纳米银与银离子对土壤微生物及酶活性的影响
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摘要
为研究纳米银和银离子对土壤微生物的影响,采用土壤培养方式,对不同剂量纳米银(10、50、100 mg·kg~(-1))和银离子(1、5、10 mg·kg~(-1))暴露下黄褐土、砖红壤中可培养微生物数量及土壤酶活性(脲酶、荧光素二乙酸酯水解酶、蔗糖酶、过氧化氢酶)进行研究,并采用纯培养方法对纳米银和银离子暴露下的大肠杆菌(Escherichia coli)、金黄色葡萄球菌(Staphylococcus aureus)凋亡情况进行检测,对纳米银释放的银离子毒性进行评估。结果表明,随着纳米银剂量的增加,土壤可培养微生物数量显著减少,脲酶和过氧化氢酶活性降低,蔗糖酶、荧光素二乙酸酯水解酶(FDA酶)活性没有显著变化;银离子处理中微生物数量明显减少,但土壤酶活性被激活。10 mg·L~(-1)纳米银暴露1 h后大肠杆菌、金黄色葡萄球菌凋亡率、死亡率增高;随着培养时间的延长,纳米银缓慢释放银离子,并促进大肠杆菌的凋亡。综上分析,纳米银能够抑制土壤可培养微生物生长和酶活性,其中脲酶、过氧化氢酶对纳米银较为敏感,蔗糖酶、FDA酶受纳米银的影响较小;纳米银的毒性一方面是其本身的特异抗菌性,也有部分来自缓慢释放的银离子。
In order to evaluate the microbial toxicity of silver nanoparticles(AgNP) and silver ions in soils, the incubation experiments were conducted to investigate the effect of nanosilver(0, 10, 50, 100 mg·kg~(-1)) and silver acetate(1, 5, 10 mg·kg~(-1)) on microbial population and enzyme activities(urease, fluorescein diacetate hydrolase, invertase and catalase) in yellow cinnama and laterite. The apoptotic cells of Escherichia coli and Staphylococcus aureus exposed to nanosilver and silver ions were also determined by the pure culture test, and silver ions released from nanosilver were evaluated. The results showed that with the increase of nanosilver content, the microbial quantities decreased significantly, the activities of urease and catalase decreased, but there was no significant changein invertase and fluorescein diacetate(FDA) hydrolase activities. The population of microorganisms in the silver acetate treatment significantly decreased, while the soil enzyme activities were activated. The apoptosis rate and mortality of Escherichia coli and Staphylococcus aureus increased after exposure to 10 mg·L~(-1) nanosilver for 1 hour. As the culture time prolonged, nanosilver slowly released silver ions and promoted the apoptosis of E. coli In summary, nanosilver can inhibit the growth of culturable microorganisms and the activity of enzyme in soil. Urease and catalase are sensitive to nanosilver but invertase and FDA hydrolase are less affected by nanosilver. The toxicity of nanosilver is from its own specific antibacterial properties, also partly from the slow release of silver ions.
In order to evaluate the microbial toxicity of silver nanoparticles(AgNP) and silver ions in soils, the incubation experiments were conducted to investigate the effect of nanosilver(0, 10, 50, 100 mg·kg~(-1)) and silver acetate(1, 5, 10 mg·kg~(-1)) on microbial population and enzyme activities(urease, fluorescein diacetate hydrolase, invertase and catalase) in yellow cinnama and laterite. The apoptotic cells of Escherichia coli and Staphylococcus aureus exposed to nanosilver and silver ions were also determined by the pure culture test, and silver ions released from nanosilver were evaluated. The results showed that with the increase of nanosilver content, the microbial quantities decreased significantly, the activities of urease and catalase decreased, but there was no significant changein invertase and fluorescein diacetate(FDA) hydrolase activities. The population of microorganisms in the silver acetate treatment significantly decreased, while the soil enzyme activities were activated. The apoptosis rate and mortality of Escherichia coli and Staphylococcus aureus increased after exposure to 10 mg·L~(-1) nanosilver for 1 hour. As the culture time prolonged, nanosilver slowly released silver ions and promoted the apoptosis of E. coli In summary, nanosilver can inhibit the growth of culturable microorganisms and the activity of enzyme in soil. Urease and catalase are sensitive to nanosilver but invertase and FDA hydrolase are less affected by nanosilver. The toxicity of nanosilver is from its own specific antibacterial properties, also partly from the slow release of silver ions.
引文
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[2]Anjum N A,Gill S S,Duarte A C,et al.Silver nanoparticles in soil-plant systems[J].Journal of Nanoparticle Research,2013,15(9):1-26
[3]Rico C M,Majumdar S,Duarte-Gardea M.Interaction of nanoparticles with edible plants and their possible implications in the food chain[J].Journal of Agricultural&Food Chemistry,2011,59(8):3485-3498
[4]Kahru A,Dubourguier H C,Blinova I,et al.Biotests and biosensors for ecotoxicology of metal oxide nanoparticles:A minireview[J].Sensors,2008,8(8):5153-5170
[5]林先贵,陈瑞蕊,胡君利.土壤微生物资源管理、应用技术与学科展望[J].生态学报,2010,30(24):7029-7037Lin X G,Chen R R,Hu J L.The management and application of soil microbial resources and the perspectives of soil microbiology[J].Acta Ecologica Sinica,2010,30(24):7029-7037(in Chinese)
[6]Du W,Sun Y,Ji R,et al.Ti O2and Zn O nanoparticles negatively affect wheat growth and soil enzyme activities in agricultural soil[J].Journal of Environmental Monitoring,2011,13(4):822-828
[7]Peyrot C,Wilkinson K J,Desrosiers M,et al.Effects of silver nanoparticles on soil enzyme activities with and without added organic matter[J].Environmental Toxicology&Chemistry,2014,33(1):115-125
[8]王秋双,戚兴超,申天琳,等.纳米银与石墨烯对土壤微生物及土壤酶的影响[J].环境科学学报,2017,37(8):3149-3157Wang Q S,Qi X C,Shen T L,et al.Effect of silver nanoparticles and graphene on soil microorganisms and enzyme activities[J].Acta Scientiae Circumstantiae,2017,37(8):3149-3157(in Chinese)
[9]Chen X,Schluesener H J.Nanosilver:A nanoproduct in medical application[J].Toxicology Letters,2008,176(1):1-12
[10]Liu J Y,Sonshine D A,Shervani S,et al.Controlled release of biologically active silver from nanosilver surfaces[J].ACS Nano,2010,4(11):6903-6913
[11]Morones J R,Elechiguerra J L,Camacho A,et al.The bactericidal effect of silver nanoparticles[J].Nanotechnology,2005,16(10):2346-2353
[12]Fabrega J,Fawcett S R,Renshaw J C,et al.Silver nanoparticle impact on bacterial growth:Effect of p H,concentration,and organic matter[J].Environmental Science&Technology,2009,43(19):7285-7290
[13]吴源.纳米银的生物效应及毒性作用机制[D].合肥:中国科学技术大学,2010:81-83Wu Y.Biological effects of silver nanoparticles and the mechanistic clue for toxicity[D].Hefei:University of Science and Technology of China,2010:81-83(in Chinese)
[14]Sharma V K,Yngard R A,Lin Y.Silver nanoparticles:Green synthesis and their antimicrobial activities[J].Advances in Colloid and Interface Science,2009,145(1-2):83-96
[15]Said J,Dodoo C C,Walker M,et al.An in vitro test of the efficacy of silver-containing wound dressings against Staphylococcus aureus and Pseudomonas aeruginosa in simulated wound fluid[J].International Journal of Pharmaceutics,2014,462(1-2):123-128
[16]Dwyer D J,Winkler J A.Identification and Characterization of Programmed Cell Death Markers in Bacterial Models[M].Necrosis:Humana Press,2013:145-159
[17]胡伟,李越中,张禹清,等.细菌的细胞程序性死亡[J].微生物学报,2002,42(2):255-258Hu W,Li Y Z,Zhang Y Q,et al.Programmed cell death in bacteria[J].Acta Microbiologica Sinica,2002,42(2):255-258(in Chinese)
[18]Xu H,Qu F,Xu H,et al.Role of reactive oxygen species in the antibacterial mechanism of silver nanoparticles on Escherichia coli O157:H7[J].Biometals,2012,25(1):45-53
[19]Fadok V A,Voelker D R,Campbell P A,et al.Exposure of phosphatidylserine on the surface of apoptotic lymphocytes triggers specific recognition and removal by macrophages[J].Journal of Immunology,1992,148(7):2207-2216
[20]Gottschalk F,Sonderer T,Scholz R W,et al.Modeled environmental concentrations of engineered nanomaterials(TiO2,Zn O,Ag,CNT,Fullerenes)for different regions[J].Environmental Science&Technology,2009,43(24):9216-9222
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[22]李振高,骆永明,滕应.土壤与环境微生物研究法[M].北京:科学出版社,2008:395-419Li Z G,Luo Y M,Teng Y.Soil and Environmental Microbiological Research[M].Beijing:Science Press,2008:395-419(in Chinese)
[23]Kaiser J P,Roesslein M,Diener L,et al.Cytotoxic effects of nanosilver are highly dependent on the chloride concentration and the presence of organic compounds in the cell culture media[J].Journal of Nanobiotechnology,2017,15(1):1-11
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[27]Yang Y,Quensen J,Mathieu J,et al.Pyrosequencing reveals higher impact of silver nanoparticles than Ag+on the microbial community structure of activated sludge[J].Water Research,2014,48:317-325
[28]Kraas M,Schlich K,Knopf B,et al.Long-term effects of sulfidized silver nanoparticles in sewage sludge on soil microflora[J].Environmental Toxicology&Chemistry,2017,36(12):3305-3313
[29]Starnes D L,Lichtenberg S S,Unrine J M,et al.Distinct transcriptomic responses of Caenorhabditis elegans to pristine and sulfidized silver nanoparticles[J].Environmental Pollution,2016,213:314-321
[30]Xiu Z M,Zhang Q B,Puppala H L,et al.Negligible particle-specific antibacterial activity of silver nanoparticles[J].Nano Letters,2012,12(12):4271-4275
[31]Benn T M,Westerhoff P.Nanoparticle silver released into water from commercially available sock fabrics[J].Environmental Science&Technology,2008,42(11):4133-4139
[32]Sedlak R H,Hnilova M,Grosh C,et al.Engineered Escherichia coli silver-binding periplasmic protein that promotes silver tolerance[J].Applied&Environmental Microbiology,2012,78(7):2289-2296
[33]Lee Y J,Kim J,Oh J,et al.Ion-release kinetics and ecotoxicity effects of silver nanoparticles[J].Environmental Toxicology&Chemistry,2012,31(1):155-159
[34]Beer C,Foldbjerg R,Hayashi Y,et al.Toxicity of silver nanoparticles-Nanoparticle or silver ion?[J].Toxicology Letters,2012,208(3):286-292
[35]Budd R C.Activation-induced cell death[J].Current O-pinion in Immunology,2001,13(3):356-362
[36]Min J S,Kim K S,Kim S W,et al.Effects of colloidal silver nanoparticles on sclerotium forming phytopathogenic fungi[J].Plant Pathology Journal,2009,25(4):376-380
[37]Buttke T M,Sandstrom P A.Oxidative stress as a mediator of apoptosis[J].Immunology Today,1994,15(1):7-10