| The efficiency of nitrogen utilization in ruminant is lower than monogastric animals. The reason may be following:nitrogen in feed excreted from animal feace and urine; degradation of high quality protein in diet; microbial protein recycling in the rumen. In the rumen micro-ecosystem, the phagocytic behavior of protozoa, autolysis phenomena, as well as the high deamination activity may exacerbate the invalidity of the rumen nitrogen cycle and reduce nitrogen utilization efficiency. Terefore, the study was conducted combined with micro-ecological environment, microbial nitrogen flow, microbial activity, microbial flora changes and number changes in the microbial system to investigate the law and mechanism of the composition of different diets on nitrogen flow of rumen protozoa and bacteria from the dynamic point of view.Chapter1Application of Spectrofluorimetry in Studying Phagocytosis of Rumen Protozoa on BacteriaThis study was conducted to examine the feasibility of spectrofluorimetry used to study the phagocytosis of rumen protozoa on bacteria. Four healthy Xuhuai goats with permanent ruminal cannulas were selected as experimental animals to obtain rumen protozoa and bacteria. The fluorescence intensity of bacteria labeled by DAPI was tested to study the phagocytosis of rumen protozoa on bacteria. The result showed that there was linear correlationship between the concentration of bacteria concentration labeled by DAPI and their fluorescence intensity (R2=0.9952). The grazing rate of protozoa was370.89cells/(cellh) for Xuhuai goats fed the diet with forage concentration ratio of7:3. There was volatility on grazing rate of rumen protozoa on bacteria of Xuhuai goats. Grazing quantity reached saturation on60min and the largest grazing quantity was370.89cells/cell. It was concluded that spectrofluorimetry could be used in studying grazing rate of rumen protozoa on bacteria.Chapter2Effect of Concentrate to Forage Ration on Nitrogen Turnover between Protozoa and Bacteria in the Rumen of GoatsIn this study,four Xuhuai goats installed with permanent rumen fistula were selected as experimental animals. Diets with different forage to concentrate ratio (0:100、30:70、50:50、70:30^100:0)were used as substrates to determine the effect of different forage to concentrate ratio on microbial turnover rate of nitrogen. Methods of DAPI fluorescence labeling and dynamic tracer technique were used in this study. The results showed that the forage to concentrate ratio affected rumen patterns,but the changes were within the normal range. Compared with DAPI fluorescence labeling method,the dynamic tracer technique can be more objective to study the process of phagocytosis of protozoa. With the change of concentrate to forage ratio,nitrogen turnover rates of protozoa, bacteria,between protozoa and bacteria showed the same rule. With the increase of the concentrate to forage ratio,they increased first and then decreased. The highest Nitrogen turnover rate of protozoa was in group C,up to9.067μg/mL h; The highest Nitrogen turnover rate of bacterial was in group D,up to5.122μg/mL h; The highest Nitrogen turnover rate between bacteria and protozoa was in group B,up to2.29pg N/(cell h). Microbial nitrogen turnover rate fluctuated with the increase of forage to concentrate ratio.There existed significantly positive correlationship of Nitrogen turnover rate s between protozoa and total microbe. Regression analysis showed that there existed strong cubic relationshipship between microbial nitrogen and treatment:Yi=0.007+0.016X-0.036X2+0.019X3, R2=0.929; Y2=0.002+0.005X+0.003X2-0.005X3, R2=0.965; Y3=0.008+0.034X-0.090X2+0.058X3, R2=0.984(Yp Y2, Y3means the Nitrogen turnover rate of protozoa,bacteria and total microbial nitrogen). There existed significantly or highly significant positive correlationship of Nitrogen turnover rate s between total nitrogen in rumen fluid and amino acid nitrogen. The highest correlationship was in group B (r=0.960). SSCP profile varied with forage to concentrate ratio. There was a certain different role of treatment on the growth of R. amylophilus> B. fibrisolvens and P. ruminicola.Chapter3Effect of NSC/SC Ratio on Nitrogen Turnover between Protozoa and Bacteria in the Rumen of GoatsIn this study,four Xuhuai goats installed with permanent rumen fistula were selected as experimental animals. Diets with different NSC/SC ratio (0.5,1,1-5,2,2.5)were used as substrates to determine the effect of different NSC/SC ratio on microbial turnover rate of nitrogen. The results showed that the NSC/SC ratio affected rumen patterns,but they were within the normal range. The Nitrogen turnover rates of protozoa were12.883、17.882、17.860、14.602、17.797ug/mL h respectively, and the lowest one was in group A. The Nitrogen turnover rate s of bacteria were6.172、7.539、3.977、6.578、4.990μg/mL h respectively. The lowest one was in group C. and the highest one was in group B. The Nitrogen turnover rate s between protozoa and bacteria were0.73、0.73、1.79、.94、1.62pgN/(cellh). With the increase of the NSC/SC ratio,the Nitrogen turnover rate s of total microbe enhanced gradually. There existed positive correlationship of Nitrogen turnover rate s between bacteria and total microbe,but they did not get the significant level. There existed significant or highly significant positive correlationship of Nitrogen turnover rate s between total environment and ammonia nitrogen.Otherwise,there existed significant positive correlationship of Nitrogen turnover rate s between total nitrogen in rumen fluid and amino acid-nitrogen in group A and E. Regression analysis showed that there were cubic relationshipship among protozoal nitrogen, microbial nitrogen and NSC/SC ratio: Y,=0.011+0.022X-0.019X2+0.004X3, R2=0.715; Y2=0.016-0.01X+0.008X2-0.002X3, R2=0.994(Y1,Y2means the Nitrogen turnover rate of protozoa and total microbial nitrogen). The activity of peptidase and protease were highest in group B. With the increase of the NSC/SC ratio,the density of protozoa/bacteria ratio enhanced. SSCP profile varied with NSC/SC ratio. There was different role of treatment on the growth of R. amylophilus, B. fibrisolvens and P. ruminicola.Chapter4Effect of Different Nitrogen Compounds on Nitrogen Turnover between Protozoa and Bacteria in the Rumen of GoatsIn this study,four Xuhuai goats installed with permanent rumen fistula were selected as experimental animals. Diets with different nitrogen compounds (NH4CI; Pro-Arg-Met; Pro, Arg, Met; mixture of above)were used as substrates to determine the effect of different kind of nitrogen on microbial turnover rate of nitrogen. The results showed that the Nitrogen turnover rate s of protozoa were4.075、3.600、4.483、5.958μg/mL h respectively. The lowest one was in group B, and the highest one was in group D. The Nitrogen turnover rate s of bacteria were2.820、2.769、2.137、1.893μg/mL h respectively. The lowest one was in group D, and the highest one was in group A. The Nitrogen turnover rate s between protozoa and bacteria were1.48、2.09、1.17、1.87pg N/(cell h) respectively. The lowest one was in group C,and the highest one was in group D. The Nitrogen turnover rate s of total microbial nitrogen were5.630、4.627、4.771、5.765μg/mL h respectively. The lowest one was in group B,and the highest one was in group D. There existed positive correlationship of Nitrogen turnover rate s between protozoa and total microbial nitrogen,and group A,B and C got the significant level. There existed highly significant positive correlationship of Nitrogen turnover rate s among total nitrogen in rumen fluid and ammonia nitrogen, amino acid nitrogen in group A. There existed highly significant positive correlationship of Nitrogen turnover rate s between total nitrogen in rumen fluid and amino acid nitrogen in group C. There existed highly significant positive correlationship of Nitrogen turnover rate s among total nitrogen in rumen fluid, ammonia nitrogen, soluble protein nitrogen and amino acid nitrogen in group D. The activity of protease and peptidase in protozoa was highest in group D. The activities of amylase,xylanase and filter paper cellulose enzyme were highest in group A. SSCP profile varied with different nitrogen compounds.There was different role of treatment on the growth of R. amylophilus, B. fibrisolvens and P. ruminicola. Chapter5Effect of Different Single Amino Acid on Nitrogen Turnover between Protozoa and Bacteria in the Rumen of GoatsIn this study,four Xuhuai goats installed with permanent rumen fistula were selected as experimental animals. Diets with different single amino acid(Pro,Arg,Met,Leu,Phe)were used as substrates to determine the effect of different amino acid on microbial turnover rate of nitrogen. The results showed that amino acid affected rumen patterns,but they were within the normal range. The Nitrogen turnover rate s of protozoa were3.304、4.017、2.890、4.431、5.024μg/mL h respectively. The lowest one was in group C,and the highest one was in group E. The Nitrogen turnover rate s of bacteria were5.347、4.944、5.165、3.944、5.637μg/mL h respectively. The lowest one was in group C,and the highest one was in group E. The Nitrogen turnover rate s between protozoa and bacteria were2.33、2.53、1.79、1.61、0.72(pg N/cell h) respectively. The lowest one was in group D,and the highest one was in group B. The Nitrogen turnover rate s of total microbial nitrogen were5.490、7.439、5.909、4.56、7.397μg/mL h respectively. The lowest one was in group C,and the highest one was in group B. There existed positive correlationship of Nitrogen turnover rate s between protozoa and total microbial nitrogen,and only group E did not get the significant level. There existed highly significant positive correlationship of Nitrogen turnover rate s among total nitrogen in rumen fluidand ammonia nitrogen.urea nitrogen in group A. There existed highly significant positive correlationship of Nitrogen turnover rate s between total nitrogen in rumen fluidand ammonia nitrogen in group B. There existed significant positive correlationship of Nitrogen turnover rate s among total nitrogen in rumen fluidand ammonia nitrogen,soluble protein nitrogen in group C. There existed significant or highly significant positive correlationship of Nitrogen turnover rate s between total nitrogen in rumen fluidand soluble protein nitrogen in group D and E. The activity of protease in protozoa was highest in group E. The activity of amylase and filter paper cellulose enzyme had no difference (P>0.05). The activities of pectase and xylanase were high in group B.SSCP profile varied with different amino acid treatment.There was different role of treatment on the growth of R. amylophilus, B. fibrisolvens and P. ruminicola.Chapter6Effect of Different Protozoal Controller on Nitrogen Turnover between Protozoa and Bacteria in the Rumen of GoatsIn this study,four Xuhuai goats installed with permanent rumen fistula were selected as experimental animals. Diets with different protozoa controller (ginaton, yucca schidigera, saponin and control)were used as substrates to determine the effect of different kind of protozoal controller on microbial turnover rate of nitrogen. The results showed that different treatment affected rumen patterns. The Nitrogen turnover rate s of protozoa were5.578、8.178、12.489、9.078 μg/mL h respectively. The highest one was in group C. The Nitrogen turnover rate s of bacteria were1.800、1.267,1.611%3.156μg/mL h respectively. The values in groups added with protozoa controllers were lower than the control group. The Nitrogen turnover rate s between protozoa and bacteria were2.86%3.05%2.68%4.06pg N/(cell h) respectively. The values in groups added with protozoal controllers were lower than the control group. The Nitrogen turnover rate s of total microbial nitrogen were6.089%9.000%13.144%7.744μg/mL h respectively.The highest one was in group C. There existed significant or highly significant positive relationship of Nitrogen turnover rate s between protozoa and total microbial nitrogen. There existed positive correlationship of Nitrogen turnover rate s between total nitrogen in rumen fluidand amino acid nitrogen. They got significant level in group A,B and D.Highly significant level was got in group C.There existed highly significant positive correlationship of Nitrogen turnover rate s between total nitrogen in rumen fluidand amino acid nitrogen in group C (r=0.986). The ratio of protozoa/bacteria was highest in group D. SSCP profile varied with treatment.There was different role of treatment on the growth of R. amylophilus,B. fibrisolvens and P. ruminicola. |
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