| 高通量测序研究慢性肾衰竭对宠物犬肠道菌群多样性的影响及基因功能预测分析 |
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| 引用本文： | 陆江,朱道仙,赵学刚,刘莉.高通量测序研究慢性肾衰竭对宠物犬肠道菌群多样性的影响及基因功能预测分析[J].畜牧兽医学报,2020,51(10):2590-2598. |
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| 作者姓名： | 陆江 朱道仙 赵学刚 刘莉 |
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| 作者单位： | 1. 江苏农牧科技职业学院宠物科技学院, 泰州 225300;2. 江苏农牧科技职业学院动物医学院, 泰州 225300 |
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| 基金项目： | 江苏高校“青蓝工程”资助项目（00000217009）；江苏农牧科技职业学院院级项目（NSF201706）；江苏省大学生创新创业训练计划项目（201812806015Y） |
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| 摘 要： | 旨在通过高通量测序研究慢性肾衰竭（chronic renal failure，CRF）对宠物犬肠道菌群多样性的影响，并预测分析菌群的基因功能。本研究选取30只2岁健康犬，随机等分为慢性肾衰竭模型组（CRF，肾动脉结扎法制备）、假手术对照组（Sham）和健康对照组（HCG），常规饲养，试验周期56 d。试验过程中，定期检测血肌酐（Scr）、血尿素氮（BUN）和尿蛋白/肌酐比（UP/C），无菌采集新鲜粪便进行细菌16S rDNA测序，分析慢性肾衰竭对犬肠道菌群结构、多样性与功能的影响，并以CRF组差异菌群为自变量进行主成分Logistic回归分析，构建菌群标志物综合指数（FMI）用于慢性肾衰竭预测。结果显示：1）从试验后第28天开始，CRF组Scr、BUN和UP/C分别显著高于自身试验前、HCG组和Sham组（P<0.05）。2） CRF组不同时间点肠道菌群有显著差异，第56天时的Chao 1指数、Shannon指数均降低，拟杆菌门（Bacteroidetes）、变形杆菌门（Proteobacteria）和放线菌门（Actinobacteria）的相对丰度增加，厚壁菌门（Firmicutes）数量减少，与0 d和28 d比较差异显著（P<0.05），与同时间点HCB组及Sham组比较差异显著（P<0.05）。3）组间菌群LEfSe分析发现，CRF组富集了20个物种，主要包括假单胞菌属（Pseudomonas）、埃希氏菌属（Escherichia）和变形杆菌属（Proteus）等，这些物种与肠道其他菌属间多呈负相关关系；差异物种基因主要富集在氨基酸代谢、糖生物合成和代谢以及碳水化合物代谢等代谢途径。用这些差异物种构建的FMI，其ROC曲线AUC为0.788，可作为犬CRF的肠道微生物标志物。以上结果表明，慢性肾衰竭可以使犬肠道菌群多样性降低、菌群结构失调以及菌群功能改变，且CRF患犬肠道中富集的菌群可作为该病的肠道微生物标志物，以FMI预测效果最好。
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| 关 键 词： | 犬 慢性肾衰竭 肠道菌群 16S rDNA测序 菌群多样性 基因功能预测 肠道微生物标志物 |
| 收稿时间： | 2020-03-19 |
High Throughput Sequencing to Study the Effect of Chronic Renal Failure on the Diversity and Gene Function Prediction of Gut Microbiota in Dogs |
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| LU Jiang,ZHU Daoxian,ZHAO Xuegang,LIU Li.High Throughput Sequencing to Study the Effect of Chronic Renal Failure on the Diversity and Gene Function Prediction of Gut Microbiota in Dogs[J].Acta Veterinaria et Zootechnica Sinica,2020,51(10):2590-2598. |
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| Authors: | LU Jiang ZHU Daoxian ZHAO Xuegang LIU Li |
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| Institution: | 1. Department of Pet Science and Technology, Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China;2. Department of Animal Medicine, Jiangsu Agri-animal Husbandry Vocational College, Taizhou 225300, China |
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| Abstract: | This study aimed to investigate the effect of chronic renal failure(CRF) on the gut microbiota diversity and predict the gene function of the flora. A total of 30 2-year-old dogs were selected and randomly divided into the chronic renal failure model group (CRF, established by renal artery ligation), sham operation control group (Sham) and healthy control group (HCG). All animals were fed normally during the 56 days of test period. The serum creatinine (Scr), blood urea nitrogen (BUN) and urine protein / creatinine ratio (UP/C) were detected regularly during the experiment. The effects of chronic renal failure on the structure, diversity and function of gut microbiota were analyzed according to the result of bacterial 16S rDNA sequencing from fresh feces collected without contamination. The flora markers index (FMI) was constructed based on the principal component Logistic regression analysis of different microbiota in CRF group to predict the development of chronic renal failure. The results showed that: 1) The levels of Scr, BUN and UP/C in CRF group from the start of the 28th day of the experiment were significantly higher than those of HCG group and Sham group (P<0.05), and significantly higher than that of the first day of CRF group (P<0.05). 2) Compared with those before chronic renal failure (CRF group at day 0 and 28), HCG group and Sham group, the Chao 1 diversity and Shannon diversity of gut microbiota in CRF group at day 56 were significantly lower (P<0.05), while the relative abundance of Bacteroidetes, Proteobacteria and Actinobacteria significantly increased and the number of Firmicutes significantly decreased (P<0.05). 3) LEfSe analysis showed that 20 species were enriched in CRF group, mainly including Pseudomonas, Escherichia, Proteus and so on, and most of them had negative correlation with other intestinal bacteria. Functional prediction revealed that genes of those different species were mainly enriched in amino acid metabolism, sugar biosynthesis and metabolism and carbohydrate metabolism in CRF group. The area under ROC curve (AUC) of the FMI constructed with those species enriched was 0.788, which could be used as the intestinal microbial marker for CRF in dogs. In summary, chronic renal failure can reduce the diversity of intestinal microbiota, lead to the imbalance of bacterial structure and change of bacterial function. Moreover, the enriched gut microbiota in CRF group can be used as the intestinal microbial marker of CRF in dogs， and the best prediction effect can be obtained by FMI. |
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| Keywords: | dog chronic renal failure gut microbiota 16S rDNA sequencing gut microbiota diversity gene function prediction intestinal microbial marker |
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