植物乳杆菌LR-39对犬的促生长及肠道菌群调节作用

doi:10.15933/j.cnki.1004-3268.2021.03.020

河南农业科学 ›› 2021, Vol. 50 ›› Issue (3): 148-156.DOI: 10.15933/j.cnki.1004-3268.2021.03.020

所属专题：动物肠道菌群专题

植物乳杆菌LR-39对犬的促生长及肠道菌群调节作用

陈则东¹，朱建平¹，郭书玉²，卢炜¹，刘蕾²

（1.江苏农牧科技职业学院宠物科技学院，江苏泰州 225300；2.西华大学食品与生物工程学院，四川成都 610039）

收稿日期:2020-09-09 出版日期:2021-03-15 发布日期:2021-03-15
通讯作者: 卢炜（1981-)，男，江苏泰州人，副教授，本科，主要从事宠物营养与食品开发。E-mail:900597@qq.com 刘蕾(1988-)，女，山西长治人，副教授，博士，主要从事食品微生物及功能产品开发。E-mail:112131536@qq.com
作者简介:陈则东(1985-)，男，江苏宿迁人，讲师，硕士，主要从事动物益生菌产品开发。E-mail:714238040@qq.com
基金资助:
中国特色高水平高职学校和专业建设项目（教职成函〔2019〕14号）；江苏农牧科技职业学院校级科研项目(NSF201807)

Effects of Lactobacillus plantarum LR-39 on Growth and Intestinal Microbiota Regulation of Dogs

CHEN Zedong¹,ZHU Jianping¹,GUO Shuyu²,LU Wei¹,LIU Lei²

(1.Department of Pet Science and Technology,Jiangsu Agri-Animal Husbandry Vocational College,Taizhou 225300,China;2.School of Food and Biological Engineering,Xihua University,Chengdu 610039,China)

Received:2020-09-09 Published:2021-03-15 Online:2021-03-15

摘要/Abstract

摘要： 为了评价从发酵泡菜中分离得到的植物乳杆菌LR-39对犬的促生长、肠道菌群调节作用，并明确其遗传背景，选取10只3.5月龄的比格犬，分为对照组与试验组，对照组饲喂基础犬粮，试验组在每日饲喂基础犬粮基础上补饲1×10⁹ cfu LR-39，试验期28 d。试验结束时，测定并比较各组平均日增质量、料重比及营养物质消化率等指标差异；对犬肠道菌群进行高通量测序，比较肠道菌群在门、种水平上的相对丰度及特征微生物差异；采用PICRUSt技术，解析并比较肠道微生物功能基因的表达水平差异；对LR-39进行全基因组序列测序，绘制该菌株全基因草图并进行序列COG及CAZy注释分析。结果显示，试验组平均日增质量极显著（P<0.01）高于对照组，粗蛋白、粗脂肪、钙的消化率显著（P<0.05）高于对照组，料重比显著（P<0.05）低于对照组。在门水平上，试验组梭杆菌门（Fusobacteria）（P<0.05）相对丰度显著高于对照组，放线菌门（Actinobacteria）相对丰度显著（P<0.05）低于对照组；效应大小线性判别分析（LEfSe）结果显示，试验组肠道菌群特征菌属是梭杆菌属（Fusobacterium），而对照组特征菌属分别是丹毒丝菌属（Bulleidia）、链型杆菌属（Catenibacterium）、真杆菌属（Eubacterium）、布劳特氏菌属（Blautia）、粪杆菌属（Faecalibacterium）；在种水平上，试验组瑞士乳杆菌（Lactobacillus helveticus）相对丰度极显著（P<0.01）高于对照组。肠道功能基因差异分析表明，试验组细胞内运输、细胞外结构基因表达相对丰度显著高于对照组（P<0.05），次级代谢物基因表达相对丰度显著（P<0.05）低于对照组。全基因组测序结果显示，LR-39菌株全长247 554 bp，无质粒，GC含量为44.26%，碳水化合物代谢相关基因丰度最高，氨基酸代谢相关基因其次，而脂代谢相关基因较低，具有一定比例拮抗机制相关基因，酶系主要包括糖苷水解酶、糖基转移酶、碳水化合物酯酶、碳水化合物结合分子、辅助性酶类。综上，LR-39能够促进犬的生长，改变犬肠道菌群结构，调节肠道菌群功能基因表达，具有开发益生菌产品的潜质。

关键词: 植物乳杆菌, 犬, 促生长, 肠道菌群, 高通量测序, 全基因组测序

Abstract: To evaluate the growth-promoting and intestinal flora regulation effects of Lactobacillus plantarum LR-39 isolated from fermented pickles,ten 3.5-month-old beagle dogs were divided into control group and experimental group. The control group was fed with basic dog food,while the experimental group was daily given LR-39 with the basic dog food for 28 days.The average daily gain,feed weight ratio,nutrient digestibility and other indicators were measured at the end of the test. Based on highthroughput sequencing,the relative abundance of intestinal flora at the phylum level and the differences in the characteristic microbes of each group of intestinal flora were analyzed. PICRUSt was performed to further analyze the expression difference of microbial functional genes in the intestinal tract.The whole genome sequencing of LR-39 and subsequent COG and CAZy annotation analysis were performed.The results of the growth-promoting test showed that the average daily gain of the experimental group was significantly(P<0. 01) higher than that of the control group;the feed weight ratio of the experimental group was significantly(P<0.05) lower than that of the control group.The digestibility of crude protein,crude fat and calcium in the experimental group was significantly(P<0.05) higher than those of the control group.High-throughput sequencing results showed that at the phylum level,the relative abundance of Fusobacteria in the experimental group was significantly(P<0.05) higher than that in the control group,while the relative abundance of Actinobacteria was significantly(P<0.05) lower than that in the control group.LDA effect size(LEfSe) analysis showed that the characteristic bacterial generus of the intestinal flora in the experimental group was Fusobacterium,while the characteristic bacterial genera of the control group were Bulleidia, Catenibacterium,Eubacterium,Blautia,and Faecalibacterium.At the species level,the relative abundance of Lactobacillus helveticus in the experimental group was significantly(P<0.01)increased.Intestinal functional gene difference analysis showed that the relative expression abundance of intracellular transport and extracellular structure genes in the experimental group was significantly(P<0.05) higher than that in the control group,and the relative expression abundance of secondary metabolite gene was significantly(P<0.05) lower than that in the control group.Whole genome sequencing showed that the strain had a full length of 247 554 bp, no plasmid, with GC content of 44.26%.The abundance of genes related to carbohydrate metabolism was the highest,followed by amino acid metabolism related genes,the abundance of genes related to lipid metabolism was lower,and the genes related to antagonistic mechanism had a certain proportion.The enzyme system mainly included glycoside hydrolases, glycosyltransferases, carbohydrate esterases, carbohydrate binding molecules and auxiliary enzymes.In summary,feeding LR-39 can promote the growth of dogs,change the structure of the intestinal flora,regulate the expression of functional genes in intestinal flora,and it has the potential to develop into probiotic products.

Key words: Lactobacillus plantarum, Dog, Growth-promoting, Intestinal flora, High-throughput sequencing, Whole genome sequencing

中图分类号:

S852.61

陈则东, 朱建平, 郭书玉, 卢炜, 刘蕾. 植物乳杆菌LR-39对犬的促生长及肠道菌群调节作用[J]. 河南农业科学, 2021, 50(3): 148-156.

CHEN Zedong, ZHU Jianping, GUO Shuyu, LU Wei, LIU Lei. Effects of Lactobacillus plantarum LR-39 on Growth and Intestinal Microbiota Regulation of Dogs[J]. Journal of Henan Agricultural Sciences, 2021, 50(3): 148-156.

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植物乳杆菌LR-39对犬的促生长及肠道菌群调节作用

Effects of Lactobacillus plantarum LR-39 on Growth and Intestinal Microbiota Regulation of Dogs

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