24 h, at aerobic atmosphere. After the incubation period, colony morphologies were assessed visually (for size, mucoid nature and lactose-fermentation) and these data were recorded. Colonies were considered small if their side was below 3 mm, or large if their size was >3 mm [33 (link)].

Urine samples were cultured using a calibrated pipette to deliver 10 μl and 100 μl of samples onto Columbia agar (Merck, Germany) supplemented with 5% sheep blood and onto MacConkey agar (Merck, Germany). The blood agar plates were incubated aerobically, and the MacConkey agar plates were incubated aerobically. All samples were incubated at 37°C for 24 h until adequate growth was present. Primary plates were carefully inspected for colonies of E. coli, which were plated onto sheep blood agar plates; these plates were incubated at 37°C for 24 h. Suspected colonies were transferred to the Eosin Methylene Blue agar (EMB agar, Merck, Germany) plates and incubated at 37°C for 24 h. Metallic green colonies with typical E. coli morphologies of the EMB agar plates were identified as E. coli using standard techniques, including indole, Methyl Red–Voges-Proskauer (MR-VP), Triple Sugar Iron agar (TSI), and citrate biochemical testing and analysis with an API-20E system (BioMérieux, Marcy l'Etoile, France) [16 ].