Establishment of Real-Time TaqMan-Fluorescence Quantitative RT-PCR Assay for Detection and Quantification of Porcine Lipoprotein Lipase mRNA

Porcine lipoprotein lipase （LPL） cDNA was cloned as the standard for real-time quantifying LPL mRNA and the TaqMan-fluorescence quantitative PCR assay for detection was established. The total RNA extracted from Longissimus dorsi of porcine was reverse-transcribed to cDNA. LPL cDNA was ligated with pGM-T vector and transformed into Escherichia coli TOP10. Plasmid DNA extracted from positive clones was verified by PCR amplification and sequenced. LPL was amplified by real-time fluorescence quantitative PCR from the plasmid DNA. The concentration of DNA template purified was detected by analyzing absorbance in 260 nm and then the combined plasmid was diluted to series as standard for fluorescence quantitative PCR （FQ-PCR）. The method of LPL mRNA real-time PCR was well established, which detected as low as 103 with the linear range 10^3 to 10^10 copies. The standard curves showed high correlations （R2 = 0.9871）. A series of standards for real-time PCR analysis have been constructed successfially, and real-time TaqMan-fluorescence quantitative RT-PCR is reliable to quantitatively evaluate FQ-PCR mRNA in L. dorsi of porcine.

Use of the N-alkanes to Estimate Intake, Apparent Digestibility and Diet Composition in Sheep Grazing on Stipa brevifl ora Desert Steppe

The application of n-alkane as markers to estimate herbage intake, apparent digestibility and species composition of diet consumed by grazing sheep was studied. Six local Mongolian sheep were used to determine dry matter （DM） intake, apparent DM digestibility and species composition of diet during summer, autumn and winter. Animals were orally dosed twice daily with n-alkane gelatin capsules containing 60 mg C32-alkane as an external marker. Diet composition was estimated by comparing the odd-chain n-alkanes pattern profile （C27-C31） of the consumed plant species with the n-alkanes fecal concentrations of grazing animals, using a non-negative least squares algorithm called EATWHAT software package. The alkane pair C32：C33 and C33 alkane were used to estimate DM intake and diet apparent DM digestibility, respectively. The results showed that daily dry matter intake of the sheep were 1.77, 1.61 and 1.18 kg d-1 in summer, autumn and winter, respectively. Apparent DM digestibility, crude protein （CP）, metabolizable energy （ME） and neutral detergent fiber （NDF） intake of diet consumed by sheep decreased significantly （P〈0.01） from summer to winter, with no evident changes in ADF and ADL intake. Diet composition indicated Artemisia frigida Willd was the most dominant diet component, contributed 79.68, 68.12 and 86.26% of sheep＇s diets in summer, autumn and winter, respectively. Cleistogenes songorica Ohwi and Convolvulus arnmannii Desr were the important components of the diet. Although Stipa breviflora Griseb is one of the main plant species in the study area, the sheep rarely choosed it. The study indicated that CP and ME in diet consumed by sheep were deficient in winter. Therefore, appropriate supplementation strategies should be indispensable during this period.

Effects of Non-fiber Carbohydrate to Neutral Detergent Fiber Ratios on Ruminal pH Dynamics in Dairy Goats

Introduction Subacute ruminal acidosis(SARA)has become common and important economic and health issues for high-yielding dairy cows.Rumen pH takes a central position in the pathogenesis of SARA.So monitoring of rumen pH is play important role in recognizing,quantifying and subsequently controlling this disorder. The objective of the

The Effect of Different Doses of Composite Artemisia annua Extract on Ruminal Fermentation of Dairy Goats in vitro

An in vitro study was conducted to determine the effect of different doses of Composite Artemisia annua extract(CAE) on rumen microbial fermentation.CAE was a crude extract from the stem and leaves of Composite Artemisia annua extracted with ethanol.The doses of CAE supplementation with frozen-dry form were:control(no addition)