| Chicken colibacillosis is a relatively common bacterial infectious disease in poultry.It is caused by pathogenic avian Escherichia coli.It is mainly manifested as inflammatory lesions in various organs.In severe cases,it can cause systemic inflammation and sepsis.At present,the common method for the treatment of chicken colibacillosis is still antibiotic treatment,but due to the problem of drug resistance and antibiotic residues in chicken,the use of antibiotics is increasingly restricted.Therefore,Chinese medicine is gradually applied in the veterinary field due to its green environmental protection and non-residue characteristics.Valued.Pomegranate Peel Powder is included in "Taiping Shenghui Recipe".It is a traditional Chinese prescription.It mainly treats dysentery,red and white,and diarrhea.It is commonly used in the folk to treat diarrhea caused by various reasons.It is not widely used as a veterinary medicine.The possibility of pomegranate peel powder as a veterinary medicine was explored.In this paper,by determining the minimum inhibitory concentration of the pomegranate peel extract,the orthogonal design was used to optimize the extraction process of the Pomegranate Peel Powder and determine the antibacterial effect of the Pomegranate Peel Powder.The results show that:the best process parameters The extraction time is 1h,the number of extractions is once,the extraction temperature is 50℃,the ratio of material to liquid is 1:15,and the volume fraction of ethanol is 45%.Under these conditions,the minimum inhibitory concentration of the pomegranate peel powder was 6.25mg/mL,and the extraction rate was 25.9±0.35%.This experiment confirmed that the pomegranate peel extract has a therapeutic effect on chicken-derived E.coli diseases by establishing a colibacillosis suckling mouse model and administering treatment.At the same time,HE staining was performed on the intestinal tissue of suckling mice.The intestinal tissue of the mice in the pomegranate peel extract group was significantly healthier than that of the model group,which further demonstrated the ability of porcine peel powder against chicken-derived E.coli.In order to explore the curative effect and compatibility mechanism of Pomegranate Peel Powder,the chemical components and targets of the two traditional Chinese medicines were studied through the Traditional Chinese Medicine System Pharmacology Database and Analysis Platform(TCMSP),and a composite target network,protein interaction(PPI)network and Target pathway network,explore the mechanism of action of Pomegranate Peel Powder.Through analysis,a total of 11 components and 258 targets related to three traditional Chinese medicines were screened out.The PPI network contains 107 targets,including key targets such as APP,NMUR2,IL8L1,ADRA2C,ADRA2A,and CHRM2.There are 18 gene ontology(GO)entries,including 13 biological process entries,4 molecular function entries and 1 cell component entry.In addition,there are 6 KEGG pathways,neuroactive ligand-receptor interaction pathways,calcium signaling pathways,NOD-like receptor signaling pathways,Toll-like receptor signaling pathways,Salmonella infections,and regulation of actin cytoskeleton.The research results preliminarily verify the basic medicine of Pomegranate Peel Powder.UPLC was used to analyze the effective components of Pomegranate Peel Powder,and it was determined that the elution system was acetonitrile-0.1%phosphoric acid aqueous solution,and the elution gradient conditions were 0min:0%acetonitrile,2min:6%acetonitrile,4min:9%acetonitrile,6min:12%acetonitrile,12min:25%acetonitrile,the best detection wavelength is 280nm.On this basis,quantitative analysis of the main chemical components of the pomegranate peel powder and methodological investigation showed that the linear relationship,precision,stability,and repeatability were good.The recovery rates of gallic acid,punicarin and ellagic acid were respectively They are:98.83±1.9%,99.09±1.9%,101.38±1.8%.The content of ellagic acid was 13.4±0.20μg/mg,the content of punicalagin was 158±1.89μg/mg,and the content of ellagic acid was 21±0.18μg/mg.The experiment also investigated the antibacterial mechanism of the pomegranate peel powder by measuring the influence of the pomegranate peel extract on the total leakage rate of Escherichia coli and the conductivity of the culture medium,as well as the scanning electron microscope observation of the E.coli under the action of the pomegranate peel powder extract.The pomegranate peel powder can kill E.coli by destroying the cell membrane of E.coli.Physiological effects and related mechanisms have laid the foundation for further research on its mechanism of action. |
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