Effects of Silver Nanoparticles and Ions Exposure on the Soil Invertebrates Folsomia candida and Enchytraeus crypticus.

The aim of the study was to assess the effects of silver nanoparticles (AgNPs1 = 2.7 d·nm, AgNPs2 = 6.5 d·nm) and silver nitrate (AgNO3) on Enchytraeus crypticus and Folsomia candida using toxicity tests (OECD Guideline 220, 232). A 28-day chronic toxicity study was performed to evaluate the reproduction and mortality rate. E. crypticus reproduction was more sensitive to AgNO3 with a 28dEC50 of 86.40 (62.52-119.4) mg·kg-1 dry weight (d.w.) compared to AgNPs1 (28dEC50 = 119.3 (60.4-235.6) mg·kg-1 d.w). Similarly, the reproduction of F. candida was inhibited the most by AgNO3 with a 28dEC50 of 126.2 (104.2-152.9) mg·kg-1 d.w. followed by AgNPs1 (28dEC50 = 158.7 (64.05-393.2) mg·kg-1 d.w.) and AgNPs2 (28dEC50 = 206.4 (181.9-234.1) mg·kg-1 d.w.). No mortalities were observed for tested soil invertebrates exposed to AgNPs at concentrations up to 166 mg·kg-1 d.w. of AgNPs1 and 300 mg·kg-1 d.w. of AgNPs2, respectively. It was found that silver ions are more toxic in comparison with AgNPs.

Effect of Gold Nanoparticles and Ions Exposure on the Aquatic Organisms.

An increase in the production and usage of gold nanoparticles (AuNPs) triggers the necessity to focus on their impact on ecosystems. Therefore, the purpose of this study was to investigate the acute toxicity of AuNPs and ionic gold (Au (III)) to organisms representing all trophic levels of the aquatic ecosystem, namely producers (duckweed Lemna minor), consumers (crustacean Daphnia magna, embryos of Danio rerio) and decomposers (bacteria Vibrio fischeri). The organisms were exposed according to a standardized protocol for each species and endpoints. The AuNPs (1.16 and 11.6 d.nm) were synthesized using citrate (CIT) and polyvinylpyrrolidone (PVP) as capping agents, respectively. It was found, that Au (III) was significantly more toxic than AuNPs PVP and AuNPs CIT. AuNPs showed significant toxicity only at high concentrations (mg/L), which are not environmentally relevant in the present time, but a cautious approach is advised, due to the possibility of interactions with other contaminants.

Development of New Silver Nanoparticles Suitable for Materials with Antimicrobial Properties.

Silver nanoparticles are the most important nanoparticles in connection with the antimicrobial effect. Nowadays, the green synthesis of various types of nanoparticles is rapid, effective and produce less toxic nanoparticles often with specific properties. In our experiment we have developed and described in details various types of silver nanoparticles synthesized chemically or by the green synthesis. Nine different silver nanoparticles were synthesized, three by citrate method at different pHs (8; 9; 10), four using gallic acid at alkaline pHs (10; 11), and two by green synthesis using green tea and coffee extracts, both at pH 9. Characterisation of silver nanoparticles was performed using dynamic light scattering, scanning electron microscopy, and ultraviolet-visible absorption spectroscopy. Silver nanoparticles prepared by green synthesis showed the highest antioxidant activity and also ability for quenching of free radicals. Antibacterial activity of silver nanoparticles was determined on bacterial cultures such as Staphylococcus aureus and Escherichia coli. Silver nanoparticles synthesized using green tea and coffee extracts showed the highest antibacterial activity for both bacterial strains. Minimal inhibition concentration for both strains was found to be 65 µM at each silver nanoparticle synthesized using green synthesis.

Evaluation of platinum nanoparticles ecotoxicity using representatives of distinct trophic levels of aquatic biocenosis.

OBJECTIVES: The aim of this study was to clarify the influence of three different sizes of platinum nanoparticles on aquatic ecosystem and assess the toxic effect in term of particle size. Tests were conducted on organisms representing all trophic levels of the aquatic ecosystem, namely producers (duckweed Lemna minor), consumers (water fleas Daphnia magna) and decomposers (bacteria Vibrio fischeri). DESIGN: Experiments were carried out methodologically in accordance with the following standards: OECD 221 guideline (Lemna sp. Growth Inhibition test), OECD 202 guideline (Inhibition of the mobility of Daphnia magna) and ISO 11348-2 (Inhibitory effect of platinum nanoparticles on the light emission of Vibrio fischeri). RESULTS: The most toxic have been the smallest sized platinum nanoparticles for all tested organisms. The highest toxicity of all tested samples (Pt1, Pt2, Pt3) was observed in bacteria (30´EC50 = 135.47; 167.94; 254.64 µg.L-1), respectively. The lowest toxicity was recorded for Daphnia (48hEC50 = 405.74; 413.24; 514.07 µg.L-1), respectively. CONCLUSION: The ecotoxicity of platinum nanoparticles varies considerably according to the test organisms and particle size.

Ecotoxicity of selected antibiotics for organisms of aquatic and terrestrial ecosystems.

OBJECTIVES: The aim of this study was to assess the ecotoxicity of selected antibiotics (i.e. penicillin G, vancomycin and tetracycline) using ecotoxicological tests. Tests were conducted on organisms representing all trophic levels of the aquatic ecosystem, namely producers (green freshwater algae Pseudokirchneriella subcapitata), consumers (water fleas Daphnia magna) and decomposers (bacteria Vibrio fischeri). The effect of antibiotics on the representative of edaphon was measured by testing the inhibition of the reproduction of springtails Folsomia candida and earthworms Eisenia fetida. DESIGN: Methodologically, the procedure was carried out in accordance with the following standards: OECD 201 (Fresh water algal growth inhibition test), OECD 202 (Inhibition of the mobility of Daphnia magna), ISO 11348-2 (Inhibitory effect of antibiotics on the light emission of Vibrio fischeri), OECD 232 (Inhibition of reproduction of Collembola Folsomia candida) and OECD 222 (Inhibition of reproduction of Eisenia fetida). RESULTS: In aquatic organisms the highest level of toxicity was shown by tetracycline to algae (72hEC50 = 1.82 mg.l-1) and daphnia (48hEC50 = 8.16 mg.l-1). The least toxic for all test organisms was penicillin G. The results of the tests performed on the representative of edaphon, Folsomia candida, showed that its reproduction was most inhibited by penicillin G (28dEC50 = 328 mg.kg-1) and least by tetracycline (28dEC50 = 2560 mg.kg-1). Similar results were observed in Eisenia fetida (56dEC50 = 348 mg.kg-1 for penicillin G and 56dEC50 = 2735 mg.kg-1 for tetracycline. CONCLUSION: The ecotoxicity of antibiotics differed significantly depending on the test organism and testing conditions.

Doxorubicin interactions with bovine serum albumin revealed by microdialysis with on-line laser-induced fluorescence detection at subpicogram level.

Doxorubicin (DOX) is an effective antitumor drug employed for treatment of a wide range of cancers types such as neuroblastoma, osteosarcoma, breast and esophageal carcinomas. On the other hand, the cumulative dose is restricted (300-550 mg/m(2)) and its amount administered to a patient has to be closely controlled due to its cardiotoxicity. To understand the mechanisms of the DOX side effects as well as to reveal the ways how to reduce its adverse impact on cardiomyocytes, the interactions with particular components of the blood and tissues have to be studied in greater detail. In this work, microdialysis technique was optimized to extract DOX from samples and subsequently monitor its interaction with BSA. Finally, the microdialysis probe was connected on-line to the LIF detector to ensure the real-time detection. The best flow rate was 1 µL/min and after 120 min of microdialysis 28% of the DOX was dialyzed out from the sample. The results from investigation of the DOX-BSA interaction indicate that the interaction occurs in less than 30 min, causing marked decrease in the amount of DOX extracted by microdialysis.

Fullerene as a transporter for doxorubicin investigated by analytical methods and in vivo imaging.

Carbon nanomaterials, including fullerenes, exhibit not only unique structure and electronic properties but also a significant potential to serve as radical scavengers and/or anti-oxidants. Their conjugation with anticancer drugs such as doxorubicin (DOX) may help to balance severe negative side effects of these cytostatics and also improve the delivery of the drug taking advantage of the enhanced cellular uptake, selectivity to cancer cells, and pH regulated release. In this study, the fullerene (C60) surface was oxidized by concentrated nitric acid, which enabled simple DOX-fullerene conjugation based on π-π stacking and hydrophilic interactions with carboxylic groups. The strength of this noncovalent binding is pH dependent. At a low pH, the amino group of DOX is protonated, however at a higher pH, the amino group is deprotonated, resulting in stronger hydrophobic interactions with the fullerene walls. CE and HPLC were employed for characterization of resulting complexes. The cell toxicity of the conjugates was evaluated using Staphylococcus aureus and finally they were administered into the chicken embryo to assess the applicability for in vivo imaging.

Impact of platinum group elements on the soil invertebrate Enchytraeus crypticus.

OBJECTIVES: The platinum group elements (PGE) platinum (Pt), palladium (Pd), and rhodium (Rh) are used in automobile catalytic converters, from which they have been emitted into the environment to an increasing degree over the last 20 years. Despite the bioavailability of these metals to plants and animals, studies determining the effects of PGE on organisms are extremely rare. Enchytraeids are ecologically relevant soil organisms, due to their activity in decomposition and bioturbation in many soil types worldwide. DESIGN: The experiments were carried out as described in the OECD Guideline 220 [CSN EN ISO 16387--Soil quality--Effects of pollutants on Enchytraeidae (Enchytraeus sp.)--Determination of effects on reproduction]. The reproductive effects of platinum (PtCl4), palladium (PdCl2) and rhodium (RhCl3) were examined. The concentrations of PGE tested were as follows: 5, 10, 25, 50 and 100 µmol x L(-1) PdCl2; 50, 100, 150, 200 and 250 µmol x L-1 PtCl4/RhCl3. The EC50 (medium effective concentration) was determined after 28 days of exposure. The inhibition of the reproduction of PGE-exposed enchytraeids was compared against controls. RESULTS: Values of 28dEC50 of PtCl4, PdCl2 and RhCl3 amounted to 161.9 µmol x L(-1), 70.0 µmol x L(-1) and 246.6 µmol x L(-1), respectively. We can confirm that the relative order of toxicities is Pd (II) >Pt(IV) >>Rh(III). CONCLUSION: To the best of our knowledge, this is the first study to use Enchytraeus crypticus as an indicator species to assess the risk of soil contamination by platinum, palladium and rhodium. Results of this study contribute important data on the ecotoxicity of a rarely studied elements.

Study of the influence of platinum, palladium and rhodium on duckweed (Lemna minor).

OBJECTIVES: Road traffic pollutants and the residues of cytostatics that are widely used in anti-cancer therapy are a significant sources of platinum group elements (PGE; Pt, Pd and Rh) in environment. These metals can migrate into sewage and thus pollute surface waters. The purpose of our study was to evaluate the effect of PtCl4 on the antioxidant and enzymatic activity of duckweed (Lemna minor), a bioindicator of the aquatic environment. METHODS: The study was performed using a 7-day conventional test based on the OECD 221 (CSN EN ISO 20079)--Lemna sp. Growth Inhibition Test. We also conducted a microbiotest to analyse the effects of PtC4, PdCl2 and RhCl3 on the morphology and vegetative growth of colonies of this plant and compared their inhibitory effects during the microbiotest. RESULTS: We observed inhibition of colony growth and clear morphological changes. Antioxidant and enzymatic activities increased with platinum doses increased. The 168hEC50 of PtCl4 was 12.16 µM (95% confidence interval = 9.88-14.44) and the 168hEC50 of PdCl2 was 50.39 (95% confidence interval = 23.83-76.96). The greatest inhibition of growth by RhCl3 was observed at 25 µM. CONCLUSIONS: The obtained results suggest that L. minor phytotoxicity tests should be widely used in the biomonitoring.

Lead toxicosis of captive vultures: case description and responses to chelation therapy.

BACKGROUND: Lead, a serious threat for raptors, can hamper the success of their conservation. This study reports on experience with accidental lead intoxication and responses to chelation therapy in captive Cinereous (Aegypius monachus) and Egyptian (Neophron percnopterus) Vultures. RESULTS: Soil contamination by lead-based paint sanded off the steel aviary resulted in poisoning of eight Cinereous and two Egyptian Vultures. A male Egyptian Vulture developed signs of apathy, polydipsia, polyuria, regurgitation, and stupor, and died on the next day. Liver, kidney and blood lead concentrations were 12.2, 8.16 and 2.66 µg/g, respectively. Laboratory analyses confirmed severe liver and kidney damage and anaemia. Blood Pb levels of Pb-exposed Cinereous Vultures were 1.571 ± 0.510 µg/g shortly after intoxication, decreased to 0.530 ± 0.165 µg/g without any therapy in a month and to 0.254 ± 0.097 µg/g one month after CaNa(2)EDTA administration. Eight months later, blood lead levels decreased to close to the background of the control group. Blood parameters of healthy Pb-non-exposed Cinereous Vultures were compared with those of the exposed group prior to and after chelation therapy. Iron levels in the lead-exposed pre-treatment birds significantly decreased after chelation. Haematocrit levels in Pb-exposed birds were significantly lower than those of the controls and improved one month after chelation. Creatine kinase was higher in pre-treatment birds than in the controls but normalised after therapy. Alkaline phosphatase increased after chelation. A marked increase in the level of lipid peroxidation measured as thiobarbituric acid reactive species was demonstrated in birds both prior to and after chelation. The ferric reducing antioxidant power was significantly lower in pre-treatment vultures and returned to normal following chelation therapy. Blood metallothionein levels in lead-exposed birds were higher than in controls. Reduced glutathione dropped after CaNa(2)EDTA therapy, while oxidised glutathione was significantly lower in both pre- and post-treatment birds. A chick in an egg produced by a Cinereous Vulture female two months after lead toxicosis died on day 40 of artificial incubation. Lead concentrations in foetal tissues were consistent with levels causing avian mortality. CONCLUSIONS: The reported blood parameters and reproduction impairment in captive birds may have implications for professionals dealing with lead exposure in wild birds.

Impact of platinum group elements on the soil invertebrate Folsomia candida.

OBJECTIVES: Trace concentrations of the platinum group elements (PGE; Pt, Pd and Rh) are nowadays an irreplaceable part of environmental analysis and assessment. These rare elements are used as effective substances in automotive catalysts to reduce pollution by emissions originating from fuel combustion. Due to their harmful potential, it is necessary to monitor their content and behaviour in different samples. Effect assessment using ecotoxicological bioassays with organisms at different trophic levels can provide valuable pieces of information on the risk of chemical substances in the ecosystem. DESIGN: The experiments were carried out as described in the OECD Guideline 232 [CSN ISO 11267 - Soil quality - Inhibition of reproduction of Collembola (Folsomia candida) by soil pollutants]. The reproductive effects of palladium (PdCl2) and rhodium (RhCl3) were examined. Concentrations of PGE tested were as follows: 5, 10, 25, 50 and 100 µmol.L-1. The EC50 (medium effective concentration) was determined after 28 days of exposure. Inhibition of reproduction of PGE-exposed collembolans was compared against controls. RESULTS: Values of 28dEC50 of PdCl2 and RhCl3 amounted to 21.0 µmol.L-1 and 266.22 µmol.L-1, respectively. We can confirm that the relative order of toxicities is Pd (II) > Pt(IV) >> Rh(III). CONCLUSION: To the best of our knowledge, this is the first study to use Folsomia candida as an indicator species to assess the risk of soil contamination by palladium and rhodium. However, more toxicity data for various species are needed to evaluate the environmental risks of PGEs in soils.

Molecular biology of beta-estradiol-estrogen receptor complex binding to estrogen response element and the effect on cell proliferation.

Group of estrogen pollutants, where the highest estrogen activity is reported at estradiol, is characterized by the fact that even at very low concentrations have potential to cause xenoestrogenic effects. During exposure of excessive amounts of estradiols may be produced undesirable effects resulting in the feminization of males of water organisms. The presence of estradiols in drinking water implies also a risk for the human population in the form of cancers of endocrine systems, abnormalities in reproduction or dysfunctions of neuronal and immune system. Currently, the research is focused mainly to uncover the relationship between the estrogen receptors binding affinity with an estrogen response element and estradiol. In this review we summarized facts about molecular biological principles of ß estradiol-estrogen receptor complex binding with estrogen response element and its successive effect on cancer genes expression.

Yew poisoning of olive baboons (Papio anubis) in captivity: laboratory diagnosis.

OBJECTIVES: Toxic effects of the yew have been known since ancient times. Yew toxicity is due to the content of cyanogenic glycosides and a mixture of alkaloids known as taxines. Taxine B is probably responsible for the most part of adverse effects in poisoned organisms. This particular taxoid is common in body fluids of the yew-poisoned. The present study is engaged with laboratory examination to confirm substances that lead to fatality of a pair of olive baboons (Papio anubis) following ingestion of yew seeds. When both cage mates (male and female) died suddenly, poisoning was suspected because many berries had fallen into the cage from a nearby fruiting yew tree (Taxus baccata) during the windy night before. METHODS: The analysis was performed using electrospray ionization quadrupole time-of-flight mass spectrometry. A flow injection analysis/mass spectrometry setting was prepared for this purpose. RESULTS: The above mentioned mass spectrometry analysis of taxoids confirmed poisoning by taxanes. The presence of taxin B/isotaxin B was confirmed in all investigated samples. Apparently in urine and bile there were concentrations ranging 150-220 ng.mL-1 and in blood serum concentrations 25-30 ng.mL-1. CONCLUSION: It follows from the results obtained that we confirmed that baboons were deadly intoxicated by yew fruits.

Impact of platinum on the soil invertebrate Folsomia candida.

OBJECTIVES: Regarding the environmental pollution, platinum group elements (PGE) are in the centre of interest of current research. These rare elements are used as effective substances in automotive catalysts to reduce pollution by emissions originating from fuel combustion. Due to their harmful potential, it is necessary to monitor their content and behaviour in different samples. Comprehensive studies on PGE behaviour and effects are still lacking. Their distribution in the food chain and data on bioaccumulation has not been described so far. METHODS: We focused on reproductive effects of platinum (PtCl4), in particular. Our study is based on a collembolan laboratory breed, test optimalization and validation according to the OECD 232 standards [CSN ISO 11267 - Soil quality - Inhibition of reproduction of Collembola (Folsomia candida) by soil pollutants]. The concentrations of PtCl4 tested were as follows: 5, 10, 25, 50 and 100 µM. The EC50 was determined after 28 days of testing. RESULTS: The results were evaluated using the inhibition of reproduction compared with controls. The EC50 was determined after the 28-day test. The value of 28dEC50 of the boric acid test was estimated at 120 mg/kg and the measured 28dEC50 of PtCl4 was 200.4 µM. CONCLUSION: The presented data can be considered as a step forward in the assessment of the potential risk of platinum in the terrestrial environment. However, more toxicity data for various species are needed to evaluate the environmental risk of platinum in soils.

Comparison of the effect of platinum on producers in aquatic environment.

OBJECTIVES: An enhanced worldwide application of platinum group elements (PGE), in particular platinum, has been observed during recent decades. An increased concentration of PGE was determined in collected samples of great amount of aqueous ecosystems.The aim was to compare phytotoxic effect of platinum (PtCl4) by performing two different bioassays on green algae Pseudokirchneriella subcapitata and macrophyte duckweed, Lemna minor. MEDTHODS: The algal experiment (Pseudokichneriella subcapitata) followed OECD 201, the concentration row for PtCl4 was: 0.05; 0.01; 0.25; 0.5; 1 µM. The duckweed (Lemna minor) experiment was conducted according to OECD 221, employed PtCl4 concentrations were: 5; 10; 25; 50; 100 µM. Plants were cultivated as a microbiotest, using micro-volumes. RESULTS: The results of the algal test showed significant growth inhibition of the final biomass. The values of 72hEC5(µ), 72hEC10(µ), 72hEC20(µ) counted on a basis of average specific growth rate (µ) were 0.31 µM, 0.58 µM and 1.12 µM of PtCl4, respectively. The values, obtained on a basis of the area under the growth curves (A), were 0.04 µM (72hEC5(A)), 0.24 µM (72hEC10(A)) and 0.64 µM (72hEC20(A)). The experiment with duckweed showed 50% of growth inhibition and the values of 168hEC50(µ) were 19.55 µM and 168hEC50(A) 13.63 µM of PtCl4. CONCLUSION: The fronds of duckweed showed strong adverse effect of platinum influence (chlorosis, necrosis). The algal test and the estimation of 72hEC5(A) appears to be the most sensitive.

Paramagnetic antibody-modified microparticles coupled with voltammetry as a tool for isolation and detection of metallothionen as a bioindicator of metal pollution.

Low-molecular mass proteins rich in cysteines called metallothioneins (MT) can be considered as markers for the pollution of the environment by metals. Here, we report on suggestion for an automated procedure for the isolation of MT followed by voltammetric analysis. Primarily, we optimized the automated detection of MT using an electrochemical analyser. It was found that the most sensitive and repeatable analyses are obtained at a temperature of 4 °C for the supporting electrolyte. Further, we optimized experimental conditions for the isolation of MT by using antibody-linked paramagnetic microparticles. Under the optimal conditions (4 h long interaction between the microparticles and MT), the microparticles were tested on isolation of various amounts of MT. The lowest isolated amount of MT by antibody-linked paramagnetic microparticles was 5 µg ml(-1) of MT (50 ng). The automated procedure of MT isolation was further tested on isolation of MT from guppy fish (Poecilia reticulata) treated with silver(i) ions (50 µM AgNO(3)). The whole process lasted less than five hours and was fully automated. We attempted to correlate these results with the standard method for MT isolation. The correlation coefficient is 0.9901, which confirms that results are in good agreement. Moreover, the concentration of silver ions in tissues of fish treated with Ag(i) ions was determined by high performance liquid chromatography with electrochemical detection.

Comparison of various easy-to-use procedures for extraction of phenols from apricot fruits.

Phenols are broadly distributed in the plant kingdom and are the most abundant secondary metabolites of plants. Plant polyphenols have drawn increasing attention due to their potential antioxidant properties and their marked effects in the prevention of various oxidative stress associated diseases such as cancer. The objective of this study was to investigate a suitable method for determination of protocatechuic acid, 4-aminobenzoic acid, chlorogenic acid, caffeic acid, vanillin, p-coumaric acid, rutin, ferulic acid, quercetin, resveratrol and quercitrin from apricot samples. A high-performance liquid chromatograph with electrochemical and UV detectors was used. The method was optimized in respect to both the separation selectivity of individual phenolic compounds and the maximum sensitivity with the electrochemical detection. The lowest limits of detection (3 S/N) using UV detection were estimated for ferulic acid (3 µM), quercitrin (4 µM) and quercetin (4 µM). Using electrochemical detection values of 27 nM, 40 nM and 37 nM were achieved for ferulic acid, quercitrin and quercetin, respectively. It follows from the acquired results that the coulometric detection under a universal potential of 600 mV is more suitable and sensitive for polyphenols determination than UV detection at a universal wavelength of 260 nm. Subsequently, we tested the influence of solvent composition, vortexing and sonication on separation efficiency. Our results showed that a combination of water, acetone and methanol in 20:20:60 ratio was the most effective for p-aminobenzoic acid, chlorgenic acid, caffeic acid, protocatechuic acid, ferulic acid, rutin, resveratrol and quercetin, in comparison with other solvents. On the other hand, vortexing at 4 °C produced the highest yield. Moreover, we tested the contents of individual polyphenols in the apricot cultivars Mamaria, Mold and LE-1075. The major phenolic compounds were chlorgenic acid and rutin. Chlorgenic acid was found in amounts of 2,302 mg/100 g in cultivar LE-1075, 546 mg/100 g in cultivar Mamaria and 129 mg/100 g in cultivar Mold. Generally, the cultivar LE-1075 produced the highest polyphenol content values, contrary to Mold, which compared to cultivar LE-1075 was quite poor from the point of view of the phenolics content.

Protein-based electrochemical biosensor for detection of silver(I) ions.

Silver(I) ions are extremely toxic to aquatic animals. Hence, monitoring of these ions in the environment is needed. The aim of the present study was to suggest a simple biosensor for silver(I) ions detection. The suggested biosensor is based on the modification of a hanging mercury drop electrode (HMDE) by the heavy metal binding protein metallothionein (MT) for silver(I) ions detection. Metallothionein accumulated for 120 s onto the HMDE surface. After rinsing the electrode, the biosensor (MT modified HMDE) was prepared prior to detection of silver(I) ions. The biosensor was immersed in a solution containing silver(I) ions. These ions were bound to the MT structure. Furthermore, the electrode was rinsed and transferred to a pure supporting electrolyte solution, in which no interference was present. Under these experimental conditions, other signals relating to heavy metals naturally occurring in MT were not detected. This phenomenon confirms the strong affinity of silver(I) ions for MT. The suggested biosensor responded well to higher silver(I) ion concentrations. The relative standard deviation for measurements of concentrations higher than 50 microM was approximately 2% (n = 8). In the case of concentrations lower than 10 microM, the relative standard deviation increased to 10% (n = 8). The detection limit (3 signal/noise) for silver(I) ions was estimated as 500 nM.

Complexes of glutathione with heavy metal ions as a new biochemical marker of aquatic environment pollution.

Reduced glutathione (GSH) plays a number of key roles in many biochemical pathways. This peptide is highly reactive and forms conjugates with other molecules via its sulfhydryl moiety. The interactions of the common heavy metal pollutant Cd(II) with GSH were determined by using the Brdicka reaction to evaluate whether this technique would be suitable as a biomarker. After GSH interaction with Cd(II) ions, two characteristic changes in the measured voltammogram were observed: Cat2 signal height decreased, and a new signal called P1 was found. The observed signal probably relates to the formation of a GSH-heavy metal ion complex adsorbed on the surface of the working electrode. When the interaction of GSH with cisplatin was studied, the same characteristic changes in the voltammogram were observed, which confirmed our hypothesis. Moreover, changes in the height of P1 and Cat2 signals with increasing time of GSH interaction with Cd(II) ions and/or cisplatin were also investigated. Cat2 peak height decreased proportionally with increasing time of interaction. This decrease can be explained by shielding of free sulfhydryl moiety by heavy metal ions, so it cannot catalyze the evolution of hydrogen from the supporting electrolyte. In addition, we found that, with increasing time of the interaction, the P1 signal was enhanced and shifted to more positive potentials for both Cd(II) ions and cisplatin.

Heavy metals and metallothionein in vespertilionid bats foraging over aquatic habitats in the Czech Republic.

There has been growing interest in the study and conservation of bats throughout the world. Declines in their absolute numbers in recent decades are due, in part, to the fact that insectivorous bats may bioaccumulate toxic pollutants. The purpose of the present study was to quantify heavy metal concentrations in kidney, liver, and pectoral muscle samples in relation to metallothionein (MT) levels. In total, 106 bats belonging to 11 European species (i.e., Myotis myotis, Myotis daubentonii, Myotis brandtii, Myotis nattereri, Myotis emarginatus, Myotis mystacinus, Pipistrellus pipistrellus, Pipistrellus nathusii, Pipistrellus pygmaeus, Nyctalus noctulla, Eptesicus serotinus) were used for the study. The highest MT levels were found in Pipistrellus pipistrellus. High MT levels were also found in juvenile bats and aquatic-insect-foraging species. Cadmium was found only in the liver and kidney of Myotis myotis, except for a solitary finding in Pipistrellus pipistrellus. Myotis myotis juveniles had significantly higher liver and kidney Zn concentrations than adults. Interestingly, the liver Pb concentration was approximately two times higher in females than in males of Myotis myotis. The same gender difference was found for kidney Zn concentration in Pipistrellus pipistrellus. The present study confirms exposure of vespertilionid bats to toxic heavy metals (Pb, Cd) in the Czech Republic and provides data on the essential element Zn and the metal-binding protein MT in bats foraging over aquatic, aquatic and terrestrial, and terrestrial habitats.