Choice of chelating antidotes for acute cadmium intoxication∗

During acute oral intoxication by cadmium compounds, gastrointestinal epithelial damage contributes to immediate toxicity. However, secondary systemic toxicity may develop due to intestinal uptake of cadmium. This review presents an evaluation of the effects of chelators on the acute toxicity of cadmium after parenteral or oral exposure and on the intestinal uptake of cadmium. This review shows:

1. Chelating agents may affect the acute toxicity of cadmium in a variety of ways depending on the exposure route for cadmium and administration route for the chelator.

2. With regard to survival, systemic toxicity of absorbed cadmium is of major importance, as intraperitoneal administration of chelators could eliminate or reduce mortality due to orally administered cadmium chloride.

3. Lipophilicity of chelators and their cadmium complexes may result in extensively augmented intestinal uptake. However, hydrophilic chelators may efficiently reduce the intestinal cadmium uptake.

4. For hydrophilic chelators, the stability of the cadmium complex is an important determining factor of efficacy.

5. The optimal oral antidote towards orally administered cadmium are the BAL analogs, especially DMSA, while the optimal intraperitoneal antidotes towards orally or intraperitoneally administered cadmium are the higher members of the polyaminopolycarboxylate family, especially TTHA.

6. When administered simultaneously (DMSA orally and TTHA intraperitoneally), these chelators synergistically reduce the whole‐body retention of cadmium.

In conclusion, chelation treatment in acute oral cadmium intoxication should first prevent/reduce intestinal damage and uptake by rapid oral administration of a chelating antidote and then alleviate systemic toxicity due to absorbed cadmium and enhance renal/biliary cadmium excretion by parenteral administration of a chelating antidote.     

Alkylation of metals and the activity of metal‐alkyls†

The biochemical basis for resistance to metal ion toxicity is emerging though it is complicated by the different resistance mechanisms. Several strategies for resistance to toxic metal ions have been identified:

1. The development of energy driven efflux pumps which keep toxic element levels low in the interior of the cell. Such mechanisms have been described for Cd(II) and As(V).

2. Oxidation (e.g. AsO^2‐ to AsO₄ ^3‐) or reduction (e.g. Hg²⁺ to Hg⁰) can enzymatically and intracellularly convert a more toxic form of an element to a less toxic form.

3. The biosynthesis of intracellular polymers which serve as traps for the removal of metal ions from solution such as traps have been described for cadmium, calcium, nickel and copper.

4. The binding of metal ions to cell surfaces.

5. The precipitation of insoluble metal complexes (e.g. metal sulfides and metal oxides) at cell surfaces.

6. Biomethylation and transport through cell‐membranes by diffusion controlled processes.

In this short review I shall discuss the implications of biomethylation as a detoxification mechanism for microorganisms as well as for certain higher organisms.     

Chromium binding by calf thymus nuclei and effects on chromatin

1. No binding of chromium was detected after incubation of calf thymus nuclei with hexavalent chromium up to 0.5 mM.

2. Chromium was readily taken up and tightly bound after incubation with trivalent chromium.

3. In a DNA‐filter binding assay, increasing amounts of chromium and DNA were bound with increasing chromium trichloride concentrations incubated with the nuclei.

4. Treatment with proteinase K abolished the increase in DNA retention induced by trivalent chromium.

5. It is concluded that trivalent chromium is the ultimate genetoxic agent after chromate uptake by living cells.

     

Environmental and economic analysis of aluminium recycling through life cycle assessment

Aluminium is the most-used metal in the world after steel, with a wide range of applications in the industrial field owing to its physical and mechanical properties. The aim of this study was environmental and economic analysis of secondary aluminium. In particular, this study demonstrated that aluminium recycling offers many advantages to both consumers and industry:

• Increased energy savings compared to primary aluminium production;

• Recovery and reuse of raw materials for future production avoids consumption of non-renewable resources;

• Reduction in landfill waste and consequent environmental damage;

• Reduction in bauxite mining which, in turn, will reduce the socio-economic impact on populations of those countries, mainly underdeveloped, that contain mines;

• Economic advantages for Italy, the first European producer of secondary aluminium, which lacks bauxite mines and has high energy costs.

This study used the Life Cycle Assessment (LCA) method, the modified Eco-indicator 99 method for damage calculation, and the SimaPro calculation code. The study examined secondary aluminium production by ICMET, based in Reggio Emilia, Italy. The economic and environmental evaluation utilised environmental information supplied by the company.     

A study of the kinetic in transition phase of the anaerobic fluidised bed reactor with immobilised cells in microsand

Anaerobic fluidised bed reactor is an instrument very much adapted in the experimental study of the kinetics of the complex biological process. In this experimental work the efficiency of the treatment of an anaerobic fluidised bed reactor with immobilised cells is examined in transition phase. The results of the experiments can be summarised as follows:

• a zero order elimination rate of the substrate is presented;

• the order of the reaction changes after a period of significant substrate elimination and follows a homographic equation

• the non‐dissociated part of the substrate ions is degradable.

     

Titanium dioxide nanoparticles induce bacterial membrane rupture by reactive oxygen species generation

Nano-titania is widely used in the food industry due to its efficient antimicrobial activity. However, the mechanism of microbial toxicity of nano-titania is poorly known. Here, nano-TiO₂ has been fabricated by microwave-irradiation chemistry, a new method, and then tested for antimicrobial activity. Mutagenicity of nano-TiO₂ was evaluated using Salmonella typhimurium histidine-auxotrophic strains. The reactive oxygen generation test was performed using 2,7-dichlorofluorescein diacetate dye. To test membrane permeabilization, E. coli cultures were grown in nutrient broth at an optical density of 0.3–0.5 at 610 nm, harvested by centrifugation at 11,000g for 10 min, washed and resuspended in 0.5 % NaCl solution. We also analyzed superoxide formation and membrane integrity, and we used scanning electron microscopy. Results show that nano-TiO₂ has a minimum inhibitory concentration of 15 µg/mL, and a minimum bactericidal concentration of 20 µg/mL for E. coli. The bacterial inner wall was ruptured, and cytoplasmic content was released after 5 min of treatment in a dose-dependent manner. Notably, superoxide formation was not observed, which establishes the fact that reactive oxygen generation and alteration of membrane integrity, as well as permeability, is the major mechanism of antimicrobial activity of nano-TiO₂.     

Lethal critical body residues as measures of Cd,Pb, and Zn bioavailability and toxicity in the earthworm<Emphasis Type="Italic">Eisenia fetida</Emphasis>

Background

Earthworm heavy metal concentrations (critical body residues, CBRs) may be the most relevant measures of heavy metal bioavailability in soils and may be linkable to toxic effects in order to better assess soil ecotoxicity. However, as earthworms possess physiological mechanisms to secrete and/or sequester absorbed metals as toxicologically inactive forms, total earthworm metal concentrations may not relate well with toxicity.

Objective

The objectives of this research were to: i) develop LD₅₀s (total earthworm metal concentration associated with 50% mortality) for Cd, Pb, and Zn; ii) evaluate the LD₅₀ for Zn in a lethal Zn-smelter soil; iii) evaluate the lethal mixture toxicity of Cd, Pb, and Zn using earthworm metal concentrations and the toxic unit (TU) approach; and iv) evaluate total and fractionated earthworm concentrations as indicators of sublethal exposure.

Methods

Earthworms (Eisenia fetida (Savigny)) were exposed to artificial soils spiked with Cd, Pb, Zn, and a Cd?Pb?Zn equitoxic mixture to estimate lethal CBRs and mixture toxicity. To evaluate the CBR developed for Zn, earthworms were also exposed to Zn-contaminated field soils receiving three different remediation treatments. Earthworm metal concentrations were measured using a procedure devised to isolate toxicologically active metal burdens via separation into cytosolic and pellet fractions.

Results and Discussion

Lethal CBRs inducing 50% mortality (LD₅₀, 95% CI) were calculated to be 5.72 (3.54–7.91), 3.33 (2.97–3.69), and 8.19 (4.78–11.6) mmol/kg for Cd, Pb, and Zn, respectively. Zn concentrations of dead earthworms exposed to a lethal remediated Zn-smelter soil were 3-fold above the LD₅₀ for Zn and comparable to earthworm concentrations in lethal Zn-spiked artificial soils, despite a 14-fold difference in total soil Zn concentration between lethal field and artificial soils. An evaluation of the acute mixture toxicity of Cd, Pb, and Zn in artificial soils using the Toxic Unit (TU) approach revealed an LD₅₀ (95% CI) of 0.99 (0.57–1.41) TU, indicating additive toxicity.

Conclusions

Total Cd, Pb, and Zn concentrations in earthworms were good indicators of lethal metal exposure, and enabled the calculation of LD₅₀s for lethality. The Zn-LD₅₀ developed in artificial soil was applicable to earthworms exposed to remediated Zn-smelter soil, despite a 14-fold difference in total soil Zn concentrations. Mixture toxicity evaluated using LD₅₀s from each single metal test indicated additive mixture toxicity among Cd, Pb, and Zn. Fractionation of earthworm tissues into cytosolic and pellet digesis yielded mixed results for detecting differences in exposure at the sublethal level.

Recommendation and Outlook

CBRs are useful in describing acute Cd, Pb, and Zn toxicity in earthworms, but linking sublethal exposure to total and/or fractionated residues may be more difficult. More research on detoxification, regulation, and tissue and subcellular partitioning of heavy metals in earthworms and other invertebrates is needed to establish the link between body residue and sublethal exposure and toxicity.

     

Die Wirkung von Photooxidantien auf Sommerweizen (Triticum aestivum L. Star)

The effect of short-time fumigation with the peroxidic photooxidants ozone, peroxyacetyl nitrate (PAN), and hydrogen peroxide (H₂O₂) on the gas exchange of wheat leaves was investigated. Two types of experiments are to be distinguished:

1. Fumigation is carried out at atmospheric CO₂ partial pressure (350 μbar) and the assimilation and transpiration rates are followed simultaneously.
2. The alteration of CO₂-response curves by fumigation is determined; from this, characteristic photosynthetic parameters can be calculated.

With this method, statements about the reaction of both photosynthesis as a whole and specific reaction sites within the most important photosynthetic processes are possible. Ozone in concentrations of 260–280 ppb causes a primary perturbation of the photosynthetic apparatus, whereby the carboxylation rate is affected to nearly the same extent as the electron-transport rate.PAN (120 ppb) causes a reduction in the assimilation rate, primarily due to the closing of the stomata. H₂O₂ affects the gas exchange only in concentrations of above 1 ppm, whereby the reduction of the assimilation rate represents an almost pure stomatal effect. All three fumigants cause an increase in the respiration rate, which may point to an activation of detoxification and repair mechanisms.     

Synthesis and in vitro biological evaluation of 3, 4-annelated coumarin-pyrimido-pyrimidine systems

3,4-Annelated coumarin-pyrimido-pyrimidine systems (7-imino-8-mercapto-7H-5-oxa-7a,9,12-triaza-benzo[a]anthracen-6-one 2 and 8-hydroxy-7-imino-10-methyl-7H-5-oxa-7a,9,12-triaza-benzo[a]anthracen-6-one 3) were synthesized by the action of 4-amino-2-mercapto-pyrimidine, 2-amino-4-hydroxy-6-methylpyrimidine, respectively, on 4-chlorocoumarin-3-carbonitrile 1. The synthesized compounds were examined for in vitro antibacterial activity against 4 bacterial cultures: Staphylococcus aureus, Streptococcus pneumonia, Escherichia coli, and Aeromonas Salmonicida. The antibacterial activity of synthesized compounds 2 and 3 was compared to antibacterial activity of novobiocin as a standard drug.     

Langfristige in situ-messungen zum sauerstoffgaswechsel benthischer meerespflanzen in einem kontinuierlich registrierenden durchflußsystem

Most in situ gas-exchange measurements in aquatic organisms are still performed in closed systems (bottle method, bell-jar experiments). The disadvantages of these methods are that insufficient water movement (turbulence) or relatively long-lasting exposure periods result in under- or oversaturation phenomenas. To avoid these disadvantages as far as possible, an open, continuously recording flow-through system, employing O₂-electrodes, was devised and tested. Examples are provided of O₂-exchange recording in Fucus vesiculosus from the Baltic Sea.

---

---

Aus dem SFB 95, Kiel.     

纳米TiO2与重金属Cd对铜绿微囊藻生物效应的影响

为了更全面地评价纳米TiO2的生物效应,尤其是纳米TiO2与其他环境污染物的联合作用,以铜绿微囊藻为受试生物,探讨了不同浓度的纳米TiO2,以及纳米TiO2与Cd联合作用对藻生长的影响。根据叶绿素a及藻胆蛋白的含量变化,低浓度的纳米TiO2溶液(0～50mg.L-1)可以促进藻的生长,当纳米TiO2的浓度大于50mg·L-1时,藻细胞的生长有所抑制,生长减慢,并呈剂量-效应关系;当纳米TiO2与Cd离子同时存在时,由于纳米TiO2对Cd离子的吸附作用,水中游离态Cd离子浓度降低,Cd离子对藻的毒性明显降低。因此,纳米TiO2的生态毒性和环境效应不容忽视,同时,应重视纳米材料及与其他环境污染物质共同作用后的生物效应。     

Biomonitoring von Stickstoffimmissionen

Background and aim

Air pollution caused by oxidized and reduced nitrogen is distributed over wide areas of Europe at a high level. As an alternative or complement to physical measurements and modelling calculations, biomonitoring with plants provides techniques to assess amounts and effects of pollution from oxidized and reduced nitrogen compounds (depositions and concentrations). Many of the previously implemented techniques are based on well-proven standardised methods, e.g. documented in VDI guidelines, modified more or less for a biomonitoring of atmospheric nitrogen pollution. This paper gives a review of the techniques for a biomonitoring of atmospheric nitrogen pollution, their possibilities as well as their limitations.

Main features

-diversity of the ground vegetation

-nitrogen accumulation in vascular plants

-exposure of vascular plants

-mapping of epiphytic lichens, bryophytes and algae

-nitrogen accumulation in lichens and bryophytes

-exposure of lichens and bryophytes

Results and Discussion

Important response parameters are nitrogen concentrations in plant tissue (shoot, needle, leave) and biodiversity of plant species. These responses of vascular plants in many cases are influenced by other local varying conditions, in particular the soil. The exposure of vascular plants over a short period provides a standardised quantification of the total atmospheric nitrogen input in a model ecosystem. The enrichment of nitrogen in the plant tissue of bryophytes and lichens from the ground vegetation is closely correlated with the amount of nitrogen deposition. The diversity of epiphytic lichens and the response of exposed Hypogymnia physodes is more sensitive to ammonia than to nitrous oxide, whereas with the diversity of epiphytic bryophytes or the abundance of algae no significant correlation with atmospheric N pollution were found.

Conclusions

Some bioindication techniques provide a causeeffect related, partly standardised biomonitoring of nitrogen pollution. In some cases varying experiences with some biomonitors like the biodiversity of the ground vegetation, exposure of lichens, moss bags and grass cultures stress the need for more research in this subject.     

Abschätzung des umweltchemischen und ökotoxikologischen Verhaltens von Stoffen durch computergestützte Analyse von Struktur und Verhalten sowie von Struktur und Wirkung

Risk assessment of chemicals requires comprehensive data material, which often is not available. To provide substitutes for the lacking experimental ecotoxicological and physico-chemical data, a software-system (SAR-System) has been developed comprising more than 90 estimation models for relevant endpoints. The approach is based on quantitative structure-activity relationships (QSAR). Two major aspects characterize the SAR-System:

1. The implemented models were tested for their validity and application range.
2. The QSARs are accessible by a menu-driven programme package.

The following endpoints are included: Thysico-Chemical Data: 1-Octanol/water-partition-coefficient log P_ow, vapour pressure, water solubility, pK_a-value, boiling point Biological Data: Toxicity towards fish, daphnia, tetrahymenae, algae, bacteria and mammals, mutagenicity. Distribution: soil sorption, bioconcentration, Henry-Constant, Mackay (Level I). Degradation: photodegradation, biodegradability.     

Microbiological evaluation of 4-(6-chloro-7H-purin-8-ylamino)-3-nitro-chromen-2-one with a comparison to antibacterial activity of standard drugs

Nucleophilic substitution of 4-chloro-3-nitro-2-oxo-2H-chromen-2-one 3 by 6-chloro-7H-purin-2-ylamine (a), yielded the corresponding 4-(6-chloro-7H-purin-8-ylamino)-3-nitro-chromen-2-one 4. The chemical structure of synthesized compound was characterized using IR and NMR spectra, and elemental analyses. The purified synthesized compound 4 was tested at concentrations 1, 3, 5 mg/ml for its antibacterial activity against three bacterial cultures: Staphylococcus aureus, Escherichia coli, and Bacillus cereus. The antibacterial activity of synthesized compound was compared to antibacterial activity of standard antibiotics cephalexine and streptomycin.     

Comparative effects of five chelating agents on testicular toxicity in mice induced by acute exposure to cadmium

The aim of this study was to develop new antidotes for cadmium (Cd) since this metal is known to produce mammalian toxicity. N-p-hydroxymethylbenzyl-D-glucamine dithiocarbamate (HBGD), N-benzyl-D-glucamine dithiocarbamate (BGD), diethyldithiocarbamate (DDTC), 2,3-dimercaptopropanol (BAL), and ethylenediamine-tetraacetic acid (EDTA) were studied for their ability to inhibit the adverse effects induced by Cd on mouse testes. The parameters examined included concentrations of Cd, calcium (Ca), iron (Fe), and zinc (Zn) in testes, lipid peroxidation (LPO) levels in testes, lactate dehydrogenase (LDH) activity in serum and reproductive ability of male mice. Mice injected intraperitoneally (ip) with CdCl₂ (2.5?mgCd?kg^?1) after 30?min or 24?h, were then injected ip with chelating agents (400?µmol?kg^?1). Cd increased the concentrations of testicular Ca, Cd, Fe, Zn, and LPO levels as well as the activity of LDH in serum. HBGD and BGD effectively prevented the increase in above indices, and improved the reproductive ability weakened by exposure to Cd. The results suggested that HBGD and BGD are more effective detoxificants in the case of testicular toxicity in mice induced by acute exposure to Cd.     

Growth and survival of Mytilus edulis larvae exposed to low levels of dibutyltin and tributyltin

Two studies were conducted to observe effects of dibutyltin (DBT) and tributyltin (TBT) on larvae of Mytilus edulis for an exposure period of 25 d. Endpoints for evaluation were shell growth and mortality measured at 33 d. Larvae were cultured in a new laboratory assay chamber in a recirculating static test. The control, 2, 20, and 200 g/l DBT-treated populations had mean shell lengths of 527, 523, 417, and 180 m, respectively. Survival was 1% for the 200 g/l DBT-treated population, but ranged from 73 to 83% for controls, 2, and 20 g/l treatments. The no-observed-effect concentration (NOEC) was 2 g/l for DBT, while the lowest-observed-effect concentration (LOEC) was 20 g/l. The chronic toxicity value was 6.3 g/l. In the TBT bioassay, mean shell lengths for the control, 0.006, 0.050, and 0.130 g/l-treated populations were 565, 437, 385, and 292 m, respectively. Control survival was 74%, whereas TBT-treated populations survival ranged from 52 to 58%. The NOEC for TBT was 0.006 g/l TBT and the LOEC was 0.050. A chronic toxicity value of 0.017 g/l was calculated. The results of this study indicated that the toxicity of DBT was less than that of TBT. It was concluded that shell length was inversely related to exposure level in both DBT and TBT bioassays. In this study, we have observed TBT effects at lower exposure levels in the laboratory than previously reported, and also report the first data for DBT effects on mussel larvae.     

Effects of sublethal concentrations of mercuric chloride on ammonium-limitedSkeletonema costatum

The effect of sublethal additions of mercuric chloride on the marine diatomSkeletonema costatum (Grev.) Cleve grown in NH₄-limited chemostats and batch cultures was assessed. In short-term Hg exposure experiments (up to 5 h), the effect of Hg on ammonium uptake rates was studied by simulatneously perturbing the culture with 5 M NH₄ Cl and Hg concentrations ranging from 0.04 to 5.52 nM HgCl₂. The threshold of Hg toxicity occurred between 1.8 and 3.7 nM, based on a decrease in ammonium uptake rates. When the NH₄-limited culture was starved of ammonium for 30 h, the threshold of Hg toxicity decreased about an order of magnitude to 0.2 nM. In long-term Hg exposure experiments (679.5 h), NH₄-limited continuous cultures were semi-continuously exposed to 0.37 and 3.68 nM HgCl₂. After 4 days, the cell density in the Hg-treated chemostats began to drastically decline. After about 16 days these populations recovered, even though Hg additions continued. At the end of the experiment (26 days), cell densities had reached the levels observed at the beginning of the experiment. The reason for the recovery is unknown, but several possibilities are discussed. Ammonium uptake rates determined during the time-course of this long-term Hg exposure, indicated that these NH₄-limited cultures exhibited a significant loss in their ability to take up ammonium at low concentrations (e.g. 1 M). Thus, mercury pollution may seriously decrease the ability of a species to utilize the limiting nutrient during periods of seasonal nutrient limitation.     

Metabolic adaptation of the intertidal worm Sipunculus nudus to functional and environmental hypoxia

The scope of anaerobic metabolism of Sipunculus nudus L. was assessed from the maximal activities of some enzymes of the intermediary metabolism and from the concentration of some metabolites accumulated during enhanced muscular activity and during prolonged experimental hypoxia.

1. Maximal enzyme activities demonstrate that the scope of anaerobic glycolysis, as indicated by maximal activities of glycogen phosphorylase (0.84 U g^-1 fresh wt), far exceeds the aerobic capacity, which is assumed not to surpass the activity of succinate dehydrogenase (0.09 U g^-1 fresh wt). Three pyruvate reductase activities (alanopine-, strombine- and octopine dehydrogenase) can possibly terminate anaerobic glycolysis.
2. During muscular activity, energy is provided by the degradation of phospho-L-arginine and by anaerobic glycolysis. Octopine is the major endproduct during functional anaerobiosis while the formation of strombine is less pronounced.
3. During exposure to a nitrogen atmosphere, several anaerobic endproducts are found to accumulate. Anaerobic glycolysis is terminated by strombine synthesis. This opine accumulates in concentrations much higher than octopine. In addition the concentrations of succinate, propionate and acetate are found to increase in tissues, and/or in the coelomic fluid and the incubation water.
4. The relative contribution of energy by the different anaerobic metabolic pathways are estimated during functional and environmental hypoxia.

     

HPLC analysis of nonprotein thiols in planktonic diatoms: pool size,redox state and response to copper and cadmium exposure

A sensitive method was developed to analyze low molecular weight thiols involved in metal homeostasis and detoxification in phytoplankton. The aims of this study were to (1) separate and measure all relevant thiols in a single HPLC run, (2) measure redox states of the thiols and (3) identify specific responses of thiols (pools, redox) to heavy metals by testing diatoms with different metal tolerances (Ditylum brightwellii, Phaeodactylum tricornutum, Skeletonema costatum andThalassiosira pseudonana). Copper or cadmium were dosed at a maximum tolerable, species-dependent level, to exponential phase cells growing in artificial medium (14 salinity). Loss of cell viability was monitored by the decrease of fluorescein fluorescence after a 24-h metal exposure. Thiols in extracts of exposed cells and controls were labeled with monobromobimane. Picomoles of cysteine, glutathione, -glutamylcysteine and phytochelatins (PCs) were detected and separated by reversed-phase high performance liquid chromatography. Cysteine increased in all species after metal exposure. The species with the largest glutathione pools in the control (P. tricornutum) synthesized the largest PC pools upon metal exposure, however, at a 40% loss of glutathione. A considerable increase of glutathione was observed inD. brightwellii upon metal exposure. However, it produced little PC (and only with Cu). In controls ( 3 pM Cu²⁺), PC₂ was detectable inS. costatum, P. tricornutum andT. pseudonana. Oxidized thiol fractions were recovered by the reductants DTT and TCEP, both performing identically. Compared to the other thiols, cysteine had low redox ratios. InD. brightwellii glutathione and PC redox ratios were lower than inP. tricornutum, S. costatum andT. pseudonana. It was expected that Cu-induced oxidative stress would decrease the thiol redox ratios, however, this was not observed.This is Communication No. 2172 of NIOO.