Isolation, Identification and Nitrogen Removal Effect of Nitrifying Bacteria in Sediment and Water of Pacific White Shrimp Litopenaeus vannamei Culture Ponds
ZHANG Dajuan1,2, ZHANG Shulin1,2, DAI Wei1,2, KONG Xin1, WANG Zebin1, BI Xiangdong1,2
1.Department of Fishery Science, Tianjin Agricultural University, Tianjin 300384, China; 2.Tianjin Key Laboratory of Aqua-Ecology and Aquaculture, Tianjin 300384, China
Abstract：Five strains of bacteria were isolated and purified from water and sediment in Pacific white leg shrimp Litopenaeus vannamei culture ponds using dilution-plate method and streak plate method, after Gram staining, and identified as Pseudomonas mendocin, Methylobacterium aminovorans, Rhizobium radiobacter, P. stutzeri and Microbacterium oxydans by morphological observation, physiological and biochemical characteristics and 16S rDNA sequence analysis to investigate the component of bacteria of water and sediment in the Pacific white leg shrimp culture pond. The water from the shrimp ponds was filtered into 250 mL conical flasks and inoculated with the five strains (with final density of 2 ×107 cfu/mL) and cultured at 30 ℃ and 120 r/min for 48 hours to study their nitrification function and ammonia nitrogen removal rates.The ammonia nitrogen removal efficiency was shown to be all above 85%, 88.83% in P. mendocin and 91.73% in P. stutzeri; the removal efficiency of nitrite nitrogen were all above 90%, 95.08% in P. mendocin and 97.39% in P. stutzeri; and the removal efficiency of nitrate was above 90%, 90.49% in P. mendocin and 91.22% in P. stutzeri. The findings indicated that Pseudomonas has higher removal efficiency of ammonia nitrogen and nitrite nitrogen, and has potential application in treatment of shrimp effluent.
张达娟, 张树林, 戴伟, 孔欣, 王泽斌, 毕相东. 凡纳滨对虾养殖池塘硝化细菌的分离鉴定及脱氮效果研究[J]. 水产科学, 2020, 39(2): 265-270.
ZHANG Dajuan, ZHANG Shulin, DAI Wei, KONG Xin, WANG Zebin, BI Xiangdong. Isolation, Identification and Nitrogen Removal Effect of Nitrifying Bacteria in Sediment and Water of Pacific White Shrimp Litopenaeus vannamei Culture Ponds. 水产科学, 2020, 39(2): 265-270.
[1]刘淑集,吴成业.水产饲用益生菌的机理及功效[J]. 天津农业科学,2010,16(5):58-62. [2]臧淑梅.氨氮在水产养殖中的产生、危害及控制[J].黑龙江水产,2012(2):38-39. [3]张庆华,封永辉,王娟,等.地衣芽孢杆菌对养殖水体氨氮、残饵降解特性研究[J].水生生物学报,2011,35(3):498-503. [4]黄翔鹄,李长玲,郑莲,等.亚硝酸盐氮对凡纳滨对虾毒性和抗病相关因子影响[J].水生生物学报,2006,30(4):466-471. [5]郑佳佳,沈涛,傅罗琴,等.一株硝化反硝化菌的筛选鉴定及反硝化特性研究[J].水生生物学报,2011,36(1):161-167. [6]Li P, Zhang S, Liu D L.Study progress of bacterial aerobic denitrification[J]. Journal of Microbiology,2005,25(1):60-64. [7]葛辉.异养硝化与好氧反硝化细菌的筛选、培养及综合除氮研究[D].苏州:苏州科技学院,2011. [8]张峰峰,周可,谢凤行,等.一株异养硝化菌AD-28氨氮去除特性研究[J].水产科学,2019,38(1):92-97. [9]姚艳玲,袁春营,崔青曼,等.一株去除亚硝态氮细菌分离鉴定及特性研究[J].水产科学,2018,37(5):689-693. [10]谢骏,方秀珍,郁桐炳.池塘单循环中各种细菌与理化因子的相关性研究[J].水生生物学报,2002,26(2):180-187. [11]刘慧玲,张战锋,李长玲,等.光合细菌对罗非鱼鱼苗养殖水质及抗病力的影响[J].渔业现代化,2009,36(2):47-51. [12]尹文林,沈锦玉,潘晓艺,等.复合硝化菌制剂对水质改良的应用效果[J].水产养殖,2010,31(9):12-17. [13]宋蓉.乳酸菌对养殖水体亚硝酸盐控制机理的研究[D].武汉:华中农业大学,2012. [14]王京伟.微生物对养殖水体水质调控作用的研究[D].太原:山西大学,2007. [15]沈萍,范秀容,李广武.微生物学实验[M].北京:高等教育出版社,2004. [16]洪庆华,石璐,孙井梅,等.革兰氏染色三步法应用实验探讨[J].实验室探索与研究,2010,29(11):15-17. [17]王芳,叶宝兴,宋瑛琳,等.微生物学实验中革兰氏染色两种方法的比较[J].试验室研究与探索,2007,26(8):123-124. [18]国家环境保护总局《水和废水监测分析方法》编委会.水和废水监测分析方法[M].北京:中国环境科学出版社,2002. [19]郑平,徐向阳,胡宝兰.新型生物脱氮理论与技术[M].北京:科学出版社,2004:55-74. [20]王景峰,金敏,谌志强,等.1株好氧脱氮菌的筛选与脱氮特性研究[J].环境科学,2011,32(8):2409-2413. [11]曾庆武.梁运祥,葛向阳.反硝化细菌的分离筛选及其反硝化特性的初步研究[J].华中农业大学学报:自然科学版,2008,27(5):616-620. [22]魏巍,黄廷林,苏俊峰,等.1株贫营养好氧反硝化菌的分离鉴定及其脱氮特性[J].生态环境学,2010,19(9):2166-2171. [23]Kim M, Jeong S Y, Yoon S J, et al. Aerobic denitrification of Pseudomonas putida AD-21 at different C/N ratios[J].Journal of Bioscience and Bioengineering,2008,106(5):498-502. [24]邢国伟,李彦芹,李凤超,等.一株反硝化细菌与光合细菌对养殖海水的净化效果[J]. 生物技术通报,2014(3):151-154. [25]Cladera A M, Sepúlveda-Torres L del C,Valens-Vadell M, et al. A detailed phenotypic and genotypic description of Pseudomonas strain OX1[J]. Systematic & Applied Microbiology,2006,29(5):422-430. [26]邓斌.施氏假单胞菌SC221_M的反硝化特定及其调节草鱼养殖水体水质的研究[D].杭州:浙江大学,2014. [27]孔庆鑫,李君文,王新为,等.一种新的好氧反硝化菌筛选方法的建立及新菌株的发现[J].应用环境生物学报,2005,11(2):222-225. [28]林娜,郭楚玲,柯林,等.富营养化池塘中好氧反硝化细菌的分布及脱氮研究[J].中国科技论文在线,2010,5(5):369-372,376. [29]杨俊忠,倪砚,许尚营,等.一株高效好氧反硝化菌的分离及特性[J].微生物学通报,2010,37(11):1594-1599. [30]吴小丹.一株极端环境微生物生理特性研究以及菌株鉴定[D].武汉:华中师范大学,2011. [31]Chen J, Xiang L, Jun L, et al. Degradation of environmental endocrine disruptor di-2-ethylhexyl phthalate by a newly discovered bacterium, Microbacterium sp. strain CQ0110Y[J]. Applied Microbiology & Biotechnology,2007,74(3):676-682. [32]张通,吴琼,刘柯澜,等.微杆菌对乌海高硫煤中有机硫脱除的研究[J].煤炭学报,2009,34(7):957-960. [33]王亚南,彭志英.海水养虾场底泥中产H2S的细菌特性[J].华南理工大学学报:自然科学版,2004,32(8):89-92. [34]黄兴如,张彩文,张晓霞.根瘤菌在污染土壤修复中的地位和作用[J].中国土壤与肥料,2016(5):5-10. [35]董晓霞,刘兆辉,李志禄,等.豆科牧草对滨海盐渍土壤盐分特性和肥力影响的研究[J].安徽农业科学,2008,36(14):6060-6062. [36]Wani P A, Khan M, Zaidi A. Effect pf metal-tolerant plant growth-promoting Rhizobium on the performance of per grown in metal-amended soil[J]. Archives of Environmental Contamination and Toxicology, 2008,55(1):33-42. [37]Ahmad D, Mehmannavaz R, Damaj M. Isolation and characterization of symbiotic N2-fixing Rhizobium meliloti from soils contaminated with aromatic and chloroaromatic hydrocarbons:PAHs and PCBs[J]. International Biodeterioration and Biodegradation,1997,39(1):33-43. [38]孙彦波,高志茹,赵贤俊.嗜甲基菌处理甲醇废水[J].今日科苑,2007(12):110-110. [39]马立周,李江华,房峻,等.1株产丝氨酸甲基转移酶菌株的分离鉴定及其培养条件优化[J].食品与发酵工业,2008,34(5):18-23.
2020, Vol. 39 
