纳米银对反硝化无色杆菌的毒性作用

为研究纳米银(AgNPs)对反硝化无色杆菌(Achromobacter denitrificans)的毒性作用,选择室内培养方式,探究纳米银对Achromobacter denitrificans的生长抑制、氨化作用、同化吸收NH₄+-N、细胞膜表面结构和活性氧生成的影响。研究发现,纳米银可抑制细菌生长,抑制效果与浓度和暴露时间呈正相关。添加10 mg/LAgNPs在硝化培养基中,Achromobacter denitrificans生长抑制率12h后达38.2%,而添加1 mg/LAgNPs时生长抑制率仅为11.5%。不同AgNPs暴露浓度条件下,4~6 h后生长抑制率均趋于稳定。暴露培养后的细菌生化活性降低,AgNPs投加浓度从1 mg/L提高到10 mg/L时,NH+₄-N生成速率从2.77 mg/(L·h)降低至2.07 mg/(L·h),降低了25.3%;NH₄+-N同化速率从5.52 mg/(L·h)降低至1.71 mg/(L·h),降低了69.1%。pH是影响毒性作用的重要因素,弱酸(pH 5.0)与弱碱(pH 9.0)均不利于细菌生存。通过毒性作用机理分析可知,纳米银可导致细胞膜表面凹陷破裂,膜内物质泄漏流出,细胞内发生活性氧的累积。     

纳米银与镉的复合毒性对拟南芥根系形态及叶片生理指标的影响

为研究纳米银（AgNPs）与镉（Cd²⁺）对拟南芥的复合毒性,本文以不同浓度的PVP coated-AgNPs和Cd²⁺处理拟南芥幼苗一周后,测定了拟南芥幼苗主根长度,根毛数量,叶片中光合色素和可溶性蛋白含量及抗氧化系统相关酶活性.结果表明,高浓度（5mg/L） Cd²⁺能显著抑制根长,提高可溶性蛋白含量及SOD（超氧化物歧化酶）和POD（过氧化物酶）的活性.在高浓度Cd²⁺（5mg/L）中加入AgNPs能显著增强Cd²⁺对根长的抑制作用,而高浓度的Cd²⁺能缓解AgNPs对根毛的抑制作用;在低浓度Cd²⁺（0.1mg/L和1mg/L）中加入AgNPs能显著增强Cd²⁺对色素含量抑制作用;在含有Cd²⁺的处理中,SOD和POD在1mg/L AgNPs下活性最高;以上结果说明Cd²⁺和AgNPs对拟南芥幼苗根系形态及叶片生理指标都存在交互影响.     

Molecular toxicity and defense mechanisms induced by silver nanoparticles in Drosophila melanogaster

The widely use of silver nanoparticles (AgNPs) as antimicrobial agents gives rise to potential environmental risks. AgNPs exposure have been reported to cause toxicity in animals. Nevertheless, the known mechanisms of AgNPs toxicity are still limited. In this study, we systematically investigated the toxicity of AgNPs exposure using Drosophila melanogaster. We show here that AgNPs significantly decreased Drosophila fecundity, the third-instar larvae weight and rates of pupation and eclosion in a dose-dependent manner. AgNPs reduced fat body cell viability in MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assays. AgNPs caused DNA damage in hemocytes and S2 cells. Interestingly, the mRNA levels of the entire metallothionein gene family were increased under AgNPs exposure as determined by RNA-seq analysis and validated by qRT-PCR, indicating that Drosophila responded to the metal toxicity of AgNPs by producing metallothioneins for detoxification. These findings provide a better understanding of the mechanisms of AgNPs toxicity and may provide clues to effect on other organisms, including humans.     

纳米银对聚磷菌吸磷和释磷的影响及毒性效应

利用硼氢化钠还原硝酸银,并使用聚乙烯醇（PVA）作为分散剂,制备出分散良好、粒径为（14±3）nm的纳米银颗粒,考察了其对聚磷菌（Microlunatus phosphovorus）好氧吸磷和厌氧释磷的影响,以及产生的毒性效应.结果表明,在好氧状态下,7mg/L的纳米银能够完全抑制聚磷菌的生长（P <0.01）,达到10mg/L时才能完全抑制聚磷菌的吸磷能力（P=0.01）;在厌氧状态下,大于20mg/L的纳米银才使聚磷菌释磷能力受到部分抑制（P <0.05）.活性氧簇（ROS）和扫描电子显微镜（SEM）的检测结果表明,纳米银使细菌体内ROS水平降低,部分细菌菌体表面塌陷,这说明,纳米银不但可以毒害聚磷菌菌体表面,还可以降低菌内ROS水平.     

Sunlight-driven reduction of silver ion to silver nanoparticle by organic matter mitigates the acute toxicity of silver to Daphnia magna

Due to the unique antibacterial activities, silver nanoparticles (AgNPs) have been extensively used in commercial products. Anthropogenic activities have released considerable AgNPs as well as highly toxic silver ion (Ag⁺) into the aquatic environment. Our recent study revealed that ubiquitous natural organic matter (NOM) could reduce Ag⁺ to AgNP under natural sunlight. However, the toxic effect of this process is not well understood. In this work, we prepared mixture solution of Ag⁺ and AgNPs with varied Ag⁺% through the sunlight-driven reduction of Ag⁺ by NOM and investigated the acute toxicity of the solutions on Daphnia magna. Formation of AgNPs was demonstrated and characterized by comprehensive techniques and the fraction of unconverted Ag⁺ was determined by ultrafiltration-inductively coupled plasma mass spectrometry determination. The formation of AgNPs enhanced significantly with the increasing of solution pH and cumulative photosynthetically active radiation of sunlight. The toxicity of the resulting solution was further investigated by using freshwater crustacean D. magna as a model and an 8 hr-median lethal concentration (LC₅₀) demonstrated that the reduction of Ag⁺ by NOM to AgNPs significantly mitigated the acute toxicity of silver. These results highlight the importance of sunlight and NOM in the fate, transformation and toxicity of Ag⁺ and AgNPs, and further indicate that the acute toxicity of AgNPs should be mainly ascribed to the dissolved Ag⁺ from AgNPs.     

Interaction of silver nanoparticles with mediterranean agricultural soils: Lab-controlled adsorption and desorption studies

The production of silver nanoparticles (AgNPs) has increased tremendously during recent years due to their antibacterial and physicochemical properties. As a consequence, these particles are released inevitably into the environment, with soil being the main sink of disposal. Soil interactions have an effect on AgNP mobility, transport and bioavailability. To understand AgNP adsorption processes, lab-controlled kinetic studies were performed. Batch tests performed with five different Mediterranean agricultural soils showed that cation exchange capacity and electrical conductivity are the main parameters controlling the adsorption processes. The adsorption kinetics of different sized (40, 75, 100 and 200?nm) and coated (citrate, polyvinylpyrrolidone and polyethyleneglycol (PEG)) AgNPs indicated that these nanoparticle properties have also an effect on the adsorption processes. To assess the mobility and bioavailability of AgNPs and to determine if their form is maintained during adsorption/desorption processes, loaded soils were submitted to leaching tests three weeks after batch adsorption studies. The DIN 38414-S4 extraction method indicated that AgNPs were strongly retained on soils, and single-particle inductively coupled plasma mass spectrometry confirmed that silver particles maintained their nanoform, except for 100?nm PEG-AgNPs and 40?nm citrate-coated AgNPs. The DTPA (diethylenetriaminepentaacetic acid) leaching test was more effective in extracting silver, but there was no presence of AgNPs in almost all of these leachates.     

纳米银与石墨烯对土壤微生物及土壤酶的影响

采用室内暗培养试验分别探究了纳米银与石墨烯对土壤微生物及土壤酶的不同影响.将不同剂量的纳米银(0、10、100、150 mg·kg~(-1))与高纯石墨烯(0、10、100、1000 mg·kg~(-1))分别与等量棕壤充分混匀,然后进行暗培养.在第3、7、15、30和60 d时取样,测定土壤脲酶、土壤碱性磷酸酶、土壤脱氢酶和土壤过氧化氢酶的活性及土壤细菌、真菌和放线菌的数量,并在培养期间测定土壤呼吸速率及CO2累积量.结果表明,所有纳米银处理均抑制土壤的呼吸作用,并且剂量越高,抑制作用越明显;而石墨烯处理未对土壤呼吸产生显著影响.10 mg·kg~(-1)纳米银处理下,土壤真菌数量在整个培养期内均显著低于对照,土壤细菌在第60 d时也被显著抑制,但土壤放线菌数量无变化;与对照相比,100和150 mg·kg~(-1)的纳米银处理显著降低了土壤细菌、真菌、放线菌的数量.10和100 mg·kg~(-1)的石墨烯处理下,土壤细菌、真菌、放线菌数量则均无显著变化.1000 mg·kg~(-1)的石墨烯显著增加了土壤中细菌与真菌的数量,却对土壤放线菌数量无影响.纳米银处理显著抑制土壤脲酶、脱氢酶活性,却对土壤过氧化氢酶与磷酸酶活性基本无影响.10和100 mg·kg~(-1)石墨烯处理对土壤脲酶有一定的促进作用,1000 mg·kg~(-1)石墨烯处理对土壤过氧化氢酶和脱氢酶有一定的促进作用,而不同剂量的石墨烯在培养后期均对碱性磷酸酶产生抑制作用.总体来说,纳米银在一定程度上对土壤酶及土壤微生物结构产生了负面影响,而石墨烯对土壤酶及土壤微生物结构的影响不明显.     

抗生素和纳米银对大肠杆菌耐药性的联合效应

以大肠杆菌（E.coli）为模式生物,研究了磺胺类抗生素（SAs）和纳米银（AgNPs）在单一或联合作用下对RP4质粒接合转移效应及E.coli突变效应的影响,并对联合作用方式进行了判别,基于分子对接技术和线性回归分析,探讨了两种效应的机制及其与毒性效应的关系.结果表明,在较低浓度范围内,所测12种SAs以及AgNPs在单一或联合暴露下对接合转移和突变均有促进作用,12组SAs-AgNPs混合物对接合转移频率促进率峰值的最大值和最小值分别为105.32%和46.96%,对突变体促进率峰值的最大值和最小值分别为1410.25%和238.38%.此外,SAs和AgNPs联合暴露对RP4质粒接合转移效应主要表现为协同作用,对E.coli突变效应主要表现为拮抗作用,且接合转移效应、突变效应与毒性效应之间具有良好的相关性.     

抗生素和纳米银对大肠杆菌耐药性的联合效应

以大肠杆菌（E.coli）为模式生物,研究了磺胺类抗生素（SAs）和纳米银（AgNPs）在单一或联合作用下对RP4质粒接合转移效应及E.coli突变效应的影响,并对联合作用方式进行了判别,基于分子对接技术和线性回归分析,探讨了两种效应的机制及其与毒性效应的关系.结果表明,在较低浓度范围内,所测12种SAs以及AgNPs在单一或联合暴露下对接合转移和突变均有促进作用,12组SAs-AgNPs混合物对接合转移频率促进率峰值的最大值和最小值分别为105.32%和46.96%,对突变体促进率峰值的最大值和最小值分别为1410.25%和238.38%.此外,SAs和AgNPs联合暴露对RP4质粒接合转移效应主要表现为协同作用,对E.coli突变效应主要表现为拮抗作用,且接合转移效应、突变效应与毒性效应之间具有良好的相关性.     

Effects of silver nanoparticles with different dosing regimens and exposure media on artificial ecosystem

Due to the wide use of silver nanoparticles (AgNPs) in various fields, it is crucial to explore the potential negative impacts on the aquatic environment of AgNPs entering into the environment in different ways. In this study, comparative experiments were conducted to investigate the toxicological impacts of polyvinylpyrrolidone-coated silver nanoparticles (PVP-AgNPs) with two kinds of dosing regimens, continuous and one-time pulsed dosing, in different exposure media (deionized water and XiangJiang River water). There were a number of quite different experimental results (including 100% mortality of zebrafish, decline in the activity of enzymes, and lowest number and length of adventitious roots) in the one-time pulsed dosing regimen at high PVP-AgNP concentration exposure (HOE) compared to the three other treatments. Meanwhile, we determined that the concentration of leached silver ions from PVP-AgNPs was too low to play a role in zebrafish death. Those results showed that HOE led to a range of dramatic ecosystem impacts which were more destructive than those of other treatments. Moreover, compared with the continuous dosing regimen, despite the fact that higher toxicity was observed for HOE, there was little difference in the removal of total silver from the aquatic environment for the different dosing regimens. No obvious differences in ecological impacts were observed between different water columns under low concentration exposure. Overall, this work highlighted the fact that the toxicity of AgNPs was impacted by different dosing regimens in different exposure media, which may be helpful for assessments of ecological impacts on aquatic environments.     

纳米银在环境水样和实验介质中的理化性质和毒性效应

纳米银在自然环境中的理化性质可能严重影响其环境效应,本文的目的是为了比较纳米银在自然水样和实验介质条件下的理化性质和毒性效应.实验表征了粒径4 nm表面包裹物为油胺的纳米银,在不同介质条件下的粒径和团聚性,并进一步研究了不同环境介质条件下纳米银暴露对枯草芽孢杆菌的毒性效应.透射电镜照片显示油胺包裹的4 nm纳米银在环境介质中发生明显团聚.油胺包裹的4 nm纳米银暴露会抑制培养基中细菌的生长速率,降低生理盐水溶液中细菌的存活率.纳米银在实验条件下对细菌的毒性呈一定剂量效应关系,但是在环境水样中剂量效应关系不明显.低浓度纳米银在环境水样中的毒性比其在实验介质中弱.纳米银的透析袋暴露实验中,细菌的存活率有显著提升,揭示纳米银与细菌的直接相互作用对毒性效应有重要作用.与实验条件的培养基和生理盐水介质相比,环境水样中的纳米银更趋向以团聚形式存在,与细菌的相互作用几率减小,因此纳米银在环境介质中对细菌的毒性会降低.     

Effects of nitrate concentration on biological hydrogen production by mixed cultures

The effects of nitrate on fermentative hydrogen production and soluble metabolites from mixed cultures were investigated by varying nitrate concentrations from 0 to 10 g N/L at 35°C with an initial pH of 7.0. The results showed that the substrate degradation rate, hydrogen production potential, hydrogen yield, and average hydrogen production rate initially increased with increasing nitrate concentrations from 0 to 0.1 g N/L, while they decreased with increasing nitrate concentrations from 0.1 to 10 g N/L. The maximum hydrogen production potential of 305.0 mL, maximum hydrogen yield of 313.1 mL/g glucose, and maximum average hydrogen production rate of 13.3 mL/h were obtained at a nitrate concentration of 0.1 g N/L. The soluble metabolites produced by the mixed cultures contained only ethanol and acetic acid (HAc) without propionic acid (HPr) and butyric acid (HBu). This study used the Modified Logistic model to describe the progress of cumulative hydrogen production in batch tests. A concise model was proposed to describe the effects of nitrate concentration on average hydrogen production rate.     

Thiolated arsenicals in arsenic metabolism: Occurrence, formation, and biological implications

Arsenic (As) is a notoriously toxic pollutant of health concern worldwide with potential risk of cancer induction, but meanwhile it is used as medicines for the treatment of different conditions including hematological cancers. Arsenic can undergo extensive metabolism in biological systems, and both toxicological and therapeutic effects of arsenic compounds are closely related to their metabolism. Recent studies have identified methylated thioarsenicals as a new class of arsenic metabolites in biological systems after exposure of inorganic and organic arsenicals, including arsenite, dimethylarsinic acid (DMAV), dimethylarsinous glutathione (DMAIIIGS), and arsenosugars. The increasing detection of thiolated arsenicals, including monomethylmonothioarsonic acid (MMMTAV), dimethylmonothioarsinic acid (DMMTAV) and its glutathione conjugate (DMMTAVGS), and dimethyldithioarsinic acid (DMDTAV) suggests that thioarsenicals may be important metabolites and play important roles in arsenic toxicity and therapeutic effects. Here we summarized the reported occurrence of thioarsenicals in biological systems, the possible formation pathways of thioarsenicals, and their toxicity, and discussed the biological implications of thioarsenicals on arsenic metabolism, toxicity, and therapeutic effects.     

商品氯氰菊酯对斜生栅藻生长的影响及其生理生化效应

研究了商品氯氰菊酯对斜生栅藻(Scenedesmus obliquus)生长及生理生化指标的影响,以了解除虫菊酯农药对水生态系统的影响.在氯氰菊酯暴露下,藻细胞生长受到明显抑制,氯氰菊酯对斜生栅藻生长的72h半效应浓度(EC50)为2.37mg/L.藻细胞所有生理生化指标对氯氰菊酯响应迅速,12h到达最大促进或者抑制效果,48h后趋于平稳.其中12h处理,可溶性糖和可溶性蛋白含量上升,中等浓度组的促进作用最强;超氧化物歧化酶(SOD)活性则呈现出低浓度促进、高浓度抑制效应.氯氰菊酯能促进藻细胞膜脂氧化产物丙二醛(MDA)的产生,且呈现出明显的剂量—效应关系,MDA含量可以作为监测氯氰菊酯污染的生物标记指标之一.研究结果表明,SOD活性抑制以及膜脂氧化是氯氰菊酯对斜生栅藻的重要致毒机理,但自然环境中的氯氰菊酯水平不会对淡水水体浮游藻类的生长产生抑制作用.     

纳米银对小球藻光合作用和呼吸作用的影响

使用硼氢化钠还原硝酸银,聚乙烯醇(PVA)作为分散剂,制备出粒径为(7 ± 3)nm的纳米银,分别使用计数法和溶解氧法,研究了纳米银对小球藻(Chlorella vulgaris)生长、光合作用和呼吸作用的影响,并调查了对叶绿素a的抑制状况.结果显示,黑暗条件下加入3mg/L的纳米银,基本抑制了小球藻的呼吸作用,当暴露于4mg/L的纳米银时,小球藻生长的抑制率为93%;而光照条件下加入10mg/L纳米银时,才能抑制其光合作用,此时对小球藻生长的抑制率达到90%.光照时,叶绿素a在10mg/L纳米银的作用下,抑制率达到77%.研究表明了纳米银对小球藻呼吸作用有很强的抑制作用,对光合作用的影响可能通过抑制叶绿素a的合成或破坏叶绿体的结构来完成;光照能够明显减弱纳米银的毒性.     

Influence of stabilizers on the antimicrobial properties of silver nanoparticles introduced into natural water

Physical, chemical and biochemical properties of silver nanoparticles(AgNPs) depend to a great extent on their size, shape, size distribution, and stabilizers located on their surface. This study focused on two typical stabilizers, namely citrates(cit), low molecular ions protecting nanoparticles by electrostatic repulsion, and polyvinylpyrrolidone(PVP), a hydrophilic, neutral, high molecular polymer protecting nanoparticles by steric stabilization. Natural bacterioplankton was collected from a eutrophic, downtown lake and exposed to five concentrations(0.1–5 mg/L) of AgNPs-PVP and AgNPs-cit. Responses were monitored after 1, 3, 5 and 7 days of exposure, by evaluating the survival rate of bacteria, their respiratory activity, and the general activity of extracellular esterases. A significantly better(greater) survival rate of bacterioplankton was observed in water with an addition of AgNPs-cit. The inhibition of extracellular esterases was observed only in samples containing AgNPs-PVP. The inhibitory effect increased proportionally to the concentration of AgNPs-PVP applied. Within the studied concentration range, there was no statistically significant inhibition of bacterioplankton respiratory activity by AgNPs-PVP and AgNPs-cit.     

Single and joint effects of petroleum hydrocarbons and cadmium on the polychaete Perinereis aibuhitensis Grube

Using the concentration gradient and combined pollutant exposure method, the single and joint effects of petroleum hydrocarbons （PHCs） and cadmium （Cd） on polychaete Perinereis aibuhitensis Grube, an ecologically keystone species in estuarine and coastal environment, have been investigated. The results indicate that the toxicity of PHCs to P. aibuhitensis is stronger than that of Cd to the organism. There are positive correlations between the mortality of worms and the exposed concentration of single Cd or PHCs in solution. Similarly, the accumulation of Cd or PHCs in worms increased with increasing Cd- or PHC-exposed concentrations. All the correlation relationships can be described using unitary quadratic equations （Y or Z = aX^2 ＋ bX ＋ c）. It is calculated, on the basis of these expressions, that the median lethal dose （LC50） ofP aibuhitensis exposed to a single Cd or PHCs is 793.4-13567.3 and 28.0-119.9 μg/L, respectively. The exposed time has some stimulative effect on the two pollutants and on the mortality of the worms. Thus, even a low concentration of a single Cd or PHCs may have strong toxic effects on the worms when the exposed time becomes longer. The accumulation of Cd or PHCs in worms differs with an increase in exposure time at the given exposed concentration of a single Cd or PHCs. Noticeably, the accumulation of PHCs in worms decreases with an increase in exposure time at the given high concentration of PHCs in solution. The joint effect of PHCs and Cd on P. aibuhitensis is very complicated and changes with the exposed concentrations of the two pollutants. At the given concentration of PHCs, the joint toxicity of the two pollutants on the worms changes from synergism to antagonism with an increase in Cd concentration. The accumulation of Cd in the worms significantly decreases with the addition of PHCs to exposure solution.     

四环素类抗生素对淡水绿藻的毒性作用

四环素类抗生素是目前世界上应用最为广泛的抗生素之一,也是我国畜禽养殖业中使用量最大的兽药和饲料添加剂.由于该类物质不易被生物体吸收且当前的污水处理设施对抗生素不能彻底去除,故大量抗生素及其代谢产物最终进入地表水环境,其对水生态系统和人体健康的潜在威胁值得关注.本研究以水生态系统初级生产者淡水绿藻为受试生物,系统考察四环素、金霉素和强力霉素对蛋白核小球藻、斜生栅藻细胞膜通透性和生长抑制作用的影响.结果表明,抗生素暴露96 h后,强力霉素在整个作用浓度范围内使供试淡水绿藻细胞膜通透性降低;而金霉素和四环素低浓度暴露可以增大藻细胞膜通透性,此后随着作用浓度的增大转为降低.3种抗生素对淡水绿藻生长抑制率大致呈现强力霉素>四环素>金霉素的顺序趋势.斜生栅藻对四环素类抗生素的毒性响应较蛋白核小球藻更为敏感.     

不同类型城市人工湿地水体汞的分布特征

为了解城市人工湿地水体中汞的时空分布及甲基化特征,以便探究其潜在的汞暴露生态风险,在重庆市选择4个不同类型的人工湿地,于2017年3月～2018年3月,按季度采集垂直剖面水样,分析总汞(THg)和甲基汞(MeHg)的分布.结果表明:城市人工湿地由于景观布局、功能设置不同,其汞分布特征有一定差异. 4个湿地水体THg均高于背景水域,但远低于有污染历史的水体;湿地中部水体THg均略高于进、出水口,表明城市湿地对水体THg有截留作用.除彩云湖湿地出水口MeHg浓度高于入水口外,其余3个湿地出水口MeHg均低于入水口;湿地中水体MeHg呈随水深增加而升高的趋势,且MeHg占THg的比例(MeHg/THg)均高于其他水域,说明城市湿地具有净汞甲基化的作用. 4个城市湿地水体THg春秋两季高,夏季略有降低,冬季最低;水体MeHg冬季最低,而其他三季差异不大但远高于冬季,约为冬季的3倍.本研究明晰了城市湿地中汞的时空分布和甲基化规律,探究了人类活动对湿地的干扰程度和湿地的响应特征以及湿地汞对下游流域的影响,从控制潜在汞暴露风险的角度,为人工湿地建设提供建议.