gilamemis BSH-3T Nitrogen source Enzyme activity (IU) Cell mass (mg/ml) Beef extract 160.8[+ or -]3.4 0.82[+ or -]0.013 Malt extract 152.4[+ or -]4.2 0.76[+ or -]0.023 Peptone 158.2[+ or -]5.2 0.78[+ or -]0.031
Tryptone 156.4[+ or -]6.7 0.74[+ or -]0.035 Urea 123.2[+ or -]3.4 0.43[+ or -]0.024 Yeast extract 220.2[+ or -]5.2 1.40[+ or -]0.034 Table 4.
Pneumococcal isolates were saved at -40AdegC in Skimmed milk
tryptone glucose glycerol (STGGA) medium.
All the diluted samples were cultured on two plates of different culture media, inorganic salts starch agar (ISP4) media supplemented with cycloheximide (100pg/ml) and colistin (5pg/ ml) for Actinomycetes; and
tryptone soya agar supplemented with cycloheximide (30pg/ml) and penicillin (100pg/ml) for Eubacteria.
For the determination of total aerobic mesophilic bacteria, dilutions of the water samples were made in
tryptone salt and then 1 mL of each dilution was incorporated in plate count agar medium (Biokar Diagnostics) and incubated at 30[degrees]C for 72 h.
The media, including Luria Bertani broth (LB) (
tryptone, 10.0 g/L; yeast extract, 5.0 g/L; NaCl, 10.0 g/L),
tryptone soy broth (TSB) (
tryptone, 17.0 g/L; soytone, 3.0 g/L; glucose, 2.5 g/L; NaCl, 5.0 g/L; [K.sub.2]HP[O.sub.4], 2.5 g/L), and modified yeast extract broth (YSG) (glycerol, 5.0 mL/L; yeast extract, 5.0 g/L; 1 M MgS[O.sub.4], 5.0 mL/L; [(N[H.sub.4]).sub.2]S[O.sub.4], 2.0 g/L; 1 M K[H.sub.2]O[P.sub.4], 5.0 mL/L; 1 M [K.sub.2]HO[P.sub.4], 5.0 mL/L; and 1 M [Na.sub.2]S[O.sub.4], 10.0 mL/L) were used in comparative studies to find the optimal nutrient medium for maximizing the antifungal activity.
Cultures with these confirmed nonpathogenic or pathogenic strains were stored at -80[degrees]C in modified
tryptone soy broth (mTSB) (Land Bridge Technology Co., Ltd., Beijing, China) supplemented with 3% NaCl and adjusted to pH 8.0 [+ or -] 0.2) with addition of 25% (v/v) glycerol (National Pharmaceutical Group Corporation, Shanghai, China).
Antimicrobial assay: Before inoculation, all Staphylococcus spp isolates were transferred from the stock cultures to
Tryptone Soya Agar (TSA) (Oxoid-CM0131) and incubated overnight at 37 AdegC.
For experimental use, freeze-dried cultures of the organisms were grown on
tryptone soya agar (Oxoid) at 37[degrees]C.
The plate was incubated overnight at 4 [degrees]C and subsequently washed once with 0.05% PBST, and blocking solution (1%
tryptone, 1% BSA, 3% sucrose (150 [micro]L/well)) in PBS for 1 h at 37 [degrees]C was then added.
Axenic cultures of Tetrahymena pyriformis were obtained from Carolina Biological Supply Company (Carolina) and maintained in Carolina's growth medium (proteose peptone, 5 g/l;
tryptone, 5 g/l; [K.sub.2]HP[O.sub.4], 1.1 mmol/l).
Samples were stored in cryotubes containing 0.8 mL of TSB (
Tryptone Soya Broth, Himedia) and 0.2 mL Glycerin at -20 [degrees]C.
sullae were grown on the yeast extract mannitol agar (YMA) solid medium (mannitol, 10 g/liter; [KH.sub.2][PO.sub.4], 0.5 g/liter; [MgSO.sub.4].7[H.sub.2]O, 0.2 g/liter; NaCl, 0.1 g/liter; yeast extract, 0.5 g/liter; agar, 15 g/liter), on the Tryptone-yeast (TY) broth medium (
Tryptone, 5 g/liter; yeast extract, 3 g/liter; [CaCl.sub.2].[H.sub.2]O, 0.87g/liter), or Tryptone- yeast agar (TYA) solid medium (TY supplemented with 12g of agar in liter) at 28[degrees]C for 24h.