本试验旨在研究不同品系德州驴(三粉驴和乌头驴)血浆生化指标和肠道菌群结构的差异。随机选择3~5岁、体重260~300 kg健康空怀三粉驴母驴和乌头驴母驴各6头,颈静脉采集血液,全自动生化分析仪测定血浆生化指标;无菌采集直肠内容物,通过Illumina MiSeq平台进行16S rDNA扩增子测序,并对高通量测序数据进行生物信息学分析、关联分析和表型预测。结果表明:1)三粉驴血浆中总胆固醇(TC)含量、碱性磷酸酶(ALP)活性显著高于乌头驴(P<0.05),甘油三酯(TG)、总蛋白(TP)含量和谷草转氨酶(AST)、肌酸激酶(CK)活性显著低于乌头驴(P<0.05)。2)α多样性结果显示,三粉驴直肠内容物菌群Ace指数、Chao指数和Shannon指数极显著低于乌头驴(P<0.01);β多样性结果显示,三粉驴和乌头驴直肠内容物菌群整体结构之间区分明显,组内样品聚类程度较高。3)三粉驴和乌头驴直肠内容物微生物在门水平没有差异,均以厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidota)为优势菌门;在属水平上,三粉驴直肠内容物毛螺菌科_XPB1014(Lachnospiraceae_XPB1014)和阿克曼氏菌属(Akkermansia)相对丰度显著高于乌头驴(P<0.05),而普雷沃氏菌属(Prevotella)和考拉杆菌属(Phascolarctobacterium)相对丰度显著低于乌头驴(P<0.05)。4) Spearman相关分析结果显示,Prevotella相对丰度与血浆中AST活性和TG含量呈极显著正相关(P<0.01),与ALP活性和TC含量呈极显著负相关(P<0.01);Phascolarctobacterium相对丰度与血浆中TC含量呈显著负相关(P<0.05);Akkermansia相对丰度与血浆中ALP活性和TC含量呈极显著正相关(P<0.01),与AST活性和TG含量呈极显著负相关(P<0.01);Lachnospiraceae_XPB1014相对丰度与血浆中ALP活性和TC含量呈显著或极显著正相关(P<0.05或P<0.01),与AST活性呈极显著负相关(P<0.01)。5) BugBase表型预测结果显示,三粉驴直肠微生物群的潜在致病性显著低于乌头驴(P<0.05),而包含的可移动元件显著高于乌头驴(P<0.05)。由此可见,三粉驴和乌头驴血浆生化指标以及直肠菌群组成、相对丰度和功能预测都存在差异。从血浆生化指标和肠道菌群结构的角度分析,三粉驴在生产和抗病性能方面或更优于乌头驴。
The aim of this study was to investigate the difference of plasma biochemical parameters and intestinal microflora structure between different strains of Dezhou donkey (Sanfen donkeys and Wutou donkeys). Six healthy and nonpregnant Sanfen female donkeys and Wutou female donkeys aged 3 to 5 years and weighed 260 to 300 kg were randomly selected, the blood was collected from jugular vein, and the plasma biochemical parameters were determined by an automatic biochemical analyzer; the rectum content was aseptic collected, the high-throughput amplicon sequencing for 16S rDNA was performed using the Illumina MiSeq platform, and the bioinformatics analysis, correlation analysis and phenotypic prediction were performed on high-throughput sequencing data. The results showed as follows: 1) the total cholesterol (TC) content and alkaline phosphatase (ALP) activity in plasma of Sanfen donkeys were significantly higher than those of Wutou donkeys (P<0.05), and the triglyceride (TG) and total protein (TP) contents and aspartate transaminase (AST) and creatine kinase (CK) activities in plasma were significantly lower than those of Wutou donkeys (P <0.05). 2) The α diversity results showed that the ACE index, Chao index and Shannon index in rectal content microflora of Sanfen donkeys were significantly lower than those of Wutou donkeys (P<0.01); the β diversity results showed that the rectal content microflora overall structure was obviously segregated between Sanfen donkeys and Wutou donkeys, and the clustering degree of samples in the group was high. 3) There was no difference at the phyla level in rectal content microflora between Sanfen donkeys and Wutou donkeys, and Firmicutes and Bacteroidota were the dominant bacteria; at the genus level, the relative abundances of Lachnospiraceae_XPB1014 and Akkermansia in rectal content of Sanfen donkeys were significantly higher than those of Wutou donkeys (P<0.05), while the relative abundances of Prevotella and Phascolarctobacterium were significantly lower than those of Wutou donkeys (P<0.05). 4) The Spearman correlation analysis results showed that the Prevotella relative abundance was significantly positively associated with plasma AST activity and TG content (P<0.01), and was significantly negatively associated with plasma ALP activity and TC content (P<0.01); the Phascolarctobacterium relative abundance was significantly negatively associated with plasma TC content (P<0.05); the Akkermansia relative abundance was significantly positively associated with plasma ALP activity and TC content (P<0.01), and was significantly negatively associated with plasma AST activity and TG content (P<0.01); the Lachnospiraceae_XPB1014 relative abundance was significantly positively associated with plasma ALP activity and TC content (P<0.05 or P<0.01), and was significantly negatively associated with plasma AST activity (P<0.01). 5) The BugBase phenotypic prediction results showed that the potentially pathogenic of Sanfen donkeys was significantly lower than that of Wutou donkeys (P<0.05), and the contains mobile elements was significantly higher than that of Wutou donkeys (P<0.05). In conclusion, there are differences in the plasma biochemical parameters and the bacterial composition, relative abundance and function prediction in rectum between Sanfen donkeys and Wutou donkeys. From the perspective of plasma biochemical parameters and intestinal microflora structure, the Sanfen donkeys are better than the Wutou donkeys in terms of production performance and disease resistance.
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