东北梅花鹿肠道纤维素降解微生物的分离筛选

食草动物依赖胃肠道微生物分解利用天然木质纤维素物质。为获得高效纤维素降解菌，本文将东北梅花鹿粪便样品，经过研磨，重悬于羧甲基纤维素钠（CMC-Na）作为唯一碳源的富集培养基中，进行富集培养和筛选，用革兰氏碘液染色显示纤维素水解圈，通过纤维素水解圈直径（D）和菌落直径（d）比值，初步判断纤维素降解菌降解纤维素能力．筛选出106株纤维素降解微生物，包括72株细菌、22株真菌和12株放线菌．细菌16S rDNA和真菌ITS序列测序结果显示，这些纤维素降解菌主要集中分布于变形菌门（Proteobacteria）、厚壁菌门（Firmicutes）、放线菌门（Actinobacteria）和子囊菌门（Ascomycota）．本文提供了一种快速有效的反刍动物肠道纤维素降解微生物的筛选方法，为东北梅花鹿肠道微生物高通量测序结果提供实验数据支持． 

Isolation of cellulolytic microorganisms from sika deer feces

Herbivores rely on gastrointestinal microorganisms to decompose and utilize natural lignocellulosic materials．Here cellulose-degrading microorganisms were isolated from feces of sika deer (Cervus nippon hortulorum) to screen for highly efficient cellulose-degrading microorganisms．The feces samples were crushed, re-suspended in enrichment medium with sodium carboxymethyl cellulose (CMC-Na) as the sole carbon source．The sample suspension was diluted and spread on selection medium．Cellulose hydrolysis circle was measured by Gram’s iodine staining．Ratios of D (diameter of cellulose hydrolysis circle) to d (colony diameter) were used to determine cellulose degradation ability of the isolated microorganisms．Altogether 106 cellulose-degrading microorganisms were obtained, including 72 bacteria, 22 fungi and 12 actinomycetes．Bacterial 16S rDNA and fungal internal transcribed spacer (ITS) sequencing revealed that the isolated microorganisms were mainly distributed in Proteobacteria, Firmicutes, Actinobacteria and Ascomycota．