Abstract：
Dietary anti-interleukin (IL)-10 antibodies may protect broiler performance during coccidiosis by inhibiting Eimeria host-evasion pathways; however, anti-IL-10\'s effects on microbial communities during coccidiosis and secondary Clostridium perfringens (necrotic enteritis) challenge is unknown. The study objectives were to assess the jejunal microbiota of broilers fed anti-IL-10 during E. maxima ± C. perfringens challenge. Two replicate studies using Ross 308 chicks placed in wire-floor cages (32 cages/ replicate study; 20 chicks/ cage) were conducted, with chicks assigned to diets ± 0.03% anti-IL-10 for 25 d. In both replicate studies, challenge-designated chicks were inoculated with 1 × 108Salmonella Typhimurium colony forming units (CFU) at placement. On d14, S. Typhimurium-inoculated chicks were gavaged with 15,000 sporulated Eimeria maxima M6 oocysts and half the E. maxima-challenged chicks received 1×108C. perfringens CFUs on d 18 and 19. Six chicks/ treatment were euthanized for distal jejunum content collection at baseline (d 14), 7 d post-inoculation (pi) with E. maxima/ 3 dpi with C. perfringens (peak) or 11 dpi with E. maxima/ 7 dpi with C. perfringens (post-peak) for 16S rRNA gene amplicon sequencing. Sequences were quality screened (Mothur V.1.43.0) and clustered into de novo operation taxonomical units (OTU; 99% similarity) using the SILVA reference database (v138). Alpha diversity and log-transformed relative abundance data were analyzed in SAS 9.4 with replicate study, diet, challenge, and timepoint main effects plus associated interactions (P ≤ 0.05). Few baseline changes were observed, but E. maxima ± C. perfringens challenge reduced Romboutsia and Staphylococcus relative abundance 4- to 800-fold in both replicate studies (P ≤ 0.008). At peak challenge with secondary C. perfringens, feeding anti-IL-10 instead of the control diet reduced Clostridium sensu stricto 1 relative abundance 13- and 1,848-fold in both replicate studies (P < 0.0001); however, OTUs identified as C. perfringens were not affected by dietary anti-IL-10. These results indicate that anti-IL-10 does not affect the jejunal microbiota of unchallenged broilers, while coccidiosis or necrotic enteritis challenge generally contributed to greater microbiota alterations than diet.
摘要：
饮食抗白细胞介素（IL）-10抗体可能通过抑制艾美耳球虫宿主逃避途径来保护球虫病期间的肉鸡性能；然而，抗IL-10对球虫病和继发性产气荚膜梭菌（坏死性肠炎）攻击期间微生物群落的影响尚不清楚。研究目的是评估在最大大肠杆菌±产气荚膜梭菌攻击期间饲喂抗IL-10的肉鸡的空肠微生物群。使用Ross308只小鸡放置在铁丝底笼中进行了两次重复研究（32笼/重复研究；20只小鸡/笼），小鸡被分配给±0.03%的抗IL-10饮食25d。在两个重复研究中，在放置时，用1×108鼠伤寒沙门氏菌菌落形成单位（CFU）接种攻击指定的小鸡。在d14，接种鼠伤寒沙门氏菌的雏鸡被15,000个孢子化的最大艾美耳球虫M6卵囊灌胃，一半的E.maxima攻击的雏鸡接受1×108C。产气荚膜CFU在d18和19。在基线（第14天）对六只小鸡/治疗进行安乐死以收集远端空肠内容物，接种后7d（pi）用E.maxima/3dpi用产气荚膜梭菌（峰）或用E.maxima/7dpi用产气荚膜梭菌（峰后）进行16SrRNA基因扩增子测序。使用SILVA参考数据库（v138）对序列进行质量筛选（MothurV.1.43.0），并将其聚类为从头操作分类单位（OTU；99％相似性）。在SAS9.4中通过重复研究分析了Alpha多样性和对数转换的相对丰度数据，饮食，挑战,和时间点主要效应加上相关相互作用（P≤0.05）。很少观察到基线变化，但在两项重复研究中，最大产气荚膜梭菌挑战使Romboutsia和葡萄球菌的相对丰度降低了4至800倍(P≤0.008)。在二级产气荚膜梭菌的高峰挑战中，在两个重复研究中，饲喂抗IL-10而不是对照饮食使严格的梭菌相对丰度降低了13和1,848倍（P<0.0001）；然而，鉴定为产气荚膜梭菌的OTU不受饮食抗IL-10的影响。这些结果表明，抗IL-10不影响未攻击的肉鸡的空肠微生物群，而球虫病或坏死性肠炎挑战通常导致比饮食更大的微生物群变化。
