This organism was cultured under solid-state fermentation for 72 h using wheat bran as the substrate. After 72 h, crude enzyme was extracted from the culture medium. The fibrinolytic enzyme was purified from the crude sample by various steps: ammonium sulfate precipitation, dialysis, ion exchange chromatography, and casein-agarose affinity chromatography. All purification steps were performed at 4°C until otherwise stated. The crude enzyme was precipitated at 70% saturation of ammonium sulfate, and the protein was collected by centrifugation (10,000×g for 10 min).
On the second day of incubation, the plate was removed from the incubator and placed over a hot plate heating Iodine solids. The smoke of the Iodine stained the plate to display the presence or absence of a halo around the bacteria 2.12 Lipid Hydrolysis This test was done by making a single line streak inoculation on a tributyrin agar plate and allowing incubation. After the incubation period, the plate was observed for the presence or absence of a halo around the bacteria. 2.13 Gelatin Liquefaction A gelatin deep was deep stabbed and incubated. After incubation the tubes were placed in 4ºC for 30 minutes.
3.1 The isolation of Aeromonas from several pond waters, healthy fish, and infected fish The isolate of A. hydrophila grown for 24 hours at 37C on Rimler-Shoots+novobiocin medium should show bright yellow color with white edge. Figure 1 shows the control of isolate A. hydrophila ATCC 7699 grown on RS+novobiocin medium. Isolate selection on RS medium resulted in 95 isolates, presumed to be A. hydrophila, which would run the phenospecies test (morphology and biochemistry), based on the protocol of SNI 7303 (2009), plus one control isolate the A. hydrophila ATCC 7699 obtained from Microbiologic Co. Figure 1. The Isolate of A. hydrophila ATCC 7699 Grown on the Rimler-Shoots Agar Medium + Antibiotic Novobiocin at 37C for 24 Hours 3.2 The
TPR71H. From this study we found that the optimized parameters for the production of CGTase by Bacillus sp. TPR71H were Soluble Starch 3%, Yeast Extract 0.5%, K2HPO4 0.1%, Inoculum Level 3.5%, Inoculum Age 24h, Incubation Period 36h, rpm 220, Incubation Temperature 32°C and the pH of 7.5. M. Manoj Saravana Guru (2012) et. al.. Cyclodextrin glucosyltransferase (CGTase) producing alkalophilic bacteria were isolated from the water samples collected from Marneri pond of Tirunelveli, Tamil Nadu by serial dilution and plating method.
Using chromogenic media which is based on essential pathogenic virulence factors of listeria has been administered recently . Detection of Listeria monocytogenes in Listeria agar with Ottaviani and Agosti(ALOA) is based on detection of phosphatidylinositol-specific phospholipase C(PI-PLC) activity which Listeria monocytogene and some strains of L.inanovii hydrolysis the l-α-phosphatidylinositol and produce a fatty acid and form a opaque halo around the colonies [130, 131]. The other media which are similar to ALOA plate include: BCM listeria (Biosynth, Switzerland), OCLA (Oxoid, UK), CHROMagar_ Listeria (BD e Diagnostic Systems, USA) and OAA (bioMérieux, France). Rapid L’mono_ (Bio-Rad Laboratories, USA), could distinguish between hemolytic and non hemolytic listeria based on fermentation of xylose. L.ivanovii could ferment the xylose and produce blue colonies with yellow halo but L.monocytogene is non hemolytic and could not ferment the xylose and produce blue colonies without a halo.
The melanin synthesis using homogentisic acid as a precursor of was first reported in Vibrio cholerae, Hyphomonas species and Shewanella colwelliana (Kotob et al., 1995). The synthesis of melanin and its characterization such as solubility, free radical nature was initially studied in Proteus mirabilis (Agodi et al.,1996). A novel marine bacterium Alteromonas strain MMB-1, was isolated from the Mediterranean Sea and its melanin synthesis ability was studied using L-tyrosine as a precursor previously (Solano et al., 1997). The melanin pigment from Burkholderia cepacia was formerly reported for escaping monocyte respiratory burst activity by scavenging superoxide anion (Zughaier et al.,1999). The extra cellular melanin from Shewanella algae BrY was reported previously to serve as the sole terminal electron acceptor.
Smaller and smaller organelles may be sendimented by successive centrifugation at increasing speed. 1ml of the homogenate (H) was pipetted into a clean microfuge tube labelled P1. The tube labelled ‘P1’ was centrifuged at 1,000g for 10 minutes to sediment the first pellet (P1). The supernatant was carefully removed with an automatic pipette and placed in a micro centrifuge tube labelled ‘P2’. The P2 tube was then placed in the refrigerated centrifuge at a speed of 15,000g for 30-60 minutes at 4°C.
► 2.5-5% is messenger RNA RNA analysis- Electrophoresis- 1-Nucleic acids are are negatively charged,which makes them migrate to the positive pole in the electric field. 2-The separation process depends on the voltage used,the properties of the gels,as well as the charge and shape of the molecule. 3-Agarose gel electrophoresis and polyacrylamide gel electrophoresis used for high resolution. Agarose gel electrophoresis- 1 - it is used to separate DNA fragments that vary in size .Agarose is a polysaccharides obtsained from marine red algae .it is added to an electrophoresis buffer and then dissolved by heating it. 2 - The presence of many hydroxyl ngroup enable hydrogen brides to form,which lends firmness to the large pored gel matrix.
Lactobacillus lactis was inoculated in various media containing different vegetal source instead of beef extracts and was compared to the MRS culture medium as standard. These were incubated at 37oC for 72 hours wherein the growth was recorded every 24 hours using an undiluted, 10-4, 10-5 and 10-7 CFU/ml. The results of cultivation showed that the alternative test media using seed powders demonstrated better growth of colonies than the Man Rogosa Sharpe (MRS) cultre medium. Orange juice was used as an enrichment means for the enumeration of Alicyclobacillus acidoterrestis. In this process the inoculum was suspended in a medium containing orange juice and inoculated to different media such as K Agar, Yeast Starch Glucose Agar, Alicyclobacillus Agar, and Bacillus acidoterrestis Thermophilic Agar using duplicates for each run (Anjos et al, 2014).
MATAG inflorescence were collected from Department of Agriculture Terengganu’s farm. The samples were stored in ice-box to maintain its freshness prior to culture. B. Optimization of Sterilization Procedure The rachillae bearing inflorescence were cut off from the spadices and then were washed under run-ning tap water for 15 to 20 minutes to remove dirt and debris. The male inflorescence was isolated from rachillae and subjected to three different method of sterilization (as described in Table 1). C. Media Compositions and Growth Conditions The basal media used for this experiment were Y3 media (Eeuwens, 1976).