Effects of Dietary Rutin on Rumen Microbial Community Diversity and Composition of Hu Sheep
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摘要:
目的 研究日粮中添加芦丁对湖羊瘤胃细菌菌群多样性及组成的影响。 方法 选择3月龄左右、健康且体重相近的湖羊36只，随机分为3个处理，每个处理12只（公母各半，分栏饲养）。对照组（CON组）饲喂基础日粮，试验组R50和R100分别在基础日粮中添加50、100 mg·kg−1（湖羊体重基础）的芦丁。试验期为70 d，预试期14 d，正试期56 d。 结果 1）3组共产生446个OTUs（Operational taxonomic units），其中共有OTU为396个，占总OTU的88.79%，所有样品共注释了14个门（Phylum）、18个纲（Class）、21个目（Order）、34个科（Family）和99个属（Genus）。2）相对于CON组，R100组的ACE指数、Shannon指数、Chao1指数显著降低（P＜0.05）。但CON和R50组中Alpha多样性均无显著差异（P＞0.05）。3）通过PCoA分析（Principal co-ordinates analysis）、NMDS分析（Non-metric multi-dimensional scaling analysis）、UPGMA聚类方法（Unweighted pair group method with arithmetic mean clustering analysis）和Anosim分析可知，R100组显著影响微生物群落结构。4）在门水平，拟杆菌门（Bacteroidetes）和厚壁菌门（Firmicutes）为两大优势菌门。相比CON组，R100组髌骨细菌门（Patescibacteria）的相对丰度显著提高，迷踪菌门（Elusimicrobia）的相对丰度显著降低（P＜0.05），但R50组Elusimicrobia的相对丰度显著高于CON组和R100组（P＜0.05）。5）在属水平，优势菌属均为普雷沃菌属_1（Prevotella_1）。在已鉴定丰度排名前30的属水平上，R100组新月形单胞菌属_1（Selenomonas_1）和琥珀酸菌属（Succiniclasticum）的相对丰度显著低于CON组（P＜0.05），但与R50组差异不显著（P＞0.05）。 结论 添加100 mg·kg−1的芦丁能够影响湖羊瘤胃微生物多样性。 Abstract:Objective Effects of rutin supplementation in forage on the rumen microbial community diversity and composition of Hu sheep were studied. Method Thirty-six healthy 3-month-old Hu sheep of similar body weight were randomly divided into 3 groups of 12 animals (half male and female) each for the treatments. The sheep in the control group (CON) were fed a basal diet, while those in two treatment groups on a daily diet supplemented with rutin at a rate of 50 mg (R50) or 100 mg (R100) per kg of animal body weight. The feeding test lasted for 70 d that included 14 d of pre-test and 56 d of actual test. Result (1) Of the 446 OTUs collected in the sheep rumens, 396 (constituted 88.79% of total) were commonly shared by the sheep in the 3 groups that annotated into 14 phyla, 18 classes, 21 orders, 34 families, and 99 genera. (2) The ACE, Shannon, and Chao1 indices of the sheep in the R100 group were significantly lower than those of CON (P ＜0.05), while the alpha diversity not significantly different between CON and R50 groups (P＞0.05). (3) The results of the principal co-ordinates analysis (PCoA), the non-metric multi-dimensional scaling analysis (NMDS), the unweighted pair group method with arithmetic mean clustering analysis (UPGMA), and the anosim analysis showed that the microbial community was significantly altered by the R100 treatment (P＜0.05). (4) Bacteroidetes and Firmicutes were two dominant phyla. R100 significantly increased the relative abundance of Patescibacteria but decreased that of Elusimicrobia (P ＜0.05). On the other hand, R50 significantly raised the relative abundance of Elusimicrobia over CON or R100 (P＜0.05). (5) At genus level, the dominant microbes were Prevotella_1 . The relative abundance of Selenomonas_1 and Succiniclasticum in the sheep fed with R100 were significantly lower than CON (P＜0.05) but not significantly different from R50 (P＞0.05). Conclusion Addition of rutin to forage at a rate of 100 mg·kg−1 of sheep body weight significantly affected the rumen microbial diversity in Hu sheep . -
Key words:
- Hu sheep /
- rutin /
- rumen /
- microorganism
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图 6 湖羊瘤胃微生物群落结构Anosim分析
Figure 6. Anosim analysis on microbial community structure in rumen of Hu sheep
表 1 瘤胃液细菌群Alpha多样性分析
Table 1. Alpha diversity analysis on microbial community in rumen fluid
项目
ItemsCON组
CON GroupR50组
R50 groupR100组
R100 groupACE 指数
ACE index405.40±7.47 a 395.08±3.34 a 372.51±9.07 b 赵氏指数
Chao1 index409.07±7.63 a 398.95±4.10 ab 377.80±12.46 b 香农指数
Shannon index6.52±0.17 a 6.69±0.14 a 5.92±0.23 b 辛普森指数
Simpson index0.97±0.01 ab 0.98±0.00 a 0.95±0.01 b 注：同行数据后的大写字母不同表示差异极显著（P＜0.01），小写字母不同表示差异显著（P＜0.05），相同或无字母表示差异不显著（P＞0.05）。下表同。
Note：Different uppercase letters on the same line indicate extremely significant differences (P＜0.01), lowercase letters indicate significant differences
(P＜0.05), while the same or no letters mean no significant difference (P＞0.05). The same as below.
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表 2 芦丁对湖羊瘤胃微生物相对丰度的影响（门水平）
Table 2. Effect of rutin on microbial relative abundance in rumen of Hu sheep (at phylum level) （单位：%）
项目
ItemsCON组
CON groupR50组
R50 groupR100组
R100 group拟杆菌门 Bacteroidetes 52.84±3.03 59.18±3.02 62.76±6.39 厚壁菌门 Firmicutes 35.47±1.87 31.56±1.39 29.67±5.23 变形菌门 Proteobacteria 5.80±2.10 1.97±1.22 1.67±0.75 互养菌门 Synergistetes 1.68±0.97 2.70±1.53 0.77±0.53 广古菌门 Euryarchaeota 1.43±1.15 0.67±0.13 2.12±1.52 螺旋体门 Spirochaetes 0.54±0.21 1.65±0.70 1.08±0.41 放线菌门 Actinobacteria 1.33±1.32 0.61±0.29 0.85±0.42 纤维杆菌门 Fibrobacteres 0.36±0.09 0.31±0.06 0.37±0.14 髌骨细菌门 Patescibacteria 0.17±0.06 b 0.30±0.06 ab 0.56±0.18 a 迷踪菌门 Elusimicrobia 0.06±0.04 b 0.80±0.40 a 0.00±0.00 c 软壁菌门 Tenericutes 0.27±0.14 0.22±0.11 0.07±0.03 Kiritimatiellaeota 0.05±0.01 0.03±0.01 0.03±0.01 蓝藻菌门 Cyanobacteria 0.00±0.00 0.01±0.00 0.05±0.04 k__Bacteria_Unclassified 0.02±0.02 0.01±0.01 0.02±0.02
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表 3 芦丁对湖羊瘤胃微生物相对丰度的影响（属水平）
Table 3. Effect of rutin on microbial relative abundance in rumen of Hu sheep (at genus level)
（单位：%） 项目
ItemsCON组
CON groupR50组
R50 groupR100组
R100 group普雷沃菌属_1 Prevotella_1 20.28±5.92 26.34±3.35 26.79±7.98 理研菌科_RC9_gut_group
Rikenellaceae_RC9_gut_group12.16±2.99 11.05±2.13 18.60±4.48 f__F082_Unclassified 3.47±0.41 5.83±1.64 5.09±1.07 f__Muribaculaceae_Unclassified 4.83±1.60 5.55±1.93 1.98±1.35 新月形单胞菌属 Selenomonas_1 5.10±1.40 a 2.56±0.40 ab 1.51±0.52 b f__Prevotellaceae_Unclassified 2.67±0.53 2.61±0.52 3.65±1.55 瘤胃球菌属_2 Ruminococcus_2 0.80±0.19 2.11±1.31 5.11±3.26 f__Veillonellaceae_Unclassified 2.53±0.81 3.14±0.98 2.10±0.77 Ruminococcaceae_NK4A214_group 2.73±0.73 2.14±0.42 2.26±0.63 Christensenellaceae_R-7_group 1.95±0.11 1.66±0.31 2.89±1.99 Quinella 1.86±0.78 2.17±0.86 2.46±1.05 [Eubacterium]_coprostanoligenes_group 3.99±3.71 0.56±0.16 1.94±1.02 琥珀酸弧菌属 Succinivibrio 3.46±1.52 0.81±0.52 1.21±1.02 f__Bacteroidales_RF16_group_Unclassified 3.11±0.87 a 0.89±0.27 b 1.32±0.29 b Prevotellaceae_UCG-003 2.29±0.97 1.48±0.39 1.45±0.21 Fretibacterium 1.54±0.97 2.55±1.52 0.72±0.52 Prevotellaceae_UCG-001 1.30±0.43 1.91±0.79 1.49±0.46 Lachnospiraceae_ND3007_group 0.94±0.54 1.37±0.49 2.15±0.95 甲烷短杆菌属
Methanobrevibacter1.38±1.16 0.61±0.13 2.11±1.52 f__Lachnospiraceae_Unclassified 1.21±0.15 AB 1.87±0.42 A 0.52±0.14 B 瘤胃球菌属_1 Ruminococcus_1 1.15±0.37 1.41±0.28 0.78±0.14 密螺旋体属_2 Treponema_2 0.53±0.21 1.65±0.70 1.08±0.41 Veillonellaceae_UCG-001 1.43±0.52 1.04±0.24 0.49±0.15 f__Bifidobacteriaceae_Unclassified 1.32±1.32 0.61±0.30 0.85±0.42 琥珀酸菌属 Succiniclasticum 1.67±0.61 a 0.73±0.25 ab 0.26±0.12 b f__Bacteroidales_BS11_gut_group_Unclassified 0.50±0.21 1.00±0.39 0.78±0.54 Succinivibrionaceae_UCG-002 1.58±0.71 0.61±0.52 0.00±0.00 厌氧弧菌属 Anaerovibrio 0.98±0.27 0.73±0.23 0.48±0.28 Ruminococcaceae_UCG-002 0.42±0.19 0.66±0.13 0.65±0.17 Lachnospiraceae_NK3A20_group 0.58±0.18 0.61±0.06 0.55±0.09
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