Development of Protocol for Production of Primary Antibody against Ovalbumin Protein in Chicken for Detection of the Protein through Western Blotting

Ovalbumin, a major protein of egg white plays many roles including providing nutrition to the developing embryo, acting as coagulating agent, folliculogenesis and angiogenesis in chicken and other animals. This protein is expressed mainly in magnum and then deposited over the yolk of the oocyte/zygote. Hence, it is important in formation of egg and is an essential target to measure. We cloned chicken ovalbumin CDS in pAcGFP-C1 vector and has been initially expressed in chicken primary magnum cell culture. The ovalbumin protein tagged with 6x Histidine was purified from cell culture and used for production of primary antibody in rat. The ovalbumin protein along with freund’s adjuvant was injected to the rat, booster was given, and finally, hyper-immune sera was collected from rat. The antisera was purified for isolation of IgG. The IgG was used as primary antibody for Western blotting. Through Western blotting, ovalbumin protein isolated from chicken magnum was detected and the protocol was established to detect chicken ovalbumin protein.

Impact of Ni(II), Zn(II) and Cd(II) on biogassification of potato waste.

A study was conducted on anaerobic digestion of potato waste and cattle manure mixture, inoculated with 12% inoculum and diluted to 1:1 substrate water ratio at 37 +/- 1 degrees C. Initially pH of substrate was found to be 4.5 to 5.0. Lime and sodium bicarbonate solutions were employed to adjust the pH to 7.5. Biogas production continued up to 10 and 7 days, when lime and sodium bicarbonate solutions were used to adjust the pH, respectively. Biogassification potential was studied in response to different ratio of waste and cattle manure. Biogas production rate was higher when potato waste and cattle manure were used in 50:50 ratio. Effect of two different concentrations (2.5 and 5.0 ppm) of three heavy metals viz. (Ni (II), Zn (II) and Cd (II)) on anaerobic digestion of substrate (potato waste--cattle manure, 50:50) was studied. At 2.5 ppm, all the three heavy metals increased biogas production rate over the control value. The percentage increase in biogas production over the control was highest by Cd, followed by Ni and Zn. In all the treatments, methane content of biogas increased with increase in time after feeding. Various physico-chemical parameters viz. total solids, total volatile solids, total organic carbon and chemical oxygen demand considerably declined after 7 days of digestion and decline was greater in presence of heavy metals as compared to control. The physico-chemical parameters revealed maximum decrease in the presence of 2.5-ppm concentrations of heavy metals with the substrate. Among all the three heavy metals employed in the study, Cd++ at 2.5 ppm was found to produce maximum biogas production rate. The use of three heavy metals to enhance biogas production from potato and other horticultural waste is discussed.