Study On The Protective Effect And Mechanism Of Chinese Medicine Combined With Probiotics On Escherichia Coli In Chicken Induced Diarrhea In Mice

Isolation of pathogenic Escherichia coli and Salmonella from different samples of the Yanbian hennery,Gram staining,biochemical test and serological identification,the interactions of Chinese medicine,probiotics and pathogenic bacteria were analysis by the application of Oxford cup method,continuous dilution analysis and orthogonal experimental design analysis.With the most pathogenic Escherichia coli strains of mice diarrhea model,They were divided to control group,model group,Chinese medicine group,probiotics group and Traditional Chinese medicine combined probiotics,during the treatment of daily intragastric administration before weighing and diet,intragastric administration after continuous observation of 4 h,recorded the clinical symptoms,mortality rate,diarrhea rate,cure rate,diarrhea index and loose stool grade,After 3 days,They were collected the weight,diet,and the eyeball blood for Hematology analysis,autopsy eyes analysis of various viscera changes,took heart and liver and lung and spleen and kidney,weighed and calculate viscera index,intestinal contents were collected in sterile for bacterial counts,take appropriate stool for occult blood test,intestinum tenue tissue for HE staining,immunohistochemical staining and scanning electron microscopy analysis of pathologic histology and NF-κB p65 protein expression and ultrastructure changes,testd the serum and intestinal homogenate of MPO content,serum TNF-α and IL-1β and IL-6 contents and intestine homogenate were detected by ELISA,TNF-α and IL-6 mRNA expressions in the blood and intestinal tissue were detceted by RT-PCR.We successfully separated the chicken pathogenic Escherichia coli 28 strains and Salmonella 25 strains.The strongest pathogenicity Escherichia coli isolates the D4.The bacteriostatic effect of crude extract of dandelion on the isolated strain was dose dependent.The effects of total flavonoids from Radix Astragali on the inhibition of Bacillus subtilis,Escherichia coli and other isolates were not stable.Astragalus polysaccharide could inhibit all the tested strains.The inhibitory effect of Bacillus subtilis on Salmonella isolates was stronger than that of Escherichia coli.The results of orthogonal test showed that the optimal ratio of A1B2C2D2 was the best for inhibiting Escherichia coli.D4.The test successfully established the bacterial diarrhea in mice model and the relevant mechanism.Model group compared with the blank group extremely depressed,diarrhea in mice rate is 100%,autopsy viscera had obvious pathological changes.In the other groups also improved to different extent in all respects.Chinese medicine group,the probiotic group,traditional Chinese medicine combined probiotics could improve model group diarrhea index,the diet content,weight,hematology indexes of pathological changes,traditional Chinese medicine combined probiotic group works best.Compared with model group of liver,spleen,viscera index can Chinese traditional medicine group significantly lowered(P<0.05),the traditional Chinese medicine combined probiotic group can significantly reduced(P<0.01).Fecal occult blood test and intestinal contents results showed that the bacterial count of traditional Chinese medicine combined probiotic group could obviously improve the mice diarrhea.Model group small intestine tissue and serum MPO,TNF-α,IL-6,IL-1β content and small intestine tissue NF-κB p65 protein high expression,only the traditional Chinese medicine combined probiotic group was significantly lower(P<0.05).Histologic lesions in the model group,including small intestinal villus loss serious,intestinal significant degeneration,mucosal inflammatory cells infiltration,Chinese medicine group,the probiotic group,traditional Chinese medicine combined probiotic group outcomes were different degree and the traditional Chinese medicine combined probiotic group resume is best.Scanning electron microscopy results showed that the model groups small intestinal villus height dehydration atrophy,serious damage,microvilli falls off phenomenon obviously,traditional Chinese medicine group and probiotics could only improve,combination of traditional Chinese medicine probiotic group highly repaired.RT-PCR results showed that the model group in the blood,small intestine tissue TNF-α and IL-6 mRNA is highly expressed,Traditional Chinese medicine combined probiotics can obviously reduced,showed a significant difference(P<0.05).The test results showed that the traditional Chinese medicine combined with probiotics of bacterial control bacterial diarrhea mice NF-κB pathway,inhibited the high expression of NF-κB p65 protein in intestinal tissue,the expression of IL-6 and TNF-α mRNA in the blood and intestinal tissue were reduced,the content of TNF-αand IL-6 and IL-1β in the serum and intestinal tissue were lowered,improved the indexes of the body the mice,it plays an important role in protecting bacterial diarrhea in mice.