在制浆造纸过程中,白水的循环回用加重了纸机微生物污染的问题,促进了纸机腐浆的形成,影响了纸机的正常运转。此次研究用滤膜抽滤富集细菌菌体,对白水样品直接提取DNA,通过16S rDNA序列分析,构建16S rDNA文库,对芬欧汇川(UPM)常熟纸厂PM1纸机白水中的细菌多样性进行了研究。获得的61个16S rDNA序列可分为28个可操作分类单元(OTUs),分属于6个不同门类,优势菌种顺序为变形菌门(Proteobacteria)(77.05%)、厚壁菌门(Firmicutes)(11.48%)、放线菌门(Actinobacteria)(4.92%)、异常球菌-栖热菌门(Deinococcus-Thermus)(3.28%)、拟杆菌门(Bacteroidetes)(1.64%)、蓝藻门(Cyanobacteria)(1.64%)。造纸白水具有较丰富的细菌多样性,以变形菌为优势菌群,占库容总量的77.05%,其中尤以β-变形菌群为主,占库容总量的45.90%,而首次在中国纸机鉴定出的水生施莱格尔氏菌(Schlegelella aquatica)为克隆文库的优势菌株,另外还含有2个菌种属于异常球菌-栖热菌门,该菌门为红色腐浆形成菌种。
Abstract
Paper machines offer a suitable environment for microbial growth, which results in slime problems during the process of papermaking. Investigation of the bacterial diversity in whitewater which recycled in paper mill UPM was carried out in this paper. Total DNA were directly extracted from the whitewater sample. 16S rDNA were amplified directly from purified DNA by PCR with universally bacteria-specific rDNA primers and cloned. Sixty-one bacteria were identified by partial sequencing of their 16S rDNAs. Twenty-eight operational taxonomic units(OTUs)were clustered in the following phyla including Proteobacteria(77.05%), Firmicutes(11.48%), Actinobacteria(4.92%), Deinococcus-Thermus(3.28%), Bacteroidetes(1.64%),Cyanobacteria(1.64%). Bacterial diversity was high in the whitewater sample. The results showed a high proportion of Schlegelella aquatica in the white water samples. It was the first time that Schlegelella aquatica strain had been identified from a paper machine in China. We also identified the red-slime formation bacteria Deinococcus-Thermus in whitewater sample.
{{custom_sec.title}}
{{custom_sec.title}}
{{custom_sec.content}}
参考文献
[1] 黄淋佳,黄创锋,杨伟和,等. 造纸沉积物的产生和防治[J].广东化工, 2009, 36(7):306-307.
Huan L J, Huang C F,Yang W H, et al.Prevention and control of deposits in papermaking process[J].Guangdong Chemical Industry,2009, 36(7):306-307.
[2] Blanco M A, Negro C, Gaspar I, et al. Slime problems in the paper and board industry[J]. Appl Microbiol Biotechnol, 1996, 46(3):203-208.
[3] Lacorte S, Latorre A, Barceló D, et al. Organic compounds in paper-mill process waters and effluents[J]. TrAC Trends in Analytical Chemistry, 2003, 22(10):725-737.
[4] Lahtinen T, Kosonen M, Tiirola M, et al. Diversity of bacteria contaminating paper machines[J]. Journal of Industrial Microbiology and Biotechnology, 2006, 33(9):734-740.
[5] Desjardins E, Beaulieu C. Identification of bacteria contaminating pulp and a paper machine in a Canadian paper mill[J]. Journal of Industrial Microbiology and Biotechnology, 2003, 30(3):141-145.
[6] Ekman J. Bacteria colonizing paper machines[D]. Helsinki:Department of Food and Environmental Sciences Faculty of Agriculture and ForestryUniversity of Helsinki, 2011.
[7] 赵丰,焦虎美,时留新.造纸网下白水所含细菌的分离和鉴定[J].南京林业大学学报:自然科学版, 2011,35(3):115-119.
Zhao F, Jiao H M, Shi L X.Separation and identification of microorganisms from the white water in the wire pit[J]. Journal of Nanjing Forestry University:Natural Sciences Edition, 2011,35(3):115-119.
[8] 焦虎美, 时留新.腐浆中细菌的鉴定及其生物膜活性研究[J]. 南京林业大学学报:自然科学版,2012,36(2):77-81.
Jiao H M, Shi L X.Identification of bacterias from the slime and the study on their biofilm activity[J]. Journal of Nanjing Forestry University:Natural Sciences Edition, 2012,36(2):77-81.
[9] Pace N R. A molecular view of microbial diversity and the biosphere[J]. Science,1997,5313: 734-740.
[10] 国家环境保护总局.水和废水监测分析方法[M]. 4版. 北京:中国环境科学出版社, 2002.
[11] Zhou J, Bruns M A, Tiedje J M. DNA recovery from soils of diverse composition[J]. Applied and Environmental Microbiology, 1996, 62(2):316-322.
[12] Granhall U, Welsh A, Throbäck I N, et al. Bacterial community diversity in paper mills processing recycled paper[J]. Journal of Industrial Microbiology & Biotechnology, 2010, 37(10):1061-1069.
[13] Chou Y J, Sheu S Y, Sheu D S, et al. Schlegelella aquatica sp. nov., a novel thermophilic bacterium isolated from a hot spring[J]. International Journal of Systematic and Evolutionary Microbiology, 2006, 56(12):2793-2797.
[14] Kolari M, Nuutinen J, Rainey F A, et al. Colored moderately thermophilic bacteria in paper-machine biofilms[J]. Journal of Industrial Microbiology and Biotechnology, 2003, 30(4):225-238.
[15] Julie D, Beaulieu C, Villemur R. Composition of the bacterial biota in slime developed in two machines at a Canadian paper mill[J]. Canadian Journal of Microbiology, 2011, 57(2):91-104.
[16] Prince V, Simao-Beaunoir A M, Beaulieu C. Amplified ribosomal DNA restriction analysis of free-living bacteria present in the headbox of a Canadian paper machine[J]. Canadian Journal of Microbiology, 2009, 55(7):810-817.
[17] 董玉玲,朱万斌,郭鹏,等.一组小麦秸秆好氧分解菌系的构建及组成多样性[J]. 环境科学, 2010, 31(1):249-254.
Dong Y L,Zhu W B,Guo P, et al. Characteristics and diversity of a microbial community decomposing wheat straw under aerobic conditions[J].Environmental Science,2010, 31(1):249-254.
基金
收稿日期:2013-04-25 修回日期:2013-11-13
基金项目:国家林业局“948”项目(2012-4-09); 江苏高校优势学科建设工程资助项目(PAPD)
第一作者:吴芳芳,硕士生。*通信作者:时留新,副教授。E-mail: shiliuxin@njfu.edu.cn。
引文格式:吴芳芳,程强,时留新,等. 造纸白水中的细菌多样性及系统发育树分析[J]. 南京林业大学学报:自然科学版,2014,38(2):123-127.
PDF(1937984 KB)
