高风险四环素抗性基因在人工湿地中分布和去除的季节变化

doi:10.13287/j.1001-9332.202210.036

摘要/Abstract

摘要： 畜禽养殖废水是抗生素抗性基因(ARGs)的重要贮存库,环境风险不容忽视。本研究考察了夏、冬两季养猪废水中高环境风险四环素抗性基因(TRGs)在水平潜流人工湿地中的分布和去除情况,通过外源添加四环素(TC)和铜离子(Cu²⁺)探究了养猪废水中抗生素与重金属单一和复合污染对湿地出水中TRGs丰度的影响。结果表明: 养猪废水中3种高环境风险TRGs(tetM、tetO、tetW)均有检出,人工湿地可有效消减废水中的TRGs,夏、冬两季时出水中TRGs的绝对丰度较进水分别降低1.1～2.4和1.7～2.9个数量级。TRGs在湿地土壤中的丰度呈现出水端低于进水端、植物非根际低于根际、冬季低于夏季的特征。与对照相比,养猪废水中TC、Cu²⁺单一和复合污染在夏、冬两季均会导致湿地出水中TRGs相对丰度的增加。人工湿地作为一种生态处理工艺可用于控制畜禽养殖废水中的ARGs向环境中扩散。

关键词: 畜禽养殖废水, 人工湿地, 四环素抗性基因, 高环境风险, 铜离子

Abstract: Livestock wastewater is an important reservoir of antibiotic resistance genes (ARGs), with high environmental risks. We investigated the seasonal variations of distribution and removal of swine wastewater originated high-risk tetracycline resistance genes (TRGs) in horizontal subsurface flow constructed wetlands. The effects of exogenous addition of tetracycline (TC) and copper ion (Cu²⁺) on the abundance of TRGs in effluent with single and combined pollution of antibiotic and heavy metal were studied. The results showed that all the three high-risk TRGs (tetM, tetO and tetW) were detected in swine wastewater. Wetlands could effectively reduce the ARGs, with the absolute abundance of TRGs in effluent being decreased by 1.1-2.4 and 1.7-2.9 orders of magnitude in summer and winter compared with the influent, respectively. The abundance of TRGs in wetland soils showed the characte-ristics that the outflow side was lower than the inflow side, the non-rhizosphere area was lower than the rhizosphere area, and lower in winter than in summer. In summer and winter, single and combined pollution of TC and Cu²⁺ in swine wastewater would increase the abundance of TRGs in effluent compared with that in the control. The constructed wetland is suitable for controlling the environmental diffusion of ARGs in livestock wastewater.

Key words: livestock wastewater, constructed wetland, tetracycline resistance genes, high-risk, copper ion (Cu²⁺)

范增增, 赵伟, 杨新萍. 高风险四环素抗性基因在人工湿地中分布和去除的季节变化[J]. 应用生态学报, 2022, 33(11): 2997-3006.

FAN Zeng-zeng, ZHAO Wei, YANG Xin-ping. Seasonal variation of distribution and removal of high-risk tetracycline resistance genes in constructed wetland.[J]. Chinese Journal of Applied Ecology, 2022, 33(11): 2997-3006.

参考文献

[1] 周启星, 罗义, 王美娥. 抗生素的环境残留、生态毒性及抗性基因污染. 生态毒理学报, 2007, 2(3): 243-251
[2] 邹威, 金彩霞, 魏闪, 等. 华北地区不同规模畜禽养殖场粪便中抗生素抗性基因污染特征. 农业环境科学学报, 2020, 39(11): 2640-2652
[3] Wu XF, Wei YS, Zheng JX, et al. The behavior of tetracyclines and their degradation products during swine manure composting. Bioresource Technology, 2011, 102: 5924-5931
[4] 陈小桐, 安静, 杨合, 等. 辽宁省畜禽养殖业抗生素排放特征及减排路径分析. 环境科学学报, 2021, 41(7): 2896-2904
[5] Liu MM, Zhang Y, Yang M, et al. Abundance and distribution of tetracycline resistance genes and mobile elements in an oxytetracycline production wastewater treatment system. Environmental Science & Technology, 2012, 46: 7551-7557
[6] Liu L, Liu YH, Liu CX, et al. Accumulation of antibio-tics and tet resistance genes from swine wastewater in wetland soils. Environmental Engineering Management Journal, 2016, 15: 2137-2145
[7] 何帅雄, 陈梅雪, 强志民. 畜禽养殖废水生物处理剩余污泥臭氧减量过程中的重金属释放. 环境工程学报, 2014, 8(1): 55-61
[8] Feng L, Casas ME, Ottosen LDM, et al. Removal of antibiotics during the anaerobic digestion of pig manure. Science of the Total Environment, 2017, 603: 219-225
[9] Jia S, Zhang XX, Miao Y, et al. Fate of antibiotic resistance genes and their associations with bacterial community in livestock breeding wastewater and its receiving river water. Water Research, 2017, 124: 259-268
[10] Sun YM, Shen YX, Liang P, et al. Multiple antibiotic resistance genes distribution in ten large-scale membrane bioreactors for municipal wastewater treatment. Bioresource Technology, 2016, 222: 100-106
[11] Shao SC, Hu YY, Cheng JH, et al. Research progress on distribution, migration, transformation of antibiotics and antibiotic resistance genes (ARGs) in aquatic environment. Critical Reviews in Biotechnology, 2018, 38: 1195-1208
[12] Liu L, Liu YH, Wang Z, et al. Behavior of tetracycline and sulfamethazine with corresponding resistance genes from swine wastewater in pilot-scale constructed wetlands. Journal of Hazardous Materials, 2014, 278: 304-310
[13] Tao CW, Hsu BM, Ji WT, et al. Evaluation of five antibiotic resistance genes in wastewater treatment systems of swine farms by real-time PCR. Science of the Total Environment, 2014, 496: 116-121
[14] Zhang AN, Li LG, Yin X, et al. Choosing your battles: Which resistance genes warrant global action? BioRxiv, 2019, 2019: 784322
[15] Borin M, Politeo M, De Stefani G. Performance of a hybrid constructed wetland treating piggery wastewater. Ecological Engineering, 2013, 51: 229-236
[16] Liu L, Chen SR, Xu KQ, et al. Influence of hydraulic loading rate on antibiotics removal and antibiotic resis-tance expression in soil layer of constructed wetlands. Chemosphere, 2021, 265: 129100
[17] 程羽霄, 吴丹, 陈铨乐, 等. 潮汐-复合流人工湿地系统优化及对抗生素抗性基因的去除效果. 环境科学, 2021, 42(8): 3799-3807
[18] 郑加玉, 刘琳, 高大文, 等. 四环素抗性基因在人工湿地中的去除及累积. 环境科学, 2013, 34(8): 3102-3107
[19] Liu L, Liu CX, Zheng JY, et al. Elimination of veterinary antibiotics and antibiotic resistance genes from swine wastewater in the vertical flow constructed wetlands. Chemosphere, 2013, 91: 1088-1093
[20] Wang WD, Zheng J, Wang ZQ, et al. Performance of pond-wetland complexes as a preliminary processor of drinking water sources. Journal of Environmental Sciences, 2016, 39: 119-133
[21] Aminov RI, Garrigues-Jeanjean N, Mackie RI. Molecular ecology of tetracycline resistance: Development and validation of primers for detection of tetracycline resis-tance genes encoding ribosomal protection proteins. Applied and Environmental Microbiology, 2001, 67: 22-32
[22] Luo Y, Mao DQ, Rysz M, et al. Trends in antibiotic resistance genes occurrence in the Haihe River, China. Environmental Science & Technology, 2010, 44: 7220-7225
[23] Duan QJ, Shang SQ, Wu YD. Rapid diagnosis of bacterial meningitis in children with fluorescence quantitative polymerase chain reaction amplification in the bacterial 16S rRNA gene. European Journal of Pediatrics, 2009, 168: 211-216
[24] 王丽梅, 罗义, 毛大庆, 等. 抗生素抗性基因在环境中的传播扩散及抗性研究方法. 应用生态学报, 2010, 21(4): 1063-1069
[25] de la Fuente Cantó C, Simonin M, King E, et al. An extended root phenotype: The rhizosphere, its formation and impacts on plant fitness. The Plant Journal, 2020, 103: 951-964
[26] Tong XN, Wang XZ, He XJ, et al. Effects of antibiotics on microbial community structure and microbial functions in constructed wetlands treated with artificial root exudates. Environmental Science: Processes & Impacts, 2020, 22: 217-226
[27] Liu X, Guo X, Liu Y, et al. A review on removing antibiotics and antibiotic resistance genes from wastewater by constructed wetlands: Performance and microbial response. Environmental Pollution, 2019, 254: 112996
[28] 陈迪, 杨勇, 郑祥, 等. 人工湿地对病原微生物去除的研究进展. 农业环境科学学报, 2013, 32(9): 1720-1730
[29] Karim MR, Glenn EP, Gerba CP. The effect of wetland vegetation on the survival of Escherichia coli, Salmonella typhimurium, bacteriophage MS-2 and polio virus. Journal of Water & Health, 2008, 6: 167-175
[30] 杨芳, 陶然, 杨扬, 等. 人工湿地中抗生素抗性大肠杆菌和抗性基因的去除与分布. 环境工程学报, 2013, 7(6): 2057-2062
[31] Yuan QB, Guo MT, Yang J. Monitoring and assessing the impact of wastewater treatment on release of both antibiotic-resistant bacteria and their typical genes in a Chinese municipal wastewater treatment plant. Environmental Science: Processes & Impacts, 2014, 16: 1930-1937
[32] Li JN, Cheng WX, Xu LK, et al. Antibiotic-resistant genes and antibiotic-resistant bacteria in the effluent of urban residential areas, hospitals, and a municipal wastewater treatment plant system. Environmental Science Pollution Research, 2015, 22: 4587-4596
[33] Silva J, Castillo G, Callejas L, et al. Frequency of transferable multiple antibiotic resistance amongst coliform bacteria isolated from a treated sewage effluent in Antofagasta, Chile. Electronic Journal of Biotechnology, 2006, 9: 533-540
[34] 赵伟, 范增增, 杨新萍. 水平潜流人工湿地对畜禽养殖废水中特征污染物的去除. 环境科学, 2021, 42(12): 5865-5875
[35] 谢冰, 董亮. 芦苇人工湿地底泥微生物群落结构的PCR-SSCP技术研究. 农业系统科学与综合研究, 2007, 23(2): 205-211
[36] 曹曼曼, 王飞, 周北海, 等. 铁尾矿芦苇根际微生物和根内生菌群落分布及其限制性因子解析. 环境科学, 2021, 42(10): 4998-5009
[37] 张翠萍, 李淑英, 王蓓, 等. 六氯苯胁迫下2种湿地植物根际土壤微生物数量与酶活性变化. 生态与农村环境学报, 2018, 34(2): 177-183
[38] 李科德, 胡正嘉. 芦苇床系统净化污水的机理. 中国环境科学, 1995, 15(2): 140-144
[39] 朱芸芸, 李敏, 赵暾, 等. 冬季野鸭湖湿地植物根际微生物群落结构分析. 环境科学与技术, 2016, 39(9): 171-176
[40] 张俊亚, 魏源送, 陈梅雪, 等. 畜禽粪便生物处理与土地利用全过程中抗生素和重金属抗性基因的赋存与转归特征研究进展. 环境科学学报, 2015, 35(4): 935-946
[41] Pal C, Bengtsson-Palme J, Kristiansson E, et al. Co-occurrence of resistance genes to antibiotics, biocides and metals reveals novel insights into their co-selection potential. BMC Genomics, 2015, 16: 1-14
[42] Poole K. At the nexus of antibiotics and metals: The impact of Cu and Zn on antibiotic activity and resistance. Trends in Microbiology, 2017, 25: 820-832
[43] Huang X, Liu CX, Li K, et al. Performance of vertical up-flow constructed wetlands on swine wastewater containing tetracyclines and tet genes. Water Research, 2015, 70: 109-117
[44] Berg J, Thorsen MK, Holm PE, et al. Cu exposure under field conditions coselects for antibiotic resistance as determined by a novel cultivation-independent bacterial community tolerance assay. Environmental Science & Technology, 2010, 44: 8724-8728
[45] Xu YB, Hou MY, Li YF, et al. Distribution of tetracycline resistance genes and AmpC β-lactamase genes in representative non-urban sewage plants and correlations with treatment processes and heavy metals. Chemosphere, 2017, 170: 274-281
[46] He X, Xu Y, Chen J, et al. Evolution of corresponding resistance genes in the water of fish tanks with multiple stresses of antibiotics and heavy metals. Water Research, 2017, 124: 39-48
[47] Liu L, Xin Y, Huang X, et al. Response of antibiotic resistance genes in constructed wetlands during treatment of livestock wastewater with different exogenous indu-cers: Antibiotic and antibiotic-resistant bacteria. Bioresource Technology, 2020, 314: 123779