International Journal of Biotechnology Applications

147-154

Authors: GUERMOUCHE M.A., BENSALAH F., GRAY N

In contaminated soils, the efficiency of natural attenuation or engineered bioremediation largely depends on the biodegradation capacities of the total microflorae. In the present study, the biodegradation capacities of various bacteria towards petroleum-hydrocarbons were determined under laboratory conditions. The purpose of the study was to isolate and characterize petroleum-degrading bacteria from contaminated soil obtained from a refinery in Arzew, Algeria. A collection of 15 bacterial isolates were obtained by enrichment cultivation from oil-contaminated soil and an indigenous microbial consortium was developed by assembling four species of bacteria which could degrade different fractions of the petroleum hydrocarbons. 16S rRNA gene analysis was used to identify members of the consortium and oil biodegradability was analyzed by Thin Layer Chromatography (TLC) with Flame Ionization Detection (FID) that performs quantitative compositional analysis of oil samples. The Iatroscan TLC/FID system measured the relative percentages of the four major fractions of petroleum i.e. saturates aromatics, resins and asphaltenes. Results indicated that the constructed consortium which comprised the genera Pseudomona, Shewanella, Enterobacter and Serratia used the hydrocarbons as sole sources of carbon where biodegradation was defined by an initial rapid decrease in the saturate and aromatic fractions from 56.44% and 34.72% to 51.77% and 27.77% respectively, coinciding with an increase in the asphaltene fraction. The resin content remained relatively constant throughout the project. Alkane hydroxylase genes (alkB) were positively amplified in the Pseudomonas isolate by the polymerase chain reaction (PCR) method using degenerate primers. This functional gene was used as a marker to assess the catabolic potential of the bacteria for alkane pollutant biodegradation. The selected bacterial consortium looks promising for its application in bioremediation technologies.

134-136

Authors: LAKSHMI NARASU MANGAMOORI, KALYANA CHAKRAVARTHY BONALA

Extra cellular lipase production by the bacterial strain, Bacillus tequilensis was investigated in submerged shake flask fermentation. This work was intended to optimize culture conditions of Lipase production Bacillus tequilensis with submerged fermentation using 1% (w/v) carbon source, Nitrogen source and different oils along with physical parameters like pH, temperature, inoculum concentration, agitation speed. The optimum pH and temperature for Lipase production in submerged fermentation were found to be 7.0 and 30°C. Maximum activity was observed with 1% inoculum concentration, at incubation period of 48hrs and agitation speed of 200rpm.

118-129

Authors: SATISH INGALE, SHRIVASTAVA S.K

Proximate, nutritive and anti-nutritive analyses of some new varieties of oilseeds were determined. Oilseeds studied include Sunflower (Helianthus annuuus L.) variety LSF-11 & LSF-8, Safflower (Carthamus tinctorius L.) variety PBNS-12 & PBNS-40 and Groundnut (Arachis hypogaea L.) variety JL-24. The proximate parameter such as moisture, crude fibre, total lipid, crude protein, carbohydrate, ash calcium phosphorus and energy ranged from 3.627-7.393, 0.488-3.411, 25.699-46.224, 15.91-25.20, 21.26-48.93, 2.577-4.866, 0.087-0.150, 0.15-0.41 and 490.651-601.856 g/100g respectively. The predominant fatty acid was found to be Linoleic acid. The protein solubilizations at different pH ranging from 0.5 to 13.5, the maximum seed proteins were extracted at pH 12. The amino acid analysis reveled that the oilseeds were superior with respect to glutamic, arginine, methionine and proline. The anti-nutritional factors like cyanogenic glucoside, tannin, oxalate and hemagglutinin activity contents ranged from 3.458-4.818, 0.412-0.651, 0.079-0.180 and 1:16-1:8 g/100g respectively. Trypsin inhibitor was not found in these varieties. The nutritive values were determined for feed utilization, nitrogen utilization, protein efficiency ratio and feed efficiency ratio ranged between 5.260-6.552g, 0.232-0.296g, 1.368-1.509 and 0.345-0.365 respectively. Results from these studies have revealed that the potential for use of sunflower, safflower and groundnut for enhancement of these nutrients/chemical constituents.

97-101

Authors: SRINU NAIK S., PYDI SETTY Y

The objective of this study is to investigate the optimal parameter levels for removal of nitrates from wastewater using Pseudomonas stutzeri in a fluidized bed bioreactor by Taguchi method and validating the predicted values. In this study the influence of airflow rate, temperature, carbon source, PP beads (Poly Propylene) and pH at different levels was investigated was made using Taguchi’s L16(4)5 orthogonal array technique. The nitrate removal efficiency after optimization is more than 96% with airflow rate at level 2 (A2), temperature at level 3(B3), carbon source at level 4(C4), PP beads at level 2(D2) and pH at level 2(E2). The analysis of variance (ANOVA) test is used to determine the percent contribution of each parameter and found that airflow rate contribution is the highest and temperature provides the least contribution.

106-109

Authors: PYDI SETTY Y., SRINU NAIK S

Nitrate is a major pollutant present in effluent wastewater from nitrogenous fertilizer, explosives, paper mills, pulp mills, and also from municipal waste. Nitrate is harmful to both mankind and animal and also to the environment. The World Health Organization has set a limit of 10 mg/L NO3- for human consumption and 100 mg/l NO3- for animals. Water above these limits requires denitrification. Nitrates causes cancer, blue-bay syndrome, hypertension and thyroid hypertrophy. Experimentation on biological denitrification was carried out in a fluidized bed bio-reactor using synthetic wastewater with polypropylene beads as supporting media for the growth of microorganism. Synthetic wastewater is taken into the reactor for biological treatment and air was fed from bottom with solids being fluidized at the top due to low density. Experiments were performed for water of different initial nitrate concentrations with Pseudomonas stutzeri microorganism by varying the parameters like airflow rate, temperature, carbon source, poly propylene beads and pH in the range 2 lpm to 3.5 lpm, 20°C to 35°C, 70 mg/L to 85 mg/L, 5 mg/L to 25 mg/L and 6 to 8 respectively. The optimum parameters at which maximum denitrification is noticed are found to be 2.5 lpm (air flow rate), 30°C (temperature), 85 mg/L (carbon source), 15 mg/L (polypropylene beads) and 7 (pH). More than 95% removal of nitrates is noticed from the experimental work. The objective of the study is to investigate the effect of operating parameters like airflow rate, temperature, carbon source, polypropylene beads and pH.

72-79

Authors: SHANMUGAM GIRIJA, KILANKAJE ASHAKIRAN, MAHADEVAN CHIDAMBARESWARAN, VAITHIYANATHAN GOVINDASAMY, VELU SIVANKALYANI

Agrobacterium mediated transformation through somatic embryogenesis was achieved using cotyledon explants of tomato cv. Shalimar (Solanum lycopersicum L.). Somatic embryos were induced directly from the cotyledon explants on MS medium supplemented with 2, 4-D (5mg/l) for the first time, and also through the callus produced from cotyledon explants on MS liquid medium supplemented with 2, 4-D (2mg/l). The maturation of somatic embryo globular to torpedo was presumed by using sorbitol (0.4 M) and ABA (0.25mg/l). The competency of induced somatic embryos for agrobacterium mediated transformation was confirmed by using GUS reporter gene in pBI121 vector and the npt II with PCR. The transformant was selected using kanamycin (100mg/l). Seventy percentage of transformation efficiency was obtained from the transformed somatic embryo.