Department of Animal Science, The Ohio State University, Columbus, OH.
Department of Poultry Science, University of Arkansas, Fayetteville, AR.
Poult Sci. 2020 Mar;99(3):1254-1266. doi: 10.1016/j.psj.2019.10.017. Epub 2020 Jan 30.
Pioneer colonization of the gastrointestinal tract (GIT) by bacteria is thought to have major influence on neonatal tissue development. Previous studies have shown in ovo inoculation of embryos with saline (S), species of Citrobacter (C, C2), or lactic acid bacteria (L) resulted in an altered microbiome on day of the hatch (DOH). The present study investigated GIT proteomic changes at DOH in relation to different inoculations. Embryos were inoculated in ovo with S or ∼10 cfu of C, C2, or L at 18 embryonic days. On DOH, the GIT was collected, and tissue proteins were extracted for analysis via tandem mass spectrometry. A total of 493 proteins were identified for differential comparison with S at P ≤ 0.10. Different levels were noted in 107, 39, and 78 proteins in C, C2, and L groups, respectively, which were uploaded to Ingenuity Pathway Analysis to determine canonical pathways and biological functions related to these changes. Three members of the cytokine family (interleukin [IL]-1β, IL6, and Oncostatin M) were predicted to be activated in C2, indicated with Z-score ≥ 1.50, which suggested an overall proinflammatory GIT condition. This was consistent with the activation of the acute-phase response signaling pathway seen exclusively in C2 (Z-score = 2.00, P < 0.01). However, activation (Z-score = 2.00) of IL-13, upregulation of peroxiredoxin-1 and superoxide dismutase 1, in addition to activation of nitric oxide signaling in the cardiovascular system of the L treatment may predict a state of increased antioxidant capacity and decreased inflammatory status. The nuclear factor erythroid 2-related factor 2 (NRF2)-mediated oxidative stress response (Z-score = 2.00, P < 0.01) was predicted to be upregulated in C which suggested that chicks were in an inflammatory state and associated oxidative stress, but the impact of these pathways differed from that of C2. These changes in the proteome suggest that pioneer colonizing microbiota may have a strong impact on pathways associated with GIT immune and cellular development.
细菌对胃肠道（GIT）的先驱性定植被认为对新生儿组织发育有重大影响。先前的研究表明，在胚胎期向胚胎中用生理盐水（S）、柠檬酸杆菌（C、C2）或乳酸菌（L）种接种会导致孵化日（DOH）时微生物组发生改变。本研究调查了与不同接种相关的 DOH 时 GIT 的蛋白质组变化。胚胎在 18 天胚龄时用 S 或约 10cfu 的 C、C2 或 L 进行卵内接种。在 DOH 时，收集 GIT 并提取组织蛋白进行串联质谱分析。与 S 相比，在 P≤0.10 时共鉴定出 493 种差异蛋白。在 C、C2 和 L 组中，分别有 107、39 和 78 种蛋白的水平不同，这些蛋白被上传到 Ingenuity Pathway Analysis 以确定与这些变化相关的经典途径和生物学功能。细胞因子家族的三个成员（白细胞介素[IL]-1β、IL6 和 Oncostatin M）被预测在 C2 中被激活，其 Z 分数≥1.50，表明 GIT 存在整体炎症反应状态。这与仅在 C2 中观察到的急性期反应信号通路的激活一致（Z 分数=2.00，P<0.01）。然而，在 L 处理中，IL-13 的激活（Z 分数=2.00）、过氧化物酶 1 和超氧化物歧化酶 1 的上调以及心血管系统中一氧化氮信号的激活可能预示着抗氧化能力增加和炎症状态降低的状态。核因子红细胞 2 相关因子 2（NRF2）介导的氧化应激反应（Z 分数=2.00，P<0.01）被预测在 C 中上调，这表明小鸡处于炎症状态和相关的氧化应激状态，但这些途径的影响与 C2 不同。蛋白质组的这些变化表明，先驱定植的微生物群可能对与 GIT 免疫和细胞发育相关的途径有重大影响。
