Sabaraoud agar

Serial two-fold dilutions of the KB were made; then, 0.1 mL aliquots from these dilutions were pipetted onto MRS agar plates [77 ]; specific Acetobacter agar (Oxoid, UK); Sabaraoud agar (Oxoid) for isolation of lactic acid bacteria; acetic acid bacteria; yeasts, respectively. The agar plates were incubated at 35 °C for either 48 h for bacteria or 4 days for yeasts. Pure and single colonies of the obtained microbes were picked up by sterile needles and inoculated into Brain Heart Infusion broths (BHI broth, Oxoid). After 24 h of incubation at 35 °C, 100 µL aliquots of microbial suspensions were loaded aseptically onto API kits (BioMérieux, France) that were then used for identification of the microorganisms isolated as given by the manufacturer’s instructions. Both bacteria and fungi isolated were checked for their Gram stain and cell morphology using a light microscope [2 ,3 (link),78 ,79 ].

Serial two-fold dilutions of the KFB were made; then 0.1 mL aliquots from these dilutions were pipetted onto specific Acetobacter agar (Oxoid, Basingstoke, UK); MRS agar plates [63 ,64 (link)]; Sabaraoud agar (Oxoid) for isolation of acetic acid bacteria; lactic acid bacteria; yeasts respectively. The agar plates were incubated at 35 °C for either 48 h for bacteria or 4 days for yeasts. Pure and single colonies of the obtained microbes were picked up by sterile needles and inoculated into Brain Heart Infusion broths (BHI broth, Oxoid). After 24 h of incubation at 35 °C, bacterial identification was carried out automatically by Vitek 2 system (Vitek^® 2: Healthcare|Biomérieux, France). The isolated bacterial species were checked for their Gram stain and cell morphology using a light microscope [65 ]. Yeasts were identified via morphological examination under light microscope and API-yeasts kits (BioMérieux, France) as given by the manufacturer’s instructions [66 ].