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饲养方式对藏猪小肠菌群结构的影响

Effects of feeding model on small intestinal flora structure of Tibetan pigs

  • 摘要: 为了探究饲养方式对藏猪小肠微生物多样性的影响,试验选取5头体重均匀的健康断奶藏猪,白天放牧,夜间归牧后补饲少量玉米;另外选取5头体重均匀的健康断奶藏猪舍饲饲养,饲喂玉米-豆粕型日粮。两种饲养方式饲养试验藏猪至7月龄屠宰,采集每只试验猪的十二指肠、空肠和回肠的内容物,PCR扩增16S rRNA基因序列,采用高通量测序技术测序,测序数据进行微生物操作分类单元(OTUs)、Alpha指数(包括Chao1、Observed species、Shannon、Simpson、Faith′s PD和Pielou′s evenness指数)、主成分分析(PCA)以及分类水平方面的分析。结果表明:16S rRNA基因测序序列的长度均在1 400~1 500 bp之间。放牧和舍饲藏猪获得的OTUs数量不同,放牧藏猪的Chao1、Observed species、Shannon、Simpson、Faith′s PD和Pielou′s evenness指数均高于舍饲藏猪,但均差异不显著(P>0.05),放牧和舍饲藏猪小肠菌群结构存在差异。变形菌门(Proteobacteria)、厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes)为藏猪的主要菌门,放牧藏猪小肠中变形菌门的相对丰度低于舍饲藏猪,而厚壁菌门和拟杆菌门高于舍饲藏猪,但均差异不显著(P>0.05)。伯克氏菌科(Burkholderiaceae)、丛毛单胞菌科(Comamonadaceae)、肠杆菌科(Enterobacteriaceae)、普雷沃氏菌科(Prevotellaceae)和噬几丁质菌科(Chitinophagaceae)等10种菌科为藏猪的优势菌科,其中放牧藏猪的肠杆菌科和梭菌科(Clostridiaceae)显著高于舍饲藏猪(P<0.05)。青枯菌属(Ralstonia)、嗜糖假单胞菌属(Pelomonas)、埃希氏菌属(Escherichia)、普雷沃氏菌属(Prevotella)、月形单胞菌属(Selenomonas)、苍白杆菌属(Ochrobactrum)、慢生根瘤菌属(Bradyrhizobium)、梭菌属(Clostridium)、乳杆菌属(Lactobacillus)和弯曲菌属(Campylobacter)为藏猪的主要菌属,放牧藏猪小肠中的埃希氏菌属和梭菌属显著高于舍饲藏猪(P<0.05),而弯曲杆菌属显著低于舍饲藏猪(P<0.05);在藏猪小肠中鉴定到了48个菌种,其中放牧藏猪小肠中反刍月形单胞菌(Selenomonas bovis)、Sharpea azabuensis的相对丰度显著低于舍饲藏猪(P<0.05),而普氏菌(Prevotella copri)、发酵氨基酸球菌(Acidaminococcus fermentans)、猪肠链球菌(Streptococcus hyointestinalia)和黏膜乳杆菌(Lactobacillus mucosae)显著高于舍饲藏猪(P<0.05)。说明饲养方式会影响藏猪小肠的菌群结构。

     

    Abstract: The purpose of this experiment was to study the effects of feeding model on small intestinal microbial diversity of Tibetan pigs.Five healthy weaned Tibetan pigs with uniform body weight were selected in the experiment, and were grazed during the day and supplemented with a small amount of corn after grazing at night. Another five healthy weaned Tibetan pigs with uniform body weight were selected for house feeding, and were fed-corn soybean meal diet. The experimental Tibetan pigs were raised in two feeding methods until slaughter at the age of seven months.The contents of duodenum, jejunum and ileum of each test pig were collected after slaughter, 16 S rRNA gene sequences were amplified by PCR, sequenced by high-throughout sequencing technology, and the sequencing data were analyzed in terms of microbial operational taxonomic units(OTUs), Alpha indexes(Chao1,Observed species, Shannon, Simpson, Faith′s PD and Pielou′s evenness index), principal component analysis(PAC) and taxonomic levels analysis. The results were as follows: the sequenced sequences of 16 S rRNA genes were all between 1 400-1 500 bp in length. The number of OTUs in grazing Tibetan pigs and housed Tibetan pigs were different, the Alpha indexes of Chao1, Observed species, Shannon, Simpson, Faith′s PD and Pielou′s evenness in grazing Tibetan pigs were higher than those in housed Tibetan pigs, but there were no significant difference(P>0.05).There were differences in intestinal bacterial structure between grazing and housed Tibetan pigs.Proteobacteria, Firmicutes and Bacteroidtes were the dominant bacterial phyla in Tibetan pigs.The relative abundance of Proteobacteria in grazing Tibetan pigs was lower than that in housed Tibetan pigs, while Firmicutes and Bacteroidetes in grazing Tibetan pigs was higher than that in housed Tibetan pigs, but the difference was not significant(P>0.05). Burkholderiaceae, Comamonadaceae, Enterobacteriaceae, Prevotellaceae and Chitinophagaceae were the dominant bacterial families in Tibetan pigs, the relative abundance of Enterobacteriaceae and Clostridiaceae in grazing Tibetan pigs were significantly highter than housed Tibetan pigs(P<0.05).Ralstonia, Pelomonas, Escherichia, Prevotella, Selenomonas, Ochrobactrum, Bradyrhizobium, Clostridium, Lactobacillus and Campylobacter were the dominant bacterial genera in Tibetan pigs. The relative abundant of Escherichia and Clostridium in grazing Tibetan pigs were significantly higher than that in housed Tibetan pigs(P<0.05), while Campylobacter was significantly lower than that in housed Tibetan pigs(P<0.05). In addition, 48 bacterial species were identified in small intestine of Tibetan pigs, among which the relative abundance of Selenomonas bovis and Sharpea azabuensis in grazing Tibetan pigs was significantly lower than that in housed Tibetan pigs(P<0.05),while the relative abundance of Prevotella copri, Acidaminococcus fermentans, Streptococcus hyointestinalis and Lactobacillus mucosae in grazing Tibetan pigs was significantly higher than that in housed Tibetan pigs(P<0.05). This result indicated that the different feeding model could effect the structure of small intestinal microbial community of Tibetan pigs.

     

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