Agar plate preparation

SD-CAA selective growth media and agar plates prepared in Subheading 2.2.3. 

Acetyl-1-cyclohexene Ethanone, 1-(1-cyclohexen-1-yl)- (932-66-1), 76, 203 Acetylenes, terminal, cyanation of, 75,148 Agar plate preparation, 76, 79... 

API agar plate prepared according to Allred (6). reducing colonies become black on this medium. 

LB-arapicillin/X-gal/IPTG agar plates Prepared as in step 4 except that 160 pL of X-gal solution and 200 pL of IPTG are added along with ampicillin. 

Solutions of the substrate have been prepared, for example, in acetone or diethyl ether, and added to or spread over the surface of agar plates either before or after inoculation (Sylvestre 1980 Shiaris and Cooney 1983). 

A spore suspension was prepared by resuspending the spores from the agar plate in 15 mL physiological aqueous solution (8.5 g NaCl, 1 g peptone, and 1 g Tween 80 in 1 L distilled water, sterilized at 121 °C for 20 min) with a Drigalski spatula. The spore suspension was diluted to a concentration of approximately 2.5 x lO spores/mL and stored in a 50 mL Falcon tube at 4 °C. 

Thirty microliters of the suspension is spread on one quarter of a square LB agar plate containing 50 pg/mL of kan-amycin and incubated at 37°C overnight. Thus four kinds of samples are on one plate and 24 plates are needed per a 96-sample set. In addition, glycerol stocks are prepared by blending 100 pL of the cell suspension with 50 pL of Cell Stock Buffer in a 96-well format. 

Another test that has been adapted by the Center for Veterinary Drug Residues, Saskatoon, Canada, to screen muscle samples from imported carcasses is the cube inhibition test (CIT) (83). Tissue preparation is similar to that in the German three-plate test in that tissue cubes are placed on seeded media. The organism is Bacillus subtilis, two different agar plates are used, and incubation is carried out at 37 C for 18-24 h. 

Referring to US Patent 6,015, 832 (Table 1), an emulsion was prepared from the disclosure and the activity tested against P. ageruinosa and Escherichia coli bacteria. The results are reported in Table 4.2 from plate tests and an image of each agar plate is appended (Fig. A4.1). The emulsions were prepared again for reproducibility, and the tests were repeated (without dilution) in Table 4.3 with similar results. 

Samples for cell viability tests were taken three times during the different batch and fed-batch fermentations. A sample of 1 mL was diluted 104-105 times and 0.1 mL of diluted sample was applied to an agar plate. Five to six plates were prepared for every sample, and the plates were incubated for 24 h at 30°C. The total cell concentration in the reactor sample was determined with microscopy using a Biirker counting chamber. The viability, expressed as the fraction of cells able to form colonies, was calculated by dividing the concentration of colony-forming cells by the total cell concentration. 

Two key steps involved in the traditional methods are antiserum preparation and the agglutination tests. Sera for O determination are produced by immunization of rabbits with cultures that have been heated at 100°C for 2 h. Broth cultures or agar plate suspensions heated at 100°C for 1 h are used as antigens for typing. With these two procedures, bacterial agglutination is a very simple and sensitive method for qualitative O antigen determination. 

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