动物营养学报 >

2019 , Vol. 31 >Issue 11: 5310 - 5320

DOI: https://doi.org/10.3969/j.issn.1006-267x.2019.11.048

饲料科学与技术 Feed science and technology

坚强芽孢杆菌CX10益生性评价

  • 马玉俊 ,
  • 梁子敬 ,
  • 张凯 ,
  • 李杰 ,
  • 马小军 ,
  • 王磊 ,
  • 张康 ,
  • 张景艳 ,
  • 李建喜
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  • 1. 中国农业科学院兰州畜牧与兽药研究所, 兰州 730050;
    2. 甘肃农业大学动物医学院, 兰州 730070
马玉俊(1993-),男,甘肃临夏人,硕士研究生,研究方向为兽医内科学。E-mail:2451276640@qq.com

收稿日期: 2019-04-03

  网络出版日期: 2019-11-19

基金资助

国家自然科学基金项目（31602101）；中兽医与临床科技创新工程（CAAS-ASTIP-2015-LIHPS）；甘肃省农业生物技术研究与应用开发项目（GNSW-2016-12）；甘肃省科技支撑计划项目（1604NKCA069-03）

收起

Probiotic Evaluation of Bacillus firmus CX10

  • MA Yujun ,
  • LIANG Zijing ,
  • ZHANG Kai ,
  • LI Jie ,
  • MA Xiaojun ,
  • WANG Lei ,
  • ZHANG Kang ,
  • ZHANG Jingyan ,
  • LI Jianxi
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  • 1. Lanzhou Institute of Husbandry and Pharmaceutical Science, Chinese Academy of Agricultural Sciences, Lanzhou 730050, China;
    2. College of Veterinary Medicine, Gansu Agricultural University, Lanzhou 730070, China

Received date: 2019-04-03

  Online published: 2019-11-19

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摘要

本试验通过灌胃小鼠不同剂量的坚强芽孢杆菌CX10（BF-CX10）后测定小鼠体重、脏器指数、血清酶活性和免疫细胞因子表达以及粪便菌群结构及其多样性，对BF-CX10进行益生性评价。试验选用50只18~20 g雌性BALB/c小鼠，随机分为对照组、大肠杆菌自愈（ESH）组、低剂量BF-CX10（LBF）组、中剂量BF-CX10（MBF）组和高剂量BF-CX10（HBF）组，每组10只小鼠。对照组小鼠全程灌胃生理盐水；ESH组小鼠前3 d灌胃大肠杆菌O78，后3 d灌胃生理盐水；LBF、MBF和HBF组前3 d分别灌胃1.5×104、1.5×106和1.5×108 CFU/mL的BF-CX10，后3 d用大肠杆菌O78灌胃；灌胃量均为0.02 mL/g BW。第7天处理小鼠。结果表明：与ESH组相比，BF-CX10组小鼠体重增长稳定，肾脏指数和心脏指数显著提高（P<0.05），血清酸性磷酸酶（ACP）、碱性磷酸酶（AKP）和谷胱甘肽过氧化物酶（GSH-Px）活性显著提高（P<0.05），血清溶菌酶（LYZ）活性无显著性差异（P>0.05）；与ESH组相比，BF-CX10灌胃显著上调小鼠肾脏干扰素-γ（IFN-γ）、肿瘤坏死因子-α（TNF-α）、白细胞介素-4（IL-4）和白细胞介素-10（IL-10）mRNA相对表达水平（P<0.05），显著下调小鼠肾脏白细胞介素-12（IL-12）mRNA相对表达水平（P<0.05）。小鼠粪便菌群测序多样性表明，测得的物种数在ESH组和HBF组之间具有显著的差异（P<0.05）；物种注释结果显示，在门水平上，LBF和HBF组以拟杆菌门和厚壁菌门为优势菌群，其中厚壁菌门主要为芽孢杆菌科和乳杆菌科；菌群功能预测结果显示，HBF组功能类聚以新陈代谢为主。结果提示，BF-CX10灌胃可维持小鼠正常体增重，提高血清酶活性及免疫功能，对小鼠肠道微生物种群恢复作用显著，其益生性较好。

关键词： 坚强芽孢杆菌CX10; 益生菌评价; 微生物测序

本文引用格式

马玉俊 , 梁子敬 , 张凯 , 李杰 , 马小军 , 王磊 , 张康 , 张景艳 , 李建喜 . 坚强芽孢杆菌CX10益生性评价[J]. 动物营养学报, 2019 , 31(11) : 5310 -5320 . DOI: 10.3969/j.issn.1006-267x.2019.11.048

Abstract

The aim of this study was to evaluate the probiotics of Bacillus firmus CX10 (BF-CX10) by oral gavage in mice, measuring their body weight, viscera index, serum enzyme activity and immune cytokine expression, as well as the structure and diversity of fecal flora. A total of 50 female BALB/c mice of 18 to 20 g were randomly divided into control group, Escherichia coli self-healing (ESH) group, low-dose BF-CX10 (LBF) group, medium-dose BF-CX10 (MBF) group and high-dose BF-CX10 (HBF) group, with 10 mice in each group. Mice in the control group were given normal saline throughout the whole period, those in the ESH group were given Escherichia coli O78 on the first 3 days and normal saline on the second 3 days, and those in the LBF group, MBF group and HBF group were given 1.5×104, 1.5×106 and 1.5×108 CFU/mL BF-CX10 on the first 3 days respectively and Escherichia coli O78 on the second 3 days. The gavage dose was 0.02 mL/g BW. Mice were killed at day 7. The results show that compared with the ESH group, mice in BF-CX10 groups got the stable weight gain, and the indices of kidney and heart as well as serum activity of acid phosphatase (ACP), alkaline phosphatase (AKP) and glutathione peroxidase (GSH-Px) of mice in BF-CX10 groups were increased significantly (P<0.05), while serum lysozyme (LYZ) activity of mice in BF-CX10 groups was no significant difference (P>0.05). Compared with the ESH group, BF-CX10 gavage significantly up-regulated the mRNA relative expression levels of interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), interleukin-4 (IL-4) and interleukin-10 (IL-10) (P<0.05), and significantly down-regulated the interleukin-12 (IL-12) mRNA relative expression level (P<0.05) in mouse kidney. The diversity of mouse fecal flora sequencing showed that the measured species number had a significant difference between ESH group and HBF group (P<0.05). Species annotation results showed that at the phylum level, Bacteroidetes and Firmicutes were the dominant bacteria in LBF group and HBF group, among which the Firmicutes were mainly Bacillaceae and Lactobacillaceae. The results of bacterial function prediction showed that the functional clustering of HBF group was dominated by metabolism. The results suggest that BF-CX10 gavage can maintain normal body weight gain, increase serum enzyme activity and immune function of mice, and have a significant probiotic effect on the recovery of intestinal microbial population in mice.

Key words： Bacillus firmus CX10; probiotic evaluation; microbial sequencing

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