Effect of complexation on the accumulation and elimination kinetics of cadmium and ciprofloxacin in the earthworm Eisenia fetida

Land application of solid wastes leads to the accumulation of both metals and antimicrobials in soils. To understand the effects of metal and antibiotic interaction on their accumulation by the earthworm Eisenia fetida, uptake and elimination kinetics and subcellular distribution of cadmium (Cd) and ciprofloxacin (CIP) were determined. The kinetics was accurately described by a one-compartment first-order kinetic model. Bioaccumulation kinetics and subcellular distribution of CIP were not affected by Cd addition. However, Cd exhibited different metabolic and subcellular distribution patterns. With CIP, Cd uptake flux and elimination rate constants were about 2.2 and 9.8 times, respectively, those without CIP. In the presence of CIP, Cd redistributed from fractions D (associated with granules) and E (associated with tissue fragments and cell membranes) to fraction C (associated with cytosol). Without CIP, Cd in fraction C could not be excreted, whereas with CIP, Cd in fraction C was significantly excreted, and the excretion rate constant was consistent with that of CIP. A good relationship was found between CIP and Cd in earthworms during uptake and elimination periods (p < 0.01). Our results indicated that the Cd-CIP complex may be taken up, stored, and eliminated by earthworms.     

砷及砷的形态检测方法的研究现状

砷是一种广泛分布于自然界的微量元素,摄入过量砷元素容易引起砷的急性中毒而致死,应引起人们足够的重视。砷的毒性主要取决于它的化学形态,形态不同毒性差异很大,所以选择适当、简便、快速的测定方法是现代分析技术取得发展的必要条件。本文综述了近年来砷及其形态常用的测定方法,并且分析了各检测方法的优缺点,展望了砷及其形态检测技术的发展前景。      

Surface complexation of ferrous iron and carbonate on ferrihydrite and the mobilization of arsenic

Surface complexation models are commonly used to predict the mobility of trace metals in aquifers. For arsenic in groundwater, surface complexation models cannot be used because the database is incomplete. Both carbonate and ferrous iron are often present at a high concentration in groundwater and will influence the sorption of arsenic, but the surface complexation constants are absent in the database of Dzombak and Morel. This paper presents the surface complexation constants for carbonate and ferrous iron on ferrihydrite as derived for the double-layer model. For ferrous iron the constants were obtained from published data supplemented by new experiments to determine the sorption on the strong sites of ferrihydrite. For carbonate the constants were derived from experiments by Zachara et al., who employed relatively low concentrations of carbonate. The double-layer model, optimized for low concentrations, was tested against sorption experiments of carbonate on goethite at higher concentration by Villalobos and Leckie, and reasonable agreement was found. Sorption was also estimated using linear free energy relations (LFER), and results compared well with our derived constants. Model calculations confirm that sorption of particularly carbonate at common soil and groundwater concentrations reduces the sorption capacity of arsenic on ferrihydrite significantly. The displacing effect of carbonate on sorbed arsenate and arsenite has been overlooked in many studies. It may be an important cause for the high concentrations of arsenic in groundwater in Bangladesh. Sediments containing high amounts of sorbed arsenic are deposited in surface water with low carbonate concentrations. Subsequently the sediments become exposed to groundwater with a high dissolved carbonate content, and arsenic is mobilized by displacement from the sediment surface.     

Spectrophotometric estimation of arsenic in nitric acid extracts of soil and soil additives

The spectrophotometric determination of arsenic in solutions containing nitric acid necessitates the removal of nitrate ions without loss of arsenic. A convenient and effective method for its removal was achieved by treatment with formic acid.     

Metal compartmentation and speciation in a soil sentinel: the earthworm, Dendrodrilus rubidus

Earthworms are well-studied organisms in ecotoxicology because of their keystone ecological status and metal-accumulating capacity. However, the direct estimation of the bioreactive fractions of accumulated metal burdens remains technically elusive. In this study we exploited two physical techniques, electron probe X-ray microanalysis (EPXMA) and X-ray absorption spectroscopy (XAS), to improve understanding of the subcellular spatial distributions, ligand affinities, and coordination chemistries of Cd, Pb and Zn in a field population of the epigeic earthworm, Dendrodrilus rubidus. EPXMA and XAS analyses were performed on cryopreparations to maintain compositional fidelity; EPXMA data were analyzed by multivariate statistics. XAS provided whole-worm insights; EPXMA provided in situ, subcellular data from the major metal-sequestering tissue, the chloragog. Both techniques showed that Cd is coordinated with S; the measured Cd-S bond distance in XAS suggests a metallothionein-type ligand. The mean Cd:S molar ratio (EPXMA) of 0.36 is higher than the ratio of 0.29 estimated from published biochemical data. EPXMA and XAS data also found that Ca, Pb, and Zn are predominantly bound to one or more O-donating, probably phosphate-rich, ligands. X-ray distribution maps (EPXMA) of the hepatocyte-resembling chloragocytes revealed that the O-seeking (Ca, Pb, Zn) metals and S-seeking Cd bioaccumulate in distinct organelles. Extended X-ray absorption fine structure showed that the Pb complex is not biogenic pyromorphite, although X-ray absorption near edge structure did not eliminate the possibility. XAS provided no evidence of Pb spillage from the "sequestration compartment" within D. rubidus. However, the correspondence of Pb with Ca and P in EPXMA is not as strong as that of Zn. This is indicative either of spillover or of a second, hitherto unidentified, sequestered-Pb pool. By exploiting the complimentary techniques of EPXMA and XAS,we are closer to describing the mechanistic link between equilibrated body burdens and biomarker responses in earthworms.     

Biotransformation and accumulation of arsenic in soil amended with seaweed

For many coastal regions of the world, it has been common practice to apply seaweed to the land as a soil improver and fertilizer. Seaweed is rich in arsenosugars and has a tissue concentration of arsenic up to 100 micro/g g(-1). These arsenic species are relatively nontoxic to humans; however, in the environment they may accumulate in the soil and decompose to more toxic arsenic species. The aim of this study was to determine the fate and biotransformation of these arsenosugars in soil using HPLC-ICP-MS analysis. Data from coastal soils currently manured with seaweeds were used to investigate if arsenic was accumulating in these soils. Long-term application of seaweed increased arsenic concentrations in these soils up to 10-fold (0.35 mg of As kg(-1) for nonagronomic peat, 4.3 mg of As kg(-1) for seaweed-amended peat). The biotransformation of arsenic was studied in microcosm experiments in which a sandy (machair) soil, traditionally manured with seaweed, was amended with Laminaria digitata and Fucus vesiculosus. In both seaweed species, the arsenic occurs in the form of arsenosugars (85%). The application of 50 g of seaweed to 1 kg of soil leads to an increase of arsenic in the soils, and the dominating species found in the soil pore water were dimethylarsinic acid (DMA(V)) and the inorganic species arsenate (As(V)) and arsenite (As(III)) after the initial appearance of arsenosugars. A proposed decomposition pathway of arsenosugars is discussed in which the arsenosugars are transformed to DMA(V) and further to inorganic arsenic without appreciable amounts of methylarsonic acid (MA(V)). Commercially available seaweed-based fertilizers contain arsenic concentration between 10 and 50 mg kg(-1). The arsenic species in these fertilizers depends on the manufacturing procedure. Some contain mainly arsenosugars while others contain mainly DMA(V) and inorganic arsenic. With the application rates suggested by the manufacturers, the application of these fertilizers is 2 orders of magnitude lower than the maximum permissible sewage sludge load for arsenic (varies from 0.025 kg ha(-1) yr(-1) in Styria, Austria, to 0.7 kg ha(-1) yr(-1) in the U.K.), while a direct seaweed application would exceed the maximum arsenic load by at least a factor of 2.     

An exchange method to investigate the kinetics of Cd complexation in soil solutions

Knowledge of the dynamic speciation of metals in soil solution is necessary for soil-plant transfer models. The lability of a metal in a given context partly depends on the kinetics of interconversion between the free ion (M) and one complex (ML), governed by the association (c(k)a) and dissociation (c(k)d) rate constants of the reaction of formation of ML If only one single complex is assumed to be present in solution, a single kinetic experiment will enable estimation of ckd and cka', that is, the product of cka by the concentration of free ligand in solution. The method consists of the measurements over time of the radioactive metal remaining in solution after the labeling of a Ca resin--solution system in balanceforthe nonlabeled metal initially present. The method is described for Cd and was applied on four soil solution extracts. The order of magnitude of the estimated c(k)a'- and c(k)d-values was about 10(-3) s(-1). The advantages and drawbacks of the method are discussed.     

糙米中砷形态检测方法的对比研究

目的比较液相色谱-原子荧光光谱法(highperformance liquid chromatography atomic fluorescence spectrometry,HPLC-AFS)和液相色谱-电感耦合等离子体质谱法(high performance liquid chromatography inductively coupled plasma mass spectrometry, HPLC-ICP-MS)两种方法对糙米中砷形态的测定。方法糙米样品经0.15 mol/L硝酸溶液提取,分别采用HPLC-AFS联用仪,流动相为15 mmol/L的磷酸二氢氨缓冲溶液(pH=6.0),经CNWSep AX阴离子交换色谱柱分离检测;同时,采用HPLC-ICP-MS联用仪,流动相为含5 mmol/L己烷磺酸钠、20 mmol/L柠檬酸的缓冲溶液(pH=4.3),经安捷伦ZorbaxSB-Aq反相色谱柱分离检测。结果两种方法检测4种砷形态在5.0～100.0 ng/mL范围内线性良好,相关系数均在0.999以上; HPLC-AFS法的检出限分别为2.3、0.6、1.0和1.1 ng/mL,加标回收率在85.1%～106%之间,相对标准偏差(relative standard deviation, RSD)均小于5%; HPLC-ICP-MS法的检出限分别为0.1、0.1、0.1和0.2 ng/mL,加标回收率在82.8%～106%之间,相对标准偏差(RSD)均小于5%。结论两种方法都可以满足糙米中4种砷形态的检测,在实际情况中可根据需要选择相应的检测方法。     

不同重金属胁迫蔬菜产生金属硫蛋白的同源性检测

目的 对不同重金属胁迫蔬菜产生金属硫蛋白(金属硫组氨酸三甲基内盐)进行同源性检测。方法 用Cd-MT免疫家兔产生IgG后, 用不同浓度硫酸铜、乙酸铅、氯化钴、氯化汞等重金属胁迫萝卜幼苗、油白菜和油麦菜诱导产生金属硫蛋白, 通过免疫印迹检测其同源性。结果 Cd-MT与重金属胁迫三种蔬菜所产生的金属硫蛋白具有同源性; 不同浓度的重金属对幼苗中MTs表达量有影响。结论 Cd-MT诱导家兔产生的IgG可用于农产品中重金属污染的检测。     

Bacterial populations associated with the oxidation and reduction of arsenic in an unsaturated soil

Microbial populations responsible for the oxidation and reduction of As were examined in unsaturated (aerobic) soil columns treated with 75 microM arsenite [As(III)] or 250 microM arsenate [As(V)]. Arsenite [As(III)] was rapidly oxidized to As(V) via microbial activity, whereas no apparent reduction of As(V) was observed in the column experiments. Eight aerobic heterotrophic bacteria with varying As redox phenotypes were isolated from the same columns. Three isolates, identified as Agrobacterium tumefaciens-, Pseudomonas fluorescens-, and Variovorax paradoxus-like organisms (based on 16S sequence), were As(III) oxidizers, and all were detected in community DNA fingerprints generated by PCR coupled with denaturing gradient gel electrophoresis. The five other isolates were identified (16S gene sequence) as A. tumefaciens, Flavobacterium sp., Microbacterium sp., and two Arthrobacter sp. -like organisms and were shown to rapidly reduce As(V) under aerobic conditions. Although the two A. tumefaciens-like isolates exhibited opposite As redox activity,their 16S rDNA sequences (approximately 1400 bp) were 100% identical, and both were shown to contain putative arsC genes. Our results support the hypothesis that bacteria capable of either oxidizing As(III) or reducing As(V) coexist and are ubiquitous in soil environments, suggesting that the relative abundance and metabolic activity of specific microbial populations plays an important role in the speciation of inorganic As in soil pore waters.     

核酸适配体技术对砷的检测研究进展

砷是目前环境中污染较为严重的几种重金属元素之一,天然存在于地壳中并可以通过食物链进入人体,基于砷的毒性,人们越来越重视对环境及食品中砷的检测。目前已经建立了多种检测环境及食品中砷含量的技术,包括原子吸收光谱法（atomic absorption spectrometry, AAS）、氢化物发生原子荧光光谱法（hydride generation atomic fluorescence spectrometry, HG-AFS）、电感耦合等离子体质谱法（inductively coupled plasma mass spectrometry, ICP-MS）以及核酸适配体技术等,其中核酸适配体技术是一种新型的快速检测技术,因其具有高特异性、高亲和力和高灵敏度而被应用于生物医学、环境分析与食品工程中,并在现场快速检测方面展现出广阔的应用前景。本文主要介绍了核酸适配体技术的原理及其应用,重点综述了基于核酸适配体技术的砷离子检测方法,并对核酸适配体技术自身的发展以及在食品等方面的应用进行了展望。     

Stable earthworm serine proteases: application of the protease function and usefulness of the earthworm autolysate

The fibrinolytic enzymes from Lumbricus rubellus [Nakajima, N. et al., Biosci. Biotechnol. Biochem., 57, 1726-1730 (1993), 60, 293-300 (1996), and 63, 2031-2033 (1999)] were further characterized to exploit their catalytic functions. These enzymes are stable in solution for long periods at room temperature and strongly resistant to organic solvents, even toluene and n-hexane. The serine proteases can act on various protein substrates such as elastin and hemoglobin as well as fibrin, and also catalyzed the hydrolysis of esters such as ethyl acetate and a bioplastic, poly[(R)-3-hydroxybutyrate] film. The enzymes, in the absence of microbial degradation, contributed to the production of the earthworm autolysate possessing antioxidant ability and protease activity, whose components were similar to those of soy sauce. The extract of the earthworm autolysate could be used as a peptone substitute in media for the cultivation of microorganisms.     

In vitro model improves the prediction of soil arsenic bioavailability: worst-case scenario

There is a strong interest in developing an in vitro arsenic (As) model that satisfactorily estimates the variability in in vivo relative oral bioavailability (RBA) measurements. Several in vitro tests have been developed, but none is universally accepted due to their limited success in predicting soil As RBA. A suite of amorphous and crystalline solid As phases were chosen, utilizing a worst-case scenario (WCS) that simulated fasting children's gastric solution chemistry. The objectives of this study were to (i) determine the effects of residence time, pH, and solid-to-solution ratio on As bioaccessibility and speciation in the in vitro gastric test; (ii) provide the fundamental basis for an optimized in vitro model constrained by the WCS; and (iii) validate the optimized in vitro test with the in vivo RBA obtained with BALB/c mice. The gastric pH was the only significant (p < 0.05) factor influencing solid As bioaccessibility. Bioaccessible As retained the oxidation state after its release from the solid into the gastric solution. The optimized in vitro model adequately predicted RBA values for a suite of solid As phases typically encountered in soils, with the exception of aluminum-based solids. This study is an excellent starting point for developing an in vitro test applicable to different As-contaminated soils.     

Surface complexation of copper(II) on soil particles: EPR and XAFS studies

The interactions of transition metals with natural systems play an important role in the mobility and the bioavailability of these metals in soils. In this study, the adsorption of copper(II) onto natural soil particles was studied as a function of pH and metal concentration. The retention capacity of soil particles was determined at pH 6.2 to be equal to 6.7 mg of copper/g of solid. The Langmuir and Freundlich isotherm equations were then used to describe the partitioning behavior of the system at different pH values. A combination of EPR, extended X-ray absorption fine structure (EXAFS), and X-ray absorption near-edge structure (XANES) spectroscopies was used to probe the Cu atomic environment at the soil particles/aqueous interface. The spectroscopic study revealed that copper(II) ions are held in inner-sphere surface complexes. It also revealed that Cu was in an octahedral coordination with first-shell oxygen atoms. A weak tetragonal distortion was pointed out due to the Jahn-Teller effect, with a mean Cu-Oequatorial bond distance of 1.96 A and a Cu-Oaxial bond distance of 2.06 A. A detailed analysis of the spectroscopic data suggested that Cu(II) was bonded to organic matter coated onto the mineral fraction of soil particles.     

Transport implications resulting from internal redistribution of arsenic and iron within constructed soil aggregates

Soils are an aggregate-based structured media that have a multitude of pore domains resulting in varying degrees of advective and diffusive solute and gas transport. Consequently, a spectrum of biogeochemical processes may function at the aggregate scale that collectively, and coupled with solute transport, determine element cycling in soils and sediments. To explore how the physical structure impacts biogeochemical processes influencing the fate and transport of As, we examined temporal changes in speciation and distribution of As and Fe within constructed aggregates through experimental measurement and reactive transport simulations. Spherical aggregates were made with As(V)-bearing ferrihydrite-coated sand inoculated with Shewanella sp. ANA-3; aerated solute flow around the aggregate was then induced. Despite the aerated aggregate exterior, where As(V) and ferrihydrite persist as the dominant species, anoxia develops within the aggregate interior. As a result, As and Fe redox gradients emerge, and the proportion of As(III) and magnetite increases toward the aggregate interior. Arsenic(III) and Fe(II) produced in the interior migrate toward the aggregated exterior and result in coaccumulation of As and Fe(III) proximal to preferential flow paths as a consequence of oxygenic precipitation. The oxidized rind of aggregates thus serves as a barrier to As release into advecting pore-water, but also leads to be a buildup of this hazardous element at preferential flow boundaries that could be released upon shifting geochemical conditions.     

Spatial distribution and temporal variability of arsenic in irrigated rice fields in Bangladesh. 2. Paddy soil

Arsenic-rich groundwater from shallow tube wells is widely used for the irrigation of boro rice in Bangladesh and West Bengal. In the long term this may lead to the accumulation of As in paddy soils and potentially have adverse effects on rice yield and quality. In the companion article in this issue, we have shown that As input into paddy fields with irrigation water is laterally heterogeneous. To assess the potential for As accumulation in soil, we investigated the lateral and vertical distribution of As in rice field soils near Sreenagar (Munshiganj, Bangladesh) and its changes over a 1 year cycle of irrigation and monsoon flooding. At the study site, 18 paddy fields are irrigated with water from a shallow tube well containing 397 +/- 7 microg L(-1) As. The analysis of soil samples collected before irrigation in December 2004 showed that soil As concentrations in paddy fields did not depend on the length of the irrigation channel between well and field inlet. Within individual fields, however, soil As contents decreased with increasing distance to the water inlet, leading to highly variable topsoil As contents (11-35 mg kg(-1), 0-10 cm). Soil As contents after irrigation (May 2005) showed that most As input occurred close to the water inlet and that most As was retained in the top few centimeters of soil. After monsoon flooding (December 2005), topsoil As contents were again close to levels measured before irrigation. Thus, As input during irrigation was at least partly counteracted by As mobilization during monsoon flooding. However, the persisting lateral As distribution suggests net arsenic accumulation over the past 15 years. More pronounced As accumulation may occur in regions with several rice crops per year, less intense monsoon flooding, or different irrigation schemes. The high lateral and vertical heterogeneity of soil As contents must be taken into account in future studies related to As accumulation in paddy soils and potential As transfer into rice.     

云南地区茶叶中总砷及无机砷含量调查分析

目的 为云南茶叶的开发利用提供数据支持.方法 利用国家标准方法对云南省110份茶叶样品的总砷和无机砷含量进行测试,并就产地、包装、流通环节等进行调查分析.结果 经对采集的110个茶叶样品进行总砷检测,最高0.152 mg/kg,均值0.0325 mg/kg,仅1件检出无机砷,检测值为0.0591 mg/kg,均符合国家...