复合抗菌肽对川中黑山羊瘤胃纤毛虫种群结构的影响  

Effects of Antibacterial Peptides on Rumen Ciliate(Rumen ciliata)Community Structure in Chuanzhong Black Goat(Capra hircus)

作　　者：刘旗[1] 陈芸[1] 邓俊良[1] 杨颜铱 高爽[1] 陈憧[1] 姚淑华[1] 

机构地区：[1]四川农业大学动物医学院/动物疫病与人类健康四川省重点实验室/环境公害与动物疾病四川省高校重点实验室,温江611130

出　　处：《农业生物技术学报》2017年第10期1689-1696,共8页Journal of Agricultural Biotechnology

基　　金："长江学者和创新团队发展计划"创新团队项目(No.IRT0848);四川农业大学双支计划(No.03572070)

摘　　要：抗菌肽(antimicrobial peptide,AMP)作为新兴的饲料添加剂,在反刍动物中的应用鲜有报道,本实验应用高通量测序技术研究复合抗菌肽对山羊(Capra hircus)瘤胃纤毛虫(Ciliata)种群结构的影响。选取12只雄性4月龄川中黑山羊,随机分成4组,每组各3头。正常精料组(A),正常精料抗菌肽组(C),双倍精料组(D),双倍精料抗菌肽组(E)分别饲喂精料300、300、600和600 g/d,C和E组在精料中添加3 g/kg复合抗菌肽,正常饲喂20 d后采集瘤胃液样品,提取样品总DNA后,扩增真核生物18S rRNA V4区,扩增产物使用Illumina MiSeq平台测序。结果表明,1)共获得高质量序列471 580条,聚类后共得到1 398个运算分类单位(OTU)。2)所得OTU经物种注释99.17%被归类为真核生物界,门水平上,A组和C组相对丰度最高的为纤毛门(Ciliophora)(A:46.0%;C:48.2%),其次为分类位置未知(incertae sedis)(A:35.5%;C:42.83%),D组与E组相对丰度最高的为分类位置未知(D:49.93%;E:60.27%),其次为纤毛门(D:45.33%;E:35.23%);组间无显著差异。3)属水平分类上,共发现9个纤毛虫种属,A组与D组相对丰度最高的纤毛虫种属为多加多泡双毛属;另外,C组头毛属占总纤毛虫比例显著高于A组(P<0.05),E组头毛属与D组无显著差异(P>0.05),C组内毛属显著低于A组(P<0.05),而E组内毛属与D组无显著差异(P>0.05),E组多甲多泡双毛属显著高于C组(P<0.05),而头毛属显著低于C组(P<0.05)。5)各组在alpha多样性赵氏Chao、艾斯ACE、香浓Shannon和辛普森Simpson指数上差异不显著(P>0.05)。本实验结果表明,幼龄川中黑山羊瘤胃纤毛虫相对丰度最高的种属为多加多泡双毛属;复合抗菌肽可提高头毛属含量,降低内毛属含量,且在正常精料中较为显著。精料饲喂量从300 g/d改变为600 g/d时纤毛虫种群结构无显著变化;另外,瘤胃内真核生物多样性不受本实验下复合抗菌肽及精料量的影响。本研究结果为复合抗菌肽应用于反刍动物生产Antimicrobial peptide（AMP）, a burgeoning feed additive, is rarely reported when it apllies to the ruminant. The aim of this study was to evaluate the effects of antibacterial peptide （AMP） on rumen ciliate community structure by MiSeq high throughput sequencing technology. Twelve male Chuanzhong black goats （4-month-old） were randomly divided into 4 groups according to the similar weight. Four groups were normal concentrate group （A 300g/d）, normal concentrate＋AMP group （C 300g/d＋3 g/kg）, double concentrate group （D 600 g/d） and double concentrate＋AMP group （E 600 g/d＋3 g/kg）, The rumen fluid samples were collected after 20 d, and then the total DNA were extraced for amplification 18S rRNA（547F-V4R）, sequencing by MiSeq Illumina250. The results showed that： 1）A total of 471 580 valid 18S rDNA sequences and 1 398 operational taxonomic unit （OTU） across 12 samples were obtained. 2） The 99.17% of all OTUs belonged to eukaryotes, At phylum level, Ciliophora was the most abundant phyla in A and C groups（A： 46.0%; C： 48.2%）, followed by incertae sedis （A： 35.5%; C： 42.83%）; On the contrary, incertae sedis was the most abundant phyla in D and E groups （D： 49.93%; E： 60.27%）, followed by Ciliophora （D： 45.33%; E： 35.23%）, and no significant difference between the groups. 3） At family level, only two families （Ophryoscolecidae, Isotrichidae） were found belong to ciliates. And the differences between the groups were not obvious. 4） At genus level, a total of nine different ciliates genera were detected in every last sample. And the content of Ophryoseolex in C group were significantly higher than A group （P〈0.05）, but no significant difference between D and E （P〉0.05）; the content of Entodinium in C group are significant lower than A group （P〈 0.05）, but there have not significant difference between D and E （P〉0.05）; The content of Polyplastron in E group are significant higher than C group（P〈0.05）, and t

关 键 词：复合抗菌肽 川中黑山羊 瘤胃纤毛虫 种群结构 MiSeq测序 

分 类 号：S815.4[农业科学—畜牧学]