基于16S rRNA高通量测序技术对热应激环境下鸡肠道菌群多样性分析  

Investigation of the diversity of chicken intestinal microbiota in a thermal stressed environment based on 16S rRNA high-throughput sequencing

作　　者：李春苗[1] 吴兆林[1] 孔令琳 黄华云[1] 王钱保[1] 黄正洋[1] 薛龙岗[1] 赵振华[1] LI Chunmiao;WU Zhaolin;KONG Linglin;HUANG Huayun;WANG Qianbao;HUANG Zhengyang;XUE Longgang;ZHAO Zhenhua(Jiangsu Institute of Poultry Sciences,Yangzhou 225125)

机构地区：[1]江苏省家禽科学研究所,扬州225125

出　　处：《安徽农业大学学报》2024年第6期1004-1012,共9页Journal of Anhui Agricultural University

基　　金：现代农业产业技术体系建设专项(CARS-41-Z21);江苏省种业振兴揭榜挂帅项目(JBGS(2021)109);扬州市现代农业项目(YZ2023054);富民强村科技帮促项目(GY-BC202202)共同资助。

摘　　要：为探讨热应激对育成期至产蛋前期母鸡盲肠微生物的影响及其对家禽肠道微生物的作用机理,以苏禽3号为试验对象,对照组按照正常饲养程序进行饲养,热应激组在正常饲养的基础上,8~17周室内温度调至30℃。120日龄分别收集对照组和热应激组母鸡盲肠内容物,对其微生物的16Sr RNAV3-V4可变区进行Illumina高通量测序和生物信息学分析。结果表明:所有序列归属于15658个ASVs(amplicon sequence variant,ASV,扩增子序列变体),包含19个门、133个属。在门水平上,拟杆菌门(Bacteroidota)在热应激组和对照组占的比例均最大,分别为52.27%和49.95%。在属水平上,热应激组和对照组的优势菌群均为拟杆菌属(Bacteroides)、理研菌科RC9肠道菌属(Rikenellaceae_RC9_gut_group)、考拉杆菌属(Phascolarctobacterium)。与对照组相比,热应激组中拟杆菌属和考拉杆菌属的比例均呈上升趋势,而理研菌科RC9肠道菌属则呈下降趋势,属水平上有13个差异物种(P<0.05)。通过Alpha多样性分析可得知,热应激组的chao1指数(Chao1 richness)、香农指数(Shannon wiener diversity index)和辛普森指数(Simpson’s diversity index)均显著低于对照组水平(P<0.05),说明在热应激状态下鸡肠道内的菌群丰度和多样性均降低。通过PCo A分析,热应激组和对照组菌群结构差异显著(P<0.05);PICRUSt分析显示,热应激组与对照组之间在代谢和疾病通路有显著差异,热应激组在青霉素和头孢菌素的生物合成以及致病性大肠杆菌感染功能丰度均显著低于对照组。研究结果初步揭示了热应激状态下鸡的肠道菌群特征,热应激状态下,鸡盲肠内微生物会发生改变,调整肠道微生物结构,保护鸡肠道健康,该结果为进一步研发热应激状态下微生态制剂提供理论基础。To investigate the effects of thermal stress on cecum microbes of hens from growing to pre-natal period and the mechanisms of its effects on intestinal microorganisms of poultry,"Suqin-3"hens were selected as the experi-mental object in this study.The hens in the control group were fed according to standard feeding procedures,while the hens in the thermal stressed group experienced an indoor temperature adjustment to 30°C from weeks 8 to 17 in addi-tion to standard feeding conditions.At 120 days of age,hens’caecum contents were collected from both groups for Il-lumina high-throughput sequencing and bioinformatics analysis targeting the V3-V4 variable region of bacterial 16S rDNA gene.The results indicated that the sequences were classified into a total of 15658 ASVs,encompassing 19 phy-la and 133 genera.At the phylum level,Bacteroidetes bacteria exhibited dominance in both the thermal stressed group(52.27%)and control group(49.95%).At the genus level,prominent bacterial taxa in both groups included Bacteroides,Rikenellaceae_RC9_gut_group and Phascolarctobacterium.However,compared with the control group,the proportion of Bacteroides and Koala in the thermal stress group increased,and the RC9 intestinal bacteria of Riken family decreased,with 13 different species at the genus level(P<0.05).Through Alpha diversity analysis,it was observed that the chao1 index,Shannon Wiener diversity index,and Simpson’s diversity index of the thermal stress group exhibited a significant reduction compared to those of the control group(P<0.05),indicating a decline in microbial abundance and diversity within the chicken gut under thermal stress conditions.By PCoA analysis,the microflora structure of thermal stressed group and control group were significantly different(P<0.05).PICRUSt analysis showed that there were significant differences in metabolism and disease pathways between the thermal stress group and the control group,and the biosynthesis of penicillin and cephalosporin and the functional abundance of resistance to patho

关 键 词：鸡 盲肠 肠道微生物 高通量测序 

分 类 号：S858.31[农业科学—临床兽医学]