| At present,the animal husbandry production in our country,the production efficiency of dairy cows and the increase of milk production in order to be improved by feeder manager,so feeding of high concentrate diets is adopted,but a series of nutrition and health problems will be caused in this way,for example,subacute rumen acidosis is one of the most common nutritional and metabolic diseases.In this experiment,mid lactation Holstein cows were used as experimental animals,divided into two groups:low concentrate group and high concentrate group.Effects of histamine on the inflammatory signal pathway in the liver of the rumen of the animals under high concentrate diets is researched by ELISA,Real time Quantitative PCR and Western Blot.The relieving effect of Comprehensive regulation and control method on the effects of histamine on liver in the rumen of high concentrate diets is also be studied in this experiment.The milk goats is used in this experiment as experimental animals,which are divided into three groups:low concentrate group,high concentrate group and comprehensive control group(6:4 concentrate roughage ratio,Fine granulation,adding buffer).The aim is to put forward an improved method for the modernization of dairy farming.Experiment 1:Effects of histamine on the inflammatory signal pathway in the liver of the rumen of the ruminant animals under high concentrate diets:12 lactating Holstein cows were selected to study the effects of histamine in the rumen on the liver inflammatory signal pathway produced in the rumen of the animals under high concentrate diets.Cows were randomly divided into two groups:Low concentrate and High concentrate.The rumen fluid was collected to determine the value of pH at 0-12h after feeding through the rumen fistula.Liver tissue was collected after slaughter of experimental animals,histamine content in liver of dairy cows was determined by ELISA method,the expression of histamine receptors in liver was detected by real-time fluorescence quantitative PCR,the relative expression of NF-?B,IL-1 ?,IL-6,TNF-a in liver was detected by real-time fluorescence quantitative PCR,the expression of NF-KBp65 and p-p65 protein in liver was measured by Western Blot method.The result showed:the pH value in the rumen was significantly decreased by feeding high concentrate,and the time of the rumen pH value was less than 5.6 and 6 was significantly prolonged,the histamine content in the liver of the high concentrate group was significantly higher than that in the low concentrate group,The relative expression of histamine HI receptor in high concentrate group was significantly higher than that in low concentrate group,while no significant difference in histamine H2 receptor,the relative expression of NF-?B,IL-1 ?,IL-6,TNF-a genes in the liver of the high concentrate group were higher than those in the low concentrate group,Expression of NF-?Bp65 and p-p65 protein in liver was significantly higher than that in low concentrate group.Conclusion:the rumen pH value was significantly decreased by high concentrate diets,the histamine content in the liver was increased,and the expression of histamine H1 receptor was increased,the NF-?B signaling pathway may be activated by histamine through the histamine H1 receptor way,so then the relative expression of NF-?B,IL-1 ?,IL-6,TNF-a genes in the liver increased,Expression of NF-KBp65 and p-p65 protein in liver increased.Experiment 2:The regulation of the effects of histamine produced in the rumen on the liver inflammatory signal pathway by comprehensive regulation and control method:In this experiment,18 dairy goats in the middle of lactation were selected to do the comprehensive regulation and control experiment,the regulation of the effects of histamine produced in the rumen on the liver inflammatory signal pathway by comprehensive regulation and control method was studied in this experiment.18 dairy goats were randomly divided into three groups,Low concentrate,High concentrate and Comprehensive control.The other feeding condition were same,rumen fluid was collected at 0-10h after feeding,and the rumen pH value was tested by pH instrument,liver tissue samples was collected from dairy goats after slaughter,histamine content in liver was detected by ELISA method,the relative expression of histamine H1 receptor and histamine H2 receptor gene in liver tissue and inflammatory factor NF-?B,IL-1 ?,IL-6,TNF-a genes was detected by Real time fluorescent quantitative PCR method,the expression of NF-?Bp65 and p-p65 protein in liver was measured by Western Blot method.The result showed:the rumen pH value in the comprehensive control group was significantly higher than that in the high concentrate group,but there was no significant difference between the comprehensive regulation group and the low concentrate group,the histamine content in the liver of the comprehensive control group was significantly lower than that in the high concentrate group,and the difference was not significant compared with the low concentrate group,but higher than that in the low concentrate group,histamine H1 receptor gene relative expression of comprehensive control group was significantly lower than the high concentrate group,no difference with low concentrate group,and there was no significant difference in the relative expression of histamine H2 receptor gene between the three groups.The relative expression levels of NF-?B,IL-1?,IL-6,TNF-a genes in the comprehensive control group were lower than those in the high concentrate group and the difference was significant,with no significant difference between the comprehensive control group and the low concentrate group,the relative expression of NF-?Bp65 protein in the comprehensive control group was significantly lower than that in the high concentrate group,while the p-p65 protein in the comprehensive control group was lower than the high concentrate group but the difference was not significant.The results showed that the comprehensive regulation method can significantly regulate the rumen pH value,increase the pH value,alleviate the situation of rumen acidosis,and then reduce the production of rumen histamine,and reduce the inflammatory reaction of the liver.The ruminant animal damage by high concentrate diets is reduced by comprehensive control method,and the feeding method will be improved by this way for efficient and healthy production. |
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