Dynamic extreme values modeling and monitoring by means of sea shores water quality biomarkers and valvometry.

Water quality can be evaluated using biomarkers such as tissular enzymatic activities of endemic species. Measurement of molluscs bivalves activity at high frequency (e.g., valvometry) during a long time period is another way to record the animal behavior and to evaluate perturbations of the water quality in real time. As the pollution affects the activity of oysters, we consider the valves opening and closing velocities to monitor the water quality assessment. We propose to model the huge volume of velocity data collected in the framework of valvometry using a new nonparametric extreme values statistical model. The objective is to estimate the tail probabilities and the extreme quantiles of the distribution of valve closing velocity. The tail of the distribution function of valve closing velocity is modeled by a Pareto distribution with parameter 𝜃 t,τ , beyond a threshold τ according to the time t of the experiment. Our modeling approach reveals the dependence between the specific activity of two enzymatic biomarkers (Glutathione-S-transferase and acetylcholinesterase) and the continuous recording of oyster valve velocity, proving the suitability of this tool for water quality assessment. Thus, valvometry allows in real-time in situ analysis of the bivalves behavior and appears as an effective early warning tool in ecological risk assessment and marine environment monitoring.

Effects of soil pH on the Vicia-micronucleus genotoxicity assay.

In the field of contaminated sites and soil management, chemical analyses only bring typological data about pollution. As far as bioavailability and effects on organisms are concerned, we need ecotoxicology tools. In this domain, among many existing tests, we chose to study genotoxicity because it is a short-term endpoint with long-term consequences. The aim of this study is to assess the effects of soil pH on the results of the Vicia faba root tip micronucleus test for the two following reasons: (i) to define the pH range within which the test can be performed without modifying the soil to be tested, within the framework of the ISO standard of the test and (ii) to provides information about the effects of the pH on the genotoxic potential of soils. In this context, we modified the pH of a standard soil with HCl or NaOH and we spiked the matrix with copper (2, 4 and 8 mmol kg(-1) dry soil) or with maleic hydrazide, an antigerminative chemical (5, 10 and 20 µmol kg(-1) dry soil). We concluded that the pH had no effect on the mitotic index or micronucleus frequency in the root cells of the negative controls: extreme pH values did not induce micronucleus formation in root cells. Moreover, according to our results, the Vicia-micronucleus test can be performed with pH values ranging between 3.2 and 9.0, but in the ISO 29200 "Soil quality--assessment of genotoxic effects on higher plants--V. faba micronucleus test" we recommended to use a control soil with a pH value ranging between 5 and 8 for a more accurate assessment of chemical genotoxicity. We also found that acid pH could increase the genotoxic potential of pollutants, especially heavy metals. With hydrazide maleic spiked soil, plants were placed in a situation of double stress, i.e. toxicity caused by extreme pH values and toxicity induced by the pollutant.

Genotoxicity potential of a new natural formicide.

BACKGROUND, AIM, AND SCOPE: Assessment of environmental impacts from pesticide utilization should include genotoxicity studies, where the possible effects of mutagenic/genotoxic substances on individuals are assessed. In this study, the genotoxicity profile of the new formicide Macex® was evaluated with two genotoxicity tests, namely, the micronucleus test with mouse bone marrow and Vicia faba, and a mutagenicity test using the Ames Salmonella assay. MATERIALS AND METHODS: The bacterial reverse mutation test (Salmonella typhimurium strains TA97, TA98, TA100, TA102, and TA1535), the Vicia root tip and mouse micronucleus tests were conducted according to published protocols. RESULTS: In the range of the formicide Macex® concentrations tested from 0.06 to 1.0 g L⁻¹ (or mgkg⁻¹ in the mouse test), no genotoxicity was observed in the prokaryotic or eukaryotic test organisms. However, at Macex® concentrations of 0.5 g L⁻¹ and above a significant decrease in the mitotic index (P ≤ 0.05) in the V. faba was observed. Micronucleus formation was likewise increased in the test organism at concentrations starting at 2.0 g L⁻¹. CONCLUSIONS: These data allow us to classify this natural formicide preparation as a product with no geno-environmental-impact when applied at recommended concentrations.

Is there a relationship between early genotoxicity and life-history traits in Vicia faba exposed to cadmium-spiked soils?

The Vicia faba-micronucleus test is usually performed to assess the genotoxic potential of pure substances, effluents or water extracts from soil. It is also a relevant, early biological tool to detect mutagenic substances in crude soils. Nevertheless, the physiological meaning of such DNA damage for the plant in the long term remains to be elucidated. To know more about this, two experiments were carried out with the plant model V. faba. In a preliminary short-term experiment, seeds were exposed for five days to a soil spiked with different concentrations of CdCl(2) in order to identify the concentration inducing the highest number of micronuclei without affecting plant growth. Thereafter, a long-term experiment was performed in the soil spiked with such a concentration (i.e. 510µmol CdCl(2) per kg dry soil), in which V. faba seeds were directly sowed and allowed to grow during 151 days. As a result, Cd-spiked soil did not affect seed-emergence time nor the growth-rate of the plants for the first two months. The first signs of toxicity appeared after the 70th day of exposure. Interestingly, exposed plants produced their first flower earlier and had a longer flowering period than did control ones. Nevertheless, total flower production was less abundant in exposed plants than in control plants. Moreover, fruits appeared in control plants whereas no fruit was formed in exposed plants. At last, exposed plants showed a reduced life time. Our results suggest that the micronucleus assay can provide a predictive biomarker of long-term deleterious effects in plants.

Improvement of Vicia-micronucleus test for assessment of soil quality: a proposal for international standardization.

The Viciafaba root tip micronucleus test is one of the most employed plant genotoxicity assays, and has been used on various types of contaminated materials. This test has been standardized by AFNOR, the French member organization of ISO. However, this test is usually performed with a water extraction step but soil genotoxicity assessment would be more relevant when performed directly in the soil itself. In order to harmonize these protocols, an ISO standard for the V.faba micronucleus test in both liquid phase (exposure of plants to different liquid matrix, including soil water extracts) and solid phase (direct exposure of plants to the soil) would be very useful. In this context, we compared two exposure durations in the solid phase (48 h and 5 d) for the V.faba micronucleus test with two different well-known genotoxicants, maleic hydrazide and copper sulfate. We concluded that these two durations induced equivalent sensitivity: the micronucleus frequency was significantly increased with 5 µmol maleic hydrazide per kg dry soil and with 2 mmol copper sulfate per kg dry soil with both exposure durations. However, exposing roots to soil during 48 h is more practical. Moreover, organically and conventionally cultured seeds were employed to determine whether the seed provenance influenced the test sensitivity. Organic seeds were less sensitive to copper, possibly because copper-based treatments are permitted, and often applied, in organic farms. Therefore, in the absence of completely non-treated seeds, organically-cultured seeds did not appear to offer any advantages over conventional seeds.

Environmental impact of sunscreen nanomaterials: ecotoxicity and genotoxicity of altered TiO2 nanocomposites on Vicia faba.

Mineral sunscreen nanocomposites, based on a nano-TiO(2) core, coated with aluminium hydroxide and dimethicone films, were submitted to an artificial ageing process. The resulting Altered TiO(2) Nanocomposites (ATN) were then tested in the liquid phase on the plant model Vicia faba, which was exposed 48 h to three nominal concentrations: 5, 25 and 50 mg ATN/L. Plant growth, photosystem II maximum quantum yield, genotoxicity (micronucleus test) and phytochelatins levels showed no change compared to controls. Oxidative stress biomarkers remained unchanged in shoots while in roots, glutathione reductase activity decreased at 50 mg ATN/L and ascorbate peroxidase activity decreased for 5 and 25 mg ATN/L. Nevertheless, despite the weak response of biological endpoints, ICP-MS measurements revealed high Ti and Al concentrations in roots, and X-ray fluorescence micro-spectroscopy revealed titanium internalization in superficial root tissues. Eventual long-term effects on plants may occur.