The Material Basis And Efficacy Evaluation On The Above-ground Parts Of Codonopsis Tangshen Oliv As A Feed Additive

As a tonic medicine,Codonopsis radix has the effects of invigorating the spleen and lungs,nourishing generate fluid,which is used as Chinese herbal medicine for treatment and health care commonly.It has a long growth cycle and can only be excavated after 3-4 years.It is dualuse for food and medicine,with has large market demand and wide clinical application.The stems and leaves of the above-ground part of Codonopsis radix have been proven to contain polysaccharides,terpenes,sterols,saponins,amino acids,trace elements and so on,which are often discarded.Therefore,the use on medicine,economy and resource expansion shows that making the above-ground parts of Codonopsis radix into feed additives has great development prospects and value potential.But until now,the research on it both domestic and foreign is scarce.So it is of great practical significance and potential economic value to carry out in-depth research on the above-ground part of Codonopsis radix.The Codonopsis radix of Wushan Temple in Chongqing belongs to Codonopsis tangshen Oliv,and this group showed that compared with Codonopsis pilosulae from other origin,it is rich in total amino acids and taste amino acids,and has high polysaccharide content,and Duping Codonopsis tangshen Oliv and Hongchun Codonopsis tangshen Oliv from different production areas have high content of fat-soluble components,and have outstanding nutritional value.Sedondly,the research group's comparative analysis on the ingredients of Codonopsis Lanceolata,roots,stems,and leaves revealed that the difference on chemical composition betweeen stems and roots was little,but it has a big difference in the content of ingredients.It provided data support on the development and utilization of the above-ground parts of Codonopsis tangshen Oliv.This research takes the above-ground parts of Codonopsis tangshen Oliv of Wushan Temple as the main object,and focuses on the possible material basis of the above-groundl part of Codonopsis tangshen Oliv as a feed additive through the components of adenosine glycosides,polysaccharides,total saponins,hydrolyzed amino acids and trace elements.The effects on growth performance,immune performance and intestinal microflora was studied by adding the powder of above-ground part of Codonopsis tangshen Oliv to the ration feed of Wupi Redcrested chicken,which provided an experimental basis for further development and utilization of the above-ground parts of Codonopsis tangshen Oliv in livestock farming and the development of new green feed additives.Methonds 1 Study on the material basis of the above-ground parts of Codonopsis tangshen Oliv as feed additiveThe similarity of different batches of Codonopsis tangshen Oliv was compared and the content of adenosine glycosides was determined through establish a characteristic map of different parts of the above-ground parts of Codonopsis tangshen Oliv using high-performance liquid chromatography.The content of total polysaccharides,total saponins,hydrolyzed amino acids and trace elements was determined by chemical analysis methods.2 Efficacy evaluation of above-ground part of Codonopsis tangshen Oliv as a feed additiveProduction of powder on above-ground parts of Codonopsis tangshen Oliv: In autumn(Codonopsis tangshen Oliv has been harvested before and after the White Dew Festival),the above-ground parts of Codonopsis tangshen Oliv were collected from different origin.According to the requirements of the 2015 edition of the Pharmacopoeia,it was cut,which was dried at 55°C,and then crushed and sieved(No.4).According to different proportions of 1%,3%,and 5%,it is added to the ration feed of Wupi Red-crested chicken,and mixed evenly.The chicken was Raised for 49 days to study the effects of growth performance,immune performance and intestinal microflora for Wupi Red-crested chicken.2.1 Effects of the above-ground parts of Codonopsis tangshen Oliv on the growth performance of Wupi Red-crested chickenThe effects of the above-ground parts of Codonopsis tangshen Oliv on the growth performance of Wupi Red-crested chicken was investigated through these indicators including regular weighing,slaughter rate after slaughter,pectoral muscle rate,feed-to-weight ratio and so on.2.2 Effects of the above-ground parts of Codonopsis tangshen Oliv on the immune performance of Wupi Red-crested ChickenThe immune organ index,the content of immunoglobulin(IgG,IgM,IgA)and interleukins(IL-1,IL-2,IL-6)on the 21 st and 49 th were determined after regular slaughter to explore and evaluate the effects of the above-ground parts of Codonopsis tangshen Oliv on the immune performance of Wupi Red-crested chicken.2.3 Effects of the above-ground parts of Codonopsis tangshen Oliv on the intestinal microflora of Wupi Red-crested chickenAfter 49 days of feeding,red-crested chicken fecal samples was collected,and the diversity of microbial flora in fecal samples was studied by sequence analysis of the hypervariable regions after 16 SrDNA Amplicon Sequencing and PCR amplification using universal primers designed on conserved regions.Results 1 Study on the Material Basis of the above-ground parts of Codonopsis tangshen Oliv as a feed additiveThere were six characteristic peaks in the HPLC characteristic maps of Codonopsis tangshen Oliv from different origin,and the relative retention time was RSD%<1% for each peak.The similarities of the HPLC profiles between 16 batches of Codonopsis tangshen Oliv and 13 batches of Codonopsis tangshen Oliv were vary from 0.197 to 0.845.The average content of the adenosine glycosides on the above-ground parts of Codonopsis tangshen Oliv was 0.566mg/g,which accounted for about 39% of Codonopsis tangshen Olivis.The total polysaccharide content on the above-ground parts of Codonopsis tangshen Oliv was 3.234±0.171%,which accounted for about 10% of Codonopsis tangshen Olivis.The content of total saponins on the above-ground parts of Codonopsis tangshen Oliv was 13.584±0.717mg/g,which accounted for about 29% of Codonopsis tangshen Olivis.Both the above-ground parts of Codonopsis tangshen Oliv and DangshenCodonopsis tangshen Oliv contains 17 kinds of hydrolyzed amino acids and 7 kinds of essential amino acids.The hydrolyzed amino acid content on the above-ground parts of Codonopsis tangshen Oliv was 2.06 times that of the root,and the essential amino acids content on the above-ground parts of Codonopsis tangshen Oliv was 2.84 times that of the root.Both the above-ground parts of Codonopsis tangshen Oliv and Codonopsis tangshen Oliv contain 13 kinds of trace elements.In addition to iron,boron,molybdenum,arsenic,the other 9 kinds elements in the above-ground parts of Codonopsis tangshen Oliv exceeded Codonopsis tangshen Oliv.The top five trace elements in the aboveground parts of Codonopsis tangshen Oliv are iron(212.830±3.783 mg/kg),silicon(214.555±6.324 mg/kg),aluminum(116.864±6.276 mg/kg),selenium(180.800±10.536 mg/kg).kg)and manganese(120.440±0.336 mg/kg).2 Efficacy evaluation of above-ground part of Codonopsis tangshen Oliv as feed additive 2.1 Effect of the above-ground parts of Codonopsis tangshen Oliv on the growth performance of Wupi Red Crested ChickenThe results of 42-day weight gain study showed that compared with the blank group,the low-dose group increased by 3.59%,the medium-dose group increased by 1.31%,and the highdose group decreased by 3.37%.The results of average daily weight gain showed that compared with the blank group,the low-dose group increased by 3.80%,the medium-dose group increased by 1.35%,and the high-dose group decreased by 3.50%.The results of the feedweight ratio study showed that compared with the blank group,the low-dose group decreased by 5.63%,the medium-dose group decreased by 8.66%,and the high-dose group decreased by 12.99%.This results showed that the low-dose group had the best effects.2.2 Effects of the above-ground parts of Codonopsis tangshen Oliv on the immune performance of Wupi Red-crested chickenThe results of immune organ index on 49 d showed that compared with the blank group,the thymus index,spleen index,and bursa index increased by 23.97%,21.51%,and 15.32% respectively in the low-dose group,increased by 57.23%,19.77%,and 14.52% respectively in the middle-dose group,and increased by 33.06%,59.88%,and 17.74%,respectively in the highdose group.The results of the immunoglobulin study on 49 d showed that compared with the blank group,the contents of IgG,IgM and IgA in the low-dose group increased by 2.56%,28.33% and 13.63% respectively;increased by 20.70% and 46.02%,26.52% respectively in the middledose group,and increased by 36.22%,65.61%,45.16%.in the high-dose group.The results of Interleukin-1(IL-1)study on 49 d showed that compared with the blank group,the low-dose group,middle-dose group and high-dose group were reduced by 18.08%,27.19% and 52.80% respectively.The results of interleukin-2(IL-2)study on 49 d showed that compared with the blank group,the low-dose group,middle-dose group and high-dose group were increased by 24.71%,82.28% and 106.79% respectively.The results of Interleukin-6(IL-6)study on 49 d showed that compared with the blank group,the low-dose group,middle-dose group and high-dose group were increased by 106.05%,195.35% and 237.67% respectively.2.3 Effects of the above-ground parts of Codonopsis tangshen Oliv on the intestinal microflora of Wupi Red-crested ChickenAnnotation results of intestinal flora species showed that the dominant species at the phylum level include Firmicutes,Bacteroidetes,and Actinobacteria,the dominant species at the class level are Bacilli,Bacteroidia,and Clostridia;the dominant species at the order level are Lactobacillales,Bacteroidales and Clostridiales,the dominant species at the family level are Lactobacillaceae,Bacteroidaceae and Veillonellaceae;the dominant species at the genus level are Lactobacillus,Bacteroides and Megamonas;the dominant at the species level are Lactobacillus_aviarius,Lactobacillus_gasseri and Bacteroides_salanitronis.Based on the Venn diagram and petal diagramof OTU,the number of OTUs increased by 38.21%,25.13% and 31.55% respectvely in the low-dose group,middle-dose group,and high-dose group compared to the blank group.The species abundance analysis results at the family level showed that the relative abundances of Lactobacillaceae(Lactobacillus)in the low-dose group,middle-dose group and high-dose group were increased by 49.96 times,22.54 times and 18.39 times respectively,compared with the blank group.? diversity analysis: the ? Diversity index study showed that compared with the blank group,the observed_species,Chao1,ACE,and PD_whole_tree increased by 36.45%,39.58%,43.71%,and 55.48% respectively in the lowdose group,increased by 16.52%,18.45%,22.26%,24.12% respectively in the middle-dose group,and increased by 13.90%,42.51%,23.51%,25.14% respectively in the high-dose group.All the results on species diversity curve,dilution curve and grade clustering curve showed that the richness of species in each grouping sample from highto low are low-dose group,medium-dose group,high-dose group and blank group,of which the low-dose group had the best.species abundance.The UPGMA clustering tree results showed that the low-dose group,the middle-dose group,and the high-dose group are more abundant in clustering species than the blank group.The results of ? diversity analysis showed that the low-dose group,the middledose group,and the high-dose group were significantly different from the blank group.The difference analysis between the groups showed that the low-dose group,the middle-dose group,the high-dose group had significant differences comparing with the blank group(P < 0.01).Analysis of species differences between the groups: T-test results showed that the difference between the low-dose group and the blank group was significant(P < 0.01).The different species were mainly Firmicutes,Bacteroidetes,Proteobacteria,Synergistetes.Meta Stat results showed that in each group,Proteobacteria and Melainabacteria were significantly different(q value < 0.05);Firmicutes,Bacteroidetes and Synergistetes had hextremely significant X difference(P < 0.01).Simper results showed that compared with the blank group,differential contribution of the low-dose group,middle-dose group and high-dose group at the family level from big to small were Lactobacillaceae,Bacteroidaceae,Veillonellaceae,Rikenellaceae,Ruminococcaceae,Lachnospiraceae,Dermabacteraceae,Acidobacteriaceae,Enterobacteriaceae and Enterobacteriaceae.ConclusionsThe material basis results on the above-ground parts of Codonopsis tangshen Oliv showed that the components of above-ground parts of Codonopsis tangshen Oliv was similar to Codonopsis tangshen Oliv.The content of adenosine glycosides,total polysaccharides,and total saponins in the above-ground parts of Codonopsis tangshen Oliv were accounted for about 39%,10%,29% of Codonopsis tangshen Olivis respectively.The content of hydrolyzed amino acids and essential amino acids in the above-ground parts of Codonopsis tangshen Oliv were accounted for about 2.06 times and 2.84 times of Codonopsis tangshen Olivis.The aboveground parts of Codonopsis tangshen Oliv can promote the growth of Wupi Red-crested chicken,and the chicken weight in the low-dose group increased by 3.59% compared with the blank group.The above-ground parts of Codonopsis tangshen Oliv can enhance the immune function of Wupi Red-crested Chicken.Compared with the blank group,the thymus index increased by 23.97%,57.23% and 33.06% respectively in low,medium and high dose groups,the spleen index increased by 21.51%,19.77%,and 59.88%,and the bursal index increased by 15.32%,14.52%,and 17.74% respectively.Compared with the blank group,the content of IgG increased 2.56%,20.70% and 36.22% respectively in low,medium and high dose groups;IgM increased 28.33%,46.02% and 65.61% respectively;IgA increased by 13.63%,26.52%,and 45.16% respectively.Interleukin-1(IL-1)reduced by 18.08%,27.19%,and 52.80% respectively,interleukin-2(IL-2)increased by 24.71%,82.28% and 106.79% respectively;and the interleukin-6(IL-6)increased by 106.05%,195.35% and 237.67% respectively.The the aboveground parts of Codonopsis tangshen Oliv increased the species abundance and diversity of the intestinal flora of Wupi Red-crested chicken,and promoted the growth of beneficial intestinal flora(Lactobacillus).The number of OUTs in the low,middle and high dose groups increased by 38.21%,25.13% and 31.55% respectively compared with the blank group.At the family level,the species abundance of Lactobacillaceae(Lactobacillus)in the low-dose group,middle-dose group and high-dose group was respectively 49.96 times,22.54 times and 18.39 times that of the blank group.Experimental results showed that the low-dose group works best.As a feed additive,the above-ground parts of Codonopsis tangshen Oliv has good development prospects.