3.12.2 Oxidase Test On a filter paper or cotton bud, pick the desired colonies from the agar plate. Then, using a dropper, take the oxidase reagent and drop it on to the filter paper and observe for colour changes. The blue colour appearance indicates positive reaction whereas if there are no changes, then it’s a negative reaction. 3.12.3 Genus Verification using TCBS agar The TCBS medium, known as Thiosulfate Citrate Bile Salts Sucrose Agar is a recommended selective medium which allows the growth of bacteria belonging to the genera Vibrio. TCBS agar was prepared for about 100ml and poured into petri plates, after solidifying, pure colonies of bioluminescent bacteria was streaked.
MATERIALS AND METHODS Bacterial strains and culture conditions Two S. aureus strains were used in the present study; S. aureus 8325-4 (SigB-) and SH1000 representing a SigB+.strain. Overnight cultures were grown in Luria Broth (LB) at 37°Cwith shaking at 150 rpm. Exposure to antibiotics was carried out as detailed below. Antibiotics Ciprofloxacin were purchased from Sigma-Aldrich CO. 10 mg/ml stock solution of antibiotic were prepared freshly with 0.1N HCl and stored at -20°C. During the experiment we diluted with sterile water 1:10 and 1:100 depending on the different drug concentration.
The selected colonies were streaked onto blood agar plates for observing the hemolytic patterns. The potent strains were selected for biochemical characterization. These biochemical tests were performed in reference to Cowan and Steel’s Manual for bacterial identification (Holt et al., 1993). (a) (b) Fig 1 Gram staining showing gram positive cocci in chains; (a) bore soil (b) curd PRODUCTION OF STREPTOKINASE (SK): The potent strains producing streptokinase were cultured in 100mL of the production medium (g/100 mL: Glucose - 0.5 g, Yeast Extract - 0.5 g, KH2PO4 - 0.25 g, MgSO4·7H2O 0.04 g, NaHCO3 - 0.1 g, CH3COONa·3H2O - 0.1 g, FeSO4·7H2O - 0.002 g, MnCl2·4H2O - 0.002 g, pH 7.5) and were kept for incubated at 37C for 24 h (Baewald et al., 1975). Once incubation is done, the individual cultures were centrifuged at 10,000 rpm for 30 mins.
The membranes were over laid with mouse monoclonal anti human factor VIII antibody (abcam, U.K) and incubated for 2h at room temperature (RT) and washed with 0.2% Tween 20/PBS three times, 5 min each. Incubation was continued at RT for 1h with HRP-anti mouse secondary antibody according to the manufacture protocol (abcam, U.K). After a final wash separated proteins were visualized using ECL (Santa cruze, USA) substrate solution using chemiluminescense system (BioRad, USA). Flow cytometry Presence of FVIII in erythrocytes was detected by flowcytometry using specific monoclonal antibody (abcam, U.K) against FVIII. 5 μl of washed FVIII loaded erythrocytes diluted with 100 μl PBS incubated at 4 for 30 minutes with FVIII monoclonal antibody.
Rice sample (1kg) was steeped in sodium hydroxide solution (0.3%) at 25°C for 24 hrs to soften the endosperm. The supernatant liquid was drained off and rice samples were grounded. The grounded rice was again dispersed in sodium hydroxide solution (0.3%) and then allowed to settle for 6h. This process was repeated until the supernatant showed negative response to the protein test. Then the separated sediment was suspended in distilled water; the pH of the slurry was adjusted to 7.0 with hydrochloric acid (0.5M) and passed through muslin cloth followed by sedimentation for 6h.
Positive test is indicated by the development of cherry red colour . Quality control : Positive - Enterococcus faecalis ATCC 29212 Negative - Streptococcus agalactiae SPECIATION OF ENTEROCOCCI SUGAR FERMENTATION TESTS: For identification of species, 1% of sugars(Glucose, arabinose,raffinose,mannitol,sorbitol,sucrose,lactose ) in peptone water with Bromothymol blue (0.002%) as the indicator was used. To each tube of Sugars,1-2 drops of 18-24 hour BHI broth culture is added and incubated at 37 C overnight. Interpretation: Sugar Fermentation is indicated by the change of colour to yellow. Arginine Dihydrolase test : Moeller's decarboxylase basal broth with 1% Arginine along with an aminoacid free control were inoculated with the test strain.Both the tubes were overlaid with sterile liquid paraffin and incubated at 37 C overnight.The colour of the indicator reverting back to original (purple) indicating arginine hydrolysis was considered as positive provided the control tube remains yellow indicating fermentation.
STARCH CONCENTRATIONS:There was progressive increase in the productivity of α-amylase and glucoamylase by the isolates as the starch concentration increased. INCUBATION PERIOD:Increase in the incubation period resulted in a decrease in the production of enzyme,byproducts form result in the depletion of nutrients inhibited the growth of fungi and hence enzyme formation.  BIOFUEL PRODUCTION :-Amylase is used in ethanol production to break starches in grains into fermentable
The presence of GOS (0.3%) in the microencapsulation provided the best protection with only 3.1 and 2.9 logs reduction for L. acidophilus 5 and L. casei 01, respectively, after incubation in simulated gastric juice (pH 1.55), followed by simulate intestinal juice containing 0.6% bile salt. The aim of this study was to investigate the prebiotics effect of inulin-type fructans with different chain lengths (DP<10, ~ 12 and ≥ 23) on the morphology, particle size and survival of microencapsulated probiotics strain of Lactobacillus casei in alginate beads, alginate coated with chitosan, alginate-inulin beads and alginate-inulin coated with chitosan exposed to simulated gastrointestinal
Babasaheb Ambedkar Marathwada University, Aurangabad. Young leaves and nodal part of stem were taken as explants. Explants were surface sterilized in running tap water for 10 minute followed by sterilized distilled water for 5 minutes. Apart from this sterilizing agents such as 70% ethanol, Hgcl2 (0.3 %) were tried. The duration for surface sterilization was 5 minute by 0.3% mercuric chloride followed by three subsequent rinses with sterilized distilled water.