放牧条件下补充酵母硒对绵羊胃肠道微生物区系的影响

doi:10.16431/j.cnki.1671-7236.2023.03.011

摘要/Abstract

摘要： 【目的】探讨放牧条件下补充酵母硒对绵羊胃肠道微生物区系的影响。【方法】选择体重相近、健康的7~8月龄杜泊寒羊14只,采用单因素完全随机试验设计分成两组,每组7只。一组为对照组,不添加硒,另一组为硒添加组,硒(酵母硒)添加水平为0.2 mg/d。在自然放牧条件下进行为期30 d的饲养试验,分别于试验第15和30天采集瘤胃液和粪样,采用16S rRNA测序方法分析绵羊瘤胃以及后肠道微生物区系的变化。【结果】主坐标分析结果表明,补硒第30天后两组绵羊瘤胃及后肠道微生物在OTU和属水平上具有显著的组间差异(P＜0.05)。物种差异判别分析结果表明,饲养第15天时在门和属水平有8种瘤胃微生物具有显著的组间差异(P＜0.05),但这些物种的丰度均低于1%;饲养第30天时在门和属水平上有17种瘤胃微生物具有显著的组间差异(P＜0.05),其中相对丰度大于1%的有厚壁菌门、拟杆菌门、普雷沃氏菌属1、瘤胃球菌科NK4A214、拟杆菌目BS11;与对照组相比,硒添加组瘤胃中厚壁菌门、瘤胃球菌科NK4A214、拟杆菌目BS11的相对丰度显著提高(P＜0.05),拟杆菌门、普雷沃氏菌属1的相对丰度显著降低(P＜0.05)。饲养第15天时在属水平上有5种后肠道微生物具有显著的组间差异(P＜0.05),但这些物种的丰度均低于1%;饲养第30天时在门和属水平上有18种后肠道微生物具有显著的组间差异(P＜0.05),其中相对丰度大于1%的有厚壁菌门、拟杆菌门、拟杆菌属、瘤胃球菌科UCG-009、UCG-010、UCG-013和NK4A214、未排位的瘤胃球菌科、毛螺菌科AC2044、norank_f_Muribaculaceae;与对照组相比,硒添加组后肠道中拟杆菌门、拟杆菌属、瘤胃球菌科UCG-010和UCG-013的相对丰度显著提高(P＜0.05),厚壁菌门、毛螺菌科AC2044、瘤胃球菌科NK4A214和UCG-009、未排位的瘤胃球菌科、norank_f_Muribaculaceae的相对丰度显著降低(P＜0.05)。【结论】放牧条件下补充酵母硒可提高绵羊瘤胃微生物多样性和后肠道微生物丰富度,瘤胃中纤维降解菌的数量增加,后肠道中蛋白质和可溶性多糖降解菌的丰度提高。此外,放牧条件补饲酵母硒可促进绵羊后肠道益生菌的生长并抑制有害微生物的生长。

关键词: 酵母硒; 绵羊; 微生物区系

Abstract: 【Objective】 The purpose of this experiment was to investigate the effect of yeast selenium supplementation on the gastrointestinal microflora of sheep under grazing conditions.【Method】 Fourteen healthy 7-8 months old Dolphin sheep with similar body weight were selected and divided into two groups by single factor complete randomized trial design,with seven sheep in each group.One group was the control group without adding selenium,and the other group was the selenium adding group.The selenium (yeast selenium) adding level was 0.2 mg/d.A 30 day feeding experiment was conducted under natural grazing conditions.Rumen juice and feces samples were collected on the 15th and 30th days of the experiment,respectively.The 16S rRNA sequencing method was used to analyze the changes of rumen and hindgut microflora of sheep.【Result】 The results of principle coordinate analysis showed that rumen and hindgut microbes of sheep in two groups had significant differences in OTU and genus levels after 30 days of selenium supplementation (P＜0.05).The results of discriminant analysis of species differences showed that eight rumen microorganisms had significant differences between groups at phylum and genus levels on the 15th day of feeding (P＜0.05),but the abundance of these species was lower than 1%. At the 30th day of feeding,there were 17 rumen microorganisms with significant differences between groups at the phylum and genus levels (P＜0.05),among which Firmicutes,Bacteroidetes,Prevotella-1,Ruminococcaceae_NK4A214 and Bacteroidales_BS11_gut_group had relative abundance greater than 1%.Compared with the control group,the relative abundance of Firmicutes,Ruminococcaceae_NK4A214 and Bacteroidales_BS11_gut_group in the rumen in selenium supplementation group was significantly increased (P＜0.05),while the relative abundances of Bacteroidetes and Prevotella_1 was significantly decreased (P＜0.05).At the genus level,on the 15th day,there were significant differences in five species of posterior intestinal microorganisms between groups (P＜0.05),but the abundance of these species was lower than 1%.On the 30th day,there were 18 species of posterior intestinal microorganisms with significant differences between groups at phylum and genus level (P＜0.05),among them,the relative abundance of Firmicutes,Bacteroidetes,Bacteroidetes,Ruminococcaceae_UCG-009,Ruminococcaceae_UCG-010,Ruminococcaceae_UCG-013 and Ruminococcaceae_NK4A214,norank_f_Ruminococcaceae,Lachnospiraceae_AC2044 and norank_f_Muribaculaceae were greater than 1%.Compared with the control group,the relative abundances of Bacteroidetes,Bacteroidetes,Ruminococcaceae_UCG-010 and Ruminococcaceae_UCG-013 in intestinal tract of selenium supplementation group were significantly increased (P＜0.05).The relative abundances of Firmicutes,Lachnospiraceae_AC2044,Ruminococcaceae_NK4A214 and Ruminococcaceae_UCG-009,norank_f_Ruminococcaceae and norank_f_Muribaculaceae were significantly decreased (P＜0.05).【Conclusion】 Selenium supplementation of sheep under grazing conditions could improve rumen microbial diversity and hindgut microbial richness,increase the abundance of fiber-degraded bacteria in rumen as well as protein polysaccharide-degraded bacteria in hindgut.In addition,selenium supplementation could promote probiotics growth and inhibited the platation of harmful microorganisms in hindgut of sheep.

Key words: yeast selenium; sheep; microflora

中图分类号:

S816.7

崔双, 乌日力嘎, 阿拉达尔, 晓敏, 王剑秋, 弓剑. 放牧条件下补充酵母硒对绵羊胃肠道微生物区系的影响[J]. 中国畜牧兽医, 2023, 50(3): 952-965.

CUI Shuang, Wurlig, Alder, XIAO Min, WANG Jianqiu, GONG Jian. Effects of Yeast Selenium Supplementation on Gastrointestinal Microflora in Sheep Under Grazing Conditions[J]. China Animal Husbandry and Veterinary Medicine, 2023, 50(3): 952-965.

参考文献

[1] 李万栋,郝力壮,刘书杰,等.铁、锌、硒三种微量元素对反刍动物作用的研究[J].青海畜牧兽医杂志,2015,45(4):55-58. LI W D,HAO L Z,LIU S J,et al.Study on the effects of three trace elements of iron,zinc and selenium on ruminants[J].Qinghai Animal Husbandry and Veterinary Journal,2015,45(4):55-58.(in Chinese)
[2] 郭元晟,靳鹏.酵母硒对乌珠穆沁羊免疫球蛋白及细胞因子的影响[J].畜牧与饲料科学,2016,37(3):39-42. GUO Y S,JIN P.Effect of dietary supplementation of selenium-enriched yeast on immunoglobulin and cytokine of Ujumqin sheep[J].Animal Husbandry and Feed Science,2016,37(3):39-42.(in Chinese)
[3] ZHANG S,WU Z,HENG J,et al.Combined yeast culture and organic selenium supplementation during late gestation and lactation improve preweaning piglet performance by enhancing the antioxidant capacity and milk content in nutrient-restricted sows[J].Animal Nutrition,2020,6(2):160-167.
[4] 贺淼,张新,廖灿青,等.酵母硒对海兰褐商品代蛋鸡产蛋性能尧蛋硒沉积和硒利用率的影响[J].中国饲料,2020,5:113-117. HE M,ZHANG X,LIAO C Q,et al.Effects of different types of selenium yeast on egg-laying performance,egg selenium deposition and selenium utilization rate of Hy-line variety brown[J].China Feed,2020,5:113-117.(in Chinese)
[5] 弓剑.硒对奶牛乳腺抗氧化功能的影响及其机理研究[D].呼和浩特:内蒙古农业大学,2014. GONG J.The effect and underlying mechanism of selenium on antioxidative function in mammary gland of dairy cows[D].Hohhot:Inner Mongolia Agricultural University,2014.(in Chinese)
[6] AČG OBANOVÁ K,FAIX Š,PLACHÁ I,et al.Effects of different dietary selenium sources on antioxidant status and blood phagocytic activity in sheep[J].Biological Trace Element Research,2017,175(2):339-346.
[7] MIHALIKOVA K,GRESAKOVA L,BOLDIZAROVA K,et al.The effects of organic selenium supplementation on the rumen ciliate population in sheep[J].Folia Microbiol (Praha),2005,50(4):353-356.
[8] LIU Y,WANG C,LIU Q,et al.Effects of sodium selenite addition on ruminal fermentation,microflora and urinary excretion of purine derivatives in holstein dairy bulls[J].Journal of animal physiology and animal nutrition,2019,103(6):1719-1726.
[9] ANNE K K.Cecum microbiota in mice affected by selenium level in diet[D].Norwegian:Norwegian University of Life Sciences,2014.
[10] 索兰弟,李秉真.荧光法测定牧草中痕量硒[J].内蒙古农牧学院学报,1991,1:139-142. SUO L D,LI B Z.Determination of trace selenium in forage by fluorescence method[J].Journal of Inner Mongolia University of Agriculture and Animal Husbandry,1991,1:139-142.(in Chinese)
[11] 乌日力嘎,阿拉达尔,崔双,等.硒对热应激条件下放牧绵羊瘤胃发酵及瘤胃微生物区系的影响[J].中国饲料,2022,3:8-16. WURLIG,ALDER,CUI S,et al.Effects of selenium on rumen fermentation and rumen microflora of grazing sheep under heat stress[J].China Feed,2022,3:8-16.(in Chinese)
[12] EVANS N J,BROWN J M,MURRAY R D,et al.Characterization of novel bovine gastrointestinal tract treponema isolates and comparison with bovine digital dermatitis treponemes[J].Applied and Environmental Microbiology,2011,77(1):138-147.
[13] CHEN Y B,LAN D L,TANG C,et al.Effect of DNA extraction methods on the apparent structure of yak rumen microbial communities as revealed by 16S rDNA sequencing[J].Polish Journal of Microbiol,2015,64(1):29-36.
[14] MCCURDY D E,WILKINS K R,HILTZ R L,et al.Effects of supplemental butyrate and weaning on rumen fermentation in Holstein calves[J].Journal of Dairy Science,2019,102(10):8874-8882.
[15] O’HARAA E,KELLY A,MCCABE M S,et al.Effect of a butyrate-fortified milk replacer on gastrointestinal microbiota and products of fermentation in artificially reared dairy calves at weaning[J].Scientific Reports,2018,8(1):149-161.
[16] SAGHEDDU V,PATRONE V,MIRAGOLI F,et al.Infant early gut colonization by Lachnospiraceae:High frequency of Ruminococcus gnavus[J].Frontiers in Pediatrics,2016,4(11):57-63.
[17] MEEHAN C J,BEIKO R G.A Phylogenomic view of ecological specialization in the Lachnospiraceae,a family of digestive tract-associated bacteria[J].Genome Biology and Evolution,2014,6(3):703-713.
[18] RINNINELLA E,RAOUL P,CINTONI M,et al.What is the healthy gut microbiota composition? A changing ecosystem across age,environment,diet,and diseases[J].Microorganisms,2019,7(1):14-21
[19] SIBAI M,ALTUNTA?瘙塁 E,YILDIRIM B,et al.Microbiome and longevity:High abundance of longevity-linked muribaculaceae in the gut of the long-living rodent Spalax leucodon[J].OMICS,2020,24(10):592-601.
[20] CUI Z,WU S,LIU S,et al.From maternal grazing to barn feeding during pre-weaning period:Altered gastrointestinal microbiota contributes to change the development and function of the rumen and intestine of yak calves[J].Fronties in Microbiology,2020,3(11):485-492.
[21] ZHANG Z D,WANG C,DU H S,et al.Effects of sodium selenite and coated sodium selenite on lactation performance,total tract nutrient digestion and rumen fermentation in Holstein dairy cows[J].Animal,2020,14(10):2091-2099.
[22] 阿拉达尔.有机硒对热应激条件下放牧绵羊瘤胃发酵、瘤胃微生物区系以及肝脏功能的影响[D].呼和浩特:内蒙古师范大学,2021. ALDER.Effects of organic selenium on rumen fermentation,rumen microflora and liver function of grazing sheep under heat stress[D].Hohhot:Inner Mongolia Normal University,2021.(in Chinese)
[23] LIU X,MAO B,GU J,et al.Blautia—A new functional genus with potential probiotic properties?[J].Gut Microbes,2021,13(1):1-21.
[24] YU T,GUO J,ZHU S,et al.Protective effects of selenium-enriched peptides from Cardamine violifolia against high-fat diet induced obesity and its associated metabolic disorders in mice[J].RSC Advances,2020,10(52):31411-31424.
[25] CUI X,WANG Z,TAN Y,et al.Selenium yeast dietary supplement affects rumen bacterial population dynamics and fermentation parameters of Tibetan sheep (Ovis aries) in alpine meadow[J].Frontiers in Microbiology,2021,12(3):347-359.
[26] MAO S,ZHANG M,LIU J,et al.Characterising the bacterial microbiota across the gastrointestinal tracts of dairy cattle:Membership and potential function[J].Scientific Reports,2015,5(1):1-14.
[27] POWER S E,O’TOOLE P W,STANTON C,et al.Intestinal microbiota,diet and health[J].Scientific Reports,2014,111(3):387-402.
[28] 郭秀兰.猪肠道硬壁菌门和拟杆菌门数量的检测及其相对丰度与脂肪沉积的相关性研究[D].雅安:四川农业大学,2009. GUO X L.Detection of Firmicutes and Bacteroidetes in the pig gut and the correlation between their abundance and fat deposit[D].Ya’an:Sichuan Agricultural University,2009.(in Chinese)
[29] 陈宝剑,吴永绍,覃兆鲜,等.猪不同发育阶段肠道微生物菌群特征分析[J].中国畜牧杂志,2021,57(1):101-108. CHEN B J,WU Y S,QIN Z X,et al.Analysis of the characteristics of intestinal microflora in pigs at different developmental stages[J].Chinese Journal of Animal Husbandry,2021,57(1):101-108.(in Chinese)
[30] OBREGON-TITO A J,TITO R Y,METCALF J,et al.Subsistence strategies in traditional societies distinguish gut microbiomes[J].Nature Communications,2015,6(1):34-42.
[31] ZHOU S,LUO R,GONG G,et al.Characterization of metagenome-assembled genomes and carbohydrate-degrading genes in the gut microbiota of Tibetan pig[J].Frontiers in Microbiology,2020,11(8):442-452.
[32] NIEUWDORP M,GILIJAMSE P W,PAI N,et al.Role of the microbiome in energy regulation and metabolism[J].Gastroenterology,2014,146(6):1525-1533.
[33] FURUSAWA Y,OBATA Y,FUKUDA S,et al.Commensal microbe-derived butyrate induces the differentiation of colonic regulatory T cells[J].Nature,2013,504(7480):446-450.
[34] YANG H,XIAO Y,WANG J,et al.Core gut microbiota in Jinhua pigs and its correlation with strain,farm and weaning age[J].Journal of Microbiology,2018,56(5):346-355.
[35] LI Z,YI C,KATIRAEI S,et al.Butyrate reduces appetite and activates brown adipose tissue via the gut-brain neural circuit[J].Gut,2018,67(7):1269-1279.
[36] DE VADDER F,KOVATCHEVA-DATCHARY P,GONCALVES D,et al.Microbiota-generated metabolites promote metabolic benefits via gut-brain neural circuits[J].Cell,2014,156(1-2):84-96.
[37] 朱翠,师子彪,蒋宗勇,等.乳酸杆菌在调节肠道屏障功能中的作用[J].中国畜牧兽医,2012,9(39):118-122. ZHU C,SHI Z B,JIANG Z Y,et al.The role of Lactobacillus in regulating intestinal barrier function[J].China Animal Husbandry & Veterinary Medicine,2012,9(39):118-122.(in Chinese)
[38] 吕锡斌,何腊平,张汝娇,等.双歧杆菌生理功能研究进展[J].食品工业科技,2013,34(16):353-358. LYU X B,HE L P,ZHANG R J,et al.Research progress on physiological function of bifidobacteria[J].Food Industry Science and Technology,2013,34(16):353-358.(in Chinese)
[39] CANI P D,BIBILONI R,KNAUF C,et al.Changes in gut microbiota control metabolic endotoxemia-induced inflammation in high-fat diet-induced obesity and diabetes in mice[J].Diabetes,2008,57(6):1470-1481.