So far only a few publications have explored the development of extraction methods of cyanotoxin extracted from complex matrices. With regard to cyanobacterial microcystins (MCs), the data on the contamination of the flesh of aquatic organisms is hard to compare and very limited due to the lack of validated methods. In recent years, evidence that both free and bound fractions of toxin are found in these tissues has highlighted the need to develop effective methods of quantification. Several techniques do exist, but only the Lemieux oxidation has so far been used to investigate complex tissue matrices. In this study, protocols based on the Lemieux approach were adapted for the quantitative chemical analysis of free MC-LR and MMPB derived from bound toxin in the tissues of juvenile trout gavaged with MC-LR. Afterwards, the NF V03 110 guideline was used to characterize the protocols elaborated and evaluate their effectiveness.

The aim of this study was to estimate the bioavailability of essential (Zn, Cu) and non-essential metals (Cd, Pb) to the earthworm Lumbricus rubellus exposed to soils originating from a gradient of metal pollution in Southern Poland. Metal uptake and elimination kinetics were determined and related to soils properties. Experimental results were compared with tissue metal concentrations observed in earthworms from the studied transect. Cd and Pb were intensively accumulated by the earthworms, with very slow or no elimination. Their uptake rate constants, based on 0.01 M CaCl2-extractable concentrations in the soils, increased with soil pH. Internal concentrations of Cu and Zn were maintained by the earthworms at a stable level, suggesting efficient regulation of these metals by the animals. The estimated uptake and elimination kinetics parameters enabled fairly accurate prediction of metal concentrations reached within a life span of L. rubellus in nature.

A multiplicative stomatal conductance model was constructed to estimate stomatal O3 uptake of Fagus crenata exposed to O3 under different N loads to the soil. Our stomatal conductance model included environmental functions such as the stomatal responses of F. crenata to diurnal changes, chronic O3 stress (AOT0), acute O3 stress (O3 concentration), and nitrogen load to soil. The model could explain 62% of the variability in stomatal conductance. We suggest therefore that stomatal closure induced by O3 and N load-induced soil acidification must be taken into account in developing a stomatal conductance model for estimating stomatal O3 uptake for future risk assessment of O3 impact on Japanese forest tree species such as F. crenata.

Transformation of ring-14C-labelled tetrabromobisphenol-A (TBBPA) was studied in an oxic soil slurry with and without amendment with Sphingomonas sp. strain TTNP3, a bacterium degrading bisphenol-A. TBBPA degradation was accompanied by mineralization and formation of metabolites and bound-residues. The biotransformation was stimulated in the slurry bio-augmented with strain TTNP3, via a mechanism of metabolic compensation, although this strain did not grow on TBBPA. In the absence and presence of strain TTNP3, six and nine metabolites, respectively, were identified. The initial O-methylation metabolite (TBBPA-monomethyl ether) and hydroxytribromobisphenol-A were detected only when strain TTNP3 was present. Four primary metabolic pathways of TBBPA in the slurries are proposed: oxidative skeletal rearrangements, O-methylation, ipso-substitution, and reductive debromination. Our study provides for the first time the information about the complex metabolism of TBBPA in oxic soil and suggests that type II ipso-substitution could play a significant role in the fate of alkylphenol derivatives in the environment.

The present study demonstrates that species-specific isotope tracing is an useful tool to precisely measure Hg accumulation and transformations capabilities of living organisms at concentrations naturally encountered in the environment. To that end, a phytoplanktonic green alga Chlamydomonas reinhardtii Dangeard (Chlamydomonadales, Chlorophyceae) was exposed to mixtures of 199-isotopically enriched inorganic mercury (199IHg) and of 201-isotopically enriched monomethylmercury (201CH3Hg) at a concentration range between less than 1 pM to 4 nM. Additionally, one exposure concentration of both mercury species was also studied separately to evaluate possible interactive effects. No difference in the intracellular contents was observed for algae exposed to 199IHg and 201CH3Hg alone or in their mixture, suggesting similar accumulation capacity for both species at the studied concentrations. Demethylation of 201CH3Hg was observed at the highest exposure concentrations, whereas no methylation was detected.

To assess the effects of leaf age/layer on the response of photosynthesis to chronic ozone (O3), Cyclobalanopsis glauca seedlings, a dominant evergreen broadleaf tree species in sub-tropical regions, were exposed to either ambient air (AA) or elevated O3 (AA + 60 ppb O3, E-O3) for two growing seasons in open-top chambers. Chlorophyll content, gas exchange and chlorophyll a fluorescence were investigated three times throughout the 2nd year of O3 exposure. Results indicated that E-O3 decreased photosynthetic parameters, particularly light-saturated photosynthesis rate, stomatal conductance and effective quantum yield of PSII photochemistry of current-year leaves but not previous-year leaves. Stomatal conductance of plants grown under ambient conditions partially contributed to the different response to E-O3 between leaf layers. Light radiation or other physiological and biochemical processes closely related to photosynthesis might play important roles. All suggested that leaf ages or layers should be considered when assessing O3 risk on evergreen woody species.

Interaction between effects of hazardous chemicals in the environment and adverse climatic conditions is a problem that receives increased attention in the light of climate change. We studied interactive effects of phenanthrene and drought using a test system in which springtails (Folsomia candida Willem) were concurrently exposed to a sublethal phenanthrene level via passive dosing from silicone (chemical activity of 0.010), and sublethal drought from aqueous NaCl solutions (water activity of 0.988). Previous studies have shown that the combined effects of high levels of phenanthrene and drought, respectively, interact synergistically when using lethality as an end-point. Here, we hypothesized that phenanthrene interferes with physiological mechanisms involved in drought tolerance, and that drought influences detoxification of phenanthrene. However, this hypothesis was not supported by data since phenanthrene had no effect on drought-protective accumulation of myo-inositol, and normal water conserving mechanisms of F. candida were functioning despite the near-lethal concentrations of the toxicant. Further, detoxifying induction of cytochrome P450 and glutathione-S-transferase was not impeded by drought. Both phenanthrene and drought induced transcription of heat shock protein (hsp70) and the combined effect of the two stressors on hsp70 transcription was additive, suggesting that the cellular stress and lethality imposed by these levels of phenanthrene and drought were also additive.

Constructed wetlands remove trace organic contaminants via synergistic processes involving plant biomass that include hydrolysis, volatilization, sorption, biodegradation, and photolysis. Wetland design conditions, such as hydraulic loading rates (HLRs) and carbon loading rates (CLRs), influence these processes. Contaminant of emerging concern (CEC) removal by wetland plants was investigated at varying HLRs and CLRs. Rate constants and parameters obtained from batch-scale studies were used in a mechanistic model to evaluate the effect of these two loading rates on CEC removal. CLR significantly influenced CEC removal when wetlands were operated at HLR >5 cm/d. High values of CLR increased removal of estradiol and carbamazepine but lowered that of testosterone and atrazine. Without increasing the cumulative HLR, operating two wetlands in series with varying CLRs could be a way to improve CEC removal.

Canopy-level stomatal conductance over a warm-temperate mixed deciduous and evergreen broadleaf forest in Japan was estimated by the Penman–Monteith approach, as compensated by a semi-empirical photosynthesis-dependent stomatal model, where photosynthesis, relative humidity, and CO2 concentration were assumed to regulate stomatal conductance. This approach, using eddy covariance data and routine meteorological observations at a flux tower site, permits the continuous estimation of canopy-level O3 uptake, even when the Penman–Monteith approach is unavailable (i.e. in case of direct evaporation from soil or wet leaves). Distortion was observed between the AOT40 exposure index and O3 uptake through stomata, as AOT40 peaked in April, but with O3 uptake occurring in July. Thus, leaf pre-maturation in the predominant deciduous broadleaf tree species (Quercus serrata) might suppress O3 uptake in springtime, even when the highest O3 concentrations were observed.

Critical levels (CLEs) of atmospheric ammonia based on biodiversity changes have been mostly calculated using small-scale single-source approaches, to avoid interference by other factors, which also influence biodiversity. Thus, it is questionable whether these CLEs are valid at larger spatial scales, in a multi- disturbances context. To test so, we sampled lichen diversity and ammonia at 80 sites across a region with a complex land-cover including industrial and urban areas. At a regional scale, confounding factors such as industrial pollutants prevailed, masking the CLEs. We propose and use a new tool to calculate CLEs by stratifying ammonia concentrations into classes, and focusing on the highest diversity values. Based on the significant correlations between ammonia and biodiversity, we found the CLE of ammonia for Mediterranean evergreen woodlands to be 0.69 μg m−3, below the previously accepted value of 1.9 μg m−3, and below the currently accepted pan-European CLE of 1.0 μg m−3.