| The purpose of this research is to study the synergistic relationship between nutrition induction and the growth of rumen protozoa,and to explore the effect mechanism of nutrient on the signal transduction in the growth and chemical perception of rumen protozoa,in order to enrich the regulation techniques for improving the nitrogen utilization rate of ruminants.The experiment is divided into two parts.The first part studies the chemical perception of rumen protozoa on different concentrations of carbohydrates,proteins and fatty acids,including experiment 1 to experiment four.The second part studies the chemical perception effect of rumen protozoa on protein,glucose and glucose after adding insulin,sodium ferricyanide and walman penicillin,including experiment five to experiment seven.In experiment 1,the chemical sensing effects of rumen protozoa on 0,1,10,100 and 1000 m M concentrations of glucose solution were investigated under two conditions of fasting and feeding in sheep.The results showed that under the two conditions,the tendency of the entodiniomorphid to 0,1,10,100 m M concentration of glucose solution was not significant(P>0.05),and the tendency to 1000 m M concentration of glucose solution was significantly higher than that of other concentrations(P<0.05).The tendency of isotrichid increased first and then decreased with the increase of glucose concentration,in which the tendency of the isotrichidto 100 m M was the highest and the glucose tendency of 1000 m M decreased.In experiment two,samples were sampled after feeding to explore the chemical perception effect of rumen protozoa on 0,1,10,50,100,250,500,750,1000 m M concentration of glucose solution and xylose solution.The results showed that the effect of entodiniomorphid on the tendency of glucose and xylose was significant(P<0.05).In the glucose concentration and xylose concentration range from 1 m M to 750 m M,the tendency of the xylose and glucose in the entodiniomorphid increased slowly with the increase of concentration,and the difference of adjacent concentration was not significant(P>0.05).The tendencies of the entodiniomorphid were significantly higher than that of the concentration of glucose and xylose at the concentration of 1000 m M(P<0.05).The differences in the tendency of glucose and xylose were significantly different(P<0.05),but the tendency of isotrichid increased first and then decreased with the increase of glucose or xylose concentration.The glucose tendencies of isotrichid to 100 m M were significantly higher than those of other glucose concentrations(P<0.05),and the tendency of xylose to 500 m M of isotrichid was significantly higher than that of other xylose concentrations(P<0.05).In experiment three,samples were sampled after feeding to explore the chemical perception effect of rumen protozoa on the protein solution of 0,125,250,500,750,1000 m M concentration.The results showed that there was no significant difference in protein tendencies between entodiniomorphid and other caterpillars(P>0.05).The protein tendency of entodiniomorphid to 1000 m M was significantly higher than that of other groups(P<0.05),and the tendency of isotrichid increased with the increase of protein concentration.In experiment four,samples were sampled after feeding,the fatty acid solution was added to the beaker to make the final concentration of fatty acid in the medium of 0.5%,1%,2% and 3 hours and 6 hours after the culture,to explore the chemical perception of the rumen protozoa on the glucose solution of the saline and 1000 m M concentration.The results showed that although the inner caterpillar was inhibited by fatty acids,the chemotaxis of 1000 m M glucose did not change.However,the isotrichid was significantly affected by the inhibition of fatty acids.It is assumed that the chemosensory receptor is located on the membrane near the vestibule of the protozoa.The unsaturated fat may be toxic or may cause the protozoa to starve by blindness.In experiment five,samples were sampled after feeding and added insulin solution(Ins)to make the final concentration of insulin in the medium of 0,2.5,2.5 and 25 ?M,to explore the chemical perception of the glucose solution(Glc),1g / L soluble peptide(Pep),or Glc + Pep of the rumen protozoa on the concentration of 1000 m M.The results showed that after adding insulin,the tendency of entodiniomorphid to protein and protein + glucose decreased,and the number of entodiniomorphid in empty tube increased significantly(P<0.05).In experiment six,samples were sampled after feeding,adding sodium nitroferricyanide(SNP)to make the final concentration of sodium nitrite ferricyanide up to 0,50?M and 500?M,and to explore the chemical perception of 1000 m M concentration of glucose solution(Glc),1g / L soluble peptide(Pep),or Glc + Pep.The results showed that after adding 500 M sodium nitroprusside,the random swimming of the entodiniomorphid to the saline was reduced,and the tendency to glucose and glucose + protein was significantly increased(P<0.05).In experiment seven,samples were sampled after feeding,adding walman penicillin(Wort),the final concentration of walman penicillin reached 0,20?M and 200?M,to explore the chemical perception of the glucose solution(Glc),1g / L soluble peptide(Pep),or Glc + Pep of the rumen protozoa to 1000 m M concentration.The results showed that after the addition of 200 M wortmannin,the random swimming of the entodiniomorphid to the saline was reduced,the tendency to glucose increased significantly(P<0.05),and there was no significant difference between the glucose + protein tendency and the control group(P>0.05). |
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