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【目的】研究不同处理羊粪低温沼气发酵液的细菌群落多样性,为羊粪低温沼气发酵菌剂制备提供理论依据。【方法】对不同处理羊粪沼液总DNA进行16S r DNA V3区PCR扩增,扩增产物进行变性梯度凝胶电泳(DGGE)分离、回收、测序,BLAST软件比对分析,构建发育树;同时利用Biolog-ECO法对沼液样本进行微生物功能多样性分析。【结果】PCR-DGGE方法结果显示,在低温条件下(15℃),在发酵各阶段均存在共有优势菌群,包括梭菌属(Clostridium sp.)、拟杆菌属(Bacteroidales bacterium)、假单胞菌属(Pseudomonas sp.)、Proteiniphilum sp.等,其中菌剂III处理的4号为最优发酵试验组,主要为梭菌属(Clostridium sp.)表现有较高的丰度;Biolog-ECO法分析结果显示,在15℃培养条件下,产气高峰期处理2、3和4中,碳代谢活性相对较高,产气量也比对照组高。通过微生物群落的多样性分析发现,处理2的均匀性指数均高于其它处理组。处理3和4的微生物群落的丰富度、优势度指数相对较高。【结论】在低温条件下(15℃)的不同处理中,添加配伍III(配伍I+2 g ZHC固体菌剂)发酵效果最优。
【Objective】 The objective of this study was to study the diversity of bacterial communities in different temperature treatments of biogas fermentation of sheep manure and to provide a theoretical basis for the preparation of low-temperature biogas fermentation agent for sheep manure. 【Method】 The 16S r DNA V3 region of total DNA from different mannan stock treatments was amplified by PCR. The amplified products were separated by denaturing gradient gel electrophoresis (DGGE), recovered, sequenced and compared with BLAST software to construct the developmental tree. Biolog-ECO method was also used to analyze the microbial diversity of biogas samples. 【Result】 The results of PCR-DGGE showed that there were common dominant bacteria in all stages of fermentation, including Clostridium sp., Bacteroidales bacterium, Pseudomonas sp., Proteiniphilum sp. And so on. Among them, the No.4 treatment was the optimal fermentation test group, and the Clostridium sp. Showed the highest abundance. Biolog-ECO The results of the method showed that carbon metabolism activity was relatively high and gas production was higher than that of the control group in the treatments of 2, 3 and 4 during peak gas production at 15 ℃. According to the diversity analysis of microbial communities, the uniformity index of treatment 2 was higher than that of other treatment groups. The microbial community of treatments 3 and 4 had a higher abundance and a higher dominance index. 【Conclusion】 In different treatments at low temperature (15 ℃), the compatibility of III (compatibility I + 2 g ZHC solid fungus) was the best.