鹅油甘油二酯的体外抑菌效果及其对葡聚糖硫酸钠诱导大鼠盲肠微生物菌群结构的影响

谢玉娥; 王宝维; 葛文华; 张名爱; 岳斌; 刘国栋; 刘亚楠

doi:10.3969/j.issn.1006-267x.2020.01.047

动物营养学报 >

2020 , Vol. 32 >Issue 1: 405 - 416

DOI: https://doi.org/10.3969/j.issn.1006-267x.2020.01.047

实验方法与实验动物 EXPERIMENTAL METHOD AND ANIMAL

鹅油甘油二酯的体外抑菌效果及其对葡聚糖硫酸钠诱导大鼠盲肠微生物菌群结构的影响

谢玉娥 ,
王宝维 ,
葛文华 ,
张名爱 ,
岳斌 ,
刘国栋 ,
刘亚楠

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1. 青岛农业大学食品科学与工程学院, 青岛 266109;
2. 国家水禽产业技术体系营养与饲料功能研究室, 青岛 266109

谢玉娥(1994-),女,山东淄博人,硕士研究生,研究方向为营养与保健。E-mail:853402945@qq.com

收稿日期: 2019-07-01

网络出版日期: 2020-01-19

基金资助

国家水禽产业技术体系专项基金（CARS-43-11）；山东省农业重大应用技术创新项目（鲁财农指-2018）

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Antibacterial Effects of Goose Oil Diglyceride in Vitro and Its Effects on Cecal Microflora Structure by Dextran Sulfate Sodium Induced Rats

XIE Yu'e ,
WANG Baowei ,
GE Wenhua ,
ZHANG Ming'ai ,
YUE Bin ,
LIU Guodong ,
LIU Ya'nan

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1. Food Science and Engineering of Qingdao Agricultural University, Qingdao 266109, China;
2. National Waterfowl Industrial Technology System Nutrition and Feed Function Laboratory, Qingdao 266109, China

Received date: 2019-07-01

Online published: 2020-01-19

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摘要

本试验旨在通过体外抑菌试验和大鼠体内试验，评定鹅油甘油二酯（GDG）的体外抑菌效果及其对葡聚糖硫酸钠（DSS）诱导大鼠盲肠微生物菌群结构的影响。试验1（体外抑菌试验）：选取大肠杆菌、沙门氏菌、金黄色葡萄球菌和表皮葡萄球菌4种细菌，采用打孔法评定GDG抗菌活性，紫外分光光度计法测定GDG的抑菌率和4种细菌的生长曲线，并利用扫描电子显微镜观察GDG对4种细菌形态的影响。试验2（大鼠体内试验）：选用84只7周龄雄性SD大鼠，随机分为6组，每组7个重复，每个重复2只。正常对照组自由饮用蒸馏水；其余5组均进行DSS诱导，分为DSS模型组、GDG低剂量组、GDG中剂量组、GDG高剂量组和恩诺沙星（ENR）组。各组大鼠自由采食，正常对照组饮用蒸馏水；试验第1~7天，除正常对照组外，其他5组均饮用5% DSS水溶液；试验第8~14天，DSS模型组大鼠用灌胃针给予10 mL/kg BW蒸馏水灌胃处理，3个GDG组分别用灌胃针给予1、2和3 g/kg BW GDG灌胃处理，ENR组用灌胃针给予10 mL/kg BW ENR（5 μg/mL）灌胃处理。试验结束后，采用16S rRNA高通量测序技术对大鼠盲肠微生物菌群组成进行测定分析。结果表明：1）不同浓度的GDG对大肠杆菌、沙门氏菌、金黄色葡萄球菌和表皮葡萄球菌的生长有显著抑制作用（P<0.05），且最小抑菌浓度（MIC）分别为0.030、0.040、0.050和0.050 g/mL。2）扫描电子显微镜图像分析显示，GDG能够改变细菌细胞表面形态，通过破坏细菌细胞整体结构来抑制细菌生长繁殖。3）GDG能增加大鼠盲肠微生物的物种丰富度，优化盲肠微生物的菌群结构，具有调整肠道微生态失调的作用。由此可见，GDG对大肠杆菌、沙门氏菌、金黄色葡萄球菌和表皮葡萄球菌均具有显著抑菌作用，并有助于DSS诱导大鼠的盲肠菌群结构趋于恢复正常。

关键词： 甘油二酯; 抗菌; DSS; 大鼠; 盲肠菌群结构

本文引用格式

谢玉娥 , 王宝维 , 葛文华 , 张名爱 , 岳斌 , 刘国栋 , 刘亚楠 . 鹅油甘油二酯的体外抑菌效果及其对葡聚糖硫酸钠诱导大鼠盲肠微生物菌群结构的影响[J]. 动物营养学报, 2020 , 32(1) : 405 -416 . DOI: 10.3969/j.issn.1006-267x.2020.01.047

Abstract

The aim of this study was to evaluate the bacteriostatic effects of goose oil diglyceride (GDG) in vitro and its effects on cecal microflora structure by dextran sulfate sodium (DSS) induced rats through bacteriostasis test in vitro and in vivo test in rats. Test 1 (bacteriostasis test in vitro):four kinds of bacteria of Escherichia coli, Salmonella, Staphylococcus aureus and Staphylococcus epidermidis were selected, the antibacterial activity of GDG was tested by perforating method, the antibacterial rate and growth curve of four kinds of bacteria were measured by ultraviolet spectrophotometer, and the morphological changes of GDG on four kinds of bacteria were observed by scanning electron microscope. Test 2 (in vivo test in rats):eighty-four 7-week-old male SD rats were randomly divided into 6 groups with 7 replicates per group and 2 rats perreplicate. Rats in the normal control group were free drank distilled water; the other 5 groups were drank 5% DSS aqueous solution, and they were divided into DSS model group, GDG low dose group, GDG medium dose group, GDG high dose group and enrofloxacin (ENR) group. Rats in each group were ad libitum and rats in the control group were free drank distilled water; during days 1 to 7 of the experiment, all the other 5 groups drank 5% DSS aqueous solution except the normal control group; during days 8 to 14 of the experiment, rats in DSS model group were given 10 mL/kg BW distilled water by gavage, rats in 3 GDG groups were given 1, 2 and 3 g/kg BW GDG by gavage, respectively, and rats in ENR group were given 10 mL/kg BW ENR (5 μg/mL) by gavage. After the end of the experiment, 16S rRNA high-throughput sequencing technology was used to determine the composition of cecal microflora of rats. The results showed as follows:1) different concentrations of GDG had significant inhibitory effects on growth of Escherichia coli, Salmonella, Staphylococcus aureus and Staphylococcus epidermidis (P<0.05), and the minimum inhibitory concentration (MIC) were 0.030, 0.040, 0.050 and 0.050 g/mL, respectively. 2) Scanning electron microscope image analysis showed that GDG could change the surface morphology of bacteria, destroy the overall structure of bacterial cells to inhibit the growth and reproduction of bacteria. 3) GDG could increase the species richness of cecal microflora in rats, optimized the flora structure of caecal microflora, and played a role in regulating intestinal microecological imbalance. It is concluded that GDG has significant antibacterial effects on Escherichia coli, Salmonella, Staphylococcus aureus and Staphylococcus epidermidis, and helpe DSS induce the structure of cecum flora in rats to return to normal.

Key words： diglyceride; antibacterial; DSS; rats; caecal flora structure

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