Biomonitoring Atmospheric Heavy Metals with Lichens: Theory and Application

Recent records of environmental contamination noted a moderate decrease of SO2 pollution, whereas the burden of atmospheric heavy metals is still considerable. The present review refers to the entrapment, uptake, and accumulation of heavy metals by lichen thalli, made apparent by parameters of lichen vitality and stress. The particulate nature of airborne heavy metals is made evident by parameters referring to the entrapment of heavy-metal containing particles by lichen thalli. The mechanism of uptake of heavy metals, investigated by means of controlled experiments, refers to extracellular and intracellular uptake. The rate of absorption and the accumulation of heavy metals is dependent on morphological features of lichen thalli in addition to kind and intensity of emission sources and to nonanthropogenic factors such as climate and topography. The role of lichens as biomonitors is demonstrated by the case of lead. In contrast to data obtained by retrospective studies, using lichens as biomonitors of heavy metal pollution, performed in the 1970s, which indicated an increase of Pb as a result of the massive use of leaded gasoline, the subsequent disuse of this additive led to a decrease detected in later studies. The disparity of emission sources is illustrated by the case of Hg. Mercury is a product of anthropogenic activity in addition to its natural derivation. The dominance of the anthropogenic factors is made obvious by high levels of Hg recorded near chlor alkali plants and other industrial sites. The role of the substrate in the uptake and accumulation of heavy metals was investigated to detect its relative share. Airborne metals were, however, determinant factors in the metal content of lichen thalli. The interaction of contaminants and biomonitors has a definite physiological impact on the vitality of the biomonitors. Physiological processes of disintegration investigated in this context are degradation of cell membranes and chlorophyll, decrease of the quantum yield of photosynthesis, decrease of the photosynthetic rate, increase of stress-ethylene production, and severe ultrastructural change. Lichens exposed to heavy metal pollution exhibit changes of the spectral reflectance response, an increase of malondialdehyde (MDA), a decrease of ATP, and injury to enzymatic activities. A comparative analysis of data referring to the accumulation of airborne elements in lichens and of data referring to alterations in physiological parameters of lichen viability substantiates the validity of assessments of environmental quality.     

地衣对重金属的累积效应用于大气监测的初步研究

以采自不同污染区的3个种共18个地衣样本作为试验材料,利用原子吸收分光光度计,测定样品中铅、镉的含量,评估环境质量。结果表明:铅是其中两个样点中的重要沉积物,在各地衣样品中有不同程度的积累,离子积累的浓度与污染源的相对位置具有直接的关系。根据累积浓度不同,可以推测当地的空气质量。     

Susceptibility of Halobacteria to Heavy Metals

Sixty-eight halobacteria, including both culture collection strains and fresh isolates from widely differing geographical areas, were tested for susceptibility to arsenate, cadmium, chromium, cobalt, copper, lead, mercury, nickel, silver, and zinc ions by an agar dilution technique. The culture collection strains showed different susceptibilities, clustering into five groups. Halobacterium mediterranei and Halobacterium volcanii were the most metal tolerant, whereas Haloarcula californiae and Haloarcula sinaiiensis had the highest susceptibilities of the culture collection strains. Different patterns of metal susceptibility were found for all the halobacteria tested, and there was a uniform susceptibility to mercury and silver. All strains tested were multiply metal tolerant.     

Acute Toxicity,Respiratory Reaction,and Sensitivity of Three Cyprinid Fish Species Caused by Exposure to Four Heavy Metals

Using 3 cyprinid fish species zebra fish, rare minnow, and juvenile grass carp, we conducted assays of lethal reaction and ventilatory response to analyze sensitivity of the fish to 4 heavy metals. Our results showed that the 96 h LC₅₀ of Hg²⁺ to zebra fish, juvenile grass carp, and rare minnow were 0.14 mg L⁻¹, 0.23 mg L⁻¹, and 0.10 mg L⁻¹, respectively; of Cu²⁺0.17 mg L⁻¹, 0.09 mg L⁻¹, and 0.12 mg L⁻¹ respectively; of Cd²⁺6.5 mg L⁻¹, 18.47 mg L⁻¹, 5.36 mg L⁻¹, respectively; and of Zn²⁺44.48 mg L⁻¹, 31.37 mg L⁻¹, and 12.74 mg L⁻¹, respectively. Under a 1-h exposure, the ventilatory response to the different heavy metals varied. Ventilatory frequency (Vf) and amplitude (Va) increased in zebra fish, juvenile grass carp, and rare minnows exposed to Hg²⁺ and Cu²⁺ (P<0.05), and the Vf and Va of the 3 species rose initially and then declined when exposed to Cd²⁺. Zn²⁺ had markedly different toxic effects than the other heavy metals, whose Vf and Va gradually decreased with increasing exposure concentration (P<0.05). The rare minnow was the most highly susceptible of the 3 fish species to the heavy metals, with threshold effect concentrations (TEC) of 0.019 mg L⁻¹, 0.046 mg L⁻¹, 2.142 mg L⁻¹, and 0.633 mg L⁻¹ for Hg²⁺, Cu²⁺, Cd²⁺, and Zn²⁺, respectively. Therefore, it is feasible to use ventilatory parameters as a biomarker for evaluating the pollution toxicity of metals and to recognize early warning signs by using rare minnows as a sensor.     

Heavy Metals Bioremediation of Soil

Historical emissions of old nonferrous factories lead to large geographical areas of metals-contaminated sites. At least 50 sites in Europe are contaminated with metals like Zn, Cd, Cu, and Pb. Several methods, based on granular differentiation, were developed to reduce the metals content. However, the obtained cleaned soil is just sand. Methods based on chemical leaching or extraction or on electrochemistry do release a soil without any salts and with an increased bioavailability of the remaining metals content. In this review a method is presented for the treatment of sandy soil contaminated with heavy metals. The system is based on the metal solubilization on biocyrstallization capacity of Alcaligenes eutrophus CH34. The bacterium can solubilize the metals (or increase their bioavailability) via the production of siderophores and adsorb the metals in their biomass on metal-induced outer membrane proteins and by bioprecipitation. After the addition of CH34 to a soil slurry, the metals move toward the biomass. As the bacterium tends to float quite easily, the biomass is separated from the water via a flocculation process. The Cd concentration in sandy soils could be reduced from 21 mg Cd/kg to 3.3 mg Cd/kg. At the same time, Zn was reduced from 1070 mg Zn/kg to 172 mg Zn/kg. The lead concentration went down from 459 mg Pb/kg to 74 mg Pb/kg. With the aid of biosensors, a complete decrease in bioavailability of the metals was measured.     

Relative Sensitivity Distribution of Freshwater Planktonic Crustaceans to Trace Metals

Cladocerans and copepods are common inhabitants of ponds and lakes, but assessments of the toxic effects of chemicals on aquatic organisms are often based mainly on toxicity data for cladocerans of the genus Daphnia. The paucity of toxicity data for copepods and other species of cladocerans raised the question whether toxicity data derived primarily from daphniids can be extrapolated to protect other planktonic organisms. In this study, we address this question by systematically comparing between the sensitivities of cladocerans and copepods to the metals Cd, Cu, Pb, Ni, and Zn using the species sensitivity distributions. Our results indicate that cladocerans are consistently more sensitive than copepods to Cd and Cu. Most importantly, sensitivities of D. magna, D. pulex, and Ceriodaphnia dubia, the three most commonly tested cladocerans, to these metals are higher than those of most copepod species and fall within the range of those exhibited by other cladocerans. C. dubia, a cladoceran with relatively small body size, is more sensitive to all metals than most other cladoceran species. Our results imply that water quality criteria of trace metals derived from datasets dominated by Daphnia are likely also protective to copepods and most other cladocerans.     

几株细菌的重金属抗性水平和吸附量

从堆积时间为80～100a的铅锌矿渣中分离了6株细菌,通过测定部分16S rRNA基因序列确定了它们的系统发育地位。结果表明有3株细菌属于节杆菌属(Arthrobacter),同A．nicotinovorans和A．histidinolovorans两个种关系密切。另外3株属于壤霉菌属(Agromyces),同Ag．mediolanus具有较近的亲缘关系。总体来看,这些菌株都对检测的5种重金属有高的最低抑菌浓度(minimal inhibitory concentration,MIC)。节杆菌对Zn、Co的耐受明显高于壤霉菌。此外,这些重金属高抗性菌株也对重金属有较强的吸附能力。在环境中有单一重金属离子的情况下,冻干的节杆菌对Ph的吸附率平均达到了约400mg／g干菌体,对Cd和zn的吸附也分别达到了近177和80mg/g干菌体,具有进一步开发为重金属吸附剂的潜力。     

Role of Organic Acids in Sunflower Tolerance to Heavy Metals

Exposure of Helianthus annuus L. seedlings to Al³⁺, Cd²⁺ or Zn²⁺ resulted in a marked decrease of fresh and dry masses of the shoots and the roots. The increase of Al³⁺, Cd²⁺ or Zn²⁺ uptake was accompanied by a significant decrease of nitrate, phosphorus and K⁺ uptake. There was a significant increase of malic and citric acid contents in the shoots and roots of heavy metal-treated seedlings whereas the change in fumaric acid was insignificant. Al³⁺ and Zn²⁺ alone stimulated excretion of malic and citric acids to the rhizosphere. Addition of high concentrations of malic or citric acid alleviate to some extent the inhibitory effect of Al³⁺ and Zn²⁺ on plant growth. This revised version was published online in July 2006 with corrections to the Cover Date.     

Differential Sensitivity of Children and Adults to Chemical Toxicity

Children, particularly neonates, can be biologically more sensitive to the same toxicant on a body weight basis than adults. Current understanding of the rates of maturation of metabolism and evidence from case studies indicate that human infants up to 6 months of age typically lack the capacity to detoxify and eliminate substances as readily as adults. For most chemicals, the infant physiologic systems usually produce higher blood levels for longer periods. The newborn's metabolic capacity rapidly matures and, by 6 months of age, children are usually not more sensitive than adults based on their pharmacokinetic competence. Whether children are at greater risk from chemical exposures is another question. Drawing conclusions about the ability of the U.S. Environmental Protection Agency's intraspecies (UF_H) and database (UF_D) uncertainty factors to protect children on the basis of the modest data available is challenging. However, virtually all studies available suggest that a high percentage of the population, including children, is protected by using a 10-fold UF_H or by using a 3.16-fold factor each for toxicokinetic and toxicodynamic variability. Based on specific comparisons for newborns, infants, children, adults and those with severe disease, the population protected is between 60% and 100%, with the studies in larger populations that include sensitive individuals suggesting that the value is closer to 100%. UF_D is likewise protective when used with databases that are missing substantive studies.     

Waste Water Bacterial Isolates Resistant to Heavy Metals and Antibiotics

Sewage water of Casablanca, an industrial city in Morocco, was studied for microorganisms resistant to heavy metals. Isolates were purified and collected on agar slants to be screened for resistance to heavy metals, including mercury in vitro. The strains that showed high resistance to heavy metals were also studied for their resistance to antibiotics and aromatic hydrocarbons. Results indicated that the strains most resistant to all tested products belonged to Ps. fluorescens, Ps. aeruginosa, Klebsiella pneumoniae, Proteus mirabilis, and Staphylococcus sp. These strains exhibit high minimal inhibitory concentrations for heavy metals such as cadmium (2 mm) or mercury (1.2 mm). Growth of Ps. fluorescens and Klebsiella pneumoniae in the presence of heavy metals was also determined, and the growth curves indicated that mercury, copper, and zinc present a slight inhibitory action, while cadmium and silver could have a potent inhibitory action on growth compared with the controls. These studies also investigated growth in media containing aromatic compounds as the sole source of carbon. The results demonstrate that these strains could be good candidates for remediation of some heavy metals and aromatic compounds in heavily polluted sites. Received: 23 December 1999 / Accepted: 6 April 2000     

植物响应金属胁迫重要蛋白质的细胞定位

土壤中的Hg2+、Pb2+、Cd2+、Cu2+、Zn2+、Mn2+、Fe2+/Fe3+等金属离子对植物体具有毒害作用。植物体通过定位于细胞壁、细胞膜,以及各种细胞器中的蛋白质应答重金属胁迫过程。植物体利用细胞壁蛋白质与重金属离子络合,细胞膜系统的转运蛋白与通道调节金属离子运输、细胞器或细胞质基质中的分子伴侣帮助蛋白质折叠等机制完成抑制金属离子进入体内或在体内的解毒等过程。综述了近年来植物响应金属离子胁迫重要蛋白质的细胞定位与作用机制。     

‘新选七叶’油菜对土壤3种重金属的吸收和累积特征

以‘新选七叶’油菜为研究对象,采用土培方法,研究在不同浓度重金属(镉、铅、汞)胁迫下,油菜对重金属的吸收和积累特征,并探讨重金属在土壤—蔬菜间的迁移情况。结果表明:不同重金属在植物不同部位的分布和积累规律存在差异,镉和汞在植株上的积累规律为根叶茎,而铅在植株上的积累规律为根茎叶。油菜根部对重金属的累积能力远高于茎和叶,根部对三种重金属的吸收富集能力为镉汞铅。油菜各部位重金属含量与土壤重金属浓度呈显著正相关(r20.859)。以蔬菜可食部位重金属含量的最大限量为标准(GB 15201-1994、GB 14935-1994、GB 2762-1994),计算得出土壤中对应的镉、铅、汞临界值分别为0.362、43.62、2.892μg/g。     

Sublethal Toxicity Endpoints of Heavy Metals to the Nematode Caenorhabditis elegans

Caenorhabditis elegans, a free-living nematode, is commonly used as a model organism in ecotoxicological studies. The current literatures have provided useful insight into the relative sensitivity of several endpoints, but few direct comparisons of multiple endpoints under a common set of experimental conditions. The objective of this study was to determine appropriate sublethal endpoints to develop an ecotoxicity screening and monitoring system. C. elegans was applied to explore the sublethal toxicity of four heavy metals (copper, zinc, cadmium and chromium). Two physiological endpoints (growth and reproduction), three behavioral endpoints (head thrash frequency, body bend frequency and feeding) and two enzymatic endpoints (acetylcholine esterase [AChE] and superoxide dismutase [SOD]) were selected for the assessment of heavy metal toxicity. The squared correlation coefficients (R²) between the responses observed and fitted by Logit function were higher than 0.90 and the RMSE were lower than 0.10, indicating a good significance statistically. There was no significant difference among the half effect concentration (EC50) endpoints in physiological and behavioral effects of the four heavy metals, indicating similar sensitivity of physiological and behavioral effects. AChE enzyme was more sensitive to copper, zinc, and cadmium than to other physiological and behavioral effects, and SOD enzyme was most sensitive to chromium. The EC50 of copper, zinc, and cadmium, to the AChE enzyme in the nematodes were 0.68 mg/L, 2.76 mg/L, and 0.92 mg/L respectively and the EC50 of chromium to the SOD enzyme in the nematode was 1.58 mg/L. The results of this study showed that there was a good concentration-response relationship between all four heavy metals and the sublethal toxicity effects to C. elegans. Considering these sublethal endpoints in terms of simplicity, accuracy, repeatability and costs of the experiments, feeding is the relatively ideal sublethal toxicity endpoint of heavy metals to C. elegans.     

Bioaccumulation of Heavy Metals by Green Algae

The biosorption of metal ions (Cr⁺³, , Cu⁺², and Ni⁺²) on two algal blooms (designated HD-103 and HD-104) collected locally was investigated as a function of the initial metal ion concentration. The main constituent of HD-103 is Cladophora sp., while Spirulina sp. is present significantly in the bloom HD-104. Algal biomass HD-103 exhibited the highest Cu⁺² uptake capacity (819 mg/g). This bloom adsorbed Ni⁺² (504 mg/g), Cr⁺³ (347 mg/g), and (168 mg/g). Maximum of Ni⁺² (1108 mg/g) is taken by HD-104. This species takes up 306, 202, and 576 mg/g Cr⁺³, , and Cu⁺², respectively. Equilibrium data fit very well to both the Langmuir and the Freundlich isotherm models. The sorption process followed the Freundlich model better. Pseudo-first-order kinetic model could describe the kinetic data. Infrared (IR) spectroscopic data were employed to identify the site(s) of bonding. It was found that phosphate and peptide moieties participate in the metal uptake by bloom HD-103. In the case of bloom HD-104, carboxylate and phosphate are responsible for the metal uptake. The role of protein in metal uptake by HD-103 was investigated using polyacrylamide gel electrophoresis.     

Biosorption of Heavy Metals by Marine Algae

The ability of four different algae (three brown and one red) that have not been previously studied to adsorb Cr³⁺, Co²⁺, Ni²⁺, Cu²⁺, and Cd²⁺ ions was investigated. The metal uptake was dependent on the type of biosorbent, with different accumulation affinities towards the tested elements. The HCl-treated biomass decreased the metal biosorptive capacity particularly in the case of Cr³ adsorption with Laurencia obtusa. The extent of uptake of the different metals with the tested algae was assessed under different conditions such as pH, time of algal residence in solution with the metal, and concentration of algal biomass. The rate of uptake of the different metals was very fast in the first 2 h; thereafter the increase in metal uptake was insignificant. The amount of the metal uptake (5–15 mg range) increased steeply by increasing the weight of the biomass. An exception was L. obtusa, where a parallel increase of the uptake of different metals was observed on increasing the algal mass from 5 to 50 mg. Received: 21 December 1999 / Accepted: 24 April 2000     

早熟禾对4种重金属胁迫生长响应特征

采用砂培法研究了早熟禾对不同浓度Cu2 、Zn2 、Cd2 和Pb2 胁迫的生长响应特征.结果表明,当Cu2 、Zn2 、Cd2 的浓度达100 m g/L时,早熟禾的种子萌发率、幼苗株高均有所下降,并随着胁迫浓度的增加,下降幅度增大;Pb2 对两指标的影响不太明显.Cu2 对根系及地上生物量具有抑制作用,尤其对根系生长抑制效应极为明显,当浓度增加到600 m g/L时,与对照相比,根长最高下降了96.67%.Zn2 、Cd2 和Pb2 在浓度超过200 m g/L时对早熟禾根系及地上生物量表现出抑制效应,并随着浓度的增加抑制效应增加.4种重金属对叶绿素的影响表现出一致的规律,即当浓度小于200 m g/L时对叶绿素的合成起促进作用,当浓度超过200 m g/L时,叶绿素的含量又随着处理浓度的增加而下降.     

Differential Sensitivity of Mouse Hematopoietic Stem Cells to Merocyanine 540

In vivo and in vitro clonal assays of immature mouse blood cells showed that diffferent populations of hematopoietic progenitor cells differ considerably with respect to their sensitivity to photodynamic damages caused by the fluorescent dye Merocyanine 540. Late erythroid progenitors were the most sensitive cells followed in order of decreasing sensitivity by pluripotent stem cells, early erythroid progenitors, and granulocyte/macrophage progenitors. Only about 2%–4% of all nucleated marrow cells were stained with Merocyanine 540 which correlated well with current frequency estimates of progenitor cells in mouse bone marrow. Our findings indicate that the expression of Merocyanine binding sites is developmentally regulated and might, therefore, provide a useful molecular marker for blood cell differentiation and a basis for an effective purification of hematopoietic progenitor cells.     

Differential Sensitivity of Prefrontal Cortex and Hippocampus to Alcohol-Induced Toxicity

The prefrontal cortex (PFC) is a brain region responsible for executive functions including working memory, impulse control and decision making. The loss of these functions may ultimately lead to addiction. Using histological analysis combined with stereological technique, we demonstrated that the PFC is more vulnerable to chronic alcohol-induced oxidative stress and neuronal cell death than the hippocampus. This increased vulnerability is evidenced by elevated oxidative stress-induced DNA damage and enhanced expression of apoptotic markers in PFC neurons. We also found that one-carbon metabolism (OCM) impairment plays a significant role in alcohol toxicity to the PFC seen from the difference in the effects of acute and chronic alcohol exposure on DNA repair and from exaggeration of the damaging effects upon additional OCM impairment in mice deficient in a key OCM enzyme, methylenetetrahydrofolate reductase (MTHFR). Given that damage to the PFC leads to loss of executive function and addiction, our study may shed light on the mechanism of alcohol addiction.     

睫毛黑蜈蚣衣重金属的吸收及富集能力的研究

目的:评价睫毛黑蜈蚣衣重金属的吸收及富集能力及其在空气环境质量评价中的应用价值。方法:利用地衣移植法把睫毛黑蜈蚣衣移植到乌鲁木齐市区7个监测点,用电感耦合等离子光谱直读法检测睫毛黑蜈蚣衣体内8种重金属含量。结果:结果显示,睫毛黑蜈蚣衣重金属的吸收及富集能力较强,体内重金属含量具有显著的时空(ANOVA:F=17.42,P<0.05;F=32.12,P<0.05)变化特点。结论:该研究进一步证实睫毛黑蜈蚣衣具有较高的重金属富集能力,对大气污染较敏感,可以作为大气环境质量评价指示地衣种类。