Real-time PCR分析免疫应激对肉鸡各肠段微生物区系的影响

doi:10.3864/j.issn.0578-1752.2013.22.018

摘要/Abstract

摘要： 【目的】应用细菌的16S rRNA 序列设计双歧杆菌属(Bifidobacterium)、乳杆菌属( Lactobacillus) 、肠杆菌科(Enterobacteriaceae)和肠球菌属( Enterococcus) 的引物并对肉鸡肠道的这4类细菌菌属进行定量测定，检测免疫应激对肉鸡不同肠段微生物区系的影响。【方法】将180只AA肉公鸡随机分4个处理，每处理设5个重复，每个重复有9只。试验分为NV、CV、LPS和CYP 处理组。免疫应激组分别于18、19、20、32、33和34 d肌肉注射相应剂量的LPS和CYP，NV组和CV组于同日龄注射等体积生理盐水。分别于21、28、35和42 d，每处理组各取6只鸡，无菌操作，宰杀后取十二指肠、空肠、回肠和盲肠内容物。将待测样品中提取的DNA分别进行4类菌属进行Real-time PCR反应。【结果】与NV相比，4类细菌的基因拷贝数发生显著变化。集中在回肠和盲肠，肠杆菌属：5.03—7.88 copies/mg；乳酸杆菌属：5.48—10.34 copies/mg；双歧杆菌属：4.11—7.98 copies/mg；肠球菌属：5.61—8.79 copies/mg。双歧杆菌属含量为4种菌属中细菌含量最低，乳酸杆菌含量高，盲肠含菌最多，十二指肠含量最少，后期各菌群趋于稳定。【结论】免疫应激可引起肠道菌群失调，乳杆菌属和双歧杆菌属的数量在无免疫情况下的含量相对高，LPS和CYP对肉鸡后期的肠道菌群的无显著影响。

关键词: RT-PCR , 微生物区系 , 免疫应激 , 肉鸡

Abstract: 【Objective】The purpose of this experiment was to investigate the effects of immune stress on intestinal microflora and common bacteria changes of broiler chickens. 【Method】 One hundred and eighty 1-day old AA broilers were allocated into 4 groups. Each treatment had 5 replicate cages of 9 broilers per replicate pen. Different immune stress models, including the control group (no vaccines, NV), conventional vaccines group (CV), immune hyperthyroidism group (LPS) and immune suppression group (CYP) were set in this study. On d 21, 28, 35 and 42, each treatment group from each of 6 chickens, after slaughter, jejunum, ileum and duodenum and cecum intestinal contents were collected under sterile operation. 【Result】Gastrointestinal tract Enterobacteriaceae, Lactobacillus, Bifidobacterium and Enterococcus in broilers under immunological stress were analyzed by RT-PCR. The dynamic distribution and changes of the four bacterial species in the broiler's digestive tract were also analyzed systematically. Among the 4 genera of bacteria, changes mainly concentrated in the ileum and cecum, and the Enterobacteriaceae content was 5.03-7.88 copies/mg, Lactobacillus content was 5.48-10.34 copies/mg, Bifidobacterium content was 4.11-7.98 copies/mg, Enterococcus content was 5.61-8.79 copies/mg. The results showed that immune stress makes the four bacterial species in broiler's digestive tract change conspicuously in their genic copy when compared with CV. In which, the changes of the four bacterial species mainly represent in ileum and cecum, while the changes of days by duodenum species is not clear. 【Conclusion】 Immune stress results in the changes of the intestinal flora composition, Lactobacillus and Bifidobacterium are relatively high in NV case and LPS and CYP did not affect intestinal flora changes of chicken in late stage.

Key words: RT-PCR , intestinal microflora , immue stress , broilers

冯焱12, 杨小军2, 李武林2, 杨宁1, 姚军虎2. Real-time PCR分析免疫应激对肉鸡各肠段微生物区系的影响[J]. 中国农业科学, 2013, 46(22): 4800-4807.

FENG Yan-12, YANG Xiao-Jun-2, LI Wu-Lin-2, YANG Ning-1, YAO Jun-Hu-2. Effects of Immune Stress on the Bacterial Community in the Broiler Chickens Gastrointestinal Tract Analyzed by Real-Time PCR[J]. Scientia Agricultura Sinica, 2013, 46(22): 4800-4807.

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