Study On The Effect Of Compound Chinese Herbal Additives On Immune Performance Of Lactating Cows

Compared with antibiotics,Chinese herbal medicine is usually regarded as an ideal additive because of its almost no toxicity and no residue.The active ingredients in the extracts of Chinese herbal medicine are mostly plant secondary metabolites,which can raise the performance of animals,improve the metabolism of animals and enhance the immune function of animals.Although the research and application of Chinese herbal medicine in livestock production is more,there is little systematic research and application in the dairy cattle,and its mechanism is still unclear in many aspect.Therefore,the experimental study the effects of Chinese herbal additive supplementation on lactation performance,rumen environment,microbial flora and immune function and their mechanisms in midlactating dairy cow.It can provide a scientific basis for the development and utilization of Chinese herbal additive in dairy cow breeding.Chapter 1:Effects of Chinese herbal additives supplementation on production performance,somatic cell count and serum immunoglobulins content in dairy cows.The purpose of this study was to investigate the effects of Chinese herbal additives on production performance,somatic cell count and serum immunoglobulins content in dairy cows.Eighty Holstein cows with the same days in milking(201±19 d),milk yield was selected and randomly assigned into experimental group and control group.The experimental group was committd with the herbal additives at the dose of 30 g per day per cow(1.4 d/kg DM).The trial period was 35 d(5 d for adaptation,30 d for the trial).The extracts of Flos Lonicerae(3%-8%),Herba Leonuri(4%?10%),Eucommia ulmoides Oliv(5%?15%),dandelion(5%?15%)and Astragalus membranaceus(5%?15%)were mixed together with organic carrier(30%?60%).Cows in the control group(group A)were fed with basic diet,and the diets in the experimental group(B,C and D)were added 30,50 and 80 g/d of Chinese herbal additive in the basal diet,respectively.The results showed that:1)the main active component of the Chinese herbal additives used in this experiment is chlorogenic acid(0.090%),caffeic acid(0.012%),astragaloside(<0.01%)and Leonurine hydrochloride(<0.02%).2)Chinese herbal additives supplementation did not affect the dry matter intake of cows.But it could increase the milk yield and decrease the milk somatic cell count significantly(P<0.05).It could reduce the incidence of submastitis.3)Chinese herbal additives supplementation increased the serum IgG and IgM content significantly(P<0.05),but it had no affect on serum IgA content(P>0.05).The results showed that the Chinese herbal medicine additive did not significantly affect the feed intake,but it could increase milk yield and decrease milk somatic cell count and could improve the level of humoral immunity of dairy cows.Chapter 2:Effects of Chinese herbal additives supplementation on blood biochemical,antioxidation and immune function in dairy cows.The aim of this experiment was to study the effects of Chinese herbal additive on blood biochemical,antioxidation and immune function in dairy cows.Four lactating cows were selected by lactating days[(101±12)d],with similar weight[(603 ±48)kg],with good body condition and installed permanent rumen fistula(parity 2)multiparous Holstein cows.The trial was designed by 4*4 Latin square and divided into 4 periods.Each period was 28 d(7 d for adaptation,14 d for the trial,and 7 d for the empty).The experimental materials was the same as experiment one.The results showed that:1)the number of leukocytes and neutrophils in the three groups of B,C and D were significantly higher than those in group A(P<0.05),and all were within the normal range.The lymphocyte rate of group C was significantly lower than that of group A(P<0.05),and its in group B and D was a trend lower than that in group A(P<0.10).Platelet count in group B was significantly higher than that in group A and group D(P<0.05).2)compared with group A,the alanine aminotransferase(ALT)and aspartate aminotransferase(AST)activity in group B had a slightly decreased,while the ALT and AST activity of group C and D had a slightly increased,but the differences were not significant(P>0.05),the C and D group AST activity beyond the normal range.There were no significant differences in all other biochemical parameters(P>0.05).3)the concentration of growth hormone in B group was significantly higher than that in group A(P<0.05),and the concentration of prolactin in group B and D was significantly higher than that in group A(P<0.05).4)the content of malondialdehyde(MDA)in group B,C and D was significantly lower than that in group A(P<0.05).The activities of superoxide dismutase(SOD),glutathione peroxidase(GSH-Px)and catalase(CAT)in group B were significantly higher than those in group A(P<0.05),and there was a trend of increase in group C and D(P<0.10)compared with group A.5)the content of IgM in group B was significantly higher than that in group A(P<0.05),and the content of nitric oxide(NO)in group C and D was significantly higher than that in group A(P<0.05).The difference of IgA and IgG content in each group was not significant(P>0.05).6)the content of IL-2 and IL-4 in serum increased linearly with the addition of Chinese herbal additive(P<0.05),while the content of IL-6 decreased linearly(P<0.05).7)CD3+T lymphocyte rate of four group has no obvious difference(P>0.05),CD4+T lymphocyte rate in group B was significantly higher than that of group A(P<0.05),CD8+T lymphocyte rate of group B and C was significantly lower than group A(P<0.05),CD8+ratio of T lymphocyte with the dosage of Chinese herbal additive increased firstly and decreased afterwards,while CD4+/CD8+of group B and C was higher than that of group A(P<0.05).The results showed that the Chinese herbal additive could regulate the immune function of the immune system by regulating the number of blood cells,the activity of cytokines and improving the antioxidant capacity.Chapter 3:Effects of Chinese herbal additives supplementation on rumen environmental parameters in dairy cows.The aim of this experiment was to investigate the effects of Chinese herb additives on rumen environmental parameters in midlactating cows.The results showed that:1)Chinese herbal additive has a stabilizing effect on dairy cow rumen pH value(6.20-6.84),4 hours after feeding the rumen pH value of group B was significantly higher than that of the other 3 groups(P<0.05),2)the concentration of ammonia nitrogen increased with the increase of the dosage of Chinese herbal additives,but the difference was not significant(P>0.05).3)Chinese herbal additives had a trend to reduce the total volatile fatty acid content(P<0.10),4 h and 6 h after feeding the total volatile fatty acid content increased with the amount of Chinese herbal additives and a significant linear decrease(P<0.01).The Chinese herbal additives could increase the acetic acid content in the rumen fluid,and the content of acetic acid in the cows in groupB is significantly higher than that in the group A and the group D after 2 h feeding(P<0.05).It could also increase the content of propionic acid and butyric acid,increase the ratio of acetic acid to propionic acid,and 6 h after feeding with acetic acid/propionic acid had a trend to increase in rumen fluid(P<0.10),butyric acid content decreased with increasing dosage of Chinese herbal additives.4)with the increase of the amount of Chinese herbal medicine,the content of rumen microbial protein gradually increased,while the content of lactic acid bacteria decreased slightly,but these differences were not significant(P>0.05).The results showed that Chinese herbal additives could change the parameters of rumen environment and regulate rumen fermentation.Chapter 4:Effects of Chinese herbal additives supplementation on the structure and composition of rumen bacterial flora in dairy cows.The aim of this study was to investigate the effects of Chinese herbal additives on the structure and composition of rumen bacterial flora in midlactating cows by high throughput sequencing technique and to reveal the mechanism of the effect of Chinese herbal additives on microorganisms.The results showed that:1)16 samples were concluded 1,120,712 tags,1,095,818 valid tags,and about 97.8%of the original tags passed all quality control.The sequence length is about 253 bp.2)the addition of Chinese herbal additives did not significantly affect the richness and diversity of rumen bacteria,but the number of bacterial species in cows in group C(50 g/d)was significantly lower than that in the other 3 groups(P<0.05).3)at the phyla level,there are 10 phyla with a relative abundance above 0.1%.The main phyla are the Bacteroides,Firmicutes and proteobacteria.Group C(50 g/d)abundance of Lentisphaerae was significantly lower than that of the other three groups(P<0.05),the abundance of Cyanobacteria showed a linear increase trend(P=0.08)with the increasing of the amount of Chinese herbal additives.4)at the genus level,the relative abundance above 0.1%of bacteria was total of 35.The dominant genus are Prevotella,Treponema₂,Fibrobacter and RC9.With the increase of the amount of Chinese herbal additives,the abundance of Ruminococcus₁ in the rumen increased significantly(P<0.05),while the abundance of Butyrivibrio₂ and Bacteroides decreased significantly(P<0.05).5)by clustering and principal component analysis,group A and D,group B and C had the closer genetic distance,flora composition and structure are also more similar.The results showed that the Chinese herbal additives had no significant influence on the bacterial richness and diversity of rumen bacteria,but different amounts could change the structure and composition of bacterial flora.Chapter 5:Effects of Chinese herbal additives supplementation on the immunity of liver and mammary gland in dairy cows.The aim of this experiment was to study the effects of Chinese herbal additives supplememtation on liver and mammary gland tissue immunity,which was detected by inflammatory pathway in liver and mammary gland tissue,mRNA gene expression of cytokines and protein synthesis by RT-PCR technique.The results showed that:1)the immunohistochemical detection showed that the positive index of p38 protein of the liver had not significant affected(P<0.05)by Chinese herbal additives supplementation,while the positive index of TLR4 protein in group B,C and D were significantly lower in group A(P<0.05).It could increase the positive index of p38 protein in the breast,but the difference was not significant,no significant effect on TLR4(P>0.05).It could significantly reduce the positive index of p53 protein(P<0.05),significantly increased the positive index of Bcl-2 protein(P<0.05).2)Chinese herbal additives could significantly inhibit the mRNA expression of TNF-alpha,TGF-beta 1 and IFN-gamma.AS1,AS2 and beta-casein mRNA expression in group B(30 g/d)was significantly higher than the other 3 groups(P<0.01),and for each group of cows,mRNA expression of kappa-casein JaK2,STAT5 and mTOR had no significant effect(P>0.05).It could significantly increase the mRNA expression levels of cytokines IL-2 and IL-4 in mammary gland tissue(P<0.05),and can significantly reduce the mRNA expression of IL-6 in cows of group C(50 g/d).The results showed that the Chinese herbal additives could regulate the expression of inflammatory signaling pathway proteins and cytokines in the liver and mammary gland,and thus affect the immune function of the organism.