利用real-timepcr研究乳清粉对试验兔肠道益生菌的影响.doc

研究报告 乳清粉对实验兔肠益生菌 吕建敏(刘月环 (1. 浙江中医药大学动物实验研究中心，杭州 310053；2. 浙江省医学科学院，杭州，310013) 【摘要】目的利用分子生物学技术研究饲粮中添加乳清粉对断奶早期实验兔肠道微生物区系和益生菌的影响。方法 采用单因子实验设计，选择40日龄断奶实验兔，随机分为4组（每组12只），分别饲以乳清粉添加水平为0%、2%、5%和10%的饲粮。饲喂30 d 后，每组随机取8只兔，麻醉处死，取盲肠内容物提取细菌总DNA，首先利用PCR-DGGE技术分析兔盲肠微生物区系多样性，进而应用实时荧光定量PCR（SYBR Green I）法定量检测盲肠细菌中双歧杆菌和乳酸杆菌含量。结果1）实验兔盲肠细菌DGGE分析参数随乳清粉添加量的提高逐渐上升， 添加不同水平乳清粉均可显著增加DGGE条带数（P0.05，P0.01）2%和5%乳清粉添加水平DGGE条带数和香农指数极显著（P0.01）未添加组（ P0.05）。均匀度指数无（P0.05）。2）饲粮中添加乳清粉可提高实验兔盲肠内容物中乳酸杆菌和双歧杆菌数量，其中乳清粉组的乳酸杆菌数量均显著高于组（P0.05），10%乳清粉添加组的双歧杆菌数量显著高于未添加组（P0.05），但与其他2个乳清粉添加组无差异显著性（P0.05）。结论饲粮中添加乳清粉在实验兔饲粮中添加乳清粉可有效增加肠道益生菌数量。 【关键词】实验兔；乳清粉；变性梯度凝胶电泳（DGGE）；实时荧光定量PCR双歧杆菌；乳酸杆菌【中图分类号】R33【文献标识码】A【文章编号】1671-7856（201）-0000-00 Doi:10.3969/j.issn.1671-7856.2015.008.000 Effects of dried whey on the intestinal bacterial community and probiotics in weaned laboratory rabbits LV Jian-min1，LIU Yue-huan2 (1. Laboratory Animal Research Center, Zhejiang Chinese Medical University, Hangzhou 310053, China；2. Zhejiang Academy of Medical Sciences, Hangzhou 310013) 【Abstracts】 Objective To investigate the effects of dried whey on the intestinal bacterial community and probiotics in weaned laboratory rabbits. Methods A single factor design was employed to investigate the effects of dried whey supplemented at levels of 0%, 2%, 5% and 10%, respectively, on 48 weaned (40-day-old) laboratory rabbits. At the day 30, eight rabbits in each group were taken and sacrificed after anesthesia. The total bacterial DNA from the ceacal content of each selected rabbit was drew to analyze the bacterial community and intestinal probiotics (Bifidobacterium and Lactobacillius) population by PCR-DGGE and real-time fluorescence quantitative PCR, respectively. Results 1) The DGGE parameters of ceacal bacterial community were increased with the increasing dried whey supplemental levels. The number of DGGE band in 2%, 5% and 10% dried whey supplement groups (P0.05, P0.01), the Shannon index in 5% and 1