estimation of ruminal protein degradation parameters of various feeds using in vitro modified gas production technique

This study was conducted to determine in vitro crude protein degradation [IVDP] parameters and effective crude protein degradability [EPD] of various feeds using the modified in vitro gas production [GP] technique. Feed samples were alfalfa hay, soybean meal, soybean, rapeseed meal, sunflower meal and fish meal. Rumen fluid was collected before the morning feeding from four rumen fistulated lambs [49.4 +/- 3.5 kg, body weight]. Approximately 90 ml of buffered rumen fluid [BRF], 400 mg of feed samples and carbohydrates [maltose, xylose and starch] at four concentrations [100, 200, 300, and 400 mg] were added to screw-cap bottles. Gas production [ml] and ammonia nitrogen concentration [mg] in each bottle were measured at 4, 8, 12, 16, 24, and 30 h post incubation and IVDP was calculated via estimated intercept of linear regression between GP [as main variable, X] and ammonia nitrogen [as dependent variable, Y] using the linear regression procedure. Feed, time and feed x time interaction had significant effect on IVDP [P<0.001]. Estimated EPD values at the outflow rate of 0.06/h for alfalfa hay, soybean meal, soybean, rapeseed meal, sunflower meal and fish meal were 0.56, 0.77, 0.59, 0.45, 0.50 and 0.38, respectively

Subcloning and expression of recombinant Echinococcus granulosus antigen B, in Pqe-30 expression Vector

Echinococcosis or hydatid disease is a zoonotic infection caused by larval [metacestode] stages of cestodes belonging to the genus Echinococcus, family Taeniidae. We aimed to subclone antigen B gene in pQE-30 plasmid, its expression, and purification. We subcloned HI gene into pQE-30 expression vector. The recombinant vector was transformed into E. coli, M15 and mass cultured. The subcloned gene was expressed by IPTG. Subcloning of gene was confirmed by both PCR and enzyme digestion. Production of recombinant protein was confirmed by SDS-PAGE. Western blot analysis was carried out by both His-Tag monoclonal Ab and human serum to estimate the expressed potein in E. coli cells. Recombinant protein was purified and its specificity was proved by Western blotting. Production of this recombinant protein can increased sensitivity and specificity in serological test [ELISA]