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两种粒径纳米银对Nitrosomonas europaea的毒性效应
伍玲丽, 张晓雪, 舒昆慧, 司友斌
中国环境科学 ›› 2019, Vol. 39 ›› Issue (10) : 4401-4408.
PDF(1271 KB)
PDF(1271 KB)
两种粒径纳米银对Nitrosomonas europaea的毒性效应
Toxicity of two sizes of silver nanoparticles to Nitrosomonas europaea
为明晰不同粒径纳米银（AgNPs）对欧洲亚硝化毛杆菌（Nitrosomonas europaea）的毒性效应，采用室内培养方式，探究10nm和50nm的AgNPs对N.europaea生长、氮转化能力、细胞结构、活性氧生成和功能基因表达的影响.结果表明，AgNPs暴露抑制N.europaea生长，随着暴露时间的延长，细菌生长抑制率增加，在4h达到最大值；培养基中NH4+向NO2-转化速率减缓，N.europaea的铵态氮转化能力降低；扫描电镜（SEM）图像显示AgNPs造成部分细菌表面塌陷且有孔洞，细胞膜受损严重；透射电镜（TEM）图像显示AgNPs造成细菌内部核物质消融，细胞质膜界限模糊；流式细胞仪（FCM）检测发现AgNPs增加细胞内活性氧的生成；qRT-PCR技术对AgNPs暴露后N.europaea功能基因amoA、hao、merA表达进行测定，发现AgNPs抑制N.europaea功能基因的转录表达.综上所述，AgNPs通过与细胞膜相互作用和产生氧化应激损伤N.europaea，抑制amoA和merA的表达，进而影响铵态氮转化过程，且小粒径AgNPs的毒性强于大粒径.
The laboratory incubation experiments were conducted to study the toxicity of silver nanoparticles with different particle sizes on Nitrosomonas europaea, and the effects of two sizes of nanosilver (10nm and 50nm) on the bacterial growth, nitrogen transformation, cellular structure, reactive oxygen generation and gene expression were investigated. The results showed that nanosilver inhibited the growth of N. europaea. With the extension of exposure time, the inhibition rate of bacterial growth activity increased and reached to maximum at 4h. In the medium, the transformation rate of NH4+ to NO2-was slowed down, and the nitrogen transformation ability by N. europaea was reduced. Scanning electron microscopy (SEM) images showed that nanosilver heavily damaged the cell membrane by causing holes on the surface of bacteria. Transmission electron microscope (TEM) images showed that the nuclear material inside the bacteria was disappeared and the boundary of the cytoplasmic membrane was blurred. Flow cytometry (FCM) was employed to detect that nanosilver could generate intracellular reactive oxygen (ROS) in the cells. qRT-PCR technology was used to determine the expression of amoA, hao and merA of N. europaea after the exposure to nanosilver, and it was found that nanosilver inhibited the expression of the functional genes. In conclusion, nanosilver could interact with cell membrane, generated oxidative stress damage and inhibited the expression of functional genes amoA and merA of N. europaea, which further affected the transformation process of ammonium nitrogen. In addition, the toxicity of nanosilver with small particle size was stronger than that of large particle size.
amoA、hao、merA基因丰度 / Nitrosomonas europaea / 氨氧化 / 纳米银 / 氧化应激
Nitrosomonas europaea / ammoxidation / gene expression of amoA, hao, and merA / nanosilver / oxidative stress
[1] Reidy B, Haase A, Luch A, et al. Mechanisms of silver nanoparticle release, transformation and toxicity:A critical review of current knowledge and recommendations for future studies and applications[J]. Materials, 2013,6(6):2295-2350.
[2] Vance M E, Todd K, Vejerano E P, et al. Nanotechnology in the real world:redeveloping the nanomaterial consumer products inventory[J]. Beilstein Journal of Nanotechnology, 2015,6:1769-1780.
[3] Franci G, Falanga A, Galdiero S, et al. Silver nanoparticles as potential antibacterial agents[J]. Molecules, 2015,20(5):8856-8874.
[4] Ivask A, Elbadawy A, Kaweeteerawat C, et al. Toxicity mechanisms in Escherichia coli vary for silver nanoparticles and differ from ionic silver[J]. ACS Nano, 2014,8(1):374-386.
[5] Palza H. Antimicrobial polymers with metal nanoparticles[J]. International Journal of Molecular Sciences, 2015,16(1):2099-2116.
[6] Wu D, Fan W, Kishen A, et al. Evaluation of the antibacterial efficacy of silver nanoparticles against Enterococcus faecalis, biofilm[J]. Journal of Endodontics, 2014,40(2):285-290.
[7] Hsiao I L, Hsieh Y K, Wang C F, et al. Trojan-horse mechanism in the cellular uptake of silver nanoparticles verified by direct intra-and extracellular silver speciation analysis[J]. Environmental Science & Technology, 2015,49(6):3813-3821.
[8] Liu J, Sonshine D A, Shervani S, et al. Controlled release of biologically active silver from nanosilver surfaces[J]. ACS Nano, 2010,4(11):6903-6913.
[9] Morones J R, Elechiguerra J L, Camacho A, et al. The bactericidal effect of silver nanoparticles[J]. Nanotechnology, 2005,16(10):2346-2353.
[10] Tian X, Jiang X M, Welch C, et al. Bactericidal effects of silver nanoparticles on Lactobacilli and the underlying mechanism[J]. ACS Applied Materials & Interfaces, 2018,10(10):8443-8450.
[11] Yan M L, Li Gang H, Xue T Y, et al. Surface ligand controls silver ion release of nanosilver and its antibacterial activity against Escherichia coli[J]. International Journal of Nanomedicine, 2017,12:3193-3206.
[12] Anas A, Jiya J, Rameez M J, et al. Sequential interactions of silver-silica nanocomposite (Ag-SiO2NC) with cell wall, metabolism and genetic stability of Pseudomonas aeruginosa, a multiple antibiotic-resistant bacterium[J]. Letters in Applied Microbiology, 2013,56(1):57-62.
[13] Zheng X, Wang J, Chen Y, et al. Comprehensive analysis of transcriptional and proteomic profiling reveals silver nanoparticles-induced toxicity to bacterial denitrification[J]. Journal of Hazardous Materials, 2017,344:291-298.
[14] Rajesh S, Dharanishanthi V, Kanna A V. Antibacterial mechanism of biogenic silver nanoparticles of Lactobacillus acidophilus[J]. Journal of Experimental Nanoscience, 2014,10(15):1-10.
[15] Samarajeewa A D, Velicogna J R, Princz J I, et al. Effect of silver nanoparticles on soil microbial growth, activity and community diversity in a sandy loam soil[J]. Environmental Pollution, 2017,220:504-513.
[16] Vipindas P V, Anas A, Jasmin C, et al. Bacterial domination over archaea in ammonia oxidation in a monsoon-driven tropical estuary[J]. Microbial Ecology, 2015,69(3):544-553.
[17] Giao N T, Limpiyakorn T, Kunapongkiti P, et al. Influence of silver nanoparticles and liberated silver ions on nitrifying sludge:ammonia oxidation inhibitory kinetics and mechanism[J]. Environmental Science and Pollution Research, 2017,24(10):9229-9240.
[18] Yang Y, Wang J, Xiu Z, et al. Impacts of silver nanoparticles on cellular and transcriptional activity of nitrogen-cycling bacteria[J]. Environmental Toxicology & Chemistry, 2013,32(7):1488-1494.
[19] Yang Y, Li M, Michels C, et al. Differential sensitivity of nitrifying bacteria to silver nanoparticles in activated sludge[J]. Environmental Toxicology and Chemistry, 2014,33(10):2234-2239.
[20] Alito C L, Gunsch C K. Assessingthe effects of silver nanoparticles on biological nutrient removal in bench-scale activated sludge sequencing batch reactors[J]. Environmental Science & Technology, 2014,48(2):970-976.
[21] Wu J, Zhan M, Chang Y, et al. Adaption and recovery of Nitrosomonas europaea to chronic TiO2nanoparticle exposure[J]. Water Research, 2018,147:429-439.
[22] Yuan Z H, Li J W, Cui L, et al. Interaction of silver nanoparticles with pure nitrifying bacteria[J]. Chemosphere, 2013,90(4):1404-1411.
[23] Choi O K, Hu Z Q. Nitrification inhibition by silver nanoparticles[J]. Water Science & Technology, 2009,59(9):1769-1780.
[24] 刘美婷,余冉,陈良辉,等.典型纳米金属氧化物对氨氧化菌Nitrosomonas europaea的生物胁迫影响[J]. 中国环境科学, 2015, 35(1):190-195. Liu M T, Yu R, Chen L H, et al. Biological effects of typical metal oxide nanoparticles on Nitrosomonas europaea[J]. China Environmental Science, 2015,35(1):190-195.
[25] Jiang X, Miclaus T, Wang L, et al. Fast intracellular dissolution and persistent cellular uptake of silver nanoparticles in CHO-K1cells:implication for cytotoxicity[J]. Nanotoxicology, 2015,9(2):181-189.
[26] Michels C, Yang Y, Moreira S H, et al. Silver nanoparticles temporarily retard NO2 production without significantly affecting N2O release by Nitrosomonas europaea[J]. Environmental Toxicology & Chemistry, 2015,34(10):2231-2235.
[27] Park S, Ely R L. Candidate stress genes of Nitrosomonas europaea for monitoring inhibition of nitrification by heavy metals[J]. Applied and Environmental Microbiology, 2008,74(17):5475-5482.
[28] Radniecki T S, Stankus D P, Neigh A, et al. Influence of liberated silver from silver nanoparticles on nitrification inhibition of Nitrosomonas europaea[J]. Chemosphere, 2011,85(1):43-49.
[29] Yu R, Wu J, Liu M, et al. Physiological and transcriptional responses of Nitrosomonas europaea to TiO2 and ZnO nanoparticles and their mixtures[J]. Environmental Science & Pollution Research International, 2016,23(13):13023-13034.
[30] Mijnendonckx K, Leys N, Mahillon J, et al. Antimicrobial silver:uses, toxicity and potential for resistance[J]. Biometals, 2013,26(4):609-621.
[31] Barker L K, Giska J R, Radniecki T S, et al. Effects of short and long-term exposure of silver nanoparticles and silver ions to Nitrosomonas europaea, biofilms and planktonic cells[J]. Chemosphere, 2018,206:606-614.
[32] Arnaout C L, Gunsch C K. Impacts of silver nanoparticle coating on the nitrification potential of Nitrosomonas europaea[J]. Environmental Science & Technology, 2012,46(10):5387-5395.
[33] Hou J, You G X, Xu Y, et al. Antioxidant enzyme activities as biomarkers of fluvial biofilm to ZnONPs ecotoxicity and the integrated biomarker responses (IBR) assessment[J]. Ecotoxicology and Environmental Safety, 2016,133:10-17.
[34] Zhang L, Wu L L, Si Y B, et al.Size-dependent cytotoxicity of silver nanoparticles to Azotobacter vinelandii: Growth inhibition, cell injury, oxidative stress and internalization[J]. PLOSE One, 2018,13(12):e0209020.
[35] Jeong E, Chae S R, Kang S T, et al. Effects of silver nanoparticles on biological nitrogen removal processes[J]. Water Science & Technology, 2012,65(7):1298-1303.
[36] Manke A, Wang L, Rojanasakul Y. Mechanisms of nanoparticle induced oxidative stress and toxicity[J]. BioMed Research International, 2013,2013:1-15.
[37] Rosen B P. Bacterial resistance to heavy metals and metalloids[J]. Journal of Biological Inorganic Chemistry, 1996,1(4):273-277.
[38] Marie S L, Kathryn J, Millstone J E, et al. Emerging investigator series:It's not all about the ion:support for particle specific contributions to silver nanoparticle antimicrobial activity[J]. Environmental Science Nano, 2018,5(9):2047-2068.
[39] Choi O, Hu Z. Size dependent and reactive oxygen species related nanosilver toxicity to nitrifying bacteria[J]. Environmental Science & Technology, 2008,42(12):4583-4588.
[40] Ellegaard-Jensen L, Jensen K A, Johansen A. Nano-silver induces dose-response effects on the nematode Caenorhabditis elegans [J]. Ecotoxicology and Environmental Safety. 2012,80:216-23.
[41] Wang J, Shu K H, Zhang L, et al. Effects of silver nanoparticles on soil microbial communities and bacterial nitrification in suburban vegetable soils[J]. Pedosphere, 2017,27(3):482-490.
国家自然科学基金重点项目（41430752）；安徽农业大学研究生创新基金（2019ysj-67）
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