2株羊源芽孢杆菌的筛选及其对小鼠肠道生化指标和肠道菌群的影响

Screen of Two Bacterial Strains of Bacillus Isolated from Goat and Their Effects on Intestinal Biochemical Indexes and Intestinal Microflora of Mice

本试验旨在从川中黑山羊粪便中筛选具有广谱抑菌作用的菌株,并探究其对小鼠肠道生化指标和肠道菌群的影响。试验采用牛津杯法从粪便中筛选出具有广谱抑菌作用的菌株,分别命名为菌株fsznc-06和fsznc-09。选取60只健康的小鼠,随机分为5组,每组3个重复,每个重复4只。JD1组和JD2组分别灌服1×10^8和1×10^10CFU/mL菌株fsznc-06,DX1组和DX2组分别灌服1×10^8和1×10^10CFU/mL菌株fsznc-09,对照组(DZ组)灌服0.9%氯化钠溶液。每2 d灌服1次,灌服量均为0.3 mL/只。饲养28 d后测定小肠组织总超氧化物歧化酶(T-SOD)、碱性磷酸酶(AKP)和溶菌酶(LZM)活性,并取小肠内容物进行微生物测序和多样性分析。结果表明:本试验筛选出2株具有广谱抑菌作用的菌株,经鉴定并分别命名为解淀粉芽孢杆菌(Bacillus amyloliquefaciens)fsznc-06和短小芽孢杆菌(Bacillus pumilus)fsznc-09。与DZ组相比,JD1和DX1组小肠组织T-SOD、AKP和LZM活性显著或极显著提高(P<0.05或P<0.01),JD2组小肠组织T-SOD活性极显著提高(P<0.01),DX2组小肠组织AKP活性极显著提高(P<0.01)。各试验组小肠肠道菌群多样性均高于DZ组。各组优势物种分别为:JD1组,约氏乳杆菌(53.54%);JD2组,罗伊氏乳杆菌(44.56%);DX1组,约氏乳杆菌(45.21%);DX2组,约氏乳杆菌(41.38%);DZ组,约氏乳杆菌(44.51%)。JD1和JD2组肠道菌群主要提高了遗传信息处理方面的功能,DX1和DX2组肠道菌群主要提高了环境信息处理、新陈代谢和生物体系统方面的功能。由此可见,解淀粉芽孢杆菌fsznc-06和短小芽孢杆菌fsznc-09能提高小鼠小肠组织T-SOD、AKP和LZM活性,增加小鼠肠道菌群多样性和有益菌群相对丰度。

The purpose of this study was to screen bacterial strains with antibacterial activity from goat droppings,and explore their effects on the intestinal biochemical indexes and intestinal microflora of mice.In this study,the strains with antibacterial activity were screened from the goat droppings using the Oxford cup method and named bacterial strains fsznc-06 and fsznc-09,respectively.Sixty healthy mice were randomly divided into 5 groups with 3 replicates per group and 4 mice per replicate.JD1 and JD2 groups were administered orally with bacterial strain fsznc-06 with concentrations of 1×10^8 and 1×10^10 CFU/mL,respectively;DX1 and DX2 groups were administered orally with bacterial strain fsznc-09 with concentrations of 1×10^8 and 1×10^10 CFU/mL,respectively;the control group(DZ group)was administered orally with 0.9%sodium chloride solution.Every 2 days administered orally once,the dosage was 0.3 mL per mouse.After feeding 28 days,the activities of total superoxide dismutase(T-SOD),alkaline phosphatase(AKP)and lysozyme(LZM)in small intestine tissue were determined,and the contents of small intestine were taken for microbial sequencing and diversity analysis.The results showed that two bacterial strains with antibacterial activity were screened in this experiment,they were identified and named as Bacillus amyloliquefaciens fsznc-06 and Bacillus pumilus fsznc-09.Compared with the DZ group,the activities of T-SOD,AKP and LZM in small intestine tissue of JD1 and DX1 groups were significantly increased(P<0.05 or P<0.01),the small intestine tissue TSOD activity of JD2 group was significantly increased(P<0.01),and the small intestine tissue AKP activity in DX2 group was significantly increased(P<0.01).The diversities of intestinal microflora in small intestine of JD1,JD2,DX1 and DX2 groups were higher than those of DZ group.The dominant species in JD1,JD2,DX1,DX2 and DZ groups were Lactobacillus johnsonii(53.54%),Lactobacillus reuteri(44.56%),Lactobacillus johnsonii(45.21%),Lactobacillus johnsonii(41.38%),Lactobacillus johnsonii(44.51%),respectively.The intestinal microflora of JD1 and JD2 groups mainly improved the functions of genetic information processing,and the intestinal microflora of DX1 and DX2 groups mainly improved the functions of environmental information processing,metabolism and organismal systems.In conclusion,Bacillus amyloliquefaciens fsznc-06 and Bacillus pumilus fsznc-09 can improve the activities of T-SOD,AKP and LZM in small intestine tissue,and increase the diversity of small intestinal microflora and the relative abundance of beneficial microflora of mice.