Changes Induced by Probiotic, Enterococcus faecium, in Gut Microbiota of Post-weaning Piglet
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摘要: 通过细菌16S rRNA基因的高通量测序，对断奶仔猪的肠道菌群结构和饲喂两株单一益生菌（屎肠球菌）后肠道菌群的变化情况和规律进行分析。结果显示，断奶仔猪肠道微生态中拟杆菌门Bacteroidetes和厚壁菌门Firmicutes的细菌所占比例分别为50.5%和42.2%，为绝对优势菌群。饲喂屎肠球菌后，肠道菌群的多样性和细菌丰度均增加；厚壁菌门Firmicutes，变形菌门Proteobacteria、无壁菌门Tenericutes、酸杆菌门Actidobacteria、芽单胞菌门Gemmatimonadetes、放线菌门Actinobacteria细菌的丰度明显上升，而拟杆菌门Bacteroidetes的细菌丰度则发生了明显下降。屎肠球菌所在的肠球菌属Enterococcus的比例虽由0.07%（对照组）上升至0.12%（SF1组）和0.14%（SF2组），但在肠道菌群中所占比例仍然很小，说明屎肠球菌对仔猪肠道微生态的作用主要是影响其他细菌的丰度和多样性进而改变菌群结构。与对照组相比，SF1组的上述菌群变化幅度高于SF2组，通过与3组仔猪生长性能数据的对比，上述菌群变化的幅度与仔猪生长性能提高的程度呈现正相关。Abstract: Using the 16S rRNA gene high through-put sequencing technique, the microbial composition of the gut microbiota in post-weaning piglets was characterized. The compositional alterations induced by two strains of Enterococcus faecium were studied. The resulting dominant species in the guts were found to be of Firmicutes and Bacteroidetes phyla with contribution rates of 50.5% and 42.2%, respectively. The richness and diversity of the bacterial community increased after the E. faecium administration. The populations of 6 phyla (i.e., Firmicutes, Proteobacteria, Tenericutes, Actidobacteria, Gemmatimonadetes, and Actinobacteria) were increased, while that of Bacteroidetes significantly declined. A low abundance of Enterococcus spp. was detected in the piglet guts. However, the increase was merely 0.07% over control and 0.12 over the treatment group, SF1, and 0.14% over group SF2 after the feeding. Consequently, the primary effects induced by the probiotics, E. faecium strains, were contributed mainly to the changes on the richness and diversity, or the composition, of the gut microbes. It appeared that the positive results observed on the growth performances of the post-weaning piglets in the field experiment due to the introduction of the probiotics warned further attention.
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Key words:
- post-weaning piglet /
- gut microbiota /
- probiotics /
- Enterococcus faecium
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图 1 基于相对丰度值的菌群断奶仔猪肠道菌群分类树
注：各菌的相对丰度值根据圆圈大小和菌名下的数值表示。
Figure 1. Taxon tree of microbial groups in gut microbiota of post-weaning piglets based on their relative abundances
表 1 断奶仔猪分组状况
Table 1. Descriptions on piglet groups
组别 头数
/头性别 均重
/kg是否饲喂
屎肠球菌SF1 16 雌 6.86±0.23 是* SF2 16 雌 7.38±0.31 是** SF3(CK) 16 雌 7.11±0.28 否 注：*本株屎肠球菌由本课题组在前期研究中分离，已在中国普通微生物保藏中心(CGMCC)鉴定并保存，编号CGMCC 8296；**本株屎肠球菌采购自北京昕地美饲料科技有限公司。 
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表 2 100 K饲料基础日粮组成与营养水平
Table 2. Composition and nutrient contents of basal feed, 100K
日粮成分 含量
/%营养水平 含量 玉米 55.0 消化能/(MJ·kg-1) 12.85 豆粕 24.1 粗蛋白/% 20.00 麦麸 8.6 钙/% 0.70 鱼粉 4.0 磷/% 0.50 预混料* 4.0 赖氨酸/% 1.20 血浆蛋白粉 2.0 蛋氨酸/% 0.75 植物油 2.3 注：每kg日粮中含磷酸氢钙11 g、抗氧化剂0.2 g、罗氏多维0.4 g、防霉剂1.0 g、食盐3 g、甜味剂0.6 g、蛋氨酸0.5 g、赖氨酸2.3 g、苏氨酸0.5 g、硫酸亚铁500 mg、硫酸铜100 mg、硫酸锌450 mg、硫酸锰20 mg、1%亚硒酸钠5 mg、1%碘化钾2.75 mg。
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表 3 3组仔猪肠道微生态Alpha多样性值
Table 3. Alpha indices on 3 groups of gut microbiota in post-weaning piglets
组别 表观分
类数Chao1
指数Shannon
指数SF1 1196 1463.259 7.631 SF2 1133 1349.505 7.33 SF3 1062 1336.809 7.028
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表 4 不同组间门水平相对丰度值比较
Table 4. Relative microbe abundances at phylum level in 3 groups
门 组别 SF1/% SF2/% SF3/% 厚壁菌门Firmicutes 62.1 48.7 42.2 拟杆菌门Bacteroidetes 27.6 41.7 50.5 变形菌Proteobacteria 4.5 4.9 2.9 无壁菌门Tenericutes 2.8 1.9 1.7 酸杆菌门Acidobacteria 1.5 1.1 1.0 放线菌门Actinobacteria 0.58 0.42 0.31 芽单胞菌门Gemmatimonadetes 0.38 0.33 0.26 其他 0.49 0.92 1.02
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表 5 不同组间属水平相对丰度值比较
Table 5. Relative microbe abundances at genus level in 3 groups
门 目 科 属 组别 SF1/% SF2/% SF3/% 厚壁菌门
Bacteroidetes拟杆菌目
Bacteroidales普氏菌科Prevotellaceae 普氏菌属Prevotella 14.0 28.1 35.7 S24-7 S24-7科中未命名属 7.5 8.0 9.1 拟杆菌门
Firmicutes乳杆菌目
Lactobacillales乳杆菌科Lactobacillaceae 乳杆菌属Lactobacillus 13.6 11.0 7.3 梭菌目
Clostridiales瘤胃菌科Ruminococcaceae 瘤胃菌属Ruminococcus 6.3 4.5 3.9 瘤胃菌科Ruminococcaceae中未命名属 6.0 3.4 4.3 瘤胃菌科Ruminococcaceae中第2个未命名属 5.0 3.4 3.1 颤螺菌属Oscillospira 5.5 4.4 3.5 毛螺菌科Lachnospiraceae 毛螺菌科Lachnospiraceae中未命名属 3.1 2.0 1.8 罗斯氏菌属Roseburia 3.0 3.1 3.5 梭菌科Clostridiaceae 梭菌科Clostridiaceae中未命名属 2.9 2.2 3.0 其他 其他 其他 其他 33.0 29.9 24.8 注：数值代表各属的相对丰度值。
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表 6 无壁菌门Tenericutes、酸杆菌门Acidobacteria和放线菌门Actinobacteria在属水平的相对丰度值比较
Table 6. Relative abundances of OTUs showing distributions at genus level of Tenericutes, Acidobacteria and Actinobacteria
门 属 组别 SF1/% SF2/% SF3/% 无壁菌门Tenericutes RF39科中未命名属 2.7 1.8 1.7 其他无壁菌门细菌 0.034 0.011 0.011 酸杆菌门Acidobacteria Candidatus Koribacter属 0.17 0.16 0.07 Candidatus Solibacter属 0.22 0.12 0.06 其他酸杆菌门细菌* 1.12 0.84 0.92 放线菌门Actinobacteria OCS155科中未命名属 0.11 0.03 0.045 放线菌科Actinomycetaceae中未命名属 0.18 0.23 0.079 Micrococcaceae中未命名属 0.045 0.06 0.023 Gaiellaceae科中未命名属 0.034 0.023 0.079 其他放线菌门细菌* 0.204 0.079 0.079 注：*代表酸杆菌门和放线菌门中低于0.01%的属未被列出。
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表 7 3组间肠球菌属Enterococcus的丰度值比较
Table 7. Relative abundances of Enterococcus spp. In 3 groups
属 组别 SF1/% SF2/% SF3/% 肠球菌属Enterococcus 0.12 0.14 0.07
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表 8 14 d内仔猪生长性能状况
Table 8. Growth performance of piglets during 14-day experiment
屎肠球菌组 SF3 (CK) SF1 SF2 头数 16 16 16 采食量/(g·p-1·d-1) 347.1±18.6 299.1±20.1 328.1±23.1 日增重/(g·p-1·d-1) 231.0±20.7 205.4±19.0 197.5±21.2 饲料转化率/% 1.50±0.12 1.46±0.11 1.66±0.15 腹泻数/次 2 7 9
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