典型炼化废水微生物功能结构与主要致毒物质响应关系研究

宋佳宇; 张婷婷; 李立君; 李丹丹; 李兴春; 李昀照

doi:10.13198/j.issn.1001-6929.2023.02.04

典型炼化废水微生物功能结构与主要致毒物质响应关系研究

doi: 10.13198/j.issn.1001-6929.2023.02.04

宋佳宇^{1, 2,},
张婷婷³,
李立君⁴,
李丹丹^{1, 2},
李兴春^{1, 2},
李昀照³

1.
石油石化污染物控制与处理国家重点实验室，北京　102206
2.
中国石油集团安全环保技术研究院有限公司，北京　102206
3.
中国石油大学（北京）化学工程与环境学院，北京　102206
4.
盘锦市绿色发展服务中心，辽宁盘锦　124200

基金项目: 中国石油天然气集团有限公司科学与技术开发项目(No.2021DQ03-A4)

详细信息

作者简介:
宋佳宇（1985-），女，天津人，高级工程师，硕士，主要从事石油污染生物修复研究，songjiayu@cnpc.com.cn

中图分类号: X172
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- 被引次数: 0
出版历程
- 收稿日期: 2022-09-01
- 修回日期: 2022-12-26

Relationship between Microbial Diversity and Main Toxic Substances in Typical Refinery Wastewater

SONG Jiayu^{1, 2
,},
ZHANG Tingting³,
LI Lijun⁴,
LI Dandan^{1, 2},
LI Xingchun^{1, 2},
LI Yunzhao³

1.
State Key Laboratory of Petroleum Pollution Control, Beijing 102206, China
2.
CNPC Research Institute of Safety and Environment Technology, Beijing 102206, China
3.
College of Chemical Engineering and Environment, China University of Petroleum-Beijing, Beijing 102206, China
4.
Panjin Green Development Service Center, Panjin 124200, China

Funds: Scientific Research and Technology Development Program of China National Petroleum Corporation (No.2021DQ03-A4)

摘要

摘要: 为探明典型炼化废水处理系统(简称“系统”)生物毒性削减效果、主要致毒物质类别及微生物功能响应关系，以费氏弧菌和鼠伤寒沙门氏菌为受试生物，测试系统沿程生物急性毒性及遗传毒性，并结合毒性鉴别评价方法甄别系统主要致毒物质，同时利用高通量测序技术探究系统微生物功能结构与主要致毒物质响应关系. 结果表明：①系统沿程生物毒性总体呈下降趋势，生化处理单元进水为急性毒性微毒、遗传毒性阳性. 其中，炼油废水急性毒性总削减率为86.514%，遗传毒性总削减率为96.221%；化工废水急性毒性总削减率为53.281%，遗传毒性总削减率为62.273%. ②通过毒性鉴别评价方法(toxicity identification evaluation, TIE)结果推断，炼油废水主要致毒物质可能为阳离子金属及非极性有机物，化工废水主要致毒物质可能为阳离子金属. ③CCA分析表明，NH₃-N浓度(r=0.819，p=0.001)、Cr浓度(r=0.777，p=0.002)、TPH (total petroleum hydrocarbon，总石油烃)浓度(r=0.752，p=0.002)与生化处理前微生物群落结构均呈显著正相关，与生化处理后微生物群落结构均呈显著负相关. ④生化处理阶段微生物功能发生极显著(p≤0.01)变化. FAPROTAX数据库预测可知，化学异养、好氧化学异养、芳香化合物降解、亚硝酸盐呼吸、硝酸盐还原、硝酸盐呼吸、氮气呼吸7种与碳、氮循环相关的微生物功能丰度均较高. 研究显示：系统沿程生物毒性虽呈下降趋势，但生化处理单元进水仍为急性毒性微毒、遗传毒性阳性；毒性表征推断炼油废水主要致毒物质可能为阳离子金属及非极性有机物，化工废水主要致毒物质可能为阳离子金属；生化处理单元中与碳、氮循环相关的微生物功能丰度较高，表明生化处理单元存在潜在毒性冲击风险.
- 炼化废水 /
- 急性毒性 /
- 遗传毒性 /
- 主要致毒物质 /
- 微生物功能丰度
Abstract: In order to investigate the relationship between the biotoxicity reduction effect of a typical refinery wastewater treatment system (hereafter referred to as system), the main types of toxic substances and the response of functional microorganisms, Vibrio fischeri and Salmonella typhimurium were used as test organisms to examine the acute toxicity and genotoxicity with the process of the system. We used the toxicity identification evaluation (TIE) to identify the main toxic substances in the system, and used the high-throughput sequencing to explore the relationship between the microbial functional structure and the main toxic substances. The results showed that: (1) The biological toxicity showed a decreasing trend with the process in the system. The influent water of the biochemical treatment unit showed mild acute toxicity and positive in genotoxicity. The total reduction rate of acute toxicity of oil refinery wastewater was 86.514% and the total reduction rate of genotoxicity was 96.221%. The total reduction rate of acute toxicity of chemical wastewater was 53.281% and the total reduction rate of genotoxicity was 62.273%. (2) The TIE results indicated that the main toxic substances in oil refinery wastewater might be cationic metals and non-polar organic substances, and the main toxic substances in chemical wastewater might be cationic metals. (3) Canonical correlation analysis showed that the concentrations of ammonia (r=0.819, p=0.001), Cr (r=0.777, p=0.002) and total petroleum hydrocarbons (TPH) (r=0.752, p=0.002) were positively correlated with the microbial community structure before the biochemical treatment and negatively correlated with the microbial community structure after the biochemical treatment. (4) Microbial functions changed significantly during the biochemical treatment (p≤0.01). The FAPROTAX database predicted a high abundance of microorganisms in the carbon and nitrogen cycling in seven classes: chemoheterotrophy, aerobic chemoheterotrophy, aromatic compound degradation, nitrite respiration, nitrate reduction, nitrate respiration and nitrogen respiration. The study showed that although the biological toxicity along with the processes in the system decreased, the influent water of the biochemical treatment unit was still slightly toxic in acute toxicity and positive in genotoxicity. The characterization of the toxicity suggested that the main toxic substances in oil refinery wastewater were cationic metals and non-polar organic substances, and the main toxic substances in chemical wastewater were cationic metals. The abundance of functional microorganisms involved in carbon and nitrogen cycling in the biochemical treatment unit was high, indicating potential toxic risk in the biochemical treatment unit.
- refinery wastewater /
- acute toxicity /
- genotoxicity /
- main toxic substances /
- microbial functional abundance

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图 1 典型炼化废水处理系统主要处理单元及采样点设置

Figure 1. The main processing units of typical refinery wastewater and the setting of water sampling points

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图 2 TIE Phase I试验流程示意

注：pH₀为原水pH；该试验中pH的调节均采用NaOH (0.1 mol/L)和HCl (0.1 mol/L).

Figure 2. Experimental flow chart of TIE Phase I

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图 3 典型炼化废水处理系统各主要处理阶段生物毒性的变化趋势

Figure 3. Biotoxicity trends in main stages of the typical refinery wastewater treatment systems

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图 4 典型炼化废水各单元出水的毒性鉴别评价

Figure 4. Toxicity identification evaluation of effluent from each unit of typical refinery wastewater

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图 5 筛选后环境因子与典型炼化废水微生物群落CCA分析(属水平)

注：Before为典型炼化废水生化处理前水样，After为典型炼化废水生化处理后水样.

Figure 5. CCA of selected environmental factors and microbial communities in typical refinery wastewater (at the genus level)

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图 6 典型炼化废水生化处理前后疑似致毒环境因子与优势菌相关性热图(属水平)

注：*表示0.01≤p≤0.05，**表示0.001≤P≤0.01，***表示p≤0.001.

Figure 6. Heatmap of correlation between suspected toxic environmental factors and dominant bacteria before and after biochemical treatment of a typical refinery wastewater (at the genus level)

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图 7 典型炼化废水生化处理阶段微生物功能丰度的比对

Figure 7. Comparison of microbial functional abundance in the biochemical treatment stage in typical refinery wastewater

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表 1 典型炼化废水各处理阶段出水的常规理化指标

Table 1. Physicochemical parameters of the typical refinery wastewater from different units

进水类型	处理阶段	pH	浓度/(mg/L)
进水类型	处理阶段	pH	COD	硫化物	SS	NH₃-N	TPH
炼油废水	进水	7.19±0.15	186.32±5.96	35.24±3.33	28.75±1.58	15.74±0.15	133.27±8.28
	物理单元出水	7.44±0.22	202.76±10.24	10.56±1.05	20.17±2.75	21.38±2.24	85.15±8.01
	生化单元出水	7.50±0.35	39.33±3.88	8.37±0.67	5.87±0.66	1.72±0.57	20.33±2.93
	深度处理出水	7.17±0.52	33.24±5.27	0.35±0.045	3.22±0.15	0.15±0.033	0.35±0.019
化工废水	进水	7.23±0.65	136.57±15.29	15.28±2.66	22.74±2.78	14.01±1.66	35.99±7.45
	生化单元出水	7.52±0.25	26.03±1.74	4.37±0.64	18.26±1.39	2.37±0.67	20.16±5.28
	深度处理出水	7.19±0.39	23.34±1.05	0.59±0.037	12.15±1.07	0.24±0.046	3.52±0.68
注：《石油炼制工业污染物排放标准》(GB 31570—2015)中常规污染物理化指标排放限值分别为pH 6~9、COD浓度 60 mg/L、硫化物浓度1 mg/L、SS浓度70 mg/L、NH₃-N浓度8 mg/L、TPH浓度5 mg/L.

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表 2 工业废水生物毒性评价

Table 2. Biological toxicity evaluation of industrial wastewater

项目	毒性级别
TUa<0.40	无毒
0.40<TUa<1.00	微毒
1.00<TUa<10.00	中毒
10.00<TUa<100.00	高毒
TUa>100.00	剧毒

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表 3 典型炼化废水各主要处理阶段生物毒性负荷削减率

Table 3. Reduction rate of biological toxicity load in main treatment stages of typical refinery wastewater

进水来源	主要处理阶段	生物毒性负荷削减率/%
进水来源	主要处理阶段	急性毒性	遗传毒性
炼油废水	物理处理阶段	−4.267	72.334
	生化处理阶段	72.792	53.281
	深度处理阶段	52.462	70.762
	总计	86.514	96.221
化工废水	生化处理阶段	7.382	44.597
	深度处理阶段	28.181	31.905
	总计	21.860	62.273

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表 4 筛选后疑似致毒类环境因子的VIF值

Table 4. VIF values for suspected toxicogenic environmental factors after screening

疑似致毒环境因子	VIF值
NH₃-N	4.597
Cr	2.937
Ni	1.368
TPH	3.088

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表 5 典型炼化废水生化处理阶段微生物功能丰度对比T检验

Table 5. T-test of the functional abundance of microorganisms in the biochemical treatment stage of typical refinery wastewater

样品编号	个案数	相关性	显著性(p)
炼油废水生化处理阶段进出水(BC)	20	0.703	0.01
化工废水生化处理阶段进出水(EF)	20	0.941	<0.01

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