The α-L-arabinofuranosidase production was carried out in 250-ml Erlenmeyer flasks containing 5 g of washed dried agro residues (wheat bran, wheat straw, rice bran, rice straw, corn cobs, and maize bran) moistened with 10 ml of modified Mandels-Weber medium containing the following (g/l): urea, 0.3; ammonium sulfate, 1.4; KH2PO4, 0.3; CaCl2, 0.3; MgSO4.7H2O, 0.3; proteose peptone.
Production of protease from Bacillus brevis MWB-01 was carried out in a culture medium containing the following: 0.5% glucose, 0.75% peptone, and 5% salt solution made up of 0.5% MgSO4.7H2O and 0.1% NaCl, maintained at 37°C for 72 h in a shaking incubator (180 rpm).
E. cloacae strain SDM was cultured in a medium containing the following (g/L) at pH 7.0: glucose, 15; peptone, 10; yeast extract, 5; KCl, 5. Luria Bertani (LB) medium was used for E. coli cultivations.
Lactose fermentation was tested in a modified API 50CH medium containing the following: lactose 5 or 20 g.L−1 (Panreac, Lyon, France), tryptone 10 g.L−1, yeast extract 5 g.L−1, K2HPO4, 0.25 g.L−1, MnSO4 0.05 g.L−1, and bromocresol purple 0.17 g.L−1.
Cells were grown on basal medium containing the following (g/L): casein 1.0, NaNO3 1.0, K2HPO4 1.52, MgSO4 7H2O 0.52, and KCl 0.52, adjusted to pH 6.0, and supplemented with sunflower oil 1.0% (v/v).
Susceptibility tests were performed using the resistance proportion method for INH, RIF, SM, EMB and TH; strains were considered highly resistant if the same growth was observed on Lowenstein-Jensen medium containing the following drug concentrations, respectively: 0.2 1 μg/ml, 40 μg/ml, 4 μg/ml, 2 μg/ml and 2 4 μg/ml.
The liquid fermentation was conducted in 300 mL Erlenmeyer flasks by taking 50 mL of medium (containing the following: 54 g/L wheat bran; 3 g/L (NH4 2SO4; 2 g/L MgSO4·7H2O; 1 g/L Na2CO3; and 1 g/L Tween-80) on a swing shaker (100 rpm) at 34°C for 72 h.
The epithelial-containing tissue was enzymatically dissociated by using digestion medium containing the following reagents: Ham's F-12/DMEM (1:1) supplemented with 10% FBS, 5 g/mL insulin (Sigma-Aldrich, St . Louis MO, USA), 0.5 g/mL hydrocortisone (StemCell Tech, Vancouver, BC, Canada), 10 ng/mL cholera toxin (Sigma-Aldrich), collagenase/hyaluronidase 10× (StemCell Tech), and antibiotics.
For all cultivations, the nitrogen-limited medium contained the following components per liter: 2 g Na2HPO4⋅12H2O, 14.9 g KCl, 46.72 g NaCl, 14.5 g Tris, 2.05 g MgCl2, 3.53 g Na2SO4, 1 g (NH4 2 SO4, 1 g MgSO4⋅7H2O, and 2.5 mL of trace element solution.
Semi-selective medium contained the following: 37.5 mg carbendazim, 37.5 mg thiabendazole, 75 mg rose bengal, 17.5 mg NaCl, 50 mg each of streptomycin sulphate, chlortetracycline hydrochloride and chloramphenicol, 3 ml Triton X-100, and 17 g corn meal agar (Difco) in a liter of distilled water.
