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含或不含泰乐菌素的育肥日粮喂养的杂交肉牛和荷斯坦公牛肝脓肿脓性材料的细菌群落分析。

Bacterial community analysis of purulent material from liver abscesses of crossbred cattle and Holstein steers fed finishing diets with or without tylosin.

机构信息

Department of Clinical Sciences, Kansas State University, Manhattan, KS 66506, USA.

School of Biological Sciences, University of Nebraska-Lincoln, Lincoln, NE 68583, USA.

出版信息

J Anim Sci. 2021 Apr 1;99(4). doi: 10.1093/jas/skab076.

Abstract

Liver abscesses in feedlot cattle are polymicrobial infections. Culture-based studies have identified Fusobacterium necrophorum as the primary causative agent, but a number of other bacterial species are frequently isolated. The incidence of liver abscesses is highly variable and is affected by a number of factors, including cattle type. Holstein steers raised for beef production have a higher incidence than crossbred feedlot cattle. Tylosin is the commonly used antimicrobial feed additive to reduce the incidence of liver abscesses. The objective of this study was to utilize 16S ribosomal RNA amplicon sequence analyses to analyze the bacterial community composition of purulent material of liver abscesses of crossbred cattle (n = 24) and Holstein steers (n = 24), each fed finishing diet with or without tylosin. DNA was extracted and the V3 and V4 regions of the 16S rRNA gene were amplified, sequenced, and analyzed. The minimum, mean, and maximum sequence reads per sample were 996, 177,070, and 877,770, respectively, across all the liver abscess samples. Sequence analyses identified 5 phyla, 14 families, 98 genera, and 102 amplicon sequence variants (ASV) in the 4 treatment groups. The dominant phyla identified were Fusobacteria (52% of total reads) and Proteobacteria (33%). Of the top 25 genera identified, 17 genera were Gram negative and 8 were Gram positive. The top 3 genera, which accounted for 75% of the total reads, in the order of abundance, were Fusobacterium, Pseudomonas, and Bacteroides. The relative abundance, expressed as percent of total reads, of phyla, family, and genera did not differ (P > 0.05) between the 4 treatment groups. Generic richness and evenness, determined by Shannon-Weiner and Simpson's diversity indices, respectively, did not differ between the groups. The UniFrac distance matrices data revealed no clustering of the ASV indicating variance between the samples within each treatment group. Co-occurrence network analysis at the genus level indicated a strong association of Fusobacterium with 15 other genera, and not all of them have been previously isolated from liver abscesses. In conclusion, the culture-independent method identified the bacterial composition of liver abscesses as predominantly Gram negative and Fusobacterium as the dominant genus, followed by Pseudomonas. The bacterial community composition did not differ between crossbred and Holstein steers fed finishing diets with or without tylosin.

摘要

牛肝脓肿是一种多微生物感染。基于培养的研究已经确定坏死梭杆菌(Fusobacterium necrophorum)是主要的致病因子,但也经常分离到许多其他细菌种类。肝脓肿的发病率差异很大,受多种因素影响,包括牛的种类。用于生产牛肉的荷斯坦公牛的发病率高于杂交育肥牛。泰乐菌素是一种常用的抗菌饲料添加剂,可降低肝脓肿的发病率。本研究的目的是利用 16S 核糖体 RNA 扩增子序列分析来分析杂交牛(n = 24)和荷斯坦公牛(n = 24)脓性肝脓肿的细菌群落组成,两组均用或不用泰乐菌素喂养育肥日粮。提取 DNA,扩增、测序和分析 16S rRNA 基因的 V3 和 V4 区。所有肝脓肿样本的最小、平均和最大序列读数分别为 996、177070 和 877770。序列分析在 4 个治疗组中鉴定出 5 个门、14 个科、98 个属和 102 个扩增子序列变异(ASV)。鉴定出的主要门是梭杆菌门(Fusobacteria)(占总读数的 52%)和变形菌门(Proteobacteria)(占总读数的 33%)。在所鉴定的前 25 个属中,17 个属为革兰氏阴性菌,8 个属为革兰氏阳性菌。丰度最高的 3 个属,按丰度顺序排列,依次为梭杆菌属(Fusobacterium)、假单胞菌属(Pseudomonas)和拟杆菌属(Bacteroides)。4 个治疗组之间的门、科和属的相对丰度(表示为总读数的百分比)没有差异(P > 0.05)。由 Shannon-Weiner 和 Simpson 多样性指数分别确定的丰富度和均匀度没有差异。UniFrac 距离矩阵数据显示,ASV 没有聚类,表明每个治疗组内的样本之间存在差异。属水平的共现网络分析表明,梭杆菌属与 15 个其他属有很强的关联,并非所有这些属以前都从肝脓肿中分离出来。总之,这种非培养方法确定了肝脓肿的细菌组成主要为革兰氏阴性菌,其中梭杆菌属(Fusobacterium)是优势属,其次是假单胞菌属(Pseudomonas)。用或不用泰乐菌素喂养育肥日粮的杂交牛和荷斯坦公牛的细菌群落组成没有差异。

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