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

doi:10.11843/j.issn.0366-6964.2020.10.028

Abstract

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.

Key words: dog, chronic renal failure, gut microbiota, 16S rDNA sequencing, gut microbiota diversity, gene function prediction, intestinal microbial marker

CLC Number:

S858.2926.59

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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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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