Tryptone water

An incubated culture of Actinomyces antibioticus was prepared using a medium consisting of 1% tryptone-peptone, 0.5% starch, 0.2% K HPO, 0.2% NaCI and 0.25% agar in distilled water, grown at a temperature of approximately 25° to 35°C, the incubation being complete after 6 to 10 days. 50 liters of this incubated culture are extracted approximately six times with ether, using 20 liters of ether for each extraction. 

Reports on a few other desulfurization organisms also exists, including Nocardia [98], S. acidocaldarius [5,99], Arthrobacter [100], R. toruloides, [101], C. elegans [102], Klebsiella spp. [103], and Alcaligenes xylosoxydans strain deposited with the ATCC under PTA-4669 [104], A S. acidocaldarius species was reported to desulfurize DBT and produce sulfate [5], A similar strain was later used to study the DBT desulfurization [99] in complex media using tryptone however, the authors reported that the strain was not active when grown on sucrose. It was apparently inhibited by DBT and also affected by the interface in oil-water mixtures [105],... 

Bacto-tryptone (1 g), bacto-yeast extract (0.5 g) and NaCl (1 g) were dissolved in 95 mL of deionized water and the pH was adjusted to 7.0 with 5 m NaOH. The volume was adjusted to 100 mL with deionized water. A portion of the resulting solution (5 mL) was placed in a 20 mL test tube with a poromeric silicone plug and the rest was placed in a 500 mL Sakaguchi flask with a poromeric silicone plug. The media were sterilized by autoclaving (121 °C, 20 min). 

Tryptone (5 g), yeast extract (2.5 g) and NaCl (5 g) were dissolved in distilled water, the volume was adjusted to 500 mL and then autoclaved (20 min, 120 °C). A small portion of this Luria-Bertani (LB) medium (10 mL) was placed into a sterile 100 mL shake flask and ampicillin and chloramphenicol solutions were added (LBamp+cm) to final concentrations of 100 p,g mL and 20 p.g mL respectively. The solution was inoculated with E. coli JM109 pGro7 pJOE4072.6 and shaken overnight at 37 °C and 200 rpm. This overnight culture (2 mL) was used to inoculate 200 mL LBamp+cm in a... 

Test Procedure. The test organisms were subcultured onto Tryptone Soya Agar (TSA) slopes and incubated at 37 °C for 24 hours. After incubation for each inoculum, 6 ml sterile distilled water (SDW) was added to each slope in turn, the organisms washed off and the resultant suspension homogenised. A 1 ml aliquot of the suspension was added to 100 mis SDW and homogenised to form the inoculum. 

Swab transportation tryptone saline, peptone water, enriched buffered gelatine, buffered saline, buffered gelatine, and brain-heart infusion are generally recommended media that may be used for swab transportation. For RCS + testing agar strip TC (BIOTEST) or M air T air sampler for level I is used. 

Rogosa-type medium (60) made as follows 2% Tryptone (Difco), 0.5% yeast extract (Difco), 0.5% peptone (Difco), 0.5% glucose, 0.005% Tween 80 (Nutritional Biochemicals Corp.), and 2% agar (Difco) in a filtered or centrifuged fourfold dilution (with water) of tomato juice (containing no preservatives). The medium is adjusted to pH 5.5 with HC1 before adding agar. 

Luria-Bertani (LB) medium. Each liter contains 10 g Bacto-tryptone, 5 g Bactoyeast extract, and 10 g NaCl. Adjust to pH 7.5 with NaOH. Ampicillin. A solution of the sodium salt is prepared in water (25mg/mL) and sterilized by passage through a microfilter (e.g., a 0.22 H Millipore filter). Store the solution in a freezer. 

Pellet cells for 5 min in two 50 mL Falcon tubes (e.g., 2000 rpm in a Beckman J-6M swinging bucket centrifuge with Beckman JS-4.2 rotor). (We use sterile Falcon tubes (BD Biosciences, San Jose, CA, cat. 352070) to minimize the chance of nuclease contamination.) The supernatant is removed. Optionally, one pellet can be resuspended in equal volume of LB (1% Bacto Tryptone, 0.5% yeast extract and 0.5% NaCl in water) and 60% glycerol, divided into multiple fractions and stored at —80 °C for future use. The other pellet is resuspended in 25 mL at 37 °C LB, then add into 2 L LB containing 2 mL of 50 mg/mL Carbenicillin and 2 mL of 10 mg/mL Chloramphenicol. Grow at 37 °C until the ODgoo is close to 1,... 

For each experiment, the supernatant was placed in 15-mL conical tubes, and the pH was adjusted with either 1 N KOH or 1N HN03. The pH of the supernatant after the bioreactor run was 7.0. Simulated potato effluent (SPE) medium was used as a surfactin-free control and the pH adjusted likewise. SPE contained the following per liter of nanopure water 5 g of potato starch, 3.5 g of peptone, 3.5 g of tryptone, 0.2 g of MgS04-7H20,0.1 g of yeast extract, and 0.8 g of (NH4)2S04. 

X TY medium dissolve 20g/l Bacto-Tryptone (Difco), 10 g yeast extract (Difco), and 10 g NaCI in 1 I water, and autoclave... 

Erwinia herbicola NCIMB 12126 was obtained from the National Collection of Industrial and Marine Bacteria (Aberdeen, UK), HEPES buffer sachets and magnesium acetate were obtained from Sigma (Poole, UK), adenylate kinase assay kits were obtained from Acolyte Biomedica (Salisbury, UK), sterile tissue culture grade distilled water was obtained from Gibco (Paisley, UK), L-broth and tryptone soya agar plates were obtained from Oxoid (Basingstoke, UK). 

Escherichia coli NCIMB 10243 was grown overnight in Tryptone Soya Broth (Oxoid CM129) in an orbital shaker at 35 C. This was used to inoculate water samples containing biocides (2 mL of culture added to 18 mL sample). Final concentrations of the biocides were 200 ppm. Samples were taken from the biocide/bacteria solutions at various time-points and both ATP assays and plate counts performed. 

The plate counts at To were estimated by diluting the neat broth culture 1 in 10 with Neutralised Peptone Water (NPW) containing per L 1.0 g Bacteriological Peptone (Oxoid, L37), 8.8 g Sodium Chloride (May Baker), 3.0 g Amisol 910 (Degussa) and 30.0 g Tween 80 (BDH, 560234H) and then decimally with Phosphate Buffered Saline (Oxoid). For biocide treated samples 1 mL was diluted in 9 mL of NPW, mixed and allowed to stand for 5 min (for neutralisation of the biocide). The solution in NPW was diluted decimally (0.1 mL in 0.9 mL) in PBS. Appropriate dilutions (0.1 mL) were plated out on Tryptone Soya Agar Plates (bioMerieux) and incubated at 37 °C for 24 h. 

YT media (10 g bacto-yeast extract, 16 g bacto-tryptone, 5 g NaCl water added to 1 L and pH adjusted to 7.0 with NaOH autoclaved). 

LB (5 g bacto-yeast extract, 10g bacto-tryptone, 10g NaCl water added to 1L autoclaved). 

LB media. Dissolve 10 g tryptone, 5g yeast extract, and 10 g NaCl in deionized water to a volume of 1L and sterilize by autoclaving. 

LB-Amp plates Dissolve 10 g tryptone, 5 g yeast extract, 10 g NaCl, and 15 g agar in 1L deionized water and sterilize by autoclaving. Add lmL of l,000x (100mg/mL) ampicillin stock to cool (<50°C), mix and pour plates. 

NOTE The type of broth used is often sterile tryptone soya broth that may be presented in double strength to allow for dilution with buffer, saline, or water to simulate the process. Any suitable liquid culture medium may however be used but the ability of the broth to support growth should be demonstrated. 

Digestion by boiling in the presence pf 30% hydrogen peroxide (Jackson (8)) yielded incomplete recoveries of total soluble phosphorus from all the estuarine water samples tested and from organic media such as Difco tryptone and peptone. Moreover, the method failed to yield quantitative recovery of an orthophosphate standard carried through the test procedure. 

Tryptone DL-Tryptophane Sodium chloride Demineralized water... 

LB medium Dissolve 10 g Bacto Tryptone, 5 g ofyeast extract, and 10 g of NaCl in water and adjust the volume to 1 L. Autoclave in narrow mouth flask or bottle and store at room temperature. 

---