复合抗菌肽对川中黑山羊瘤胃纤毛虫种群结构的影响

发布时间：2018-05-08 06:18

本文选题：复合抗菌肽 + 川中黑山羊　；参考：《农业生物技术学报》2017年10期

【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(P0.05),E组头毛属与D组无显著差异(P0.05),C组内毛属显著低于A组(P0.05),而E组内毛属与D组无显著差异(P0.05),E组多甲多泡双毛属显著高于C组(P0.05),而头毛属显著低于C组(P0.05)。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(P0.05)。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中提供理论依据。
[Abstract]:As a new feed additive, antimicrobial peptide (AMP) was rarely reported in ruminants. The effects of antimicrobial peptides on the population structure of Capra hircusi Ciliata were studied by high-throughput sequencing technique. Twelve male 4-month-old Sichuan black goats were randomly divided into 4 groups with 3 goats in each group. The normal concentrate group, the normal concentrate antimicrobial peptide group, the double concentrate group and the double concentration group were fed with 300300600 and 600g / d ~ (-1) C and E respectively. The samples of rumen juice were collected after 20 days of normal feeding, respectively, in which 3 g/kg compound antimicrobial peptides were added to the concentrate. After total DNA was extracted, the 18 s rRNA V4 region of eukaryote was amplified. The amplified products were sequenced using Illumina MiSeq platform. The results showed that a total of 471,580 high quality sequences were obtained, and 1,398 operational taxonomic units (OTU. 2) were classified as eukaryotes in 99.17% of the species notes. At the gate level, the highest relative abundance of group A and group C was Ciliophora ciliophora C46.0and C48.2, followed by the unknown position of Ciliophora ciliophora, followed by the unknown position of Ciliophora certae sediscus, the highest relative abundance of group D and E was the highest relative abundance of group D and group E: 49.933.The highest relative abundance was in the group D: 49.933.The next was D45.33members of Ciliophora, followed by the group D 45.33: e: 35.23. there was no significant difference between the two groups, and the highest relative abundance was in the group D: 49.933.The next was the Ciliophora Ciliophora, followed by Ciliophora Ciliophora, D45.3330. There was no significant difference between the two groups. A total of 9 ciliates were found to have the highest relative abundance in group A and group D, and the species genus of ciliates were polycilium and polyvesicular. In addition, the percentage of cephalocilium in group C was significantly higher than that in group A and group D. There was no significant difference between group A and group D in the genus Cephalocephala, and the genus Cephalocephala in group C was significantly lower than that in group A, but there was no significant difference between group E and group D in the genus Cilium, while in group E there was no significant difference between group E and group D. There was no significant difference in the alpha diversity of Chao, Ace, Shannon and Simpson Simpson index among the three groups (P 0.05), but the genus Cephalocephala was significantly lower than that of group C (P 0.05) (P < 0.05), but there was no significant difference in the indexes of Chao, Ace, Shannon and Simpson (P 0.05). The results showed that the highest relative abundance of rumen ciliates in young Sichuan black goats was in the genus Polygonum, and the compound antimicrobial peptide could increase the content of Cephalocephala and decrease the content of Intratheca, and it was more significant in the normal concentrate. The population structure of ciliates did not change significantly from 300 g / d to 600 g / d, and the diversity of eukaryote in rumen was not affected by the amount of complex antimicrobial peptides and concentrate in this experiment. The results provide theoretical basis for the application of complex antimicrobial peptides in ruminant production.
【作者单位】：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;
【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)
【分类号】：S827.5

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