| With the increasing intensification of livestock and poultry farming at home and abroad,a large number of antibiotics are used to treat animal diseases,leading to increased bacterial resistance and animal food drug residues.Replacing antibiotics to prevent the occurrence of zoonotic diseases has become an inevitable trend in the healthy development of animal husbandry.Chinese herbal medicine preparations and micro-ecological preparations have the advantages of wide range of effects and low toxic and side effects,and have huge application prospects in replacing antibiotics as new feed additives.A total of 800 twenty-five week-old Jinding ducks were randomly divided into 8 groups with 5 repetitions per group and 20 ducks per relicate.Ducks in the control group were fed a basal diet,trial groups 1 were fed the basal diets supplemented with 1.5 g / kg complex probiotics,and the 2 to 4 groups added 1,1.5and 2 g / kg FML to the basic diet,groups 5 to 7 added 1.5 g / kg of compound probiotics to the basic diet,and then added 1,1.5 and 2 g / kg of FML,respectively.The pre-expermental period lasted for 1 week,and the experimental period lasted for8 weeks 1.To explore the influence of its use on the production performance,egg quality,antioxidant performance,lipid metabolism,immune function and intestinal microbes of laying ducks when used alone or in combination.New feed additives lay a certain theoretical foundation.1.The effects of FML and probiotics on the performance of laying ducks and egg qualityDuring the feeding test of laying ducks,the egg production rate,average daily feed intake,feed-to-egg ratio,average egg weight and broken egg rate are calculated daily in repeat units;after the feeding test,6 duck eggs are randomly selected in each repeat.The egg shape index,eggshell strength,egg yolk color,egg yolk percentage,eggshell thickness,and Huff units were measured.The results showed that compared with the control group,the average egg weight and feed egg ratio of the experimental group 1 were reduced by 14.2% and 5.1%(p<0.05)respectively.The egg production rate,average daily feed intake and broken egg rate were compared with the control group No significant difference(p>0.05);compared with the control group,the test 2,3,4,5 and 6 groups did not significantly improve the performance of laying ducks(p>0.05);compared with the control group,the test 7 group There was no significant difference in average egg weight,average daily feed intake,feed-to-egg ratio and broken egg rate(p>0.05),and egg production rate decreased by 5.8%(p<0.05).There was no significant difference in egg quality between the test groups 1,3 and 4compared with the control group(p>0.05);there was no significant difference between the test group 2 egg shape index,egg yolk color,eggshell thickness and Huff unit compared with the control group(p>0.05),eggshell strength increased by 15.9%(p<0.05),egg yolk percentage decreased by 7.3%(p<0.05);egg shape index,egg yolk percentage,eggshell thickness,Hough unit and control in groups 5,6 and 7 There was no significant difference between the groups(p>0.05),the eggshell strength was increased by 13.7%,10.0% and 13.6%(p<0.05),and the color of egg yolk in the test groups 5 and 6 increased by 3.2% and 2.7%(p<0.05)There was no significant difference in the color of egg yolk between the 7 groups in the test and the control group(p>0.05).The results showed that the addition of FML or probiotics to the basic diet had no obvious effect on improving the performance of laying ducks;the combined application of FML and probiotics had a significant enhancement effect on the eggshell strength.Test 7 group(1.5g/kg probiotics+2.0g/Kg FML)had the best effects.2.The effects of FML and probiotics on lipid metabolism of laying ducksAfter the feeding experiment,3 ducks and 3 duck eggs were randomly selected from each repeat.The blood was collected from the veins of the duck wings and the serum was separated.After the egg yolks were separated,the egg yolks were mixed and extracted 2 m L in an EP tube,and stored at-20 ?.Detection of triglyceride(TG),total cholesterol(TC),high-density lipoprotein cholesterol(HDL-C)and low-density lipoprotein cholesterol(LDL-C)in laying duck serum and egg yolk by ELISA method;collecting laying duck The liver was quick-frozen in liquid nitrogen and stored in a refrigerator at-80? for later use.The q PCR(dye method)was used to detect3-hydroxy-3-methylglutaryl coenzyme A reductase(HMG-COA)m RNA in laying ducks.The amount of expression in the liver.The results showed: 1)Compared with the control group,the serum TG and LDL-C content of the experimental group 1decreased by 14.5% and 23.9%(p<0.05),the HDL-C content increased by 17.8%(p<0.05),and the serum TC content Compared with the control group,there was no significant difference(p>0.05).There was no significant difference in serum TG,TC,HDL-C and LDL-C contents between the two groups in the experiment 2 compared with the control group(p>0.05).Serum TG,TC,LDL-C in the three groups had no significant difference compared with the control group(p>0.05),HDL-C content increased by 14.6%(p<0.05).Compared with the control group,there was no significant difference in serum TC and LDL-C content between the four groups(p>0.05),the TG content decreased by 13.4%(p<0.05),and the HDL-C content increased by 20.7%(p<0.05).Compared with the control group,the serum TG of the experimental 5,6 and 7 groups decreased by 21.4%,13.3% and 19.4%(p<0.05),TC decreased by 25.7%,35.1% and 18.9%(p<0.05),LDL-C decreased by 33.8%,47.5%and 27.4%(p<0.05),and HDL-C content increased by 28.8%,31.9% and 24.0%(p<0.05),respectively.2)Compared with the control group,the content of TG,TC and LDL-C in the egg yolk of the experimental group 1 decreased by 22.2%,10.0%and 22.6%(p<0.05),and the content of HDL-C increased by 16.7%(p<0.05);Compared with the control group,the content of TG,TC,HDL-C and LDL-C in the egg yolk of the test 2 group was not significantly different(p>0.05);compared with the control group,the content of TG,TC and HDL-C in the egg yolk of the test 3group No significant difference(p>0.05),LDL-C content decreased by 11.9%(p<0.05);compared with the control group,there was no significant difference in the yolk TG content of the four groups(p>0.05),TC and LDL-C content Reduced by20.0% and 16.7%(p<0.05),HDL-C content increased by 16.7%(p<0.05);compared with the control group,the yolk TG in the test groups 5,6 and 7 decreased by 22.2%,22.2% and 11.1 respectively %(P<0.05),TC decreased by 30.0%,20.0% and 20.0%(p<0.05),LDL-C decreased by 29.8%,36.9% and 23.8%(p<0.05),HDL-C content increased by 23.1 respectively %,23.1%,16.7%(p<0.05).3)Compared with the control group,the HMG-COA reductase m RNA expression in the test groups 1-7decreased by 8.9%,27.7%,27.7%,26.7%,35.6%,38.6%,and 43.6%(p<0.05).The results showed that: 1)The combined use of Moringa flavonoids and probiotics can effectively improve the lipid content in serum and egg yolk,and the best effect was obtained in the 6 groups(1.5 g/kg probiotics + 1.5 g/kg FML).2)The addition of moringa leaf flavonoids or probiotics to the basic diet can significantly inhibit the expression of HMG-COA reductase m RNA in the liver,and the combined effect is stronger when the two are used together.Among them,the test group 7(1.5 g/kg Probiotics + 2.0 g/kg FML)have the most obvious inhibitory effect.3.The effects of FML and probiotics on the antioxidant and immune function of laying ducksThe anti-oxidation indexes(malondialdehyde(MDA),superoxide dismutase(SOD),glutathione peroxidase(GSH-Px))and immune function indexes(IgG,IgA,IgM,duodenal s IgA and serum IL-2,IL-4,IL-6,and IFN-?)were tested;the spleens of laying ducks were collected,frozen in liquid nitrogen,and placed in a refrigerator at-80?.QPCR(dye method)was used to detect the expression of IL-2,IL-4,IL-6and IFN-? m RNA in the spleen.The results showed that: 1)Compared with the control group,the serum MDA content of the test 1 group was reduced by 24.5%(p<0.05),and the activities of SOD and GSH-Px were increased by 30.1% and 16.5%(P<0.05);test 2 and 3 The serum MDA content,SOD and GSH-Px activity of the group had no significant difference compared with the control group(p>0.05);compared with the control group,the serum MDA content of the test 4 group had no significant change(p>0.05),SOD and GSH-Compared with the control group,the Px activity increased by 23.7% and 12.5%(p<0.05);compared with the control group,the serum MDA content of the experimental groups 5,6 and 7 decreased by 29.3%,32.8% and 20.8%(p<0.05)),SOD activity increased by 40.8%,41.8% and 23.6%(p<0.05),GSH-Px activity increased by 19.6%,27.0% and 20.6%(p<0.05).2)Compared with the control group,the content of MDA in the egg yolk of the experimental group 1 decreased by 23.9%(p<0.05),and the SOD activity increased by 35.0%(P<0.05).The GSH-Px activity had no significant difference compared with the control group(p >0.05);MDA content,SOD and GSH-Px activity of egg yolk in test 2 group were not significantly different from the control group(p>0.05);MDA content and GSH-Px activity in egg yolk of test 3 group were not significant compared with control group Difference(p>0.05),SOD activity increased by 23.2%(p<0.05);compared with the control group,the MDA content in the egg yolk of the test groups 4,5,6 and 7 decreased by 16.6%,37.5%,34.1% and 25.0,respectively %(P<0.05),SOD activity increased by 30.5%,49.4%,47.8% and 32.2%(p<0.05),GSH-Px activity increased by 13.4%,21.6%,26.6% and 21.6%(p< 0.05).3)Serum IgG,IgA,IgM and s IgA content in test group 1 increased by 20.5%,36.2%,27.7%and 50.5% compared with the control group(p<0.05);serum IgG,IgA in test 2 group,IgM and duodenum s IgA levels were not significantly different from the control group(p>0.05);the serum IgG and IgM levels of the three groups were not significantly different from the control group(p>0.05),serum IgA and ten The s IgA content of the duodenum increased by 28.6% and 29.5%,respectively(p<0.05);the serum IgG,IgA,IgM and duodenal s IgA content of the test 4,5,6 and 7 groups increased significantly,and the test 4 group increased by 14.2% respectively,32.2%,26.2% and 39.6%(p<0.05),the test 5 group increased by 30.9%,50.0%,37.7% and 52.1%(p<0.05),the test 6 group increased by 29.8%,43.2%,43.0% And 53.1%(p<0.05),the test 7 group increased by 23.2%,40.8%,34.7% and 55.6%(p<0.05).4)The levels of serum IL-2and IL-6 in each test group were not significantly different from those in the control group(p>0.05);compared with the control group,serum IL-4 in groups 1,3,5,6,and7 The contents were increased by 18.3%,14.1%,16.5%,20.7% and 28.1%(p<0.05)respectively.There was no significant difference in serum IL-4 content between the test groups 2 and 4 compared with the control group(p>0.05);test 1.The serum IFN-? content of groups 2,3,4 and 7 was not significantly different from that of the control group(p>0.05).The serum IFN-? content of groups 5 and 6 in the experiments increased by 23.2% and 27.5% respectively compared with the control group (p<0.05).5)The spleen IL-2,IL-6 and IFN-? m RNA expression levels of the groups1 and 2 were not significantly different from the control group(p>0.05),and the spleen IL-4 m RNA expression levels were up-regulated by 31.5% and 29.1%,respectively.(P<0.05);the expression of IL-2 and IL-6 m RNA in the spleens of groups 3,4 and 7 was not significantly different from that of the control group(p>0.05).The expression of IL-4 m RNA was up-regulated by 38.3% and 34.2,respectively % And 40.5%(p<0.05),IFN-? m RNA expression was up-regulated by27.5%,23.7% and 30.4%(p<0.05);there was no significant difference in the expression of IL-2 m RNA in the spleen of test 5 group compared with the control group(P>0.05),IL-4,IL-6 and IFN-? m RNA expression were up-regulated by 32.4%,35.6% and 30.4%(p<0.05);compared with the control group,the spleen IL-2,IL-2,IL-4,IL-6 and IFN-? m RNA expression levels were up-regulated by 48.9%,41.9%,36.5% and 31.3%,respectively(p<0.05).The results show that: 1)Adding probiotics alone or high-dose moringa flavonoids to the basic diet can significantly increase the activity of SOD and GSH-Px in serum and egg yolk.When probiotics and moringa flavonoids are used in combination,serum and egg yolk The content of MDA was significantly reduced,and the activities of SOD and GSH-Px were significantly improved.The test group 6(1.5 g/kg probiotics + 1.5 g/kg FML)had the best effect.2)Adding probiotics alone or high-dose FML to the basic diet can significantly increase the serum IgG,IgA,IgM and s IgA content in the duodenum.When probiotics and moringa flavonoids are used together The IgG,IgA,IgM and duodenum s IgA content increase effect is more obvious,the test group 6(1.5 g/kg probiotic + 1.5 g/kg FML)has the best effect.3)The addition of FML or probiotics mainly stimulates laying ducks to up-regulate the expression of IL-4 gene m RNA in the spleen,which is consistent with the results of serum IL-4 content detection.When FML and probiotics are used together At this time,the m RNA expression levels of Th1 and Th2 cytokines are increased,but the m RNA expression levels of Th2 cytokine genes are generally better than Th1.4.The effects of FML and probiotics on the structure of intestinal flora of laying ducks Each group randomly selected 3 laying ducks for dissection,collected the contents of laying duck cecum in RNase-Free EP tubes and stored in dry ice,and sent to Shanghai Weiji Biological Technology Co.,Ltd.for genomic DNA extraction,with labels PCR amplification of specific primers,and then use Axy Prep DNA gel recovery kit to recover the target gene,and finally 16 S r DNA V4 ? V5 region high-throughput sequencing and bioinformatics analysis.The results showed that OTU cluster analysis of intestinal microflora was annotated to 5643 OTUs,including 16 phyla,24 classes,38 orders,62 families,131 genera and 106 species.At the portal level,it is mainly composed of Bacteroidetes,Firmicutes,Fusobacteria,and Proteobacteria.Compared with the control group,the 1 group of Proteobacteria was significantly Increase(P<0.05);At the class level,it is mainly composed of Fusobacteriia,Bacteroidia,Negativicutes,and Clostridia.Compared with the control group,the group of T1?T7group Clostridia,Bacteroides,Negativicutes and Clostridia have no significant difference in abundance compared with group C(p>0.05).At the level of the order,it is mainly composed of Fusobacteriales,Bacteroidales,Selenomonadales,and Clostridiales.T1?T7 group Clostridia,Bacteroides,Crescent There was no significant difference in the abundance of species of Monospora and Clostridium from group C(p>0.05).At the family level,Fusobacteriaceae,Bacteroidaceae,Prevotellaceae,Acidaminococcaceae,Porphyromonadaceae,and Ricanaceae and Ruminococcaceae,the T1?T7 group of Fusobacteriaceae,Bacteroides,Prevotidae,Aminococcaceae,Phytobacteriaceae and Rumenaceae have the same species abundance as Group C There was no significant difference(p>0.05);compared with group C,there was no significant difference in the abundance of Pseudomonasceae in T1?T5 groups(p>0.05),and the species of Pseudomonasceae in T6 and T7 groups Compared with the control group,the abundance changes increased by 40%(p<0.05).At the genus level,it is mainly composed of Bacteroides,Fusobacterium,Parabacteroides,Megamonas,Alistopes,and Phascolarctobacterium.Compared with the control group,there was no significant difference in the abundance of Bacteroides,Fusobacterium,and Monomonospora in the T1 ? T7 groups(p>0.05);the relative abundance of Parabacteroides in groups 6 and 7 increased by 50.0 compared with thecontrol group %(P<0.05);the relative abundance of species of the genus Cladosporium in the test 7 group was 33.3% higher than that in the control group(P<0.05);the relative abundance of Phascolarctobacterium in the test 4 group was reduced by 50%(p<0.05);At the species level,it is mainly composed of uncultured-bacterium,Bacteroides-barnesiae,Bacteroides-plebeius,Bacteroides-coprocola,uncultured-organism,and Bacteroides-coprophilus.Among them,the relative abundance of Bacteroides-plebeius increased significantly compared with the control group 44.4%(P<0.05).The results showed that the addition of high-dose FML(2.0 g/kg moringa flavonoids)alone in the diet resulted in a significant decrease in the relative abundance of koalas in the intestine,indicating that high-dose FML would cause egg Duck intestinal flora is dysregulated,and the combination of high-dose FML and probiotics improves the intestinal flora diversity.Conclusion: In summary,the addition of FML and probiotics in the diet has no obvious effect on the production performance of laying ducks.The combined strength of FML and probiotics in eggshell is significantly enhanced,and the use of FML alone FML or probiotics can improve the lipid metabolism,antioxidant function and immune function of laying ducks.When FML and probiotics are used together,the effect is better.Among them,the test 6 group(1.5 g/kg probiotic + 1.5 g/kg FML)had the best comprehensive effect.High doses of FML can cause dysbiosis in the intestinal flora of laying ducks,but in combination with probiotics,the diversity of intestinal flora is improved. |
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