Study On The Extraction,Utilization And Fermentation Of The Residue Of Lianhua Qingwen

The purpose of this project is to study the biological activity of Lianhua Qingwen residue and the potential of preparing functional feed additives,optimize the fermentation method of Lianhua Qingwen residue,and determine the growth performance of weaned piglets by studying the antibacterial and antiviral biological activity of Lianhua Qingwen residue and the extract of fermented drug residue in vitro and adding 1% Lianhua Qingwen residue to the diet of weaned piglets,so as to demonstrate that Lianhua Qingwen residue has antibacterial and antiviral biological activity,It is feasible to prepare functional feed additives.The main contents and results of the study are as follows:(1)The effective components of Lianhua Qingwen residue were used to determine the antibacterial activity of ethanol extract and water extract of drug residue against Haemophilus parasuis(HPS),Streptococcus suis(SS)and enterotoxic Escherichia coli(ETEC)by broth dilution method in vitro;In vitro antiviral activity assay,African green monkey kidney cells(Vero),African green monkey embryonic kidney cells(Marc 145)and porcine kidney cells(PK-15)were infected with pseudorabies virus(PRV),porcine circovirus type 2 infection(PCV2)and porcine blue ear virus(PRRSV),The morphological changes of cells were observed under light microscope,and the virus nucleic acid was detected and quantified by real-time fluorescence quantitative PCR.It was used as the evaluation index of the antiviral effect of Lianhua Qingwen extract in vitro.The results showed that the residual bioactive components in Lianhua Qingwen residue could effectively inhibit the appreciation of the three viruses;The feeding test results of weaned piglets showed that the addition of 1% Lianhua Qingwen residue in the diet of weaned piglets could significantly reduce the feed meat ratio of weaned piglets,and the drug residue was non-toxic and harmless to weaned piglets,which had the potential to be developed into animal feed or feed additive.(2)Screening and optimization of fermentation conditions of Lianhua Qingwen residue,using sterilized and non sterilized Lianhua Qingwen residue,mixing 10% drug residue,4%molasses and 5% yeast culture by-products during fermentation,adding four different amounts of compound enzymes(xylanase,cellulase and protease)of 0.5%,0%,5% and 15% and Aspergillus niger(an)and Bacillus subtilis(km51)with microbial inoculation amount of 106-107 CFU / g at 30,40 and 50 ℃,During the fermentation process,the water was maintained at 50-60% and treated for 1 day,2 days,4 days and 7 days respectively.The most suitable strain for fermentation was the mixture of lactic acid bacteria(sk3494)and yeast(SC)(1:1).The best fermentation process was to add 15% compound enzymes(xylanase,cellulase and protease)and ferment at 30 ℃ for 7 days.Compared with the original drug residue(YYZ),the crude protein content of optimized fermentation 1(4myl)and optimized fermentation 2(y4ml)increased by 2.051% and 2.489%,the crude fat content increased by 4.97% and 4.637%,and the crude fiber content decreased by 2.905% and 3.052%.The fermentation can effectively improve the crude protein and crude fat content of Lianhua Qingwen residue,reduce the crude fiber content in the drug residue,and improve the animal digestible components and nutrients of the drug residue.(3)By comparing the changes of biological activity before and after fermentation of drug residues,the ethanol extract and water extract of three kinds of drug residues after fermentation were extracted and compared with the two extracts of original drug residues.The results of in vitro antibacterial activity showed that the in vitro antibacterial effect of the extract was significantly different,which was reflected in the different ways of the same fermentation product,and the antibacterial effect of the extract on a pathogen was different,In the same way of different fermentation products,the antibacterial activity of extracts against a pathogen is different,and the antibacterial activity of the same extract against different strains is also different.On the whole,the antibacterial and bactericidal effect of ethanol extract is higher than that of water extract.Both ethanol extract and water extract have better effects on Haemophilus parasuis;The results of in vitro antiviral activity showed that the fermentation treatment could effectively enhance the antiviral activity of Lianhua Qingwen residue.Lianhua Qingwen residue had good antiviral activity against PRV,PCV2 and PRRSV before and after fermentation.Before fermentation,Lianhua Qingwen residue had the strongest anti PCV2 activity.High concentration ethanol extract could reduce the ratio of PCV2 gene in cell supernatant by 71.90%(P < 0.05)PRV and PRRSV in cell supernatant decreased by 57.35%(P < 0.05)and 7.37%(P > 0.05),respectively.