| At present,there are relatively many studies on the transportation stress of pigs,cattle,sheep and poultry at home and abroad.However,there are few research on the changes in acute phase protein caused by equine transportation stress in China.This study investigated the effects of transport stress on equine body temperature,blood routine indexes,major acute phase proteins,cytokines and heat shock proteins.In addtion,to facilitate the administration to large animals such as equines,two doses of traditional Chinese medicine compound granules are made,and the quality and safety evaluation of the granules were made with mice for equine farm application and production.Four 12-month-old Hanover F1 equines of similar weight were selected for a 15 h transportation test The whole journey was about 787 kilometers.The body temperature and blood indexes 1 h before transportation,7 h during transportation,1 h and 9 h after transportation were measured.RT-qPCR was used to detect the expression levels of HP,TNF-α and IFN-y mRNA in peripheral blood lymphocytes of equines.Detection the contents of SAA,HP,CRP,HSP70 and HSP90 in equine serum by ELISA.Then,the self-made two-dose traditional Chinese medicine compound granules were prepared by the wet preparation method,and the quality of relevant indicators was tested in accordance with the Chinese Pharmacopoeia,and the safety with a maximum dose of 10 000 mg/kg was evaluated with mice for equine farm application and production.In the experiment of the effect of transportation stress on the expression of HP,TNF-α and IFN-γmRNA in equines,compared with 1 h before transportation,the body temperature of the equine increased significantly at 7 h during transportation and 9 h after transportation(P<0.05).The body temperature increased significantly at 1 h after transportation(P<0.01),and the MCV decreased significantly at 9 h after transportation(P<0.05).There was no significant difference in other indicators.The expression levels of HP,TNF-α and IFN-y mRNA in peripheral blood lymphocytes all showed an upward trend.Compared with 1 h before transportation,the expression levels of HP and IFN-y mRNA in lymphocytes increased significantly at 1 h after transportation(P<0.001),the expression level of TNF-α mRNA in lymphocytes was significantly increased at 9 h after transportation(P<0.001).In the study of the effect of transportation stress on APPs and HSPs in equine serum,the HSP90 content in equine serum showed an upward trend.Compared with 1 h before transportation,the content of HSP90 in serum significantly increased at 7 h during transportation(P<0.05).There was no significant difference in other indicators.In the preparation and safety evaluation of granules for relieving equine transportation stress,the optimal ratio of self-made granules to select auxiliary materials and extracts was 1:1:3.5(extract:dextrin:sucrose),and the relevant index quality tests were in line with the provisions of the 2015 edition of the Chinese Pharmacopoeia.According to WHO regulations,the LD50 of this experiment was between 5 000～15 000 mg/kg,no death in the mice,and there were no obvious abnormalities in blood routine,organ index and pathological sections as well.The results indicate,transportation stress would increase equines’ body temperature,decrease the blood MCV index,increase the mRNA expression of HP,TNF-α and IFN-y in peripheral blood lymphocytes,and increase the HSP90 content in the equine serum.The quality testing of homemade granules is in accordance with the provisions of the 2015 edition of the Chinese Pharmacopoeia.Besides,according to WHO regulations,they are indeed non-toxic drugs which are safe and controllable. |
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