Tryptic soy broth

All media hlls have been performed with tryptic soy broth. 

Luminometer 3550i and Filtravette (New Horizons Diagnostics Corp., USA) were used for bioluminescent measurement. ATP-reagent (based on soluble Luciola mingrelica firefly luciferase) was developed in our laboratory. Dimethyl sulfoxide (DMSO) was from Reakhim (Russia). Nutrition broth (NB) from ICN, Tryptic Soy Broth (TSB) from Difco and Thioglycolate broth (TB) from Merck were used. Ultrapure deionized water was obtained on Milli-Q (Millipore). Broth culture of Escherichia coli LE392 was used in model experiments. 

Each Pit microbe was initially grown in 6 different nutrient broths as both shake (7 day) and still culture (21 day) to assess what growth conditions yielded the maximum biological activity. Each microbe was grown in a 100 mL nutrient broth in 250 mL Erlenmeyer flasks. The media used included PDB (DIFCO potato dextrose broth) PDBH+ ( PDB acidified to pH 2.5 by the drop wise addition of sterile 1.0 M H2SO4) Pit II (filter sterilized Berkeley Pit surface water added to an equivolume sterile solution of 5 g/L soytone and 5 g/L glucose) TSB (DIFCO tryptic soy broth) TSBH+ (TSB acidified to pH 2.5) and Myc (DIFCO mycological broth). 

Tryptic soy broth (TSB) is suggested for use in neutralization assay and for preparing the E. coli inocula to be distributed into the ground hamburger. 

Tryptic soy broth (TSB) (Difco, Franklin Lakes, NJ), supplemented with 0.5% glucose after sterilization by autoclaving. 

Preparation of culture supernatants - R haemolytica biotype A1 was grown as previously described (17). Briefly, colonies from a blood agar plate wCTe used to inoculate tryptic soy broth. This culture was incubated at 37 C for 4.5 hours. This broth was used to inoculate fresh tryptic soy broth which was incubated for 1 hour at 370 C with 5% CO2 bubbled throu it. The broth was stirred as rapidly as possible without foaming. After 1 hour of incubation bacteria were removed from the culture by ultrafiltration (Pellicon, Millipore, Inc.). Culture supernatants free of bacteria were lyophilized and stored dessicated at -20 C until loaded into hydrogels. 

Culture supernatants used to load hydrogels Tryptic soy broth alone Eluent 1 Eluent 2 Eluents... 

Bacillus licbeniformis (ATCC 9945a) was obtained from Dr. L- K. Nakamura, of the U. S. Department of Agriculture, Peoria, IL. This freeze dried sample was suspended using 0.5 mL Tryptic Soy Broth (Difco Labs). This solution was transferred to a larger volume of Tryptic Soy Broth (100 mL) and incubated on a rotary-shaker (250 RPM, 37 C) for 18 h. From this inoculum four stock culture plates were prepared using Tryptic Soy Broth supplemented with 1.5% agar. These culture plates were then incubated at 37 C for 18 h. 

Sterile Tryptic Soy Broth (100 mL) in a 250 mL Erlenmeyer flask was inoculated using an isolated mucoid colony from a stock culture plate. This culture was incubated on a rotary shaker (250 RPM, 37 C) for 18 h. Aliquots of this inoculum (1.5 mL) were introduced into 500 mL Erlenmeyer flasks each containing 100 mL of sterile medium E at pH 7.4. For a time course study, multiple culture flasks were prepared taking 100 mL aliquots of medium E from a larger volume. The cultures, inoculated in an identical manner, were incubated on a rotary shaker (250 RPM, 37°C). These cultures were then interrupted and analyzed at different time points. 

Frank J F et al. (2004) Attachmentt of Escherichia coh 0157 H7 grown in Tryptic Soy Broth and NUtrient Broth to Apple and Lettuce Surfaces as Related to Cell Hydrophobicity, Surface Charge and C sule Production. International Journal of Food Microbiolt y 96 103-109. 

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