Effect Of Compound Zhenqi Ultrafine Powder On Diarrhea Pup Mice And Application To Weaned Piglets

Piglets are prone to diarrhea due to environmental changes and pathogenic infections,which further leads to immune damage and production performance degradation,thus bringing huge economic losses to the pig industry.Traditional Chinese Medicine(TCM)can play a nature,green,and residue-free antibiotic substitution role in animal husbandry production and has good application prospects.The purpose of this study is to explore the effect of compound Zhenqi ultrafine powder,a TCM feed additive,on diarrheal pup mice and application to weaned piglets.Firstly,ultrafine grinding Ligustrum lucidum,Astragalus membranaceus,Acanthopanax senticosus,Paeonia lactiflora and Coix seed and mixing into compound Zhenqi ultrafine powder in a ratio of 2:2:2:1:1.The microscopic characteristics and the content of the main active ingredients are used to check the quality of compound Zhenqi ultrafine powder.It was found that under the optical microscope,we can see the light brown secretory cells of Ligustrum lucidum,the fiber bundle of Astragalus membranaceus,the calcium oxalate clusters of Acanthopanax senticosus,the calcium oxalate clusters and reticulated vessel of Paeonia lactiflora,and the circular starch cluster of Coix seed.The results of HPLC showed that,the content of Fructus Ligustri Lucidi glycosides in the compound Zhenqi ultrafine powder was 1.16%,Radix Astragali saponins content was 0.06%,syringin content was 0.05%,and paeoniflorin content was 3.10%,which met the standards of Chinese Pharmacopoeia and can be used as a TCM feed additive.The safety of the compound Zhenqi ultrafine powder was evaluated through toxicity tests in mice.The results found that,mice were no mortality,no abnormal clinical findings and histopathological change,and the acute oral MTD of mice was greater than 15g/kg,and their weights increased significantly(P<0.05),indicating that compound Zhenqi ultrafine powder was safe and non-toxic.Secondly,taking mice as a model to investigate the effects of compound Zhenqi ultrafine powder on digestive function,antioxidant function,and immune function.72 weaned pup mice(18-days-old)were randomly divided into 4 groups: control group,compound Zhenqi group,model group,model-compound Zhenqi group,with 18 mice in each group.The mice of the control group and model group were given 0.15 m L physiological saline peer day,while the mice of the compound Zhenqi group and model-compound Zhenqi group were given 0.15 m L compound Zhenqi ultrafine powder water decoction peer day,5 days continuously.At the age of 23-days-old,the mice of the model group and the mice of model-compound Zhenqi group were given with senna leaf decoction0.15 m L,and intraperitoneal injection of Escherichia coli 0.15 m L.The mice of the control group and compound Zhenqi group were given with sterile physiological saline 0.15 m L,and intraperitoneal injection of sterile physiological saline 0.15 m L,given 5 days continuously.After each gavage,the diarrhea and growth of mice were observed and recorded.At the age of 28-daysold,serum and small intestine of mice(duodenum,jejunum,and ileum)were collected.The histopathology and structural changes of the small intestine of mice were checked by Paraffin section and HE-staining.The ELISA method was used to detect the content of Ig G,Ig M,MDA,and SOD,GSH-Px activity in serum of mice,the content of s Ig A,IL-1β,IL-6,TNF-α and digestive enzyme activity in the small intestine.The q RT-PCR technology was used to detect m RNA expression of growth factors(IGF-1,IGF-1R,EGF),inflammatory factors(IL-1β、IL-6、TNF-α),and TLR4 signaling pathway related factors(TLR4、My D88、NF-κB)in jejunum.It was found that after feeding compound Zhenqi ultrafine powder,the diarrhea index of mice was significantly reduced(P<0.05),weight and weight gain increased significantly(P<0.05),duodenum,jejunum,and ileum villus height and V/C increased significantly(P<0.05),intestinal digestive enzymes activity and the expression of IGF-1,IGF-1R,and EGF increased significantly(P<0.05);the m RNA expressions of IL-1β,IL-6 and TNF-α,TLR4,My D88,NF-κB in intestinal of diarrhea mice after feeding compound Zhenqi ultrafine powder were significantly reduced(P<0.05).Compared with model group,the immunoglobulin contents in mice serum of the model-compound Zhenqi group increased significantly(P<0.05),the activities of SOD and GSH-Px increased significantly(P<0.05),and MDA content decreased significantly(P<0.05).The results showed that compound Zhenqi ultrafine powder can improve weaned mice’ growth performance,promote the intestinal development,enhance the digestion and absorption function,improve the immunity and antioxidant function,reduce the intestinal inflammatory reaction,and thus reduce the diarrhea rate.Finally,compound Zhenqi ultrafine powder was applied to weaned piglets.60 weaned pigs(30-days-old)were randomly divided into the control group and the compound Zhenqi group,with 30 in each group.The piglets in the control group were fed with basic diet,piglets in compound Zhenqi group were fed with compound Zhenqi ultrafine powder(compound Zhenqi ultrafine powder: basic diet =1: 200),30 days continuously.After adding ultrafine powder 15 days and 30 days,each group selected 15 piglets to collect blood and sperate serum.Using ELISA method detected the content of Ig A,Ig G,Ig M,MDA,GSH,GSG,CAT,the activity of T-SOD,GSH-Px and T-AOC level.It was found that,average daily weight gain of piglets feeding compound Zhenqi ultrafine powder was significantly higher than piglets of the control group(P<0.05),and there was no death in compound Zhenqi group.Meanwhile,the diarrhea rate of weaned piglets of compound Zhenqi group was significantly reduced(P<0.05);after 30 days of feeding compound Zhenqi ultrafine powder,the contents of Ig G,Ig M,GSH and CAT were increased significantly(P<0.05),the level of T-AOC was increased significantly(P<0.05),and the content of MDA and GSSG were significantly reduced(P<0.05),the activities of GSH-Px and T-SOD were increased significantly(P <0.05).The results showed that compound Zhenqi ultrafine powder can reduce the diarrhea rate and mortality of piglets,enhance immune function and antioxidant function of piglets.In summary,compound Zhenqi ultrafine powder prepared in this study can be used as a feed additive to reduce diarrhea rate of weaned mice and piglets,enhance the immune function and antioxidant function of mice and piglets,improve intestinal digestive enzyme activity,and reduce intestinal inflammation.The results of this study provide a scientific basis for the application of compound Zhenqi ultrafine powder in pig breeding.