Synergistic bacterial compositions and methods of production and use thereof
A technology of composition and bacteria, applied in the direction of antibacterial drugs, drug combinations, medical raw materials derived from bacteria, etc., can solve the problems of non-standardization and characterization of products
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example 1
[0170] Example 1. Construction of binary pairs in a high-throughput 96-well format. To allow high-throughput screening of binary pairs, vials of the -80°C glycerol stock were thawed and diluted to 1e8 CFU / mL. Each strain was then diluted 10X (to achieve a final concentration of 1e7 CFU / mL for each strain) into 200 μL of PBS + 15% glycerol in wells of a 96-well culture plate. Plates were then frozen at -80°C. When tested with C. difficile in CivSim, plates were removed from -80°C and thawed anaerobically at room temperature when required.
example 2
[0171] Example 2. Construction of triplets in a high-throughput 96-well format. To allow high-throughput screening of triplets, vials of the -80°C glycerol stock were thawed and diluted to 1e8 CFU / mL. Each strain was then diluted 10X (to achieve a final concentration of 1e7 CFU / mL for each strain) into 200 μL of PBS + 15% glycerol in wells of a 96-well culture plate. Plates were then frozen at -80°C. When required for analysis, when tested with C. difficile in CivSim, plates were removed from -80°C and thawed under anaerobic conditions at room temperature.
example 3
[0172] Example 3. Screening for Ecobiotics that inhibit the growth of Clostridium difficile TM Construction of CivSim assays of compositions. Overnight cultures of C. difficile are grown under anaerobic conditions in SweetB-FosIn or other suitable media for C. difficile growth. SweetB-FosIn is a complex medium composed of brain heart extract, yeast extract, cysteine, cellobiose, maltose, soluble starch, and fructooligosaccharide / chrysanthemum tang and hemoglobin, and with MOP buffer. After 24 hours of growth, the culture was diluted 100,000-fold into a complex medium suitable for growth of various anaerobic bacterial species (such as SweetB-FosIn). Next, the diluted C. difficile mixture was aliquoted into wells of a 96-well culture plate (180 μL per well). Next, 20 μL of unique binary pairs of potentially inhibitory species were added to each well at a final concentration of 1e6 CFU / mL per species. Alternatively, the assay can be tested with binary pairs of different initia...
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