瘤胃4株纤维降解细菌的分离鉴定及其纤维降解特性

doi:10.11843/j.issn.0366-6964.2016.11.018

Abstract

Abstract:

Four cellulose-degrading rumen bacteria strains were isolated from Mongolia sheep. The four strains bacteria were identified and numbered as H1, H2, N1 and N2. The growth characteristics and cellulase activity were measured to provide experimental material for subsequent experiments. Bacterial morphological, physiological and biochemical characterization, DNA sequence homology analysis were used for identification of isolated strains. Growth curve of strains were measured by using spectrophotometer. Filter paper enzyme activity, CMCase activity, salicin enzyme activity and microcrystalline cellulose enzyme activity of four strains of cellulolytic bacteria were measured by using reducing sugar method. Two strains of bacteria H1, H2 were identified as Ruminococcus flavefaciens and N1, N2 were Streptococcus. The growth curves of four strains showed that lag phase of H1, H2, N1, N2 were 3-4 h and the maximum cell concentration of H1, H2 appeared in 28 h and N1, N2 appeared in 14 h. Four strains of bacteria had high activity of filter paper enzyme. The activity of filter paper enzyme, CMCase and microcrystalline cellulose enzyme were significantly higher for H2 than for H1, N1 and N2 (P＜0.05). Filter paper enzyme activity of H1 was 0.08 μmol•(mL•min )^-1 and was lowest among four strains. Four strains bacteria with cellulose enzyme activity were isolated by using the method of roll tube technique. These four cellulolytic bacteria strains can be used for subsequent study.

CLC Number:

S826

WANG Wei-yun, LI Da-biao, YU Yong-qiang, XING Yuan-yuan, LI Zi-jian. Isolation and Identification of Four Cellulolytic Bacteria Strains from Sheep Rumen and Cellulose Degradation Characteristics[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2016, 47(11): 2294-2300.

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References

［1］孟培，译.瘤胃内微生物降解饲料的生理与生态(上)［J］.国外畜牧科技,1993,20(5):20-22.
MENG P (translate).Physiologycal aspects of digestion and metabolism in ruminants［J］. Animal Science Abroad,1993, 20(5):20-22. （in Chinese）
［2］DEHORITY B A. Rumen microbiology［M］.Nottingham University Press,2003：372.
［3］张迪,刘大程,卢德勋，等. 瘤胃内3 种主要纤维分解菌的研究进展［J］.中国畜牧兽医，2008，35(1):16-19.
ZHANG D, LIU D C, LU D X, et al. Advances in three species of cellulolytic bacteria in rumen［J］. China Animal Husbandry & Veterinary Medicine, 2008, 35(1):16-19. （in Chinese）
［4］张晓华, 谭蓓英, 刘敏雄. 一个分解纤维素的瘤胃梭菌新种［J］.微生物学报,1995,35（6）: 397-399
ZHANG X H, TAN P Y, LIU M X. Isolation and characterization of a clostridium rumenum sp.nov. from rumen［J］. Acta Microbiologica Sinica, 1995, 35（6）: 397-399.（in Chinese）
［5］刘玉承，侯先志，刘占英，等.2株瘤胃纤维降解细菌的分离鉴定［J］ .微生物学杂志,2007, 27（6）:1-4.
LIU Y C, HOU X Z, LIU Z Y, et al. Isolation and identification of two cellulose-degrading strains from rumen［J］.Journal of Microbiology, 2007,27（6）:1-4. （in Chinese）
［6］周非帆.瘤胃纤维降解细菌的分离鉴定及纤维降解特性研究［D］.呼和浩特:内蒙古农业大学,2014.
ZHOU F F. Isolation, identification and cellulose-degrading characterization of rumen cellulolytic bacteria strains［D］. Hohhot :Inner Mongolia Agricultural University,2014. （in Chinese）
［7］孙美洲，金巍，李袁飞，等. 瘤胃降解粗纤维产甲烷的厌氧真菌与甲烷菌共培养物的分离鉴定［J］.微生物学报,2014, 54(5): 563-571.
SUN M Z,JIN W,LI Y F, et al. Isolation and identification of cellulolytic anaerobic fungi and their associated methanogens from Holstein Cow［J］. Acta Microbiologica Sinica, 2014, 54(5): 563-571. （in Chinese）
［8］张宏福.动物营养参数与饲养标准［M］. 3 版.北京:中国农业出版社,2010.
ZHANG H F. Animal nutrition parameters and feeding standard ［M］. 3rd ed. Beijing：China Agriculture Press, 2010. （in Chinese）
［9］高巍, 孟庆翔.白色瘤胃球菌对秸秆细胞壁和微晶纤维素的附着比较［J］.中国畜牧杂志,2004, 40（6）:17-19.
GAO W, MENG Q X. Comparison of adhesion ability of ruminococcus albus-7 to corn stalks cell wall and crystalline cellulose［J］.Chinese Journal of Animal Science,2004, 40（6）:17-19. （in Chinese）
［10］刘占英, 侯先志, 刘玉承, 等.一株分解纤维素的肠球菌的分离鉴定［J］. 内蒙古大学学报, 2008, 39(2): 166-171.
LIU Z Y,HOU X Z,LIU Y C, et al. Isolation and identification of a strain cellulolytic enterococcus sp. CTB374-1［J］. Journal of Inner Mongolia University,2008, 39(2): 166-171. （in Chinese）
［11］东秀珠,蔡妙英.常见细菌系统鉴定手册［M］. 北京:科学出版社,2001.
DONG X Z,CAI M Y. Common bacterial systems identification manual ［M］.Beijing:Science Press,2001.(in Chinese)
［12］HOLT J G, KRIEG N R, SNEATH P H A, et al. Bergey′s manual of deteminative bacteriology［M］. Nith edition. Maryland:Williams＆Witkins Baltimore,1994.
［13］张莉莉,张苓花,史剑斐,等.利用氯化苄提取真菌基因组DNA及其分子生物学分析［J］.大连轻工业学院学报,2000,19(1):36-39.
ZHANG L L,ZHANG L H,SHI J F, et al. Isolation and molecular biological analyses of genomic DNAs from fungi using benzyl chloride［J］. Journal of Dalian Institute of Light Industry, 2000,19(1):36-39. （in Chinese）
［14］WEISSBURG W G, BARNS S M, PELLETIER D A, et al. 16S ribosomal DNA amplification for phylogenetic study［J］. J Bacteriol, 1991, 173(2):697-703.
［15］KOIKE S, KOBAYASHI Y. Development and use of competitive PCR assays for the rumen cellulolytic bacteria: F.succinogenes, R.albus and R.flavefaciens［J］. FEMS Microbiol Lett, 2001, 204(2):361-366.
［16］OHENE-ADJEI S,TEATHER R M,IVAN M, et al. Postinoculation protozoan establishment and association patterns of methanogenic archaea in the ovine rumen［J］. Appl Environ Microbiol,2007,72(14):4609-4618
［17］DENMAN S E,MCSWEENEY C S. Development of a real-time PCR assay for monitoring anaerobic fungal and cellulolytic bacterial populations within the rumen［J］.FEMS Microbiol Ecol,2006,58(3):572-582.
［18］GHOSE T K. Measurement of cellulase activities［J］. Pure Appl Chem, 1987, 59(2): 257-268.
［19］ZHANG Y H, LYND L R. Toward an aggregated understanding of enzymatic hydrolysis of cellulase: noncomplexed cellulase systems［J］. Biotechnol Bioeng, 2004, 88(7):797-824.
［20］焦金真，王芃芃，汤少勋，等.浏阳黑山羊胃肠道不同部位重要功能微生物的数量分布特征研究［J］.畜牧兽医学报，2013，44（10）：1590-1599.
JIAO J Z， WANG P P， TANG S X， et al. Quantity and distribution characteristice of function microoriganisms in gastrointestinal tract of liuyang black goats［J］.Acta Veterinaria et Zootechnica Sinica, 2013,44(10):1590-1599.(in Chinese)
［21］娜仁图雅，周非帆，杨金丽，等.绵羊瘤胃内纤维降解细菌的分离鉴定［J］.中国畜牧兽医,2014,41（5）: 142-146.
NAREN T Y，ZHOU F F， YANG J L， et al. Isolation and identification of the cellulose-degrading bacteria strains from sheep rumen［J］.China Animal Husbandry & Veterinary Medicine,2014,41（5）:142-146.(in Chinese)
［22］刘占英.绵羊瘤胃主要纤维降解细菌的分离鉴定及不同氮源对其纤维降解能力的影响［D］. 呼和浩特:内蒙古农业大学，2008.
LIU Z Y. Isolation and identification of major cellulolytic bacteria in rumen of sheep and effects of nitrogen sources in their cellulolytic activities［D］. Hohhot: Inner Mongolia Agricultural University,2008.(in Chinese)

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Isolation and Identification of Four Cellulolytic Bacteria Strains from Sheep Rumen and Cellulose Degradation Characteristics

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