Arsenic speciation in human hair: a new perspective for epidemiological assessment in chronic arsenicism

The analysis for arsenic in hair is commonly used in epidemiological studies to assess exposure to this toxic element. However, poor correlation between total arsenic concentration in hair and water sources have been found in previous studies. Exclusive determination of endogenous arsenic in the hair, excluding external contamination has become an analytical challenge. Arsenic speciation in hair appears as a new possibility for analytical assessing in As-exposure studies. This study applied a relative simple method for arsenic speciation in human hair based on water extraction and HPLC-HG-ICP-MS. The concentration of arsenic species in human hair was assessed in chronically As(V)-exposed populations from two villages (Esqui?a and Illapata) of the Atacama Desert, Chile. The arsenic concentrations in drinking water are 0.075 and 1.25 mg L(-1), respectively, where As(V) represented between 92 and 99.5% of the total arsenic of the consumed waters. On average, the total arsenic concentrations in hair from individuals of Esqui?a and Illapata were 0.7 and 6.1 microg g(-1), respectively. Four arsenic species, As(III), DMA(V), MMA(V) and As(V), were detected and quantified in the hair extracts. Assuming the found species in extracts represent the species in hair, more than 98% of the total arsenic in hair corresponded to inorganic As. On average, As(III) concentrations in hair were 0.25 and 3.75 microg g(-1) in Esqui?a and Illapata, respectively; while, the As(V) average concentrations were 0.15 and 0.45 microg g(-1) in Esqui?a and Illapata, respectively. Methylated species represent less than 2% of the extracted As (DMA(V)+ MMA(V)) in both populations. As(III) in hair shows the best correlation with chronic exposure to As(V) in comparison to other species and total arsenic. In fact, concentrations of As(total), As(III) and As(V) in hair samples are correlated with the age of the exposed individuals from Illapata (R= 0.65, 0.69, 0.57, respectively) and with the time of residence in this village (R= 0.54, 0.71 and 0.58, respectively).     

Probabilistic human health risk of PCDD/F exposure: a socioeconomic assessment

The aim of the present study was to assess the human health risks (carcinogenic and non-carcinogenic) of PCDD/F exposure for the adult population living in two different areas (industrial and residential) of Tarragona (Catalonia, Spain), as well as to compare these risks according to the socioeconomic status of the population. Two pathways were considered for PCDD/F exposure: direct (air inhalation, dermal contact and ingestion from soils and dust) and dietary. Although environmental exposure was higher for the population living in the neighborhood of the industrial area (mainly lower socioeconomic group subjects), it only accounted 1.05%. On the other hand, PCDD/F exposure through dietary intake was more important for those individuals of an upper socioeconomic level. This was due to the fact that their diet is richer in those foodstuffs containing higher PCDD/F concentrations. Consequently, taking together direct and dietary exposure to PCDD/Fs, the result is that upper socioeconomic group populations are more exposed to these toxic pollutants. In any case, the current levels of PCDD/F exposure would not mean significant carcinogenic and non-carcinogenic risks for the population living in the industrial and residential areas of Tarragona.     

Speciation of inorganic arsenic in coastal seawater from Ionian and Tyrrhenian Seas (Sicily, Italy) using derivative anodic stripping chronopotentiometry

The purpose of this paper was to use derivative anodic stripping chronopotentiometry (dASCP) as a sensitive and accurate technique, to determine the concentrations of dissolved As (III) and As (V) in coastal seawater samples from the Straits of Messina, the Ionian and the Tyrrhenian seas, and to investigate the relationship between the anthropogenic activities on the coastal areas and the concentration of dissolved inorganic arsenic in seawaters. The obtained data indicated that As (V) was the most abundant species, with concentration ranging from 26.7 to 307 nM, whereas As (III) levels were lower than 48 nM in all the samples. In particular, As (III) and As (V) levels significantly decreased from high to low anthropogenic activities zones (p < 0.00001, ANOVA), with the reference samples, from a wildlife reserve, showing the lowest values. Furthermore it was observed that human activities influenced inorganic arsenic speciation, since the zones that received high human input presented the highest As(V)/As (III) ratio.     

Effect of fasting on the pattern of urinary arsenic excretion

Millions of people in some of the poorest regions of the world are exposed to high levels of arsenic through drinking contaminated water. It has been reported that development of cancer caused by arsenic exposure in such populations is dependent on dietary and nutritional factors which can modulate arsenic metabolism. Many people in arsenic exposed regions of Bangladesh and India practice fasting for at least one month every year when they refrain from consumption of food and fluid during daylight hours. How such practices may modulate arsenic metabolism has not been previously investigated. This study investigated this issue by determining total arsenic and its species in urine samples from a group of 29 unexposed volunteers at the beginning of the fasting and at the end of approximately 12 h of fasting period. Inductively coupled plasma mass spectrometry (ICP-MS) and high performance liquid chromatography (HPLC) coupled with ICP-MS was used to measure the total arsenic and arsenic speciation in the urine samples, respectively. The mean total levels of arsenic at the beginning of fasting (18.3 microg g(-1) creatinine) and at the end of approximately 12 h of fasting (17.7 microg g(-1) creatinine) did not differ significantly (p > 0.05). However, the percentages of urinary arsenic as the methylated arsenic species methylarsonate (MA) were found to be significantly different (p < 0.05) and this species was observed more frequently at the end of fasting, although its overall concentration was similar. There were no significant differences (p > 0.05) in both the concentrations and percentages of other urinary arsenic species detected, namely arsenobetaine (AB) and dimethylarsinate (DMA). Arsenite (As(III)) and arsenate (As(V)) were also analyzed, but were not detected. We conclude that fasting for a period of 12 h results in a significant increase in the percentage of urinary arsenic as MA, and its frequency of detection in the volunteers at the end of the fasting period is almost nine fold higher. This suggests that metabolism of arsenic is altered by fasting.     

Rabbit Population Landscape-Scale Simulation to Investigate the Relevance of Using Rabbits in Regulatory Environmental Risk Assessment

This paper describes the development and testing of the ALMaSS rabbit model and its baseline, and subsequently its application to the question of lagomorph population vulnerability in environmental risk assessment (ERA). Development and testing following a pattern-oriented modelling protocol resulted in a model able to replicate local and landscape-level rabbit population patterns. We then tested how robust rabbit populations are to an (imaginary) extreme toxic stressor at a landscape level in a variety of landscapes, and to what extremes key uncertain model parameters must be pushed to cause extinctions. This was contrasted with the same (imaginary) toxic stressor applied to the already existing ALMaSS hare model. For EU risk assessment of plant protection products, these results clearly indicate that if the protection goal is population-level impacts, either in abundance and/or distribution, then the hare is a much more vulnerable species than the rabbit under all the conditions tested. Rabbits would only be more vulnerable than hares if the entire population were to be exposed simultaneously, when lower body mass would then be a critical factor. This did not occur even though the toxicant and exposure scenarios tested here were extreme and, in fragmented landscapes at scales used here, will not occur in reality from the use of plant protection products on crop fields. As well as specifically answering the question on rabbit versus hare vulnerability, this study generally illustrates the potential application of models for setting focal species for risk assessments.     

Adsorption and removal of oxo-anions of arsenic and selenium on the zirconium(IV) loaded polymer resin functionalized with diethylenetriamine-N,N,N',N'-polyacetic acid

A polymer adsorbent in which a Zr(IV)-edta complex analogue is immobilized has been prepared and applied to the removal of oxo-anions of As(III), As(V) and Se(IV). Effective retention of these anions has been demonstrated with the proposed polymer complex system. The adsorption mechanism of oxo-anions onto the Zr(IV)-chelated polymer complex has been investigated using Zr(IV)-edta as the model compound. The formation of mixed complexes with oxo-anions has been exemplified by the isolation of the carbonato complex K2[Zr(CO3)edta].3H2O, the structure of which has been confirmed by X-ray crystallography. NMR study suggests that oxo-anions that form weak or moderate conjugate acids, including those of As(III), As(V) and Se(IV), can form mixed complexes with Zr(IV)-edta using the unsaturated coordination site. However, oxo-anions of strong conjugate acids did not show any appreciable interaction with this complex. According to these observations, retention of oxo-anions on the Zr(IV)-chelated polymer complex has been interpreted by a ligand substitution reaction. The adsorption characteristics of As(III), As(V) and Se(IV) on the Zr(IV)-loaded resin have been examined with respect to the equilibrium adsorption, percentage extraction and the effect of co-existing ions. The adsorption and desorption cycles of the oxo-anions have been demonstrated using a column packed with the proposed resin without any loss of column performance, which indicates the possibility for repeated use.     

Investigation of arsenic speciation on drinking water treatment media utilizing automated sequential continuous flow extraction with IC-ICP-MS detection

Three treatment media, used for the removal of arsenic from drinking water, were sequentially extracted using 10 mM MgCl2(pH 8), 10 mM NaH2PO4(pH 7) followed by 10 mM (NH4)2C2O4(pH 3). The media were extracted using an on-line automated continuous extraction system which allowed the arsenic in each of the extraction fluids to be speciated on-line using IC-ICP-MS. The 10 mM MgCl2 preferentially extracted As(III) from each of the media. The percentage of the arsenic extracted by the MgCl2, relative to a HNO3/H2O2 digestion of the media, ranged from 0.1-2.3% for the three solids. The next sequential extraction fluid, 10 mM NaH2PO4, extracted some of the residual As(III) remaining on each of the media but the predominant species extracted was As(V). The 10 mM NaH2PO4 extracted 15.3 to 42.8% of the total arsenic relative to a total digested concentration for each of the media. The As(III) and As(V) stability studies conducted in these two extraction fluids indicated that conversion between As(III) and As(V) was not significant for the short extraction fluid sample contact time associated with the on-line continuous flow extraction cell. Finally, the 10 mM (NH4)2C2O4 extraction fluid was utilized in an off-line analysis mode because the Fe and As concentrations extracted from the media were not compatible with direct ICP-MS detection. The (NH4)2C2O4 extracted 2.9-29% As(III) for all three media and caused an oxidation of As(III) to As(V) during the extraction period for one of the three media. The sum of the arsenic from each of the three extraction fluids represented 92%, 44% and 53% of the available total arsenic for the three media, respectively. The speciation results for each media were obtained by adding all the speciation results from all three extraction fluids together and the resulting distribution of As(III)/As(V) compared well with the speciation results obtained via XANES.     

European bee-eater (Merops apiaster) populations under arsenic and metal stress: evaluation of exposure at a mining site

Two populations of the European bee-eater were studied, one living at a reference site and the other at a metal mining site. The concentration of arsenic and 11 metals (Al, Cd, Co, Cr, Cu, Fe, Hg, Mn, Ni, Pb, and Zn) was measured in feathers and regurgitated pellets collected at both sites. Cadmium, chromium, mercury, nickel, and lead were at least twofold higher in feathers of birds from the contaminated site than in the reference site, suggesting that this population was exposed to higher metal levels. Similarly, levels of aluminum, arsenic, cadmium, cobalt, chromium, iron, and lead were also at least twofold higher in pellets from the contaminated area. The obtained results suggested that the impacted population of Merops apiaster is at risk due to the exposure to some metals.     

Human health risk assessment for ingestion exposure to groundwater contaminated by naturally occurring mixtures of toxic heavy metals in the Lao PDR

This study constitutes the first systematic risk assessment in the Lao PDR of the safety of groundwater for consumption. Groundwater and hair samples were collected from seven Lao provinces to determine the quantitative health impact of heavy metals through ingestion exposure. Contamination levels for arsenic (As; 46.0 %) and barium (Ba; 16.2 %) exceeded World Health Organization (WHO) guidelines, especially in Mekong River floodplains. A USEPA assessment model for health risks from daily groundwater ingestion, with adjustments for local water consumption values, was applied to estimate the size of the population at risk for noncarcinogenic and carcinogenic health problems. As was the only element contributing to noncarcinogenic health risks in all contaminated areas. The populations of Bolikhamxai, Savannakhet, Saravane, Champasak, and Attapeu, moreover, were at risks of cancer. In addition to the As groundwater concentration factor, noncarcinogenic and carcinogenic risks were positively correlated with the average daily dose of As, exposure duration, and subject body weight. The level of As in hair correlated with groundwater consumption and average daily dose of As. 25.5 % of the population (n?=?228) showed As levels in hair above the toxicity level.     

Use of Stable Nitrogen Isotopes and Permeable Membrane Devices to Study what Factors Influence Freshwater Mollusk Survival in the Conasauaga River

Recent biological inventory data shows severe declines in freshwater mussel abundance and biodiversity in the Conasauga River Basin in Northwest Georgia, USA. Based on assessments of habitat conditions, mussel populations should be sustainable. We conducted a study of sediment and water quality to evaluate the impact of anthropogenic contamination on mussel populations. Permeable membrane devices (PMD), polar organic chemical integrative samplers (POCIS™), conventional water and sediment quality analyses, and stable nitrogen isotope ratio analyses (δ¹⁵N) of snails and sediments were used to assess sediment and water quality at target sites throughout the basin. Ambient concentrations of organic contaminants in water were well below any aquatic life criteria; concentrations of some nutrients were detected above aquatic life criteria levels. Most mussel species in the river are endangered or threatened; therefore, snails were collected for δ¹⁵N analyses. Mean δ¹⁵N values for snails collected at forested upper watershed sites (national forest areas) were significantly lower than δ¹⁵N values from snails in agricultural areas. δ¹⁵N values for raw cow manure and manure-treated soil were similar to δ¹⁵N values for snails collected in agricultural areas. Dissolved nitrate from water samples had elevated δ¹⁵N values similar to the upper range of δ¹⁵N values for snails in agricultural areas. Data, particularly stable nitrogen isotope data, indicates that a land use change from national forest land to agriculture alters nitrogen sources to the basin and snails. Implications of nutrient release on freshwater molluscan reproduction, growth, and survival are discussed.     

Occurrence and sorption properties of arsenicals in marine sediments

The content of total arsenic, the inorganic forms: arsenite (As(III)) and arsenate (As(V)), the methylated forms: monomethylarsonic acid and dimethylarsinic acid (DMA), trimethylarsenic oxide, tetramethylarsenonium ion and arsenobetaine was measured in 95 sediment samples and 11 pore water samples from the Baltic Sea near the island of Bornholm at depths of up to 100 m. As(III+V) and DMA were detected in the sediment and As(III+V) was detected in the sediment pore water. Average total As concentration of 10.6?±?7.4 mg/kg dry matter (DM) in the sediment corresponds to previously reported values in the Baltic Sea and other parts of the world. Existing data for on-site measurements of sorption coefficients (Kd) of arsenicals in marine and freshwater sediments show large variability from <1 to >1,000 L/kg. In this work, calculated sorption coefficients (Kd and Koc) for As(III+V) showed significant correlation with depth, dissolved oxygen (DO), salinity and sediment classification; for depths <70 m, salinity <11 %, DO >9 mg/L and sand/silt/clay sediments the Kd was 118?±?76 L/kg DM and for depths >70 m, salinity >11 %, DO?<?9 mg/L and muddy sediments the Kd was 513?±?233 L/kg DM. The authors recommend using the found Kd value for arsenic in marine sediments when conditions are similar to the Baltic Sea. At locations with significant anthropogenic point sources or where the local geology contains volcanic rock and sulphide mineral deposits, there may be significantly elevated arsenic concentrations, and it is recommended to determine on-site Kd values.     

Indicators of ecosystem health at the species level and the example of selenium effects on fish

Chemical monitoring of aquatic ecosystems describes the chemical exposures of aquatic biota and measures the success of pollution control. However, meeting water quality criteria cannot assure that aquatic biota are protected from the effects of unexpected chemicals, mixtures and interactions between toxicity and environmental stressors.Biological monitoring is an obvious solution since aquatic biota integrate spatial and temporal variations in exposure to many simultaneous stressors. Top predators, typical of specific ecosystems (e.g. lake trout in cold water oligotrophic lakes) indicate whether environmental criteria have been met. The presence of naturally reproducing, self-sustaining and productive stocks of edible fish demonstrates a high quality environment. If these conditions are not met, there is a clear sign of environmental degradation. Specific changes in population structure and performance may also diagnose which life stage is affected and the nature of the stressor.Unfortunately, environmental managers cannot rely solely on populations, communities or ecosystems to indicate chmical effects. The lag between identifying a problem and finding a cause may destroy the resource that we wish to protect, particularly where chemicals are persistent.A solution to this dilemma is the measurement of primary or secondary responses of individual organisms to chemical exposure. Since toxicity at any level of organization must start with a reaction between a chemical and a biological substrate, these responses are the most sensitive and earliest sign of chemical exposure and effect.Application of this idea requires research on molecular mechanisms of chemical toxicity in aquatic biota and adaptation of existing mammalian diagnostic tools. Since relevance of biochemical responses to populations and ecosystems is not obvious, there is a need to study the links between chemical exposure and responses of individuals, populations and ecosystems.The recognition of chemical problems and cause-effect relationships requires the integration of chemical and biological monitoring, using the principles of epidemiology to test the strength of relationships and to identify specific research needs. The contamination of a reservoir with selenium and impacts on fish populations provide an excellent example of this approach.     

Biogenic and toxic elements in feathers, eggs, and excreta of Gentoo penguin (Pygoscelis papua ellsworthii) in the Antarctic

Feathers, eggs, and excreta of Gentoo penguin (Pygoscelis papua ellsworthii), adults, from Livingston Island (South Shetlands), chosen as bioindicators, were used to test the quality of the Antarctic environment. Sex was not examined. The bioaccumulations of toxic trace elements (Cd, Pb, Al, and As), essential trace elements (Fe, Cu, Zn, Mn, Cr, V, Ni, and Sr), and major essential elements (Na, K, Mg, Ca, P, and S) were established. For the first time data about the element contents in Gentoo eggs is provided. Two hypotheses were tested: (1) there are differences in the metal levels among eggs and feathers; and (2) the element concentrations are highest in the excreta. The hypotheses were confirmed at 0.01-0.05 confidence levels. The concentrations of almost all trace elements were significantly higher in the feathers compared to those in the eggs. The following values of the concentrations ratio Fe/Zn were obtained: in the embryo, Fe/Zn = 1.5, and in the feathers, Fe/Zn = 0.5. The concentration of Pb in the embryo and excreta was below 0.4 μg/g, and Cd and As in eggs were below 0.05 and 0.3 μg/g, respectively. This indicates that there is no toxic risk for penguin offspring. Arsenic could be considered as a potential pollutant for Antarctic soil due to its relative high concentration in excreta, 5.13 μg/g. The present data (year 2007) were compared to the data for years 2002 and 2003. No trend of toxic element contamination was established. The concentrations of Pb, Cd, and As in representatives from the top of the food chain in the Antarctic (the present study) and Arctic (literature data) were compared. The data supports the hypothesis that there is an abnormality in cadmium levels in polar marine areas. Regarding Pb, the South Shetlands displayed 3-fold lower level compared to the Aleutians.     

A feeding inhibition based prediction of the toxic effect of dissolved metal mixtures upon Echinogammarus marinus (Crustacea: Amphipoda) at field relevant concentrations across a latitudinal gradient

Risk assessment of metals in the environment is performed mainly with toxicity evaluations on single metals, which is largely inadequate since these substances occur in mixtures. The development of models predicting combined toxic effects on the basis of the concentration-response relationships of individual compounds has emerged as an answer. In the present study, metal effects on post-exposure anorexia (the concept of FdC(50)--concentration causing 50% of feeding inhibition--is implemented) in Echinogammarus marinus, a widely distributed gammarid amphipod, were assessed and compared with modelled ones obtained through the application of the concentration addition (CA) model, which represents a reasonable worst-case scenario for the risk assessment of metal mixtures. Data were validated using in situ experiments performed along a latitudinal gradient (Iceland, Scotland and Portugal) aiming at establishing a geographic profile of autochthonous population susceptibilities to metals. For all of the metals studied concentrations in the water column at exposure sites were in good agreement with feeding inhibition levels. Models gave low to relatively high percentage agreement between predictions and experimental data. Boreal populations demonstrated higher susceptibility to single metals, but not to mixture exposures. Meridional populations denoted lower susceptibilities with higher FdC(50).     

Cross-border response of mosses to heavy metal atmospheric deposition in Southeastern Bulgaria and European Turkey

First cross-border atmospheric pollution of 11 heavy metals and toxic elements assessed by Hypnum cupressiforme was reported for a part of Southeastern Europe (Southeastern Bulgaria and European Turkey). Moss monitoring technique followed the main requirements of European Moss Survey. Moss samples were collected in April 2006 both in Bulgaria and Turkey. Concentration of Al, As, Cd, Cr, Cu, Fe, Ni, Pb, Sb, V, and Zn were determined by ICP-AES. Interlaboratory parallel calibration (exchanged four moss samples from each country), standard reference moss materials (M2 and M3) results ensured the study. ANOVA showed no differences between measured results in both laboratories at the 99% confidence level. Principle Component Analyze proved two factors: F1 group of Al, As, Cd, Cr, Fe, Ni, and V and F2 of Cu, Pb, and Zn as main atmospheric pollutants. Results obtained showed approximately Cu and Pb high concentrations around Istanbul and Burgas and Zn pollution in Istanbul district. Arsenic cross-border atmospheric pollution in the study area of Southeastern Europe was found.     

Using watershed characteristics,sediment, and tissue of resident mollusks to identify potential sources of trace elements to streams in a complex agricultural landscape

Trace elements used in animal feed additives can be introduced to aquatic environments through application of manures from animal feeding operations to agricultural land as fertilizer. The use of poultry feed additives containing arsenic (As) is of particular concern in the Shenandoah River watershed (Virginia, USA), an agricultural landscape with a high density of poultry operations. This study investigated the relationship between watershed characteristics of Shenandoah River tributaries and trace element concentrations in streambed sediment and tissue of resident mollusks, including: Asian clams (Corbicula fluminea), which are commonly used biomonitors, and pleurocerid snails (Leptoxis carinata), which are generally understudied. Results failed to support the primary hypothesis of a predictive relationship between watershed densities of poultry operations and As concentrations in sediment and mollusk tissue. However, there were statistical relationships between land use in tributary watersheds and other trace elements in sediment (Cu, Mn, Pb, Zn) and tissue (Cd, Hg, Pb). Principal components analysis of the sediment data suggested a possible geologic source of As at some sites. Tissue concentrations of As were significantly higher in snails than in clams, but clams accumulated higher concentrations of other trace elements (Cd, Cr, Hg, Pb, Se). Snails may be useful biomonitors of environmental As, but appear to be less suitable than clams for studies of landscape sources of other trace elements.     

Oxidation of Cr(III) to Cr(VI) during chlorination of drinking water

Drinking water treatment typically uses strong oxidants such as chlorine which are capable of converting Cr(III) to Cr(VI). The rates and extent of Cr(III) oxidation by chlorine are not well established. Cr(III) oxidation experiments were therefore conducted in distilled deionized water and New York City tap water dosed initially with Cr(III) and supplemented with sodium hypochlorite to increase free chlorine residual. Reaction progress was monitored using capillary electrophoresis which quenched reactions and allowed for quantification of Cr(VI). Three different forms of Cr(III) were used as reactants: a Cr(III) nitrate salt, Cr(III)-EDTA, and Cr(III) hydroxide. Rates of Cr(VI) production for all three forms of Cr(III) were rapid, on the order of hours. However, oxidation rates slowed and a plateau in Cr(VI) concentrations was reached. This resulted in less than 100% conversion of Cr(III) to Cr(VI) even at relatively high chlorine doses (10 to 100 mg L(-1) as Cl(2)). The loss of free chlorine due to a non-Cr chlorine demand, the precipitation of Cr(III) to Cr(OH)(3)(s), and the partial oxidation of Cr(III) to intermediate oxidation states (i.e. Cr(IV) and Cr(V)) were examined and eliminated as possible explanations for this behavior. Consumption of chlorine via reaction with intermediate oxidation states of Cr is therefore offered as a possible explanation for the plateau in Cr(VI) concentrations.     

Arsenic-speciation in arsenate-resistant and non-resistant populations of the earthworm,Lumbricus rubellus

Arsenic speciation was determined in Lumbricus rubellus Hoffmeister from arsenic-contaminated mine spoil sites and an uncontaminated site using HPLC-MS, HPLC-ICP-MS and XAS. It was previously demonstrated that L. rubellus from mine soils were more arsenate resistant than from the uncontaminated site and we wished to investigate if arsenic speciation had a role in this resistance. Earthworms from contaminated sites had considerably higher arsenic body burdens (maximum 1,358 mg As kg-1) compared to the uncontaminated site (maximum 13 mg As kg-1). The only organo-arsenic species found in methanol/water extracts for all earthworm populations was arsenobetaine, quantified using both HPLC-MS and HPLC-ICP-MS. Arsenobetaine concentrations were high in L. rubellus from the uncontaminated site when concentrations were expressed as a percentage of the total arsenic burden (23% mean), but earthworms from the contaminated sites with relatively low arsenic burdens also had these high levels of arsenobetaine (17% mean). As arsenic body burden increased, the percentage of arsenobetaine present decreased in a dose dependent manner, although its absolute concentration rose with increasing arsenic burden. The origin of this arsenobetaine is discussed. XAS analysis of arsenic mine L. rubellus showed that arsenic was primarily present as As(III) co-ordinated with sulfur (30% approx.), with some As(v) with oxygen (5%). Spectra for As(III) complexed with glutathione gave a very good fit to the spectra obtained for the earthworms, suggesting a role for sulfur co-ordination in arsenic metabolism at higher earthworm arsenic burdens. It is also possible that the disintegration of As(III)-S complexes may have taken place due to (a) processing of the sample, (b) storage of the extract or (c) HPLC anion exchange. HPLC-ICP-MS analysis of methanol extracts showed the presence of arsenite and arsenate, suggesting that these sulfur complexes disintegrate on extraction. The role of arsenic speciation in the resistance of L. rubellus to arsenate is considered.     

Assessing the impact of chloroacetanilide herbicides and their metabolites on periphyton in the Leyre River (SW France) via short term growth inhibition tests on autochthonous diatoms

The Leyre River is the main tributary to the Bassin d'Arcachon lagoon. Herbicides belonging to the chloroacetanilide class have been found in the river (S-metolachlor and acetochlor) as well as some of their metabolites at higher concentrations. As the environmental toxicity of these molecules is not well known, ecotoxicological tests have been carried out on river periphyton at different levels of biological diversity: from the clone of one diatom species (Nitzschia nana) to the population of the same species (several clones) up to the multi-specific species community dominated by diatoms. Moreover, tests were performed on diatoms coming from an unpolluted upstream site and from a contaminated downstream site, in order to investigate possible tolerance acquisition to pollutants. The method consisted in measuring diatom growth inhibition at different doses of each substance from the increase of chlorophyll-a concentration after 4 days. It resulted that acetochlor was clearly more toxic than S-metolachlor at all levels of biological diversity. EC(50) values estimated from the tests suggest no effect of contaminants on diatom growth or biomass in the river. The toxicity of the metabolites appeared very low compared to that of their parent compounds. No difference in tolerance to the herbicides was demonstrated between summer diatom communities from the two sites in spite of different specific compositions. However, concerning the populations of N. nana isolated in winter following the highest herbicide concentrations in the river (about 0.5 μg L(-1)), the downstream population showed a higher tolerance to acetochlor but there was no co-tolerance to S-metolachlor. Thus, it appeared that acetochlor represents the highest toxic pressure on periphyton among the other contaminants in the Leyre River.     

Variations in sensitivity of two populations of Corophium Orientale (Crustacea: Amphipoda) towards cadmium and sodium laurylsulphate

The aim of this work was to monitor the sensitivity of two populations of Corophium orientale (Crustacea: Amphipoda) living at the outfall of two rivers (Magra and Serchio), comparing their responses towards two different toxicant solutions. Sensitivity was monthly checked performing the 96h-LC₅₀ static water only test with Cd(NO₃)₂ and SDS. If no significant differences were found between the two populations, they could have been employed without distinction to perform sediment toxicity bioassays. As regard to Cd, an increasing in LC₅₀ values from summer to winter was evident in each population (Serchio River: August 2003  =  1,36 mg/l, February 2004  =  7,23 mg/l; Magra River: August  =  1,21 mg/l, April  =  5,01 mg/l). This pattern was correlated to the droop of temperatures in winter period. The responses of the two populations towards the cadmium were compared using the ANOVA. The analysis showed any significant differences between the populations (p  =  0.12). The pattern of sensitivity towards SDS for the population living on Magra River was similar to the same pattern found for Cd; as regard to the population living on Serchio River, data were not enough to describe the annual pattern. Anyway, statistical analysis was performed and no significant differences were found between the two populations (p  =  0.34).