Screening And Pharmacological Mechanism Of Forage Natural Plant Formula To Lower Somatic Cell Count In Dairy Milk

Somatic cell count(SCC)is an important index.It can be used to measure the health and quality of milk.It is also a reliable indicator to indicate the degree of breast health and diagnose the subclinical mastitis.When a cow suffers from an underlying mastitis,the SCC will increase in milk.High SCC can cause serious decline in milk production and dairy quality,which can cause loss to dairy farming.The ministry of agriculture has included 115 medicines of the same origin as feed to feed inventory.These forage natural plants have the characteristics of traditional Chinese herbal medicine as green and low toxicity.Using the regulate health function of these green natural plants to regulate the cow's breast function,reduce SCC avoids the disadvantages of high cost and long cycle of new veterinary drugs,will play a positive role in promoting the healthy development of dairy industry.This study aims to through the screening test to provide a valid forage natural plant formula to clinic to treat cow hidden mastitis,control the SCC in milk effectively,improve the quality of milk,and make a preliminary exploration on the anti-inflammatory mechanism of the compound drugs.In this study,four different kinds of natural plants were selected from the Feeding catalogue to compose four alternative formulas.Divided the cows which somatic cell count is higher than five hundred thousand/mL into five groups averagely according to the breast area.The experiment groups added the four forage natural plant formula 200 g into fodder respectively everyday for seven days.Collected milk from every group at 0,3,5,7 d.Measured the cow's SCC,milk composition from every breast area to ensure the best formula.In order to determine the clinical recommended dose of optimal forage natural plant formula and its effect on cow blood and milk.Divided the cows which somatic cell count is higher than five hundred thousand/mL into four groups averagely according to the breast area.The experiment groups added the best traditional Chinese medicine formula with differen doses into fodder respectively everyday for seven days.Collected milk from every group at 0,3,5,7 d.Measured the cow's SCC,milk composition and the milk production from every breast area to ensure the best clinical dosage.Collected blood and milk at the end of the medication,microplate method were used to detect AST,ALT in serum,Microenzyme labeling test were used to detect the concentration of ALP,LDH,NAG in whey,colorimetric method were used to detect the concentration of GSH-Px in serum,TBA test were used to detect the concentration of MDA in serum,WST-1 test were used to detect the concentration of SOD in serum and enzyme-linked immunosorbent assay(ELISA)were used to detect the concentration of TNF-?,IL-6 in serum and whey.In order to discuss the pharmacological mechanism of the best formual reduced the SCC of dairy cow.In this paper,the anti-inflammatory test of mouse ear swelling and toe swelling model were used to investigate the anti-inflammatory effect of the best formula.The vitro inflammatory model was established by using bacterial lipopolysaccharide(LPS)as an irritant during logarithmic phase of cultivated RAW264.7 cells.RAW264.7 cells were pretreated with the best traditional Chinese medicine formula with different concentrations for 4 h,inflammatory 24 h.Griess test were used to detect the concentration of NO and enzyme-linked immunosorbent assay(ELISA)were used to detect the concentration of TNF-?,IL-1?,IL-6 and IL-10,Western-blot test were used to detect the protein expression level of P65,I?B?,P-P65 and P-I?B?.The results showed that the the group ? has best effect which composed with houttuynia cordata,Codonopsis pilosula,motherwort and kirilowii.That can significantly reduce the somatic cell count in cow milk(P<0.05),improve the lactose,fat,no-fat soilds content in milk cow significantly(P<0.05).The clinical recommendation dose is 200 g/d.Pharmacological tests showed the best formula can reduce the level of AST,ALT,MDA,IL-6,TNF-? and improve GSH-PX,SOD significantly(P<0.05)in serum.It can reduce the level of AKP,LDH,NAG,IL-6,TNF-? significantly(P<0.05)in whey.The best formula can significantly reduce the swelling,swelling rate and swelling inhibition rate of the ear and toe of mice(P<0.05).There were no obvious toxic effects on RAW 264.7 cell when the concentration of traditional Chinese medicine was under 6 mg/mL within 24 h.The Traditional Chinese medicine group with 5 mg/mL,2.5 mg/mL,1.25 mg/mL can significantly inhibited the increase the content of NO,TNF-?,IL-1? and IL-6 in RAW264.7 cells after induced by LPS(P<0.01)and show dose-dependent to a certain extent,the phosphorylation level of P65 and I?B? was significantly reduced(P<0.01).Conclusion: The forage natural plant mainly composed of houttuynia cordata,Codonopsis pilosula,motherwort and kirilowii can significantly reduce the somatic cell count in cow milk,improve the milk production and dairy quality.The clinical recommendation dose is 200 g/d.The formula can significantly reduce the swelling of ear and toe of mice,can decrease the concentration of NO,TNF-?,IL-1? and IL-6.By blocking NF-?B signaling pathway,then reduce the quantity of the proinflammatory cytokine were one of the anti-inflammatory mechanism of dandelion flavonoids.