Department of Animal Science, Iowa State University, Ames, IA 50011, USA.
Department of Animal Science, Iowa State University, Ames, IA 50011, USA; Interdepartmental Microbiology Graduate Program, Iowa State University, Ames, IA 50011, USA.
Poult Sci. 2024 Sep;103(9):104001. doi: 10.1016/j.psj.2024.104001. Epub 2024 Jun 22.
Strategies to counteract interleukin (IL)-10-mediated immune evasion by Eimeria spp. during coccidiosis- like anti-IL-10 antibodies- may protect broiler chicken health and reduce incidence of secondary necrotic enteritis (Clostridium perfringens) via undetermined mechanisms. Objectives were to use sequencing techniques to evaluate jejunal microbial community composition and function in anti-IL-10-fed broilers during coccidiosis and necrotic enteritis. On d0, Ross 308 chicks were placed in 32 cages (15 chicks/ cage) for a 25-d study and randomly assigned to diets ± 0.03% anti-IL-10. Six chicks/ diet were euthanized for distal jejunum content and tissue collection on d 14 (baseline) before inoculating the remainder with saline or 15,000 E. maxima oocysts (M6 strain). Half the chicks challenged with E. maxima were challenged with C. perfringens (1×10 colony forming units) on d 18 and 19. Follow-up samples (6 chicks/treatment) were collected at 7 and 11 d postinoculation (pi) for the E. maxima-only group, or 3 and 7 dpi for the E. maxima + C. perfringens group with 3/7 dpi being designated as peak and 7/11dpi as postpeak challenge. DNA was extracted from digesta for microbiota composition analysis (16S rRNA gene sequencing) while RNA was extracted from tissue to evaluate the metatranscriptome (RNA sequencing). Alpha diversity and genus relative abundances were analyzed using the diet or challenge main effects with associated interactions (SAS 9.4; P ≤ 0.05). No baseline microbial changes were associated with dietary anti-IL-10. At peak challenge, a diet main effect reduced observed species 36.7% in chicks fed anti-IL-10 vs. control; however, the challenge effect reduced observed species and Shannon diversity 51.2-58.3% and 33.0 to 35.5%, respectively, in chicks challenged with E. maxima ± C. perfringens compared to their unchallenged counterparts (P ≤ 0.05). Low sequencing depth limited metatranscriptomic analysis of jejunal microbial function via RNA sequencing. This study demonstrates that challenge impacted the broiler distal jejunum microbiota more than anti-IL-10 while future research to characterize the microbial metatranscriptome may benefit from investigating other intestinal compartments.
抗白细胞介素 (IL)-10 抗体可中和艾美耳球虫在球虫病期间的免疫逃避策略-可能通过未确定的机制保护肉鸡健康并降低继发坏死性肠炎 (产气荚膜梭菌) 的发病率。本研究旨在使用测序技术评估抗白细胞介素-10 喂养的肉鸡在球虫病和坏死性肠炎期间空肠微生物群落组成和功能。在 d0，将罗斯 308 小鸡放置在 32 个笼子中（每个笼子 15 只小鸡）进行为期 25 天的研究，并随机分配给饮食 ± 0.03%的抗白细胞介素-10。在给其余小鸡接种生理盐水或 15,000 个柔嫩艾美耳球虫卵囊 (M6 株) 之前，于 d 14（基线）处死每组 6 只小鸡以采集远端空肠内容物和组织。用产气荚膜梭菌（1×10 个菌落形成单位）在 d 18 和 19 对一半感染柔嫩艾美耳球虫的小鸡进行攻毒。对仅感染柔嫩艾美耳球虫的小鸡在接种后 7 和 11 天（dpi）进行后续样本采集（每组 6 只小鸡），对感染柔嫩艾美耳球虫和产气荚膜梭菌的小鸡在 3 和 7dpi 进行后续样本采集，3dpi 和 7dpi 分别指定为攻毒高峰和攻毒后高峰。用 16S rRNA 基因测序分析消化物中的微生物群落组成（微生物组学分析），用 RNA 测序分析组织中的宏转录组（转录组学分析）。用饮食或攻毒主效应及其相关交互作用（SAS 9.4；P≤0.05）分析 alpha 多样性和属相对丰度。与对照组相比，抗白细胞介素-10 组小鸡在攻毒高峰时，饮食主效降低了观察到的物种 36.7%；然而，与未攻毒的小鸡相比，攻毒使感染柔嫩艾美耳球虫和产气荚膜梭菌的小鸡观察到的物种和香农多样性分别降低了 51.2-58.3%和 33.0-35.5%（P≤0.05）。测序深度低限制了通过 RNA 测序对空肠微生物功能的宏转录组学分析。本研究表明，与抗白细胞介素-10 相比，攻毒对肉鸡空肠远端微生物的影响更大，而未来对微生物宏转录组进行特征分析的研究可能受益于对其他肠道部位的研究。
