| Chicken chronic respiratory disease caused by Mycoplasma gallisepticum(MG)infection has a very high clinical infection rate,and the mortality after infection is not high.It mainly affects the production performance,such as the reduction of feed utilization rate of broilers and the decline of laying rate of hens,which has brought immeasurable economic losses to the poultry breeding industry.Antibiotic treatment is easy to produce drug resistance in clinic,and the preventive effect of vaccine is not optimistic.More and more researchers have found that traditional Chinese medicine has a good inhibitory effect on M G infection.Through the preliminary research in the laboratory,it is also confirmed that Scutellaria baicalensis has a good therapeutic effect on Mycoplasma gallisepticum.Baicalin is the main active ingredient,which is a prototype precursor drug.Baicalin is first treated by intestinal flora enzyme after oral administration(β-glucuronidase),and baicalein can be quickly reduced to baicalin by UDP glucuronosyltransferase in vivo.Scutellaria baicalensis starter is produced by microorganisms in the fermentation processβ-glucuronidase,under the action of the biological enzyme,can convert most baicalin in Scutellaria baicalensis in advance into baicalein in vitro,increase the hydrophobicity and fat solubility of the drug,and can quickly convert into baicalin to reach the target organ after entering the body.The aim of this pilot study was to improve the oral availability of Scutellaria baicalensis by microbial fermentation and provide a novel approach for the clinical therapeutic application of Scutellaria baicalensis against Mycoplasma gallisepticum.In this study,one strain was screened from three common traditional Chinese medicine fermentation strains(bacillus subtilis,lactobacillus and yeast)β-Glucuronidase strain.The screened strains were optimized according to different conditions affecting the fermentation effect,so that the content of baicalein in the fermentation agent reac hed the highest.The effect of Scutellaria baicalensis starter on bioavailability was verified by pharmacokinetics.The effect of baicalin starter on the therapeutic effect was investigated by ar tificially replicating the pathological model of M G infection.The details are as follows:(1)The method of plate counting was used to detect the viable rate of each strain.According to the"bacterial count determination method in food hygiene microbiolo gical examination",the strain was screened from three kinds of fermented bacteria in laboratory,and four strains with higher viable rate were obtained.The above strains were used for plate color development test to screen and produce one strain The strain ofβ-glucuronidase was screened again by fermentation experiment,and the fermentation effect was confirmed by the detection of baicalein in the fermentation liquid.The results showed that the enzyme producing activity of Lactobacillus salivarius was high,and the live rate of L30 was the highest,which was 3.1×10~8 CFU/m L,the maximum color diameter was 22.8 mm,and the highest baicalein content was 4.63%after fermentation test.Therefore,Lactobacillus salivarius L30 was selected as the fermentation strai n.(2)The growth,reproduction performance and enzyme production activity of microorganisms are different under different environmental conditions.In this experiment,five levels are set for single factor test through the changes of five different condit ions.The initial conditions were as follows:the concentration of traditional Chinese medicine was 1.0g/m L,the fermentation temperature was 37℃,the amount of bacteria was 5%,the rotating speed of shaking table was 160 rpm,and the fermentation time was 5 days.The content of baicalein was used as the evaluation index to optimize in turn.The optimal fermentation conditions were the concentration of traditional Chinese medicine was 1.5 g/m L,the fermentation temperature was 38℃,the amount of bacteria was 4%,the rotating speed of shaking table was 180 rpm and the fermentation time was 3 days.(3)Ninety seven day old chicks were divided into three groups:A,B and C.A group was unfermented scutellaria.B group was fermented scutellaria.The plasma samples were collected at different interval time points after administration,and the concentration of baicalin in chicken plasma was detected by HPLC method,and the data were processed by Win Nonlin pharmacokinetic software.The results showed that Cmax and AUC values of B group were higher than those of A group,and Tmax was advanced by 0.25 h compared with A group.(4)Four groups were set up to treat artificially infected MG of chickens to verify the therapeutic effect of starter.They were divided in to four groups:A,B,C,D group.A Group was the blank control group,B group was the MG infected group,C group was the unfermented group,and D group was the fermentation group.The serum inflammatory factor IL-1βwas detected on 1 d,3 d and 5 d after administration,TNF-αExpression of IL-1βin lung,TNF-αMRNA expression and MG colonization.The results showed that the inflammatory factor IL-1βin D group,TNF-αExpression of IL-1βin lung,TNF-αThe expression of m RNA and the number of M G fixed value were lower than those in C group.This study found a method that can produce The strain of glucuronidase,namely Lactobacillus salivarius,provides a new method for the treatment of Mycoplasma β-gallisepticum. |
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