Differential expression of microRNAs in the caecal content and faeces of broiler chickens experimentally infected with Eimeria.
Differential expression of microRNAs in the caecal content and faeces of broiler chickens experimentally infected with Eimeria.
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实验感染艾美球虫的肉鸡盲肠内容物和粪便中 microRNA 的差异表达。
DOI：
10.1080/03079457.2022.2076581
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发表时间：
2022
期刊：
影响因子：
--
通讯作者：
Williams J
中科院分区：
文献类型：
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作者：
Williams J
球虫病引起的。在鸡中引起显著的发病率和死亡率，因此具有重要的经济意义。死后肠病变评分仍然是最常见的诊断手段之一；因此，替代的、非侵入性的诊断和监测方法将是非常可取的。微rna （miRNAs）已被证明在人类和动物的粪便中是稳定的，但在胃肠道疾病中表达改变。我们假设miRNA在鸡盲肠内容物中是稳定的，并且在感染亚梅里亚病毒的个体和未感染的个体中可以看到不同的miRNA表达模式。最初，从感染（n= 3；感染后7天）和未感染（n= 3）的鸡盲肠内容物中提取RNA，以证明miRNA的稳定性。随后，对me的盲肠内容物进行下一代miRNA测序。高病变评分（LS） 3-4的鸡；n= 3)或低（LS1;n= 3）病理水平，和未感染对照（n= 3）。对比分析发现19个mirna在大肠杆菌的盲肠内容物中表现出显著的表达改变。感染鸡与未感染鸡之间的差异（t检验，错误发现率< 0.05）。其中8个mirna在感染中显著上调（9.8 ~ 105倍，FDR <0.05）。使用单独的生物重复进行定量PCR，以确认这些miRNA候选物在盲肠和粪便内容物中的差异调控。这项工作已经确定了一组候选miRNA，这些候选miRNA可能适合用作活动性盲肠球虫病的非侵入性粪便标记物，而无需剔除。研究HIGHLIGHTSE。tenella诱导的盲肠内容物和粪便中miRNA的差异表达。
Coccidiosis caused byEimeriaspp. incurs significant morbidity and mortality in chickens, and is thus of great economic importance.Post-mortemintestinal lesion scoring remains one of the most common means of diagnosis; therefore alternative, non-invasive methods of diagnosis and monitoring would be highly desirable. Micro-RNAs (miRNAs) have been shown to be stable in faeces of human and animal species with expression altered in gastrointestinal disease. We hypothesized that miRNA is stable in caecal content of chickens, and that differential miRNA expression patterns would be seen inEimeria-infected versus uninfected individuals. Initially, RNA was extracted fromEimeria tenella-infected (n= 3; 7 days post infection) and uninfected (n= 3) chicken caecal content to demonstrate miRNA stability. Subsequently, next-generation miRNA sequencing was performed on caecal content fromE. tenella-infected chickens with high (lesion score (LS) 3–4;n= 3) or low (LS1;n= 3) levels of pathology, and uninfected controls (n= 3). Comparative analysis identified 19 miRNAs that exhibited significantly altered expression in the caecal content ofE. tenella, infected chickens versus uninfected chickens (t-test, False Discovery Rate (FDR) < 0.05). Eight of these miRNAs showed significant up-regulation in infection (fold change of 9.8–105, FDR <0.05). Quantitative PCR was performed using separate biological replicates to confirm differential regulation in eight of these miRNA candidates in caecal and faecal content. This work has identified a panel of miRNA candidates which may be appropriate for use as non-invasive faecal markers of active caecal coccidiosis without the need for culling.RESEARCH HIGHLIGHTSE. tenellainduced differential miRNA expression in caecal content and faeces.