The Effects Of Rumen Protected Methionine On Performances And Apoptotic Regulation Of Lymphocyte Of Dairy Cows In Summer

1 The effects of rumen protected Methionine on performances and milk composition and amino acids of plasma in dairy cows16 Holstein dairy cows in their midlactation were selected for a 4×4 Latin-square design study, to investigate the influence of a rumen protected methionine(RP-Met) source at dosage of 0, 8, 12 and 16g.d^-1 on Met absorption, milk yield, milk protein and fat. The results showed that the supplementation with rumen-protected methionine increased milk protein and fat and casein. The supplementation with rumen-protected methionine had the tendency to increase of milk yield. The quantity of milk protein and fat were improved by rumen-protected methionine in high yielding cows. 9 Holstein dairy cows in their midlactation were taken samples for 44h before and after supplementing methionine. The result found that the addition of Smartamine led to a sharp increase in the level of serum Met with a peak from 12 to 16 h following the intake. The supplementation with rumen-protected methionine increased the level of serum Met significantly, and the addition of DL-Met did not achieve such response. The supplementationwith rumen-protected methionine and DL-Met had no significant effect on serum total amino acids.16 cows were selected to study the effects of 12 g.d^-1 rumen protected Methionine on the milk yield and composition in heat shocked condition. The results showed that supplementing rumen protected Methionine had the tendency to improve milk protein and fat, to decrease SCC. The supplementation with rumen-protected Methionine could improve milk yield and Methionine of plasma and decrease heat shock status in dairy cows. But it would not improve to the level of lysine and total amino acids. 2 The effects of rumen protected Methionine on the blood indexes in peripheral blood of dairy cowsSupplementing the different dosages(0,12,16 and 20 g.d^-1) with rumen protected Methionine in order to analyze the blood indexes by automatic analyzer of blood. The blood indexes included as follows: white blood cell(WBC), lymphocyte(LYMPH), granulocyte(GRAN), mid-cell(MID), LYMPH%, MID%, GRAN%, red blood cell(RBC), hemoglobin(HGB), mean corpuscular volume(MCV), hematocrit(HCT), mean corpuscular hemoglobin(MCH), mean corpuscular hemoglobin concentration(MCHC), red cell distribution width(RDW), platelet count(PLT), mean platelet volume(MPV) and platelet-crit(PCT). 16 cows were selected to study the effects of 12 g.d^-1 rumen protected Methionine on blood indexes in heat shocked condition. The blood indexes included as follows: GPT, GOT, TP, Albumin, Globin, GLU,LDH, ALP, CPK, SOD, GSH-Px, MDA. The results were showed as follows: Supplementing rumen protected Methionine could significantly decrease RBC(P<0.01). 20 g.d^-1 rumen protected Methionine could significantly HGB, and HGB had the increased tendency with the dosages. WBC and LYMPH and GRAN had the tendency to increase with supplementing rumen protected Methionine. It is significant to the dosage of 20 g.d^-1. HCT had the decreasing tendency with supplementing rumen protected Methionine. MCV and MCH and MCHC had the increasing tendency with supplementing rumen protected Methionine. RDW improved significantly in the dosage of 20 g.d^-1. The dosages of 16 g.d^-1 and 20 g.d^-1 could significantly improve PLT. MPV had the decreased tendency and PCT improved. GPT and GOT and albumin and CPK had the decreased tendency with the supplement with rumen protected Methionine. LDH had been improved significantly if supplemented rumen protected Methionine(P<0.05). Total album and globulin and ALP and GLU had the improved tendency to supplement rumen protected Methionine. Supplementing rumen protected Methionine could improve SOD and GSH-Px and decrease MDA in peripheral blood of dairy cows. The supplementation of rumen protected Methionine had the tendency to improve HGB and PLT. The types of WBC were transferred. The supplementation of rumen protected Methionine had the role on decreasing the damage of cell in heat shocked dairy cows.3 The effects of rumen protected Methionine on apoptosis of lymphocyte in heat shocked dairy cows40cows were selected in the mid-lactation to study the effects of 0,12,16 and 20 g.d^-1 rumen protected Methionine on the apoptotic rate of blood lymphocyte. The apoptotic rate of lymphocytes was analyzed by flow-cytometry. The results found that the apoptotic rate of control was 8.38%. It is the highest. Supplementing rumen protected Methionine reduced significantly the apoptotic rate of lymphocytes in summer. The apoptotic rate of 12,16 and 20 g.d^-1 reduced respectively 71%, 57.4% and 53.22% than that of control(P<0.01).The supplementation of rumen protected Methionine could control the apoptosis of lymphocytes because of heat stress.4 The effects of rumen protected Methionine on HSP70 OD and T₃ and T₄ and cortisol in the heat shocked dairy cowsDouble antibodies sandwich ELISA method for detecting HSP70 in plasma and RIA method for detecting T₃ and T₄ and cortisol. We studied the effects of rumen protected Methionine on of HSP70 ODand T₃ and T₄ and cortisol in the heat shocked dairy cows. The results showed HSP70OD improved respectively 52.61% and 51.51% and 26.47% in the treatments than that of control. T₃ and T₄ of plasma had the increased tendency in the treatments than that of control. Cortisol of plasma in the dosage of 12 g.d^-1 decreased 12.15% than that of control. But cortisol of plasma improved respectively 42.99% and 78.26% in the dosages of 16 and 20 g.d^-1 than that of control. It is not significant. The supplementation of rumen protected Methionine improved metabolism and decreased the heat stress of dairy cows.5 The effects of rumen protected Methionine on expression of related gene for apoptosis of lymphocyte in dairy cowsThe gene effects of HSF1mRNA and Bcl-2mRNA and BaxmRNA were analyzed by RT-PCR in the blood lymphocytes of dairy cows. The results showed that supplementing rumen protected Methionine could reduce the abundance of HSF1mRNA than that of control. Supplementing rumen protected Mthionine improved the abundance of Bcl-2mRNA than that of control. It is highest in the treatment of 12 g.d^-1. Supplementing rumen protected Mthionine reduced the abundance of BaxmRNA than that of control. It is lowest in treatment of 12 g.d^-1. Supplementing rumen protected Methionine improved Bcl-2mRNA/Bax-amRNA.16 cows were selected to study the effects of rumen protected Methionine on expression for apoptotic gene in heat shocked dairy cows. Bcl-2mRNA and BaxmRNA and p53mRNA were analyzed by quantificational PCR. The results showed that the abundance of Bcl-2mRNA improved 14.12% than that of control in seventh day. The abundance of BaxmRNA decreased 35.05% than that of control in seventh day. The ratio of Bcl-2mRNA/BaxmRNA was improved 92.15% than that of control(P>0.05). p53mRNA decreased 40.4% than that of control in seventh day(P<0.01). The abundance of Bcl-2mRNA improved 27.86% than that of control in 14th day. The abundance of BaxmRNA decreased 18.86% than that of control in 14th day. The ratio of Bcl-2mRNA/BaxmRNA was improved 36.36% than that of control(P>0.05). p53mRNA decreased 49.53% than that of control in 14th day(P<0.01). Supplementing rumen protected Methionine had the role on improving genic expression of Bcl-2 and controlling genic expression of Bax and p53.