2015年3月，第21卷，第1期，168-171页
March 2015,Vol.21,No.1,pp.168171 D0I: 10.16108/j.issn10067493.2014057
高校地质学报
Geological Journal of China Universities
生物甲烷气实验的外加菌源（沼液）中厌氧菌群测定
夏大平13，王振'，苏现波12，司青1 1.河南理工大学能源科学与工程学院，焦作454000
2.河南省瓦斯地质与瓦斯治理重点实验室-省部共建国家重点实验室培育基地，焦作454000
3.中原经济区煤层（页岩）气河南省协同创新中心，焦作454000
摘要：考虑到煤本源微生物的分离、纯化及培养相对因难，且耗时较长，采用沼液作为外加菌源，并在进行生物甲烷气实验之前，对所采集沼液中的主要厌氧菌群进行微生物特征及数量的测定。结果表明沼液中的厌氧发酵菌（Anaerobic fermentativebacteria）、厌氧纤维素分解菌（Anaerobic cellulose-decomposingbacteria）、硫酸盐还原菌（Sulfate-reducing bacteria）和产甲烷菌（Methanogens）4类厌氧菌群，均具有杆状、球状及链状等多种形态；染色后发现其中革兰氏阴性菌占主要优势；经最大或然数（MPN）计数后表明，沼液中所含厌氧菌的数量足以进行后续的生物甲烷气的产出实验。关键词：本源微生物；沼液；生物甲烷气；厌氧菌群；MPN计数
中图分类号：Q93-33
文献标识码：A
文章编号：1006-7493（2015）01-0168-04
DeterminationofAnaerobicBacterialConsortiainAddedMicrobial
Sources(BiogasSlurry)forBiogenicMethaneExperiment
XIA Daping'3,WANG Zhen',SU Xianbo'2, SI Qing
1. School of Energy Science and Engineering, Henan Polytechnic University, Jiaozuo 454000, China
2. State Key Laboratory Cultivation Base for Gas Geology and Gas Control (Henan Polytechnic University), Jiaozuo 454000, China
3. Collaborative Innovation Center of Coalbed Methane and Shale Gas for
Central Plains Economic Region, Jiaozuo 454000, China
Abstract: Given that indigenous microbial consortia of raw coal samples are in great difficulty being separated, purificated and cultured, which are invariably at a big cost of time, biogas slurry was considered to be added as the microbial sources, of which the morphological characteristics and the amount of dominant anaerobic bacteria were determined prior to the conduction of biogenic-methane-production experiment, The results show that four types of anaerobic microbial consortia including fermentative bacteria, cellulolytic bacteria, sulfate-reducing bacteria and methanogens were separately indentified and observed with such recognizable morphological features as rod-shaped, spherical and streptobacteria; the gram-negative bacteria were found to be at a notable advantage after gram stain; further the content of microbial consortia determined by most probable number (MPN) counting method was deemed to be enough for following biogenic-methane-production experiment.
Key words: indigenous microbes; biogas slurry; biogenic methane; anaerobic microbial consortia; MPN counting First author: XIA Daping, Ph.D. Candidate; E-mail: 569689819@qq-com
20世纪90年代，煤层气的次生生物成因观点被提出（Scottetal.，1994），此后国内外很多研究人员在实验室条件下，成功地通过微生物的厌
氧降解将煤利用生成以甲烷为主的气体（李明宅和张辉，1998；AhmedandSmith，2001；王爱宽等，2010），取得了较好的模拟生气效果。但煤样
收稿日期：2014-04-10；修回日期：20140922
基金项目：国家自然科学基金（41472127；41472129)；河南省教育厅科学技术研究重点项日（13A440335;132102210253)；山西省煤层气联合研究基金
资助项目(2013012004)和河南省高校科技创新团队支持计划（15IRTSTHN027)联合资助
作者简介：夏大平，女，1983年生，在读博士生，实验师，煤层气地质与水文地质研究方向；E-mail:569689819@qq-com