| 利巴韦林干预对肉鸡肠道微生物群落结构的影响 |
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| 引用本文： | 徐俊,侯玉洁,谢敏,王梦芝,邬磊,费丹,周瑶敏,胡丽芳.利巴韦林干预对肉鸡肠道微生物群落结构的影响[J].南方农业学报,2021,52(5):1139-1148. |
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| 作者姓名： | 徐俊 侯玉洁 谢敏 王梦芝 邬磊 费丹 周瑶敏 胡丽芳 |
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| 作者单位： | 1. 江西省农业科学院农产品质量安全与标准研究所/江西省农产品质量安全重点实验室, 南昌 330200;2. 南昌市农业科学院, 南昌 330008;3. 扬州大学动物科学与技术学院, 江苏扬州 225009 |
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| 基金项目： | 江西省重点研发计划项目（20192ACB60004）；江西省博士后科研基金项目（2015KY43） |
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| 摘 要： | 【目的】探讨矮脚黄鸡口服利巴韦林后其肠道微生物群落结构多样性的变化情况，揭示抗病毒药物对动物肠道健康的影响机制，为科学禁用抗生素及开发绿色生态饲料提供理论依据。【方法】90日龄矮脚黄鸡按10 mg/kg的剂量连续口服利巴韦林溶液7 d（试验组），以口服等量生理盐水为对照组，停药后收集4 h内的新鲜鸡粪。通过粪便基因组提取试剂盒提取鸡粪总DNA，根据细菌16S rDNA序列V3~V4可变区，设计通用引物338F和806R进行PCR扩增，然后参照Illumina MiSeq平台上机说明进行Illumina MiSeq高通量测序。【结果】从16个矮脚黄鸡粪便样品中共获得614626条原始序列（Raw reads），经质量控制后获得606862条优质序列（Clean reads），按97%的序列相似度进行OTU聚类分析获得968个OTUs，对照组和试验组共享624个OTUs。矮脚黄鸡肠道微生物群落丰富度指数（Chao1和ACE）表现为对照组显著（P<0.05，下同）或极显著（P<0.01，下同）高于试验组；在物种多样性方面，Simpson指数的组间差异不显著（P>0.05），Shannon指数则表现为试验组显著高于对照组。对照组和试验组的矮脚黄鸡粪便样品均以厚壁菌门、放线菌门、拟杆菌门和变形菌门为优势菌门，相对丰度均在5.00%以上；在属分类水平上，相对丰度大于0.10%的属共有108个，对照组和试验组均以乳杆菌属相对丰度最高，分别为34.07%和19.26%。矮脚黄鸡口服利巴韦林后，肠道中的乳杆菌属、肠球菌属、Paeniglutamicibacter、Tessaracoccus、埃希氏杆菌属、副球菌属、Gottschalkia及黄杆菌属呈显著或极显著下降趋势，而拟杆菌属、葡萄球菌属、叠球菌属、Jeotgalicoccus、Unclassified_f_rikenellaceae、Unclassifier_o_bacteroidales、Rikenellaceae_RC9_gut_group、明串珠菌属及丹毒丝菌属呈显著或极显著上升趋势。UniFrac-PCoA分析发现，第一主成分（PC1）和第二主成分（PC2）的贡献率分别为73.44%和6.99%，对照组和试验组间的微生物群落结构差异明显，彼此间可较好地区分开，但同处理组内的8个样品能很好地聚在一起，菌群相似度很高。【结论】口服利巴韦林会显著影响矮脚黄鸡肠道微生物区系，改变肠道微生物的稳态结构，进而导致其肠道健康受到损伤。
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| 关 键 词： | 矮脚黄鸡 利巴韦林 肠道微生物 丰富度 多样性 Illumina MiSeq高通量测序 |
| 收稿时间： | 2020-05-13 |
Effects of ribavirin intervention on intestinal microbial community structure in broilers |
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| XU Jun,HOU Yu-jie,XIE Min,WANG Meng-zhi,WU Lei,FEI Dan,ZHOU Yao-min,HU Li-fang.Effects of ribavirin intervention on intestinal microbial community structure in broilers[J].Journal of Southern Agriculture,2021,52(5):1139-1148. |
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| Authors: | XU Jun HOU Yu-jie XIE Min WANG Meng-zhi WU Lei FEI Dan ZHOU Yao-min HU Li-fang |
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| Affiliation: | 1. Institute of Quality Safety and Standard of Agricultural Products, Jiangxi Academy of Agricultural Sciences/Key Laboratory of Agro-product Quality and Safety of Jiangxi Province, Nanchang 330200, China;2. Nanchang Academy of Agricultural Sciences, Nanchang 330008, China;3. College of Animal Science and Technology, Yangzhou University, Yangzhou, Jiangsu 225009, China |
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| Abstract: | 【Objective】The structural diversity of the intestinal microbiological community in dwarf yellow chicken after oral ribavirin was discussed, the mechanism of the effects of antiviral drugs on animal intestinal health was revealed, and provided theoretical basis for scientific prohibition on the addition of antibiotics and the development of green ecological feed.【Method】Dwarf yellow chicken at 90 d were selected as experimental animal, and took ribavirin orally for 7 consecutive d according to the body weight dose of 10 mg/kg(treatment group), the same amount of normal saline was taken orally as control group. Manure samples were collected within 4 h after the drug stopped.The total chicken manure DNA was extracted from the fecal genome extraction kit, according to bacterial 16S rDNA sequence V3-V4 variable region, common primers 338F and 806R were designed for PCR amplification, then conducted Illumina MiSeq highthroughput sequencing with reference to the on-board Illumina MiSeq platform instructions.【Result】In total, 614626 raw reads were obtained for the bacterial 16S rRNA genes analysis. After screening these gene sequences with strict criteria, 606862 clean reads were obtained. OTU cluster analysis based on 97% sequence similarity obtained 968 OTUs. Additionally, there were 624 OTUs shared between the groups. The intestinal microbiological community richness indexes(Chao1 and ACE) in the control group were significant(P<0.05, the same below) or extremely significant(P<0.01, the same below) higher than in the treatment group. In species diversity, the Simpson index was not significant between groups(P>0.05), Shannon index in control group was significantly higher compared to treatment group. At the phylum level, Firmicutes, Actinobacteria, Bacteroidetes and Proteobacteria were the dominant phyla in both groups, with average relative abundance above 5.00%. At the genus level, there were 108 genera accounted for more than 0.10% relative abundance, and Lactobacillus accounted for the highest relative abundance in the control group and the treatment group, which were 34.07% and 19.26% respectively. After oral administration of ribavirin, Lactobacillus, Enterococcus, Paeniglutamicibacter, Tessaracoccus, Escherichia, Paracoccus, Gottschalkia and Flavobacterium significantly or extremely decreased, while Bacteroides, Staphylococcus, Methylosarcina, Jeotgalicoccus、Unclassified_f_rikenellaceae、Unclassifier_o_bacteroidales、Rikenellaceae_RC9_gut_group, Leuconostoc and Erysipelothrix significantly or extremely increased. The UniFrac-PCoA weighted principal coordinate analysis found that the primary component(PC1) and secondary component (PC2) contributed 73.44% and 6.99%, respectively. The microbial community structure difference between the control group and the treatment group was obvious, which could be better separated from each other, but the 8 samples in the treatment group could be well gathered together, and the flora similarity was very high.【Conclusion】Oral administration of ribavirin can significantly affect the intestinal microflora of dwarf yellow chicken, and change the steady-state structure of the intestinal microorganisms, which can cause the impaired of the intestinal health. |
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