饲喂高蛋白饲粮对雏鹅盲肠微生物菌群的影响
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摘要
【目的】研究饲喂不同蛋白质质量分数的饲粮对雏鹅盲肠微生物菌群结构和数量的影响。【方法】选取1日龄雁鹅72只,随机分成3组:A、B和C组分别饲喂质量分数为16%、20%和24%的粗蛋白。在第14天每组取6只雁鹅盲肠内容物,进行细菌培养、分离和计数;另每组取3只雁鹅盲肠内容物,根据细菌16S rDNA的保守性设计引物,采用IlluminaHiSeq测序平台对细菌16S rDNA的V4区基因进行测序,根据序列进行微生物物种多样性分析。【结果】A、B和C组共获得1 066 175条高质量的细菌16S rDNA序列,3组共有操作分类单元(OTU)数目为1 013个,主成分1(PC1)对检测到的总微生物的贡献率为46.64%,主成分2(PC2)的贡献率为16.46%。门水平上的优势菌群为放线菌门Actinobacteria、拟杆菌Bacteroidetes、厚壁菌门Firmicutes、变形菌门Proteobacteria和疣微菌门Verrucomicrobia等;科水平上的优势菌群为拟杆菌科Bacteroidaceae、肠杆菌科Enterobacteriaceae、毛螺菌科Lachnospiraceae、疣微菌科Ruminococcacea和理研菌科Rikenellaceae;属水平上的优势菌群为艾克曼菌属Akkermansia、拟杆菌属Bacteroides、棒状杆菌Corynebacterium和乳酸菌属Lactococcus。与A组相比,C组双歧杆菌Bifidobacterium的数量显著降低(P<0.05),C组大肠埃希菌Escherichia coli和沙门氏菌Salmonella数量显著升高(P<0.05)。【结论】饲喂高蛋白饲粮使雏鹅盲肠微生物菌群多样性和丰度发生变化,使双歧杆菌和乳酸杆菌的数量减少,使大肠埃希菌和沙门氏菌数量增加。
【Objective】To study the effects of feeding the diets at different protein levels on the structure and number of cecal microflora in goslings.【Method】Seventy-two one-day-old geese were randomly divided into three groups, including group A, B and C fed with diets containing 16%, 20% and 24% crude protein,respectively. On the 14 th day, the cecum contents of six geese in each group were used for bacterial culture,isolation and counting. The cecal contents of other three geese in each group were taken, and primer sequences were designed according to the conservation of bacterial 16 S rDNA. The V4 region genes of bacterial 16 S rDNA were sequenced based on the IlluminaHiSeq sequencing platform, and microbial species diversity was analyzed based on the sequences.【Result】A total of 1 066 175 high-quality bacterial 16 S rDNA sequences were obtained from group A, B and C. The total number of operational taxonomic unit(OTU) shared by three groups was 1 013, the contribution rate of principle component 1(PC1) to total microbes was 46.64%, and the contribution rate of principle component 2(PC2) was 16.46%. The dominant flora at the phylum level were Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria, Verrucomicrobia and etc. The dominant flora at the family level were Bacteroidaceae, Enterobacteriaceae, Lachnospiraceae, Ruminococcacea, and Rikenellaceae.The dominant flora at the genus level were Akkermansia, Bacteroides, Corynebacterium and Lactococcus.Comparing with group A, the number of Bifidobacteria in group C was significantly lower(P<0.05), and the number of Escherichia coli and Salmonella in group C were significantly higher(P<0.05).【Conclusion】Feeding high-protein diet can change the diversity and abundance of cecal microflora in goslings, reduces the number of bifidobacteria and lactobacilli, and increases the number of E. coli and Salmonella.
【Objective】To study the effects of feeding the diets at different protein levels on the structure and number of cecal microflora in goslings.【Method】Seventy-two one-day-old geese were randomly divided into three groups, including group A, B and C fed with diets containing 16%, 20% and 24% crude protein,respectively. On the 14 th day, the cecum contents of six geese in each group were used for bacterial culture,isolation and counting. The cecal contents of other three geese in each group were taken, and primer sequences were designed according to the conservation of bacterial 16 S rDNA. The V4 region genes of bacterial 16 S rDNA were sequenced based on the IlluminaHiSeq sequencing platform, and microbial species diversity was analyzed based on the sequences.【Result】A total of 1 066 175 high-quality bacterial 16 S rDNA sequences were obtained from group A, B and C. The total number of operational taxonomic unit(OTU) shared by three groups was 1 013, the contribution rate of principle component 1(PC1) to total microbes was 46.64%, and the contribution rate of principle component 2(PC2) was 16.46%. The dominant flora at the phylum level were Actinobacteria, Bacteroidetes, Firmicutes, Proteobacteria, Verrucomicrobia and etc. The dominant flora at the family level were Bacteroidaceae, Enterobacteriaceae, Lachnospiraceae, Ruminococcacea, and Rikenellaceae.The dominant flora at the genus level were Akkermansia, Bacteroides, Corynebacterium and Lactococcus.Comparing with group A, the number of Bifidobacteria in group C was significantly lower(P<0.05), and the number of Escherichia coli and Salmonella in group C were significantly higher(P<0.05).【Conclusion】Feeding high-protein diet can change the diversity and abundance of cecal microflora in goslings, reduces the number of bifidobacteria and lactobacilli, and increases the number of E. coli and Salmonella.
引文
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[13]NOROUZI H,DANESH A,MOHSENI M.Marine actinomycetes with probiotic potential and bioactivity against multi-drug resistant bacteria[J].Int J Mol Cell Med,2018,7(1):44-52.
[14]KOHL K D.Diversity and function of the avian gut microbiota[J].J Comp Physiol B,2012,182(5):591-602.
[15]HE X,MCLEAN J S,EDLUND A,et al.Cultivation of a human-associated TM7 phylotypereveals a reduced genome and epibiotic parasitic lifestyle[J].P Natl Acad Sci USA,2015,112(1):244.
[16]DALY K,PROUDMAN C J,DUNCAN S H,et al.Alterations in microbiota and fermentation products in equine large intestine in response to dietary variation and intestinal disease[J].Brit J Nutr,2012,107(7):989-995.
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[18]章文明,汪海峰,刘建新.乳酸杆菌益生作用机制的研究进展[J].动物营养学报,2012,24(3):389-396.
[19]SALVAT G,GUYOT M,PROTINO J.Monitoring Salmonella,Campylobacter,Escherichia coli and Staphylococcus aureus in traditional free-range‘Label Rouge’broiler production:A 23-year survey programme[J].JAppl Microbiol,2016,122(1):248-256.
[2]DICKS L M T,GELDENHUYS J,MIKKELSEN L S,et al.Our gut microbiota:A long walk to homeostasis[J].Benef Microbes,2018,9(1):3-20.
[3]PAN D,YU Z.Intestinal microbiome of poultry and its interaction with host and diet[J].Gut Microbes,2014,5(1):108-119.
[4]刘蓓一.扬州鹅肠道微生物多样性及其受饲粮纤维水平的调控研究[D].扬州:扬州大学,2012.
[5]章学东,钱定海,吴丽娟,等.日粮中添加苜蓿草粉对蛋鸡生产性能,蛋品质和免疫功能的影响[J].中国家禽,2008,30(16):40-41.
[6]李曼曼,丁雪东,荣雪路,等.一起皖西白鹅内脏型合并关节型痛风的病例报告[J].畜牧与兽医,2018,50(4):120-123.
[7]刘五岳.鹅的饲养标准[J].畜禽业,2003(10):11.
[8]DRAY S,DUFOUR A B.The ade4 package:Implementing the duality diagram for ecologists[J].J Stat Softw,2007,22(4):1-20.
[9]WHITE J R,NAGARAJAN N,POP M.Statistical methods for detecting differentially abundant features in clinical metagenomic samples[J].PLoS Comput Biol,2009,5(4):e1000352.
[10]张莉力,肖海蒂,刘黎莹,等.高通量测序技术分析大骨鸡盲肠微生物组成[J].中国家禽,2017,39(4):22-27.
[11]肖英平,杨彩梅,代兵,等.基于高通量测序的丁酸梭菌对肉鸡盲肠菌群结构的影响[J].浙江农业学报,2017,29(3):373-379.
[12]MU C,YANG Y,LUO Z,et al.Temporal microbiota changes of high-protein diet intake in a rat model[J].Anaerobe,2017,47:218-225.
[13]NOROUZI H,DANESH A,MOHSENI M.Marine actinomycetes with probiotic potential and bioactivity against multi-drug resistant bacteria[J].Int J Mol Cell Med,2018,7(1):44-52.
[14]KOHL K D.Diversity and function of the avian gut microbiota[J].J Comp Physiol B,2012,182(5):591-602.
[15]HE X,MCLEAN J S,EDLUND A,et al.Cultivation of a human-associated TM7 phylotypereveals a reduced genome and epibiotic parasitic lifestyle[J].P Natl Acad Sci USA,2015,112(1):244.
[16]DALY K,PROUDMAN C J,DUNCAN S H,et al.Alterations in microbiota and fermentation products in equine large intestine in response to dietary variation and intestinal disease[J].Brit J Nutr,2012,107(7):989-995.
[17]SANZ Y,NADAL I,S NCHEZ E.Probiotics as drugs against human gastrointestinal infections[J].Recent Pat Antiinfect Drug Discov,2007,2(2):148-156.
[18]章文明,汪海峰,刘建新.乳酸杆菌益生作用机制的研究进展[J].动物营养学报,2012,24(3):389-396.
[19]SALVAT G,GUYOT M,PROTINO J.Monitoring Salmonella,Campylobacter,Escherichia coli and Staphylococcus aureus in traditional free-range‘Label Rouge’broiler production:A 23-year survey programme[J].JAppl Microbiol,2016,122(1):248-256.