孙将, 李建章, 袁月祥, 闫志英, 廖银章. 高效除氨氮异养硝化细菌的分离鉴定及其脱氮条件优化[J]. 农业工程学报, 2018, 34(Z): 35-41. DOI: 10.11975/j.issn.1002-6819.2018.z.006
    引用本文: 孙将, 李建章, 袁月祥, 闫志英, 廖银章. 高效除氨氮异养硝化细菌的分离鉴定及其脱氮条件优化[J]. 农业工程学报, 2018, 34(Z): 35-41. DOI: 10.11975/j.issn.1002-6819.2018.z.006
    Sun Jiang, Li Jianzhang, Yuan Yuexiang, Yan Zhiying, Liao Yinzhang. Study on isolation and identification of heterotrophic nitrifying bacteria and optimal denitrification conditions[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2018, 34(Z): 35-41. DOI: 10.11975/j.issn.1002-6819.2018.z.006
    Citation: Sun Jiang, Li Jianzhang, Yuan Yuexiang, Yan Zhiying, Liao Yinzhang. Study on isolation and identification of heterotrophic nitrifying bacteria and optimal denitrification conditions[J]. Transactions of the Chinese Society of Agricultural Engineering (Transactions of the CSAE), 2018, 34(Z): 35-41. DOI: 10.11975/j.issn.1002-6819.2018.z.006

    高效除氨氮异养硝化细菌的分离鉴定及其脱氮条件优化

    Study on isolation and identification of heterotrophic nitrifying bacteria and optimal denitrification conditions

    • 摘要: 为去除畜禽粪便好养堆肥过程中产生的氨气,筛选具有高效除氮功能的微生物,该文通过摇瓶连续富集驯化,从垃圾渗滤液处理厂的活性污泥中筛选到1株高效除氨氮菌株Z53,通过形态特征及16SrRNA基因序列鉴定,确定其为脱氮副球菌(Paracoccus denitrificans),并通过正交试验对其脱氮条件进行优化。试验表明该菌株最适碳源为丁二酸钠,最佳C/N为15;最佳氨氮去除条件为摇床转速160 r/min、250 mL三角瓶装液量为40 mL、温度30 ℃、pH值为7.0。在此条件下接种1%的菌液,该菌株在11 h内将(191.4±0.7)mg/L的氨氮完全去除,平均去除速率为17.4 mg/(L·h)。通过氮素形态测定显示该菌株不仅具有异养硝化作用,可将氨态氮氧化为硝态氮,且无亚硝态氮积累;还具有反硝化作用,在好氧条件下消耗硝态氮生成亚硝态氮,硝氮平均去除率为16.8 mg/(L·h),在厌氧条件下反硝化作用产生N2,硝氮平均去除率为9.8 mg/(L·h)。该研究体现了菌株Z53有望用于堆肥厂中的生物滴滤装置并高效去除氨氮,从而达到降低氨气污染的目的,具有良好的开发应用前景。

       

      Abstract: Abstract: Aerobic composting of livestock and poultry manure, decomposing organic matter and releasing heat by microorganisms, can be harmless and thoroughly decomposed, as well as realize the utilization of solid waste. However, in the process of composting, a large amount of ammonia gas is generated, which pollutes the surrounding air seriously. It is essential to select microorganisms with high nitrogen removal ability. Through continuous enrichment and domestication of shake flask, a strain named Z53 with high-efficiency removal of ammonia nitrogen was isolated from the activated sluge of garbage leachate plant. Through identification of morphology characteristics as well as gene sequences of 16SrRNA, the strain Z53 was identified as Paracoccus denitrificans. The nitrogen removal conditions were optimized by orthogonal test, detecting the temperature(30, 35 or 40 ℃), pH value(7, 8 or 9) , and the liquid volume (40, 60 or 80 mL in 250 mL conical flask), and the results showed that the optimal carbon source of the strain was disodiam succinate, the best C/N ratio was 15, and the optimal removal conditions of ammonia nitrogen included shaking speed of 160 r/min, liquid volume of 40 mL in 250 mL conical flask, temperature of 30 ℃ as well as pH value 7.0. When inoculating 1% of the bacterial solution under optimal conditions, the strain Z53 removed (191.4±0.7) mg/L ammonia nitrogen completely within 11 h, and the average removal rate was 17.4 mg/(L·h). The determination of nitrogen morphology displayed that strain Z53 not only owned the ability of heterotrophic digestion (which can oxidize the ammonia nitrogen to nitrate nitrogen without accumulation of nitrite nitrogen) but also had the ability of denitrification to transform nitrate nitrogen into nitrite nitrogen under aerobic conditions. The average removal rate of nitrate nitrogen was 16.8 mg/(L·h), while under the anaerobic conditions, it could produce N2, and the average removal rate of nitrate nitrogen was 9.8 mg/(L·h). The strain Z53 could convert ammonia nitrogen into nitrate nitrogen through nitrification, and then converted nitrate nitrogen into intracellular nitrogen or gaseous product through denitrification, which had strong removal ability of ammonia nitrogen. During the high-temperature aerobic composting process in the composting plant, the microorganisms decompose the nitrogen-containing organic matter to produce large amounts of ammonia gas. In order to remove the ammonia gas, the gas in the composting plant could be collected and passed into the bio-trickling filter with inoculating strain Z53. The removal of ammonia nitrogen in this way could achieve the purpose of reducing pollution of ammonia gas. Moreover, the strain Z53 could also be inoculated into the compost material, which removed part of the ammonia nitrogen in the heap before the compost was warmed up. In addition, the nitrogen content in the compost leachate was high, and the accumulation of nitrate would lead to eutrophication of the water body. Because the strain Z53 can remove nitrate nitrogen efficiently, the strain Z53 also has a good potential of application in the denitrification of water body of leachate in the composting plant.

       

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