蚯蚓粪肥农用土壤中活体病原菌随雨水径流的溶出特性

王邦驰; 黄魁; 闫志泉; 陈群丰; 李同欢

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中国环境科学 ›› 2025, Vol. 45 ›› Issue (5) : 2569-2576.

水污染与控制

蚯蚓粪肥农用土壤中活体病原菌随雨水径流的溶出特性

王邦驰¹, 黄魁^1,2,3, 闫志泉¹, 陈群丰¹, 李同欢¹

作者信息 +

Leaching characteristics of viable pathogenic bacteria from earthworm manure-amended soil with rainwater runoff

WANG Bang-chi¹, HUANG Kui^1,2,3, YAN Zhi-quan¹, CHEN Qun-feng¹, LI Tong-huan¹

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文章历史 +

摘要

为明确病原微生物在雨水径流中的溶出规律,采用室内人工模拟降雨,冲刷蚯蚓粪施用的土壤.使用叠氮溴化丙锭(PMA)结合荧光定量PCR,探究其中活体粪便指示菌(FIB)的丰度变化.结果显示,混合土壤基质中的电导率、氨氮、硝酸盐氮、总磷(TP)的浓度在降雨后分别下降了51.34%、45.20%、99.09%和26.22%.在径流水中,氨氮、TP和化学需氧量(COD)的浓度均呈现出先上升后下降的变化趋势,且峰值都出现在前15min(含15min).总大肠菌(TC)、粪大肠菌(FC)和大肠杆菌(EC)和肠球菌(ES)四个FIB的qPCR定量结果都呈现出先上升后下降的变化趋势,并且PMA-qPCR与培养法之间有着显著的正相关性(Spearmanr=0.723,P<0.001).所有径流中的FC数量均超过了《地表水环境质量标准》中的限值(40000个/L).研究表明活性病原微生物可通过降雨冲刷进入水体,并在初期径流中大量扩散,增加其环境传播风险.

Abstract

To clarify the dissolution patterns of pathogenic microorganisms in rainwater runoff, a laboratory-simulated rainfall experiment was conducted to flush soil treated with earthworm castings. Propidium monoazide (PMA) combined with quantitative PCR (qPCR) was used to investigate changes in the abundance of viable fecal indicator bacteria (FIB). The results exhibited that after rainfall, the concentrations of electrical conductivity, ammonia, nitrate, and total phosphorus (TP) in the mixed soil matrix decreased by 51.34%, 45.20%, 99.09%, and 26.22%, respectively. In runoff water, the concentrations of ammonia, total phosphorus, and chemical oxygen demand (COD) exhibited a trend of initially rising and then falling, with peak values occurring within the first 15minutes. The qPCR quantification results for four fecal indicator bacteria—total coliforms (TC), fecal coliforms (FC), Escherichia coli (EC), and Enterococcus spp (ES)—also displayed a similar trend of increasing and then decreasing. A significant positive correlation was found between the PMA-qPCR results and the culture method (Spearman r=0.723, P<0.001). The fecal coliform counts in all runoff samples exceeded the limits specified in the "Surface Water Quality Standard" (40000CFU/L). The study indicates that viable pathogenic microorganisms can be washed into water bodies through rainfall and are widely dispersed during initial runoff, increasing the risk of their environmental transmission.

导出引用

王邦驰, 黄魁, 闫志泉, 陈群丰, 李同欢. 蚯蚓粪肥农用土壤中活体病原菌随雨水径流的溶出特性[J]. 中国环境科学. 2025, 45(5): 2569-2576

WANG Bang-chi, HUANG Kui, YAN Zhi-quan, CHEN Qun-feng, LI Tong-huan. Leaching characteristics of viable pathogenic bacteria from earthworm manure-amended soil with rainwater runoff[J]. China Environmental Science. 2025, 45(5): 2569-2576

中图分类号： X522

参考文献

[1] Sales-Ortells H, Medema G. Microbial health risks associated with exposure to stormwater in a water plaza [J]. Water Research, 2015,74:34-46.
[2] Denissen J K, Reyneke B, Waso M, et al. Human pathogenic bacteria detected in rainwater: risk assessment and correlation to microbial source tracking Markers and traditional indicators [J]. Frontiers in Microbiology, 2021,12:659784-659784.
[3] 牛新胜,巨晓棠.我国有机肥料资源及利用[J]. 植物营养与肥料学报, 2017,23(6):1462-1479. Niu X S, Ju X T. Organic fertilizer resources and utilization in China [J]. Journal of Plant Nutrition and Fertilizers, 2017,23(6):1462-1479.
[4] Aira M, Olcina J, Pérez-Losada M, et al. Characterization of the bacterial communities of casts from Eisenia andrei fed with different substrates [J]. Applied Soil Ecology, 2016,98:103-111.
[5] Hussain N, Singh A, Saha S, et al. Excellent N-fixing and Psolubilizing traits in earthworm gut-isolated bacteria: A vermicompost based assessment with vegetable market waste and rice straw feed mixtures [J]. Bioresource Technology, 2016,222:165-174.
[6] Huang K, Xia H, Li F S, et al. Optimal growth condition of earthworms and their vermicompost features during recycling of five different fresh fruit and vegetable wastes [J]. Environmental Science and Pollution Research International, 2016,23(13):13569-75.
[7] Huang K, Xia H, Cui G Y, et al. Effects of earthworms on nitrification and ammonia oxidizers in vermicomposting systems for recycling of fruit and vegetable wastes [J]. Science of the Total Environment, 2017, 578:337-345.
[8] Xie J C, Xia H, Guan M X, et al. Accelerating the humification mechanism of dissolved organic matter using biochar during vermicomposting of dewatered sludge [J]. Waste Management, 2023, 159:102-113.
[9] Li Y P, Wang J, Shao M A. Application of earthworm cast improves soil aggregation and aggregate-associated carbon stability in typical soils from Loess Plateau [J]. Journal of Environmental Management, 2021,278(P1):111504.
[10] Zhou G, Qiu X, Chen L, et al. Succession of organics metabolic function of bacterial community in response to addition of earthworm casts and zeolite in maize straw composting [J]. Bioresource Technology, 2019,280:229-238.
[11] Soobhany N, Mohee R, Garg K V. Inactivation of bacterial pathogenic load in compost against vermicompost of organic solid waste aiming to achieve sanitation goals: A review [J]. Waste Management, 2017,64:51-62.
[12] 邵凡凡,吴军虎.蚯蚓粪对黄土区坡耕地径流产沙和硝态氮流失特征的影响[J]. 水土保持学报, 2020,34(1):71-77. Shao F F, Wu J H. Earthworm casts on sediment yield and nitrate nitrogen loss with the runoff in slope farmland of loess area [J]. Journal of Soil and Water Conservation, 2020,34(1):71-77.
[13] 魏枫沂,徐俊杰,陈进,等.基于活体微生物揭示蚯蚓对污泥耐药基因转归的影响[J]. 中国环境科学, 2022,42(7):3425-3433. Wei F Y, Xu J J, Chen J, et al. Effects of earthworms on the antibiotic resistance genes of vermicompost from dewatered sludge revealed by active microbes [J]. China Environmental Science, 2022,42(7):3425–3433.
[14] Li H Y, Xin H Y, Li S F Y, et al. Multiplex PMA-qPCR assay with internal amplification control for simultaneous detection of viable legionella pneumophila, salmonella typhimurium, and staphylococcus aureus in environmental waters [J]. Environmental science & technology, 2015,49(24):14249-14256.
[15] 陶怡君,谌志筠,何秋水.叠氮溴化丙锭结合qPCR检测与区分活菌和死菌的研究进展[J]. 微生物学免疫学进展, 2020,48(6):63-68. Tao Y J, Zhan Z Y, Hei Q S. Advances in detection and differentiation of live and dead bacteria by PMA- qPCR technology [J]. Progress in Microbiology and Immunology, 2020,48(6):63-68.
[16] HJ 613-2011土壤干物质和水分的测定重量法[S]. HJ 613-2011 Determination of dry matter and water content on a mass basis-Gravimatric method [S].
[17] HJ 761-2015固体废物有机质的测定灼烧减量法[S]. HJ 761-2015 Solid waste-Determination of organic matter-Ignition loss method [S].
[18] HJ 535-2009水质氨氮的测定纳式试剂分光光度法[S]. HJ 535-2009 Water quality-Determination of ammonia nitrogen Nessler’s reagent spectrophotometry [S].
[19] HJ/T 346-2007水质硝酸盐氮的测定紫外分光光度法(试行) [S]. HJ/T 346-2007 Water quality-Determination of nitrate-nitrogenUltraviolet spectrophotometry [S].
[20] HJ/T 399-2007水质化学需氧量的测定快速消解分光光度法[S]. HJ/T 399-2007 Water quality － Determination of the chemical oxygen demand－Fast digestion-Spectrophotometric method [S].
[21] 段自豪,夏慧,黄魁,等.蚯蚓对污泥堆肥过程中活性真核微生物的影响[J]. 环境科学学报, 2023,43(4):408-416. Duan Z H, Xia H, Huang K, et al. Effects of earthworms on active eukaryotic microbial community during vermicomposting of dewatered sludge [J]. Acta Scientiae Circumstantiae, 2023,43(4):408-416.
[22] Carini P, Marsden P J, Leff J W, et al. Relic DNA is abundant in soil and obscures estimates of soil microbial diversity. Nature Microbiology, 2016,2(3),16242.
[23] Maheux A F, Boudreau D K, Bisson M-A, et al. Molecular method for detection of total coliforms in drinking water samples. Applied and environmental microbiology, 2014,80(14):4074-4084.
[24] Deogratias M E, King L A A, Eunice U, et al. Genetic relatedness of faecal coliforms and enterococci bacteria isolated from water and sediments of the Apies River, Gauteng, South Africa [J]. AMB Express, 2017,7(1):20.
[25] Camilla B, Giorgio V, Claudia P, et al. Use of the tna operon as a new molecular target for Escherichia coli detection [J]. Applied and environmental microbiology, 2007,73(19):6321-6325.
[26] Duan Z H, Zhu Y C, Xia H, et al. A novel strategy for eliminating antibiotic resistance genes during fertilization of dewatered sludge by earthworms: Vermicomposting practice using Chinese herbal residues derived from Lianhua Qingwen as a bulking material [J]. Journal of Environmental Management, 2024,349,119444.
[27] Cui G Y, Bhat A S, Li W J, et al. Gut digestion of earthworms significantly attenuates cell-free and -associated antibiotic resistance genes in excess activated sludge by affecting bacterial profiles [J]. Science of the Total Environment, 2019,691:644-653.
[28] Duan Z, Huang K, Huang W, et al. Bacterial dispersal enhances the elimination of active fecal coliforms during vermicomposting of fruit and vegetable wastes: The overlooked role of earthworm mucus [J]. Journal of Hazardous Materials, 2024,471:134280.
[29] GB 38400-2019肥料中有毒有害物质的限量要求[S]. GB 38400-2019 Limitation requirements of toxic and harmful substance in fertilizers [S].
[30] 黄福义,周曙仡聃,王佳妮,等.不同作物农田土壤抗生素抗性基因多样性[J]. 环境科学, 2021,42(6):2975-2980. Huang F Y, Zhou S Y D, Wang J N, et al. Profiling of antibiotic resistance genes in different croplands [J]. Environmental Science, 2021,42(6):2975-2980.
[31] 关孟欣.生物质炭影响污泥蚯蚓堆肥中硝化作用的微生态机制[D]. 兰州:兰州交通大学, 2022. Guan M X. Micro-ecological mechanism revealing the effect of biochar on the nitrification during vermicomposting of dewatered sludge [D]. Lanzhou: Lanzhou Jiaotong University, 2022.
[32] 吴颖,黄魁,夏慧,等.污泥四环素含量对蚯蚓堆肥中氨氧化菌群的影响[J]. 环境科学, 2019,40(6):2954-2960. Wu Y, Huang K, Xia H, et al. Effects of different concentrations of tetracycline in sludge on ammonia oxidizers during vermicomposting [J]. Environmental Science, 2019,40(6):2954-2960.
[33] 甘贤民,严友进,戴全厚,等.降雨强度和坡度对喀斯特坡耕地产流和氮磷流失的影响[J]. 环境科学学报, 2024,44(4):116-123. Gan X M, Yan Y J, Dai Q H, et al. Impact of rainfall intensity and slope gradient on runoff and nitrogen and phosphorus loss fluxes of sloping farmland in a karst area [J]. Acta Scientiae Circumstantiae, 2024,44(4):116-123
[34] 王祥,陈炜,黄国鲜,等.长江上游典型丘陵山区坡耕地径流及氮磷碳流失特征[J]. 环境工程技术学报, 2024,14(5):1589-1598. Wang X, Chen W, Huang G X, et al. Characteristics of runoff and nitrogen, phosphorus, and carbon loss in sloping cultivated lands in the typical hilly mountainous region of the Upper Yangtze River Basin [J]. Journal of Environmental Engineering Technology, 2024,14(5):1589-1598.
[35] Zhang Q, Wang Z, Wu B, et al. Identifying sediment transport capacity of raindrop-impacted overland flow within transport-limited system of interrill erosion processes on steep loess hillslopes of China [J]. Soil & Tillage Research, 2018,184:109-117.
[36] GB 3838-2002地表水环境质量标准[S]. GB 3838-2002 Environmental quality standards for surface water [S].
[37] 徐晓璇,康丽娟,叶建锋,等.景观河道病原微生物污染来源与分布特征[J]. 上海大学学报(自然科学版), 2020,26(2):275-282. Xu, X X, Kang L J, Ye J F, et al. Pollution sources and distribution characteristics of pathogenic microorganisms in land scape rivers [J]. Journal of Shanghai University (Natural Science), 2020,26(2):275-282.
[38] Ahmed W, Hamilton K, Toze S, et al. A review on microbial contaminants in stormwater runoff and outfalls: Potential health risks and mitigation strategies [J]. Science of the Total Environment, 2019, 692:1304-1321.
[39] HJ 347.2-2018水质粪大肠菌群的测定多管发酵法[S]. HJ 347.2-2018 Water quality—Determination of fecal coliform— Manifold zymotechnics [S].
[40] GB 4789.39-2013食品安全国家标准食品微生物学检验粪大肠菌群计数[S]. GB 4789.39-2013 Microbiological examination of food hygieneEnumeration of faecal coliforms [S].
[41] 赖后伟,黎京士,庞志华,等.深圳大工业区初期雨水水质污染特征研究[J]. 环境污染与防治, 2016,38(3):11-15. Lai H W, Li J S, Pang Z H, et al. The contamination characteristics of first-flush in large industrial district of Shenzhen [J]. Environmental Pollution and Contral, 2016,38(3):11-15.
[42] Paule-Mercado M, Ventura J, Memon S, et al. Monitoring and predicting the fecal indicator bacteria concentrations from agricultural, mixed land use and urban stormwater runoff [J]. Science of the Total Environment, 2016,550:1171-1181.
[43] Antonella P, Mauro M, Christian F, et al. Fecal bacteria contamination in the Adriatic Sea: Investigating environmental factors and modeling to manage recreational coastal waters [J]. Environmental pollution, 2023,338:122700.