持续偏热环境对肉鸡盲肠菌群多样性的影响

doi:10.3864/j.issn.0578-1752.2016.01.017

摘要/Abstract

摘要： 【目的】环境温度是影响家禽肠道菌群的重要因素之一，随着环境温度的改变，家禽肠道正常菌群也会受到影响，研究持续偏热环境对肉鸡肠道微生物多样性及结构的影响，为肉鸡健康养殖提供理论依据。【方法】试验选取22 d爱拔益加（AA）肉鸡144只转入环境控制舱，随机分成3个处理组（21、26和31℃），每个处理6个重复，每个重复8只鸡（公母各4只），在21℃、相对湿度60%的环境下适应7 d。29 d时，试验温度分别调整到21、26和31℃，相对湿度60%，至试验结束，共14 d。于试验第7、14天末，每处理随机选取6只（公母各3只；每重复选1只），无菌采集盲肠内容物，将同一处理组的6个样品混合均匀，迅速放入无菌的离心管中，液氮速冻，-80℃冰箱保存备用。采用16S rDNA的PCR-DGGE分子技术，结合共性和特异性条带割胶回收DNA进行克隆和测序，分析持续偏热处理7、14 d时，对盲肠内容物菌群的结构和多样性的影响。【结果】（1）26、31℃组肉仔鸡盲肠DGGE条带数（菌群丰富程度）均低于21℃组，且31℃组低于26℃组；31℃组肉仔鸡盲肠DGGE条带数，在试验14 d明显低于试验7 d；（2）不同组之间聚类图和多样性指数分析可知，26、31℃组对肉鸡盲肠菌群的影响较大，且31℃组肉鸡盲肠菌群多样性，在试验14 d明显低于试验7 d；（3）DGGE图谱条带序列分析表明，适温和偏热环境下肉鸡盲肠内的共性菌群是Alistipes timonensis和Barnesiella viscericola，21℃组肉鸡盲肠中特异性菌群是Ruminococus faecis，26、31℃组肉鸡盲肠中特异性菌群是Lutispora thermophila和Faecalibacterium prausnitzii，31℃组肉鸡盲肠中特异性菌群是Gemmiger formicilis。【结论】持续偏热处理（26和31℃）与21℃组相比，影响肉鸡肠道菌群的结构和多样性，且不同偏热程度、偏热处理时间对肉鸡影响程度不同。

关键词: 偏热环境, 肉鸡, 肠道菌群, 变性梯度凝胶电泳, 多样性

Abstract: 【Objective】The objective of this study is to provide useful information for healthy breeding on broilers. Ambient temperature is one of the important factors affecting the poultry intestinal flora; as the ambient temperature changes, poultry normal intestinal flora will be affected. This study was carried out to investigate the effects of constant moderate temperatures on structural diversity of cecal bacteria in broilers. 【Method】One hundred and forty four 22-day-old Arbor Acres (AA) broilers were assigned to three environment chambers, each chamber contained six cages with eight birds per cage (four males and four females), with each cage used as a replicate. The pre-test period lasted for 7 days and broilers were kept at 21℃ and 60% relative humidity. When broilers were 29-day-old, the temperatures of each environmental chamber were gradually regulated to 21, 26 and 31℃, respectively, while maintaining the relative humidity at 60% and both kept constant until the end of the experiment. The trial period lasted for 14 days. On day 21 and 42, one birds from each replicate were randomly selected and killed, cecal contents were aseptically collected, placed in a centrifugal tube, rapidly frozen in liquid nitrogen, and stored at -80℃. The bacterial community and diversity in the ceacal digesta of broilers at the 7 and 14 days of constant moderate temperatures was studied by using 16S rDNA-based denaturing gradient gel electrophoresis (DGGE). 【Result】 (1) The number of PCR-denaturing gradient gel electrophoresis bands of 26 and 31℃ group was lower than 21℃ group, and 31℃ group was lower than 26℃, at 31℃, broiler cecal DGGE bands at the 14 days are significantly lower than at the 7 days. (2) The dendrograms and Shannon-Weaver indices revealed that diversity of the intestinal microbial flora at 26 and 31℃ group was less than at 21℃; at 31℃,14 days are significantly lower than at the 7 days; (3) DGGE profiles stripe sequence analysis showed that Alistipes timonensis and Barnesiella viscericola were predominant in the 21, 26 and 31℃ groups, The special bacteria in 21℃, 26 and 31℃ groups were Ruminococus faecis, Lutispora thermophila and Faecalibacterium prausnitzii respectively, 31℃ group were Gemmiger formicilis.【Conclusion】Compared with 21℃, continuous treatment with constant moderate temperature (26℃ and 31℃) can significantly impact the community structure and diversity of intestinal flora in broilers, and the extent of effect varies with the different temperatures.

Key words: moderate temperatures, broiler, microbiota, denaturing gradient gel electrophoresis, diversity

彭骞骞，王雪敏，张敏红，冯京海，甄龙，张少帅. 持续偏热环境对肉鸡盲肠菌群多样性的影响[J]. 中国农业科学, 2016, 49(1): 186-194.

PENG Qian-qian, WANG Xue-min, ZHANG Min-hong, FENG Jing-hai, ZHEN Long, ZHANG Shao-shuai . Effects of Constant Moderate Temperatures on the Diversity of the Intestinal Microbial Flora of Broilers[J]. Scientia Agricultura Sinica, 2016, 49(1): 186-194.

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