Toward a more realistic estimate of exposure to chromium and nickel in soils of geogenic and/or anthropogenic origin: importance of oral bioaccessibility.

To enhance risk assessment for contaminated sites, incorporating bioavailability through bioaccessibility as a corrective factor to total concentration is essential to provide a more realistic estimate of exposure. While the main in vitro tests have been validated for As, Cd, and/or Pb, their potential for assessing the bioaccessibility of additional elements remains underexplored. In this study, the physicochemical parameters, pseudototal Cr and Ni concentrations, soil phase distribution, and oral bioaccessibility of twenty-seven soil samples were analysed using both the ISO 17924 standard and a simplified test based on hydrochloric acid. The results showed wide variability in terms of the concentrations (from 31 to 21,079 mg kg-1 for Cr, and from 26 to 11,663 mg kg-1 for Ni) and generally low bioaccessibility for Cr and Ni, with levels below 20% and 30%, respectively. Bioaccessibility variability was greater for anthropogenic soils, while geogenic enriched soils exhibited low bioaccessibility. The soil parameters had an influence on bioaccessibility, but the effects depended on the soils of interest. Sequential extractions provided the most comprehensive explanation for bioaccessibility. Cr and Ni were mostly associated with the residual fraction, indicating limited bioaccessibility. Ni was distributed in all phases, whereas Cr was absent from the most mobile phase, which may explain the lower bioaccessibility of Cr compared to that of Ni. The study showed promising results for the use of the simplified test to predict Cr and Ni bioaccessibility, and its importance for more accurate human exposure evaluation and effective soil management practices.

Floristic survey, trace element transfers between soil and vegetation and human health risk at an urban industrial wasteland.

This study aimed to determine the trace element accumulation in the soil and plants in an industrial wasteland and to estimate the extent of transfer to humans to measure the effects on and risks to vegetation and human health and find bioindicator plants representative of the levels of the main contaminants. In areas with the highest extractable trace element levels, we observed decreases in plant biodiversity explained by the disappearance of several families, favouring the coverage of tolerant species, such as Urtica dioica and Hedera helix. Trace elements were also found in the leaves of several plants, especially in a dominant species that is poorly studied, Alliaria petiolata. Indeed, this species had the highest contents of Zn (1750 mg.kg-1 DW), Ni (13.1 mg.kg-1 DW), and Cd (18 mg.kg-1 DW) found at the site and is a potential Zn bioindicator since its leaf contents were also representative of the Zn extractable contents in soil (R² = 0.94). The hazard quotient and carcinogen risk revealed that most of the site had an identified or possible risk, mainly due to Pb and As. Native species, especially A. petiolata, could be used in phytoextraction to manage and limit these human and environmental risks.

Oral bioaccessibility of PTEs in soils: A review of data, influencing factors and application in human health risk assessment.

Understanding the behavior of metal(loi)ds transported from soil to humans is critical for human health risk assessment (HHRA). In the last two decades, extensive studies have been conducted to better assess human exposure to potentially toxic elements (PTEs) by estimating their oral bioaccessibility (BAc) and quantifying the influence of different factors. This study reviews the common in vitro methods used to determine the BAc of PTEs (in particular As, Cd, Cr, Ni, Pb, and Sb) under specific conditions (particularly in terms of the particle size fraction and validation status against an in vivo model). The results were compiled from soils derived from various sources and allowed the identification of the most important influencing factors of BAc (using single and multiple regression analyses), including physicochemical soil properties and the speciation of the PTEs in question. This review presents current knowledge on integrating relative bioavailability (RBA) in calculating doses from soil ingestion in the HHRA process. Depending on the jurisdiction, validated or non-validated bioaccessibility methods were used, and risks assessors applied different approaches: (i) using default assumptions (i.e., RBA of 1); (ii) considering that bioaccessibility value (BAc) accurately represents RBA (i.e., RBA equal to BAc); (iii) using regression models to convert BAc of As and Pb into RBA as proposed by the USA with the US EPA Method 1340; or (iv) applying an adjustment factor as proposed by the Netherlands and France to use BAc from UBM (Unified Barge Method) protocol. The findings from this review should help inform risk stakeholders about the uncertainties surrounding using bioaccessibility data and provide recommendations for better interpreting the results and using bioaccessibility in risk studies.

In situ and ex situ bioassays with Cantareus aspersus for environmental risk assessment of metal(loid) and PAH-contaminated soils.

Environmental risk assessment of contaminated soils requires bioindicators that allow the assessment of bioavailability and toxicity of chemicals. Although many bioassays can determine the ecotoxicity of soil samples in the laboratory, few are available and standardized for on-site application. Bioassays based on specific threshold values that assess the in situ and ex situ bioavailability and risk of metal(loid)s and polycyclic aromatic hydrocarbons (PAHs) in soils to the land snail Cantareus aspersus have never been simultaneously applied to the same soils. The aims of this study were to compare the results provided by in situ and ex situ bioassays and to determine their respective importance for environmental risk assessment. The feasibility and reproducibility of the in situ bioassay were assessed using an international ring test. This study used five plots located at a former industrial site and six laboratories participated in the ring test. The results revealed the impact of environmental parameters on the bioavailability of metal(loid)s and PAHs to snails exposed in the field to structured soils and vegetation compared to those exposed under laboratory conditions to soil collected from the same field site (excavated soils). The risk coefficients were generally higher ex situ than in situ, with some exceptions (mainly due to Cd and Mo), which might be explained by the in situ contribution of plants and humus layer as sources of exposure of snails to contaminants and by climatic parameters. The ring test showed good agreement among laboratories, which determined the same levels of risk in most of the plots. Comparison of the bioavailability to land snails and the subsequent risk estimated in situ or ex situ highlighted the complementarity between both approaches in the environmental risk assessment of contaminated soils, namely, to guide decisions on the fate and future use of the sites (e.g., excavation, embankments, and land restoration). Integr Environ Assess Manag 2022;18:539-554. © 2021 SETAC.

From environmental bioavailability of metal(loid)s to their ecogenotoxicological effects in land snails.

To date, no study has linked the environmental and the ecogenotoxicological bioavailability of contaminants to land snails. Yet, understanding the specific ecotoxicological mechanisms from bioaccumulation to genotoxicity is necessary e.g., to build an adverse outcome pathway relevant to risk assessment. Consequently, the aim of our study is to look for relationships between accumulated concentrations of As, Cd and Hg in sub-adult snails and ecotoxicological effects at the individual (survival and growth) and molecular (genomic stability) levels. This study combines random amplified polymorphic DNA (RAPD) coupled with high-resolution capillary electrophoresis system (HRS) and micronucleus (MN) assay on haemocytes to consider various types of cytogenomic damage, such as chromosomal aberrations, breakages, adducts and mutations. The results showed alteration of the individual endpoints at higher accumulation quotients (AQs) that reflect the excess of transfers to snails, especially with decreased survival for As. In addition, genotoxic effects were observed with an increased occurrence of MN in haemocytes for the three meta(loid)s considered (R2 from 0.57 to 0.61 as a function of the meta(loid)s). No concentration-dependent decrease in genome stability was highlighted by RAPD-HRS in snails exposed to As and Cd but not Hg. Our results demonstrate the complementarity of the RAPD-HRS and the MN assay for understanding the different genotoxic mechanisms of the three metal(loid)s studied in land snails. They show a way to better assess environmental risks of contaminated soils by associating ecotoxicity, genotoxicity and bioaccumulation assays (ISO 24032), i.e., ecogenotoxicological bioavailability. Convergences highlighted here between the bioaccumulation of metal(loid)s in viscera and genotoxic effects in haemocytes constitute a way to better assess the bioavailability of contaminants in soils to the land snail and the subsequent environmental risk.

From bioavailability to risk assessment of polluted soil using snails: link between excess transfer and inhibition of sexual maturation.

An accurate assessment of the environmental risk of soils contaminated by metal(loid)s (MEs) requires quantifying exposure and knowing the toxicity of contaminants transferred to biota. For this purpose, two indices have been developed with the bioindicator Cantareus aspersus to assess exposure (SET: sum of the excess of transfer) and risk (ERITME: evaluation of the risk of the transferred metal elements) of multi-contaminated soils. If the SET and ERITME indices allow characterization of exposure and risk based on unspecific toxicity points, then the link between these indices and real effects on some toxicological endpoints, such as growth or sexual maturation, remains to be demonstrated. For this purpose, sub-adult snails were exposed for 28 days to 38 ME-contaminated soils. Relationships between the SET and/or ERITME indices and health alterations in C. aspersus were determined using Spearman correlations, linear regressions, univariate regression trees, and kinetic models. Relationships were determined between the values of the SET and ERITME indices, bioaccumulation as an indicator of ME bioavailability, and the alteration in physiological endpoints, such as the shell development used as a non-invasive indicator of sexual maturation. The results enabled the determination of three levels of risk according to the differences in reaching sexual maturity: no risk, uncertain, and proven risk depended on whether the value of ERITME was below, in, or beyond the interval [2574-22720], respectively. This study provides the first benchmarks with the SET and ERITME indices to interpret the risk of contaminated soils to snails and to relate the environmental and toxicological bioavailability of ME mixtures.

Impact of ageing and soil contaminants on telomere length in the land snail.

Telomeres (TLs) are non-coding DNA sequences that are usually shortened with ageing and/or chemical exposure. Bioindicators such as the land snail can be used to assess the environmental risk of contaminated soils. As for most invertebrates, the evolution of TLs with ageing or exposure to contaminants is unknown in this mollusc. The aims of this study were to explore the relationships between ageing, contaminant exposure, sublethal effects and TL length in the terrestrial gastropod Cantareus aspersus. TL length was investigated in haemocytes from five age classes of C. aspersus. The impact of contaminants on sub-adult snails exposed to Cd, Hg or a mixture of polycyclic aromatic hydrocarbons (PAHs) in soils for one or two months was studied. Bioaccumulation, growth, sexual maturity and TLs were measured. TL attrition was significant for the juvenile and sub-adult stages, but not later. Exposure to Cd increased the mortality (around 30%). Exposure to polluted soils inhibited growth (19-40%) and sexual maturity (6-100%). Although the health of the snails exposed to Cd, Hg and PAHs was altered, TL length in haemocytes was not disturbed, suggesting a high capacity of this snail species to maintain its TLs in haemocytes under chemical stress. These results first address TL length in snails and reveal that the relationship commonly proposed for vertebrates between TL shortening and ageing or exposure to contaminants cannot be generalized.

Telomere dynamic in humans and animals: Review and perspectives in environmental toxicology.

Telomeres (TLs) play major roles in stabilizing the genome and are usually shortened with ageing. The maintenance of TLs is ensured by two mechanisms involving telomerase (TA) enzyme and alternative lengthening telomeres (ALT). TL shortening and/or TA inhibition have been related to health effects on organisms (leading to reduced reproductive lifespan and survival), suggesting that they could be key processes in toxicity mechanisms (at molecular and cellular levels) and relevant as an early warning of exposure and effect of chemicals on human health and animal population dynamics. Consequently, a critical analysis of knowledge about relationships between TL dynamic and environmental pollution is essential to highlight the relevance of TL measurement in environmental toxicology. The first objective of this review is to provide a survey on the basic knowledge about TL structure, roles, maintenance mechanisms and causes of shortening in both vertebrates (including humans) and invertebrates. Overall, TL length decreases with ageing but some unexpected exceptions are reported (e.g., in species with different lifespans, such as the nematode Caenorhabditis elegans or the crustacean Homarus americanus). Inconsistent results reported in various biological groups or even between species of the same genus (e.g., the microcrustacean Daphnia sp.) indicate that the relation usually proposed between TL shortening and a decrease in TA activity cannot be generalized and depends on the species, stage of development or lifespan. Although the scientific literature provides evidence of the effect of ageing on TL shortening, much less information on the relationships between shortening, maintenance of TLs, influence of other endogenous and environmental drivers, including exposure to chemical pollutants, is available, especially in invertebrates. The second objective of this review is to connect knowledge on TL dynamic and exposure to contaminants. Most of the studies published on humans rely on correlative epidemiological approaches and few in vitro experiments. They have shown TL attrition when exposed to contaminants, such as polycyclic aromatic hydrocarbons (PAH), polychlorinated biphenyls (PCB), pesticides and metallic elements (ME). In other vertebrates, the studies we found deals mainly with birds and, overall, report a disturbance of TL dynamic consecutively to exposure to chemicals, including metals and organic compounds. In invertebrates, no data are available and the potential of TL dynamic in environmental risk assessment remains to be explored. On the basis of the main gaps identified some research perspectives (e.g., impact of endogenous and environmental drivers, dose response effects, link between TL length, TA activity, longevity and ageing) are proposed to better understand the potential of TL and TA measurements in humans and animals in environmental toxicology.

How contamination sources and soil properties can influence the Cd and Pb bioavailability to snails.

To better understand the fate of metals in the environment, numerous parameters must be studied, such as the soil properties and the different sources of contamination for the organisms. Among bioindicators of soil quality, the garden snail (Cantareus aspersus) integrates multiple sources (e.g. soil, plant) and routes (e.g. digestive, cutaneous) of contamination. However, the contribution of each source on metal bioavailability and how soil properties influence these contributions have never been studied when considering the dynamic process of bioavailability. Using accumulation kinetics, this study showed that the main assimilation source of Cd was lettuce (68%), whereas the main source of Pb was the soil (90%). The plant contribution increased in response to a 2-unit soil pH decrease. Unexpectedly, an increase in the soil contribution to metal assimilation accompanied an increase in the organic matter (OM) content of the soil. For both metals, no significant excretion and influence of source on excretion have been modelled either during exposure or depuration. This study highlights how the contribution of different sources to metal bioavailability changes based on changes in soil parameters, such as pH and OM, and the complexity of the processes that modulate metal bioavailability.

In situ assessment of phyto and zooavailability of trace elements: A complementary approach to chemical extraction procedures.

For an accurate risk assessment of sites contaminated by trace elements (TE), measurements of bioavailability must be performed. This is routinely achieved using the standardized 0.01M CaCl2 method. However, the suitability of chemical extractions as proxies of bioavailability is questionable. We analyzed the correlations between chemically estimated TE bioavailability and TE actually accumulated by coupling plant and snails bioindicators. Results showed a better correlation between plant TE contents and CaCl2 fraction while total soil concentration better explained snail TE contents. However in both cases chemical measures were not suitable to predict TE accumulation and bioavailability. Considering the soil properties only improve the estimation of Cr, Ni and Pb accumulation by plants while for snails, TE contents in viscera were dependent both on soil and plant contents and soil properties. It highlights the complementarities of biomonitoring methods to assess bioavailability. This dual approach allows a "physiologically defined" evaluation of bioavailability.

Chemical extractions and predicted free ion activities fail to estimate metal transfer from soil to field land snails.

This study investigates the relevance of several soil chemical extractions (calcium chloride, acetic acid, citric acid and a four-step sequential procedure) and predicted free metal ion activities in the soil solution to characterise the transfer of trace metals (Cd, Pb, and Zn) from soil to snail soft tissues over a large smelter-impacted area (Metaleurop Nord, Nord-Pas-de-Calais, France). The study was first performed on six snail species together and then specifically on Cepaea sp. and Oxychilus draparnaudi. When the six species were considered together, the accumulation of metals depended mostly on the species. When significant, total or extractable metal concentrations, or the predicted free ion activities, accounted for less than 7% of the variation of the metal concentrations in the snail tissues. Species-specific analyses showed that extractable concentrations explained approximately 25% of the variation of the metal concentrations in O. draparnaudi, and up to 8% in Cepaea snails. When using total soil concentrations and soil properties as explanatory variables, the models were generally slightly better, explaining up to 42% of the variance. The soil extraction procedures and predicted free ion activities used in this study did not accurately estimate the metal transfer from soil to snails and could not be used in risk assessment.