固定化微生物技术在小秦岭金矿区农田土壤修复中的应用

doi:10.16178/j.issn.0528-9017.20230614

摘要/Abstract

摘要：

为积极探索土壤污染防治新路径,开展固定化微生物技术在土壤原位生态修复方面的应用研究。选取豫西小秦岭金矿矿区及周边农田土壤进行重金属含量测定及分析评价,发现该地区土壤中Pb²⁺元素污染风险较大。将侧孢芽孢杆菌、放线菌及酵母菌株进行固定化混合菌制备并开展土壤原位生物修复。发现经固定化混合菌处理后土壤脲酶活性显著高于CK。以Pb²⁺钝化效率及去除率提高为目的,分别采用单因素试验及响应面法对影响土壤修复效果各因素进行分析,最终确定Pb²⁺最佳去除条件为固定化混合菌投加量10.496%,处理温度30.784 ℃,pH值6.242,Pb²⁺去除率理论值为72.754%。对此模型进行试验验证,结果与理论值接近,说明响应面分析可起到优化土壤原位修复试验参数的目的。

关键词: 固定化微生物技术, 土壤原位修复, 去除率, 单因素试验, 响应面分析

Abstract:

In order to explore new methods of soil pollution prevention, the application of immobilized microorganism technology in soil ecological remediation was studied. The heavy metal contents in the soil of Xiaoqinling gold mining area and the surrounding farmland in Western Henan Province were determined and evaluated, it was found that the risk of Pb²⁺ element pollution in the soil in this area was high. The mixed bacteria of Bacillus laterosporus, actinomycetes and yeast were immobilized to carry out soil bioremediation in situ, the results showed that the urease activities of soil treated with immobilized bacteria were significantly higher than that of CK. In order to improve the passivation efficiency and removal rate of Pb²⁺, the single factor test and response surface methodology were used to analyze the factors affecting the soil remediation effect, the optimal removal conditions of Pb²⁺ were determined finally: the dosage of immobilized mixed bacteria was 10.496%, the treatment temperature was 30.784 ℃, the pH value was 6.242, and the theoretical removal rate of Pb²⁺ was 72.754%. The model was verified by experiment, the result of experiment was close to the theoretical value, indicating that response surface analysis can optimize the soil remediation experimental parameters.

Key words: immobilized microorganism technology, soil in-situ remediation, removal rate, single factor test, response surface methodology

中图分类号:

李立军, 仲惟, 李涛. 固定化微生物技术在小秦岭金矿区农田土壤修复中的应用[J]. 浙江农业科学, 2024, 65(8): 1950-1956.

LI Lijun, ZHONG Wei, LI Tao. Application of immobilized microorganism technology on soil remediation of farmland in Xiaoqinling gold mining area[J]. Journal of Zhejiang Agricultural Sciences, 2024, 65(8): 1950-1956.

图/表 11

表1 Background value of heavy metal determination in soil of Xiaoqinling gold mining area and surrounding farmland单位: mg·kg-1

Table 1

主要参数	Hg	As	Pb	Cr	Cu	Zn
最小值	0.04	5.08	21.20	40.24	13.40	44.85
最大值	42.57	13.62	3 654.06	82.13	872.60	572.81
平均值	3.36	8.16	303.89	60.23	82.23	89.81
标准差	11.78	2.74	1 006.60	15.54	237.49	145.18
变异系数	3.50	0.34	3.31	0.26	2.89	1.62
周边土壤对照值	0.08	8.19	32.62	55.99	23.93	68.74
GB 15168-2018限值	3.40	25.00	170.00	250.00	100.00	300.00

表2 小秦岭金矿区及周边农田土壤污染评价

Table 2 Evaluation of soil pollution in Xiaoqinling gold mining area and surrounding farmland

评价指标	Hg	As	Pb	Cr	Cu	Zn
P_i	0.989	0.326	1.788	0.241	0.822	0.299
P_综合	8.881	0.449	15.251	0.288	6.198	1.367

图1 不同粒径SM-ETS固定化混合菌处理下土壤脲酶活性测定柱上无相同小写字母表示处理间差异达显著水平(P<0.05),图3~4同。

Fig.1 Determination of soil urease activity in the treatments of SM-ETS immobilized bacteria with different particle sizes

图2 不同处理时间污染土壤中Pb2+有效态含量测定

Fig.2 Determination of available content of Pb2+ in contaminated soil with different treatment time

图3 不同菌种组成比例对Pb2+去除效果对比

Fig.3 Comparison of Pb2+ removal effect of different strain composition ratio

图4 不同固定化混合菌投加量对Pb2+去除效果对比

Fig.4 Comparison of Pb2+ removal effect of different immobilized mixed bacteria dosage

图5 不同处理温度对Pb2+去除效果对比

Fig.5 Comparison of Pb2+ removal effects at different treatment temperatures

图6 不同pH值对Pb2+去除效果对比

Fig.6 Comparison of removal effect of Pb2+ under different pH values

表3 响应面试验因素与水平

Table 3 Response surface methodology factors and levels

变量	A固定化混合菌投加量/%	B处理温度/℃	C pH值
-1	5	25	4
0	10	30	6
1	15	35	8

表4 响应面试验设计方案及试验结果

Table 4 Response surface methodology design scheme and test results

试验号	A/%	B/℃	C	Y (Pb²⁺去除率)/%
1	5	25	6	34.44
2	15	25	6	41.43
3	5	35	6	46.16
4	15	35	6	50.20
5	5	30	4	35.12
6	15	30	4	44.46
7	5	30	8	42.13
8	15	30	8	47.12
9	10	25	4	41.98
10	10	35	4	36.24
11	10	25	8	37.65
12	10	35	8	54.17
13	10	30	6	69.15
14	10	30	6	72.11
15	10	30	6	73.78
16	10	30	6	70.90
17	10	30	6	74.63

表5 回归模型方差分析

Table 5 Variance analysis of regression model

项目	平方和	自由度	均方	F值	显著性
Model	3 478.7	9	386.52	78.56	^**
A	80.39	1	80.39	16.34	^**
B	122.23	1	122.23	24.84	^**
C	67.69	1	67.69	13.76	^**
AB	2.18	1	2.18	0.44
AC	4.73	1	4.73	0.96
BC	123.88	1	123.88	25.18	^**
A²	907.32	1	907.32	184.40	^**
B²	870.31	1	870.31	176.88	^**
C²	976.26	1	976.26	198.41	^**
Residual	34.44	7	4.92
Lack of Fit	15.08	3	5.03	1.04
Pure Error	19.36	4	4.84
Cor Total	3 513.14	16

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