Effects of characteristic pollutants in leather sludge leachate on soil ammonia nitrogen transformation and microbial community structures
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摘要:研究目的
查明制革污泥渗滤液中特征污染物（Cr(III)、盐分、有机质）对土壤NH4+−N转化和微生物群落结构的影响，为制革污泥的安全农用及土壤NH4+−N的污染防控提供理论依据。
研究方法通过土壤培养实验，研究不同污染物影响下土壤中NH4+−N向NO3−−N的转化规律，并利用高通量测序对比分析不同条件下的微生物群落组成和结构变化特征。
研究结果制革污泥渗滤液中特征污染组分对土壤中NH4+−N硝化过程的抑制程度排序为：Cr(III)>盐分>有机质。外源Cr(III)进入对土壤中NH4+−N的转化速率和转化量具有明显抑制作用，而土壤中盐分和有机质主要影响硝化作用的启动时间和降低硝化速率。随着土壤中Cr(III)含量由100 mg/kg增加到250 mg/kg，培养90 d后NH4+−N的转化量由94.23%降低到19.38%。高浓度Cr(III)和盐分在污染初期对土壤中微生物丰度和群落结构影响显著，随土壤老化过程中Cr(III)生物有效性降低及微生物适应性增强，不同污染特征土壤中微生物群落结构和组成逐渐趋同，硝化功能菌（Nitrosospira、Nitrosomonas和Nitrosospira）丰度增加明显。
结论冗余（RDA）分析结果表明，影响制革污染土壤中微生物群落结构演变的主要特征因子是Cr(III)（R2=0.53，P<0.01）、NH4+−N（R2=0.59，P<0.005）和NO3−−N（R2=0.53，P<0.01）。
创新点:（1）从污染组分含量变化及微生物群落响应特征两方面研究入手，研究制革污泥渗滤液中特征污染物Cr(III)、盐分和有机质对土壤NH4+−N转化过程的影响程度；（2）识别土壤中控制硝化作用的主要功能菌群及其丰度变化，查明影响制革污染土壤中微生物群落结构演变的主要特征因子，为制革污泥的安全农用及土壤NH4+−N的污染防控提供理论支撑。
Abstract:This paper is the result of soil environmental survey engineering.
ObjectiveThe objective is to identify the impact of characteristic pollutants (Cr (III), salt, organic matter) in the leachate of tannery sludge on the conversion of soil NH4+−N conversion and microbial community structure. It provides a theoretical basis for the safe agricultural use of tannery sludge and soil pollution prevention and control.
MethodsThe transformation of NH4+−N to NO3−−N in soil was studied through soil cultivation experiments under different pollution conditions, and the high−throughput sequencing is used to analyze the composition and structure characteristics of microbial community in soil.
ResultsThe characteristic pollution components in the tannery sludge leachate inhibit the nitrification process of NH4+−N in the soil, and the influence factors are sorted as follows: Cr(III)>saltness>organic matter. The exogenous Cr(III) pollution has a significant inhibitory effect on the transformation rate and amount of NH4+−N, whereas the saltness and organic matter only delay the start time of nitrification and reduce the nitrification rate in soil. With the increase of Cr(III) content from 100 mg/kg to 250 mg/kg, the transformation amount of NH4+−N decreases from 94.23% to 19.38% after 90 days of culture. The high Cr(III) and salinity in the leachate have a significant impact on the microbial community structure and distribution in the soil at the initial stage of pollution. With the decrease of the bioavailability of Cr(III) and the enhancement of microbial adaptation in soil aging process, the microbial community structure and composition in Cr(III), salinity, and organic matter contaminated soils become similar, and the abundances of nitrifying bacteria (Nitrosospira, Nitrosomonas and Nitrosospira) also have an obvious increase.
ConclusionsThe results of RDA analysis show that the main characteristic factors affecting the evolution of microbial community structure in tanning contaminated soil are: Cr(III) (R2=0.53, P<0.01), NH4+−N (R2=0.59, P<0.005) and NO3−−N(R2=0.53, P<0.01).
Highlights:(1) The influence of characteristic pollutants in leather sludge leachate on soil ammonia nitrogen transformation are sorted as follows: Cr(III)>saltness>organic matter. (2) The main characteristic factors affecting the evolution of microbial community structure in tanning contaminated soil are: Cr(III) (R2=0.53, P<0.01), NH4+−N (R2=0.59, P<0.005) and NO3−−N (R2=0.53, P<0.01).
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图 1 不同环境条件下土壤中氨氮、亚硝酸盐及硝酸盐含量变化
a—CK；b—Cr1；c—Cr2；d—OM；e—Sal；f—TS
Figure 1. Changes of NH4+–N, NO2––N and NO3––N contents in soils under different conditions
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图 2 不同环境条件下硝酸盐累积过程的Sigmodial拟合曲线
Figure 2. Sigmodial fitting curve of nitrate accumulation process under different conditions
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图 3 样本菌群分型图（a—韦恩图；b—聚类分布图；c—门水平组成图）
Figure 3. Classification chart of bacterial taxa (a–Veen diagram; b–Clustering distribution diagram; c–Classification diagram of phylum)
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图 4 属分类水平上的微生物群落组成丰度图
Figure 4. Relative abundances of genus level of bacterial taxa recovered from samples
表 1 不同实验条件下土壤中污染组分含量
Table 1 Contents of contaminated components in soil under different experimental conditions
样品
分组NH4+−N/
(mg/kg)Cr(III)/
(mg/kg)含盐量/
(mg/kg)有机质/
%CK 320.5 66.8 717 0.14 Cr1 316.3 103.5 1450 0.16 Cr2 318.0 252.8 2070 0.17 OM 319.7 69.8 885 0.42 Sal 318.4 67.2 10250 0.17 TS 268.5 184.0 10200 0.34 
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