灯盏花黄酮对腹泻仔猪肠道微生物多样性影响的研究

Effects of Erigeron flavonoids on intestinal microbial diversity in diarrhea piglets

为了研究灯盏花黄酮对腹泻仔猪肠道微生物多样性的影响,试验将20头60日龄健康仔猪分为空白组、药物组、阳性组和模型组,第1~3天除空白组外其他组仔猪按体重灌服浓度为3×10^(8) cfu/mL的沙门氏杆菌液0.5 mL/kg,第1~10天药物组仔猪按体重灌服灯盏花黄酮1 mL/kg,阳性组仔猪按体重灌服氟苯尼考1 mL/kg,空白组和模型组仔猪灌服等量的纯化水。观察仔猪临床症状,在第4,8,11天采集各组仔猪粪便进行微生物多样性分析。通过OTU序列与相应微生物数据库进行比对,得到各样本的物种聚类信息与各水平注释信息,基于聚类结果进行α多样性分析和β多样性分析,基于β多样性分析结果进行UPGMA聚类分析和Kru-Wall秩和检验分析,基于注释结果进行物种分类分析和差异物种分析。结果表明:仔猪灌服沙门氏杆菌后,出现精神沉郁、伏卧、采食和饮水减少、排黄色稀便等症状。空白组、药物组、阳性组和模型组在第4,8,11天共有的OTU数目分别为912,872,780,796条,空白组和药物组高于阳性组和模型组,在第11天空白组、药物组、阳性组和模型组特有的OTU数目分别为84,96,74,37条,药物组高于其他3组。与第4天比较,第11天药物组的Chao指数升高,而模型组下降,ACE指数变化趋势与Chao指数相同,药物组的Shannon和Simpson指数相对稳定,第8天与第4天相比模型组Simpson指数显著降低(P<0.05)。UPGMA聚类分析药物组和空白组分布相对均匀,模型组和阳性组分布相对集中。Kru-Wall秩和检验空白组和药物组菌群波动性较小,模型组菌群波动性较大。在门水平上,相对丰度最高的为厚壁菌门,其次为拟杆菌门,模型组浮霉菌门和蓝细菌相对丰度极显著高于药物组和阳性组(P<0.01),药物组互养菌门相对丰度极显著高于模型组和阳性组(P<0.01)。在属水平上,药物组普雷沃氏菌相对丰度极显著低于模型组和阳性组(P<0.01),乳酸杆菌相对丰度极显著高于阳性组和模型组(P<0.01),模型组巨型球菌相对丰度极显著高于阳性组和药物组(P<0.01),粪杆菌属相对丰度极显著低于药物组和阳性组(P<0.01)。说明灯盏花黄酮能够调节肠道菌群丰度,有利于腹泻仔猪康复,具有替代抗生素的潜力。

To study the effect of Erigeron flavonoids on the intestinal microbial diversity of piglets with diarrhea, in the experiment, 20 healthy 60-day-old piglets were divided into blank group, drug group, positive group and model group. On days one to three, except for the blank group, other groups were fed with a concentration of 3×10;cfu/mL Salmonella solution 0.5 mL/kg by body weight. From the 1 st day to the 10 th day, piglets in drug group were given Erigeron flavonoids 1 mL/kg by body weight, piglets in positive group were given florfenicol 1 mL/kg by body weight, and piglets in blank group and model group were given purified water in equal amount. Clinical symptoms of piglets were observed. Feces of each group were collected on days 4, 8 and 11 for microbial diversity analysis. By comparing the OTU sequence with the corresponding microbial database, the species clustering information and the annotation information of each level of each sample were obtained. Based on the clustering results, α diversity analysis and β diversity analysis were performed. Based on the results of β diversity analysis, UPGMA cluster analysis and Kru-Wall rank sum test analysis were performed. Perform species classification analysis and differential species analysis were performed based on the annotation results. The results showed that after feeding with Salmonella, piglets were depressed, lying down, feeding decreased, drinking water decreased, and excreting yellow feces. The total number of OTUs in the blank group, drug group, positive group and model group were 912, 872, 780, and 796 on the 4 th, 8 th, and 11 th day, respectively. The blank group and the drug group were higher than the positive group and the model group. The number of unique OTUs in the blank group, drug group, positive group, and model group were 84, 96, 74, and 37 on the 11 th day, respectively. The drug group was higher than the other three groups. Compared with the 4 th day, the Chao index of the drug group increased on the 11 th day, while the model group gradually decreased, and the change trend of ACE index was the same as Chao index. Shannon and Simpson index in drug group were relatively stable, and Simpson index in model group was significantly decreased on the 4 th day compared with the 8 th day(P<0.05). UPGMA cluster analysis showed that the distribution of the drug group and the blank group was relatively uniform, and the distribution of the model group and the positive group was relatively concentrated. Kru-Wall rank-sum test showed little fluctuation in the flora of blank group and drug group. The model group had great fluctuation. The phylum Firmicutes and Bacteroidetes were the most abundant in all sample phyla. The relative abundance of Planctomycetes and Cyanobacteria in model group was extremely significantly higher than that in drug group and positive group(P<0.01), and the relative abundance of Synergistetes in drug group was extremely significantly higher than that in model group and positive group(P<0.01). At the genus level, the relative abundance of Prevotella in the drug group was extremely significantly lower than that in the model group and the positive group(P<0.01), the relative abundance of Megasphaera in the model group was extremely significantly higher than that in the positive group and the drug group(P<0.01), and the relative abundance of Lactobacillus in the drug group was extremely significantly higher than that in the positive group and the model group(P<0.01). The relative abundance of Faecalibacterium in model group was extremely significantly lower than that in drug group and positive group(P<0.01). These results indicated that Erigeron flavonoids could regulate the abundance of intestinal flora and was beneficial to diarrhea recovery of piglets, which has the potential to replace antibiotics.