适用于猪场沼液处理的好氧反硝化菌筛选及其脱氮特性评价
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摘要
试验旨在筛选适合于猪场沼液处理的好氧反硝化菌。从活性污泥中分离获得16株好氧反硝化菌,其中菌株ZH-14的总氮(TN)和硝酸盐氮的去除效果最好,分别达到50.73%和99.99%。该菌株经过平板形态观察、功能基因和16S rDNA基因分析,鉴定为施氏假单胞菌(Pseudomonas stutzeri)。结果表明:菌株ZH-14在硝酸钾为唯一氮源培养基中生长时,48 h后硝酸盐氮与TN的降解率分别为100%和41.76%;以亚硝酸钠为唯一氮源培养时,亚硝酸盐氮降解率为99.24%;以氨氮为唯一氮源培养时,氨氮的降解率达94.1%;使用菌株ZH-14处理畜禽沼液,当其接种终浓度为10~7CFU/mL时,经过48h处理,氨氮、硝酸盐氮和化学需氧量的去除率分别为54.4%、97.7%和77.9%。综上表明,菌株ZH-14具有较强的反硝化和处理猪场沼液的能力,具有良好的应用开发前景。
The aim of the present study is to screen aerobic denitrifying bacteria for the treatment of piggery biogas liquid.Sixteen aerobic denitrifying strains were selected from actived sludge. Among them, stain ZH-14 had the best removal rate of total nitrogen(TN) and nitrate nitrogen, reaching 50.73% and 99.99%, respectively. The strain was identified as Pseudomonas stutzeri by plate morphology, functional genes and 16 S rDNA sequence analysis. The results showed that the maximum degradation rates of nitrate nitrogen and TN reached 100% and 41.76%, respectively, when it grew in the medium with potassium nitrate as the sole nitrogen source after 48 h's cultivation. When sodium nitrite was used as the sole nitrogen source for culturing strain ZH-14, the maximum degradation rate of nitrite nitrogen was 99.24% within 24 h. When strain ZH-14 was cultured with ammonia nitrogen as the sole nitrogen source, the degradation rate of ammonia nitrogen was 94.1% after 24 h. For the treatment of piggery biogas liquid, the removal rate of ammonia nitrogen, nitrate nitrogen and chemical oxygen demand were 54.4%, 97.7%, and 77.9%, respectively, after 48-h treatment of strain ZH-14 at a final inoculation concentration of 10`7 CFU/mL. The result indicates that strain ZH-14 has a good application prospect in biological denitrification and the treatment of piggery biogas liquid.
The aim of the present study is to screen aerobic denitrifying bacteria for the treatment of piggery biogas liquid.Sixteen aerobic denitrifying strains were selected from actived sludge. Among them, stain ZH-14 had the best removal rate of total nitrogen(TN) and nitrate nitrogen, reaching 50.73% and 99.99%, respectively. The strain was identified as Pseudomonas stutzeri by plate morphology, functional genes and 16 S rDNA sequence analysis. The results showed that the maximum degradation rates of nitrate nitrogen and TN reached 100% and 41.76%, respectively, when it grew in the medium with potassium nitrate as the sole nitrogen source after 48 h's cultivation. When sodium nitrite was used as the sole nitrogen source for culturing strain ZH-14, the maximum degradation rate of nitrite nitrogen was 99.24% within 24 h. When strain ZH-14 was cultured with ammonia nitrogen as the sole nitrogen source, the degradation rate of ammonia nitrogen was 94.1% after 24 h. For the treatment of piggery biogas liquid, the removal rate of ammonia nitrogen, nitrate nitrogen and chemical oxygen demand were 54.4%, 97.7%, and 77.9%, respectively, after 48-h treatment of strain ZH-14 at a final inoculation concentration of 10`7 CFU/mL. The result indicates that strain ZH-14 has a good application prospect in biological denitrification and the treatment of piggery biogas liquid.
引文
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[28]赵丹,于德爽,李津,等.菌株ZD-8的分离鉴定及其异养硝化和缺氧/好氧反硝化特性研究[J].环境科学学报,201333(11):3007-3016.
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[31]韩永和,章文贤,庄志刚,等.耐盐好氧反硝化菌A13菌株的分离鉴定及其反硝化特性[J].微生物学报,2013,53(1):47-58.
[32]黄廷林,张丽娜,张海涵,等.一株贫营养异养硝化-好氧反硝化菌的筛选及脱氮特性[J].生态环境学报,2015,24(1):113-120.
[2]王利燕,黄开耀,彭霞,等.3株真核微藻在不同浓度猪粪水沼液中生长差异及氮磷去除率研究[J].中国畜牧杂志,201753(7):98-112.
[3]张恒,孙宏,王新,等.微藻及菌藻共生系统处理畜禽养殖污水的研究进展[J].中国畜牧杂志,2017,53(8):15-20.
[4]王旦一,朱洪光.厌氧发酵液后续处理研究进展及展望[J]现代农业科技,2011(9):281-284.
[5]Renuka N,Sood A,Ratha S K,et al.Evaluation of microalgal consortia for treatment of primary treated sewage e uent and biomass production[J].J Appl Phycol,2013,25(5):1529-1537.
[6]Robertson L A,Kuenen J G.Aerobic denitrification:a controversy revived[J].Arch Microbiol,1984,139(4):351-354.
[7]王秀杰,王维奇,李军,等.异养硝化菌Acinetobacter sp.的分离鉴定及其脱氮特性[J].中国环境科学,2017,37(11):4241-4250.
[8]田雪雪,陈玉立,张圆圆,等.一株异养硝化-好氧反硝化功能菌的分离鉴定及其脱氮特性[J].环境工程学报,2017,11(2):1269-1275.
[9]Joo H S,Hirai M,Shoda M.Characteristics of ammonium removal by heterotrophic nitrification-aerobic denitrification by Alcaligenes faecalis No.4[J].J Biosci Bioeng,2005,100(2):184-191.
[10]白洁,陈琳,黄潇,等.1株耐盐异养硝化-好氧反硝化菌Zobellella sp.B307的分离及脱氮特性[J].环境科学,2018,39(10):4793-4801.
[11]王田野,魏荷芬,胡子全,等.一株异养硝化-好氧反硝化菌的筛选鉴定及其脱氮特性[J].环境科学学报,2017,37(3):945-953.
[12]刘天琪,金若菲,周集体,等.异养硝化-氧反硝化菌AND-42的脱氮特性[J].环境工程学报,2015,9(2):989-996.
[13]Kandeler E,Deiglmayr K,Tscherko D,et al.Abundance of narG,nirS,nirK,and nosZ genes of denitrifying bacteria during primary successions of a glacier foreland[J].Appl Environ Microbiol,2006,72(9):5957-5962.
[14]Granhall U,Welsh A,Throback IN,et al.Bacterial community diversity in paper mills processing recycled paper[J].J Ind Microbiol Biotechnol,2010,37(10):1061-1069.
[15]Ji B,Yang K,Zhu L,et al.Aerobic denitrification:a review of important advances of the last 30 years[J].Biotech Biopro Eng,2015,20(4):643-651.
[16]Yao S,Ni J R,Chen Q,et al.Enrichment and characterization of a bacteria consortium capable of heterotrophic nitrification and aerobic denitrification at low temperature[J].Bioresour Technol,2013,127(1):151-157.
[17]许涛,王国英,岳秀萍.Diaphorobacter sp.PDB3菌好氧反硝化脱氮特性[J].中国环境科学,2018,38(6):2321-2328.
[18]信欣,姚力,鲁磊,等.耐高氨氮异养硝化-好氧反硝化菌TN-14的鉴定及其脱氮性能[J].环境科学,2014,35(10):3926-3932.
[19]Sun Y L,Li A,Zhang X N,et al.Regulation of dissolved oxygen from accumulated nitrite during the heterotrophic nitrification and aerobic denitrification of Pseudomonas strtzeri T13[J].Appl Microbiol Biotechnol,2015,99(7):3243-3248.
[20]王有乐,雷兴龙,陈连军,等.土壤中好氧反硝化菌的分离及脱氮特性研究[J].环境工程学报,2011,8(9):1902-1906.
[21]Chen P,Li J,Li Q X,et al.Simultaneous heterotrophic nitrification and aerobic denitrification by bacterium Rhodococcus sp.CPZ24[J].Bioresour Technol,2012,116(13):266-270.
[22]Zhao B,An Q,He Y L,et al.N2O and N2 production during heterotrophic nitrification by Alcaligenes faecalis strain NR[J].Bioresour Technol,2012,116(4):379-385.
[23]张培玉,曲洋,于德爽,等.菌株qy37的异养硝化-好氧反硝化机制比较及氨氮加速降解特性研究[J].环境科学,201031(8):1819-1826.
[24]乔森,刘雪洁,周集体.异养硝化-好氧反硝化在生物脱氮方面的研究进展[J].安全与环境学报,2014,14(2):128-135.
[25]何腾霞,李振轮,徐义.耐冷亚硝酸盐型反硝化细菌Pseudomonas putidaY-12脱氮特性[J].环境科学学报,201535(10):3071-3077.
[26]张树松,樊月婷,孙兴滨,等.菌株Diaphorobacter polyhydroxybutyrativorans SL-205的反硝化特性[J].中国环境科学,2017,37(9):3532-3539.
[27]王兆阳,陈国耀,姜珂,等.1株耐冷兼性嗜碱好氧反硝化菌的分离鉴定及反硝化特性[J].环境科学,2014,35(6):2341-2348.
[28]赵丹,于德爽,李津,等.菌株ZD-8的分离鉴定及其异养硝化和缺氧/好氧反硝化特性研究[J].环境科学学报,201333(11):3007-3016.
[29]Robertson L A,Cornelisse R,de Vos P,et al.Aerobic den-itrification in various heterotrophic nitrifiers[J].Anton Leeuw1989,56(4):289-299.
[30]Kim Y J,Yoshizawa M,Takenaka S,et al.Isolation and culture conditions of a Klebsiella pneumoniae strain that can utilize ammonium and nitrate ions simultaneously with controlled iron and molybdate ion concentrations[J].Biosci Biotechnol Biochem,2002,66(5):996-1001.
[31]韩永和,章文贤,庄志刚,等.耐盐好氧反硝化菌A13菌株的分离鉴定及其反硝化特性[J].微生物学报,2013,53(1):47-58.
[32]黄廷林,张丽娜,张海涵,等.一株贫营养异养硝化-好氧反硝化菌的筛选及脱氮特性[J].生态环境学报,2015,24(1):113-120.