Hot topic: 16S rRNA gene sequencing reveals the microbiome of the virgin and pregnant bovine uterus.
Hot topic: 16S rRNA gene sequencing reveals the microbiome of the virgin and pregnant bovine uterus.
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DOI：
10.3168/jds.2017-12592
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发表时间：
2017-06
影响因子：
3.5
通讯作者：
Lucy MC
中科院分区：
文献类型：
--
作者：
Moore SG;Ericsson AC;Poock SE;Melendez P;Lucy MC
我们通过对处女和妊娠牛子宫进行16 S rRNA基因测序，检验了处女小母牛和妊娠牛子宫具有常驻微生物组的假设。采用无菌技术对10头发情期处女母牛的子宫内膜和5头妊娠母牛的羊水、胎盘组织、子叶间胎盘、宫颈腔和宫颈外表面（对照）进行取样。提取DNA，扩增16 S rRNA基因的V4高变区，并使用Illumina MiSeq技术（Illumina Inc.，加利福尼亚州圣地亚哥）。使用Qiime v1.8软件从序列生成操作分类单位（OTU），并使用Greengenes数据库分配分类。使用单变量（混合模型）和多变量（置换多变量ANOVA）程序测试组织对妊娠子宫内微生物组成的影响。在所有样品中均产生了16 S rRNA基因的扩增子，支持了未交配小母牛和怀孕母牛的子宫含有微生物组的论点。平均而言，在胎盘节、处女子宫内膜、羊水、子宫颈腔、子叶间胎盘和子宫颈外表面中分别产生53、199、380、382、525和13，589个读数，注释为16、35、43、63、48和176个OTU。处女小母牛和怀孕母牛子宫中最丰富的3个门是厚壁菌门拟杆菌门和变形菌门，它们分别占序列的约40%、35%和10%。门丰度是相似的怀孕子宫的组织之间。主成分分析、Bray-Curtis相似性指数的单向PERMANOVA分析以及Shannon多样性指数和Chao 1指数的混合模型分析表明，对照组织（宫颈外表面）的微生物组与羊水、子叶间胎盘和胎盘组织的微生物组显著不同。有趣的是，许多与产后子宫疾病相关的细菌物种（即，真孢霉属，不动杆菌属，梭杆菌属，变形杆菌属，普雷沃菌属，和消化链霉菌属（Peptostreptum spp.）也存在于处女小母牛和怀孕母牛的子宫中。目前研究的样本中存在16 S rRNA基因序列读数表明，子宫微生物组在女性达到生殖成熟时建立，并且在子宫微生物组存在的情况下建立和维持妊娠。
We tested the hypothesis that the uterus of virgin heifers and pregnant cows possessed a resident micro-biome by 16S rRNA gene sequencing of the virgin and pregnant bovine uterus. The endometrium of 10 virgin heifers in estrus and the amniotic fluid, placentome, intercotyledonary placenta, cervical lumen, and external cervix surface (control) of 5 pregnant cows were sampled using aseptic techniques. The DNA was extracted, the V4 hypervariable region of the 16S rRNA gene was amplified, and amplicons were sequenced using Illumina MiSeq technology (Illumina Inc., San Diego, CA). Operational taxonomic units (OTU) were generated from the sequences using Qiime v1.8 software, and taxonomy was assigned using the Greengenes database. The effect of tissue on the microbial composition within the pregnant uterus was tested using univariate (mixed model) and multivariate (permutational multivariate ANOVA) procedures. Amplicons of 16S rRNA gene were generated in all samples, supporting the contention that the uterus of virgin heifers and pregnant cows contained a microbiome. On average, 53, 199, 380, 382, 525, and 13,589 reads annotated as 16, 35, 43, 63, 48, and 176 OTU in the placentome, virgin endometrium, amniotic fluid, cervical lumen, intercotyledonary placenta, and external surface of the cervix, respectively, were generated. The 3 most abundant phyla in the uterus of the virgin heifers and pregnant cows were Firmicutes Bacteroidetes, and Proteobacteria, and they accounted for approximately 40, 35, and 10% of the sequences, respectively. Phyla abundance was similar between the tissues of the pregnant uterus. Principal component analysis, one-way PERMANOVA analysis of the Bray-Curtis similarity index, and mixed model analysis of the Shannon diversity index and Chao1 index demonstrated that the microbiome of the control tissue (external surface of the cervix) was significantly different from that of the amniotic fluid, intercotyledonary placenta, and placentome tissues. Interestingly, many bacterial species associated with postpartum uterine disease (i.e., Trueperella spp., Acinetobacter spp., Fusobacteria spp., Proteus spp., Prevotella spp., and Peptostreptococcus spp.) were also present in the uterus of virgin heifers and of pregnant cows. The presence of 16S rRNA gene sequence reads in the samples from the current study suggests that the uterine microbiome is established by the time a female reaches reproductive maturity, and that pregnancies are established and maintained in the presence of a uterine microbiome.
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影响因子：
3.7
作者：
Hart ML;Meyer A;Johnson PJ;Ericsson AC
通讯作者：
Ericsson AC
影响因子：
3.6
作者：
Sheldon IM;Cronin J;Goetze L;Donofrio G;Schuberth HJ
通讯作者：
Schuberth HJ
影响因子：
17.1
作者：
Aagaard K;Ma J;Antony KM;Ganu R;Petrosino J;Versalovic J
通讯作者：
Versalovic J
DOI：
10.1073/pnas.1000080107
发表时间：
2011-03-15
影响因子：
11.1
作者：
Caporaso, J. Gregory;Lauber, Christian L.;Knight, Rob
通讯作者：
Knight, Rob
影响因子：
4.4
作者：
Jeon, Soo Jin;Vieira-Neto, Achilles;Galvao, Klibs N.
通讯作者：
Galvao, Klibs N.