Preparation Of Fermentated Products Of Modified Baitouweng San By Probiotics And Its Prevention And Treatment Of Colibacillosis In Broilers

Broiler Colibacillosis is acute or chronic bacterial infection caused by pathogenic Escherichia coli.Clinical prevention and cure on broiler colibacillosis mostly depend on antibacterial agents,however the abuse of antibiotics leads to the emergence of antibiotics-resistant bacterial strains,which Economic efficiency of Poultry industry has been reduced and public security has been threatened.Traditional Chinese herbs play an important role not only the antiviral,antibacterial and modulating immunity function but also eliminating and reducing drug resistance.Traditional Chinese herbs is of great importance to solve the problem of pathogenic Escherichia coli resistance,effectively prevent and treat Broiler Colibacillosis,improve health level of livestock and poultry and protect the environment.Problems such as slow efficacy,active ingredients contained low and complex chemical ingredients also exist in natural Chinese herbal medicine that is against employed in livestock farm.Microbial fermentation especially probiotics is one of the traditional methods for processing Chinese herbs.According to recent reports in literature,the micorbial transformation can enhance the effect of Traditional Chinese herbs,reduce toxicity,adjust bioavailablity and break down large molecules to smaller ones to benefit the absorptive function.To enhance the effect of Classic Baitouweng San,Modified Baitouweng San was fermented by bio-fermentation engineering,and to study Prevention and Treatment of Broiler Colibacillosis of Fermentation Products of Modified Baitouweng San.This research paper is divided into five parts:1.Identificaiton of Probiotics strain with Biotransformation of Traditional Chinese medica and its enzyme-productionTo enhance the effect of Products of Modified Baitouweng San,based onphenotypic information and molecular biological methods,the identification was studied on probiotics strain with its synergistic effect with Traditional Chinese Medicine.At the same time,the probiotics strain the ability of producing extracellular enzymes such as xylanase,mannose and cellulose was identified.The results showed that the probiotics strain was identified as Bacillus subtilis by the morphological specificity,culture shape,physiological-biochemical characteristics and sequence analysis of 16 S rDNA,and the strain has xylanase,mannose,cellulose,pectinase,amylase and lipase activity.This paper provides a good foundation for the application of microbial transformation in studying active constituents of medical plants.2.Modified Baitouweng San fermented by bacillus subtilis and Prevention and treatment of fermentation products on E.coli infection in broiler chickensTo enhance the extraction efficiency as well as transformation of active components Modified Baitouweng San,and Modified Baitouweng San composed of Astragalus,Radix Pulsatillae,Rhizoma Coptidis,Cortex Phellodendri Chinensis and Cortex fraxini were fermented Bacillus subtilis as the bacteria fermentation.And then,the component changes of Modified Baitouweng San before and after fermentation were analysed by HPLC,and the efficacy for pathogenic Escherichia coli ML DE·2015016 in vitro and in vivo were observed in Fermentation Products of Modified Baitouweng San.In vitro test of inhibitory bacteria showed that the Fermentation Products of Modified Baitouweng San has an obvious antibacterial effect on Pathogenic Escherichia coli ML DE·2015016 superior to Modified Baitouweng San or probiotics.Experiment on AA broiler chickens challenged with Escherichia coli ML DE·2015016 strain showed that protection of prevention and treated group is better than infected control group,death rate remarkably reducing and cellular survival rate obviously rosing.So the Fermentation Products of Modified Baitouweng San and Modified Baitouweng San have protective effect on Escherichia coli DE strain.Survival and mortality rates of the Fermentation Products of Modified Baitouweng San and Modified Baitouweng San were significantly different(P<0.01).The results of prevention and treatment of Fermentation Products of Modified Baitouweng San on Broiler Colibacillosis caused by Artificial Infection werebetter than that of Modified Pulsatilla.3.Influence of fermentation products of Modified Baitouweng San on Immune Function of broiler colibacillosis caused by artificial infectionThe aim of the study was to investigate the prevention and cure immune Mechanism of Products of Modified Baitouweng San on broiler colibacillosis caused by artificial infection.Broilers were fed the Fermentation Products of Modified Baitouweng San,and Modified Baitouweng San group,probiotics group,infected control group,normal control group was designed.And then,immune organ index,immunoglobulin levels in peripheral blood,the concen lymphocyte transformation efficiency,percentage of the erythrocyte complement receptor rosette,percentage of erythrocyte immune compound rosette,cytokines content were determined.The result showed that:1)immune indices of Broiler Colibacillosis caused by Artificial Infection indicated that Fermentation Products of Modified Baitouweng San can improve spleen index,bursa F index and spleen index;2)Fermentation Products of Modified Baitouweng San group could elevate lymphocyte transformation efficiency significantly,increase the percentage of erythrocyte CsbR rosette and decrease the percentage of ICR Rosette.3)The contents of IgG?IgA and IgM,and the levels of IL-2?IL-6?IL-12?TNF-? and IFN-?,significantly higher than the other groups(P<0.01).4)Fermentation Products of Modified Baitouweng San can increase the levels of T-AOC?GSH-PX?SOD?GSH?CAT,significantly higher than the other groups(P<0.01),and induce the level of MDA,significantly lower than the other groups(P<0.01).So Fermentation Products of Modified Baitouweng San could remarkably enhance the immunity against pathogenic Escherichia coliinfection and improve the ability of total antioxygen capacity and the antiviral ability of body.4.The microecological regulative effect of fermentation products of Modified Baitouweng San onenteric bacteria flora in broiler colibacillosis caused by artificial infectionTo study the effects of Fermentation Products of Modified Baitouweng San on microbial community structure and main microbial populations in cecum of broilers,the V3 to V4 region of the 16 S rRNA gene of bacteria in faeces was sequenced using Illumina high-throughput sequencingtechnology.The results showed that Compared with the other groups,Fermentation Products of Modified Baitouweng San affected the alpha diversity indexes(Chaol index and Shannon index)and the beta diversity indexes(PCA),tended to rise(P<0.01).At the phylum level,the caecal microbiota of Broiler of Fermentation Products of Modified Baitouweng San was mainly consisted of Firmicutes,Bacteroidetes,Proteobacteria,Synergistetes,Tenericutes and Verrucomicrobia,and especially relative content of Firmicutes was numerically increased.The cluster analysis at the genus level showed low similarity of cecum microflora between Fermentation Products of Modified Baitouweng San group and the other groups,and the bacterial diversity and species abundance of cecal microbiota of Broiler can be partially influenced by Fermentation Products of Modified Baitouweng San.In conclusion,the Fermentation Products of Modified Baitouweng San can improve intestinal flora against pathogenic Escherichia coli infection.5.The application of fermentation products of Modified Baitouweng San in broiler antibiotic-free breedingTo improve Fermentation Products of Modified Baitouweng San's clinical application,the growth performance and eliminating ratio of 0-5 weeks ROSS308 broilers affected by Fermentation Products of Modified Baitouweng San were tested.The results showed that Fermentation Products of Modified Baitouweng San group could decrease average daily food intake and ratio of feed to meat,and elevate average daily liveweight gain and feed efficiency.This study provides evidence that constituents and content in Modified Baitouweng San fermented by bacillus subtilis has been changed,at the same time,the results of prevention and treatment of Fermentation Products of Modified Baitouweng San on Broiler Colibacillosis caused by Artificial Infection were better than that of Modified Baitouweng San.Fermentation Products of Modified Modified Baitouweng San could remarkably enhance the immunity against pathogenic Escherichia coliinfection ?the ability of total antioxygen capacity and the antiviral ability of body ? intestinal flora against pathogenic Escherichia coli infection ?the production performance of broilers and disease resistance.So Fermentation Products of Modified Modified Baitouweng San will bepossessed a good practical prospect.