| The study included three trials, the hydrogenation process used to study different concentrations and different proportions of dietary, rumen fermentation parameters and the methane generated in vitro effects of α-linolenic acid. The main work is as follows:Experiment1:Effect of different level of α-linolenic acid on in vitro rumen fermentation characteristics and Bio-hydrogenation.This study was conducted to determine the effects of free a-linolenic acid at different con-centrations on rumen fermentation and the intermediates of microbial bio-hydrogenation. α-linolenic acid were added to basal subatrate at level of0%(CON),1.5%(LL),3.0%(ML) and4.5%(HL) of substrate DM(w/w), respectively. Each treatments had three repetitions. To N-NH3, TVFA, acetic acid, butyric acid, ML and LL were significantly lower than CON (P<0.05) and propionate concentrations were significantly higher than CON (P<0.01). Linoleic acid (c9, c12-C18:2) concentration of CON and LL decreased with the incubation time, while the other two increased, and ML reached the maximum over CON and LL in12h (P<0.01). c9, t11,c15-CLnA, c9, t13, c15-CLnA of HL was significantly higher than the other groups at each time point (P<0.01). Conjugated linoleic acid (CLA) and conjugated linolenic acid (CLnA) levels of ML, HL were significantly higher than the other groups (P<0.01) during12h. Therefore, α-linolenic acid diets may be changed rumen fermentation pattern, promote the accumulation t11, c15-C18:2of CLA and CLnA in rumen fluid, and produce a variety of C18:1isomers, and it was dose-effect relationship between total CLnA and α-linolenic acid.Experiment Ⅱ:effects of α-linolenic acid added to the rumen Bio-hydrogenation under different dietary conditions.In this study, in vitro method to add α-linolenic acid ratio in the different diets, the researchers effect on rumen fermentation patterns and a-linolenic acid hydrogenation process. Design five dietary levels, there were8:2,6:4,5:5,4:6and2:8(w/w), respectively, each group are added3.0%α-linolenic acid (w/w). Each treatments had three repetitions.. The results showed that:at each time point, with the increase in the proportion of fine material, pH is rising, and the2:8group and the rest of the four groups was significant difference (P<0.05). Adding α-linolenic acid conditions, the effects of TVFA were not significant.6h and12h, the concentration of acetic acid has been inversely proportional with the concentrate. Propionate concentrations of each group varies with time delay,4:6group propionate concentrations at each time point were higher than other groups, and the difference was significant (P<0.01).2:8group NH3-N concentration at each time point were more than8:2,6:4group, and the difference was significant (P<0.01). With the increase in the proportion of fine material, at each time point CH4, C02and gas production showed an increasing trend. T11-C18:1ratio of coarse material throughout the incubation time increases with the increase.8:2group t11-C18:1in3h were no other significant differences in four groups (P>0.05);6h,12h, there was no significant difference between groups in CLA concentrations. c9, cl2-C18:2had a positive correlation between the proportion of coarse material, coarse material increases significantly increase the total CLnA,8:2group concentration in the whole training process compared with other significant differences between groups (P<0.05). Therefore, adding a-linolenic acid in the conditions of coarse material increase in the proportion of acetic acid in rumen fluid can be increased and TVFA content, reducing acid and NH3-N content. Increasing proportion of fine material can increase the total gas production and C02production, while reducing CH4production. High concentrate increased rumen pH by reducing TVFA. The test α-linolenic acid added to promote the hydrogenation of linolenic acid the hydrogenation of microorganisms.Experiment III:Using Real Time-PCR detection of a-linolenic acid and ratio of dietary effects on microbial hydrogenationThis experiment is intended to test one or two of the6h,12h samples, using Real Time-PCR detection of α-linolenic acid ratio and dietary effects on microbial hydrogenation for a-linolenic acid on rumen regulation and hydrogenation results process to provide a basis for reference. Two time points samples are processed to extract DNA, purified using using Real-time PCR technology, changes in the number of B. fibrisolvens, B. proteoclasticus and B. flavefaciens were monitored. The results showed that:6h B. fibrisolvens, DNA quantity and α-linolenic acid dosage B. proteoclasticus and B. flavefaciens inversely proportional, and each group significant difference (P<0.01).6h-12h, three kinds of bacterial DNA volume increases.12h, B. fibrisolvens, DNA varies with the amount of linolenic acid B. proteoclasticus add significantly decreased (P<0.01), while the amount of DNA in the ML group B. flavefaciens highest. Elevated compared with the coarse and fine,6h and12h of B. fibrisolvens, B. flavefaciens gradually increased, and8:2,6:4,2:8significant difference between groups (P<0.01),5:5and4:6group, the difference was not significant (P>0.05).6h, B. proteoclasticus reached the maximum set at5:5, and the other group, the difference was significant (P<0.01);12h, B. proteoclasticus reaches a maximum at4:6group (P<0.01). Therefore linolenic acid on B. flavefaciens toxicity is less than B. fibrisolvens. In adding a-linolenic acid conditions, the higher the proportion of dietary roughage more conducive to the growth of the three bacterial hydrogenation, and α-linolenic acid is added under conditions of pH change and B. flavefaciens and B. fibrisolvens related. |
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