Ecotoxicological Assays Research Articles

Evaluating Ecotoxicological Assays for Comprehensive Risk Assessment of Industrial Wastewaters in Republic of Korea

Evaluating Ecotoxicological Assays for Comprehensive Risk Assessment of Industrial Wastewaters in Republic of Korea

Commonwealth of Soil Health: How Do Earthworms Modify the Soil Microbial Responses to CeO2 Nanoparticles?

Soil ecotoxicological assays on nanoparticles (NPs) have mainly investigated single components (e.g., plants, fauna, and microbes) within the ecosystem, neglecting possible effects resulting from the disturbance of the interactions between these components. Here, we investigated soil microbial responses to CeO2 NPs in the presence and absence of earthworms from the perspectives of microbial functions (i.e., enzyme activities), the community structure, and soil metabolite profiles. Exposure to CeO2 NPs (50, 500 mg/kg) alone decreased the activities of enzymes (i.e., acid protease and acid phosphatase) participating in soil N and P cycles, while the presence of earthworms ameliorated these inhibitory effects. After the CeO2 NP exposure, the earthworms significantly altered the relative abundance of some microbes associated with the soil N and P cycles (Flavobacterium, Pedobacter, Streptomyces, Bacillus, Bacteroidota, Actinobacteria, and Firmicutes). This was consistent with the pattern found in the significantly changed metabolites which were also involved in the microbial N and P metabolism. Both CeO2 NPs and earthworms changed the soil bacterial community and soil metabolite profiles. Larger alterations of soil bacteria and metabolites were found under CeO2 NP exposure with earthworms. Overall, our study indicates that the top-down control of earthworms can drastically modify the microbial responses to CeO2 NPs from all studied biological aspects. This clearly shows the importance of the holistic consideration of all soil ecological components to assess the environmental risks of NPs to soil health.

Characteristics and impact of aged coal ash with slag emplaced in a karst cave: the case of Diva\u0161ka jama, Slovenia

A mixture of coal bottom ash and slag, with a fraction of fly ash (CAFAS) from steam locomotives, was placed in the cave Divaška jama to delimit and level tourist trails. Emplacement began in 1914 and carried on for several decades. The CAFAS mixed with other cave material gradually changed its structure and appearance. Currently the concentration of some elements in the CAFAS (As, Cu, Hg, Ni, Pb, Zn), and also to a lesser extent in cave sediments (Cr, Cu, Ni), indicates a possibly harmful effect on sediment-associated biota based on ecotoxicological assays. Compared to the cave sediment, the CAFAS contains distinctly different mineral phases and presents a different source of radioactivity. Microbial metabolic activity of CAFAS is low, 0.22 μl O2/gDW h, but higher than that of cave sediment. The present environmental hazards from CAFAS are estimated to be low. Whereas the emplacement of CAFAS was seen initially a long-term solution for waste disposal and management of the cave, it turned out that CAFAS enriches the underground environment with inorganic and organic compounds and disperses pollution into the cave ecosystem. After its removal from the cave, the CAFAS should be investigated thoroughly due to its susceptibility to alteration.

Effects of sheep manure in agricultural soils on the behavior of Folsomia candida and initial growth and development of Avena sativa.

The aim of this study was to evaluate the effects of sheep manure in agricultural soils on the behavior of Folsomia candida and initial growth and development of Avena sativa. For this, an Oxisol was submitted to different doses of sheep manure and was subsequently evaluated for Folsomia candida survival and avoidance behavior through standardized ecotoxicological assays, the initial performance of oats by germination test and the soil basal respiration rate by respirometry methodology. There was an increase in the basal respiration rate of the soil by the application of sheep manure and this was consistent with the increase of the doses. The survival rate and avoidance behavior of springtails were not altered and there was no change in the initial performance of oats, indicating that this manure can be used for organic fertilization of soils with low soil pollutant potential.

Genotoxic and mutagenic effects of chlorothalonil on the estuarine fish Micropogonias furnieri (Desmarest, 1823).

Chlorothalonil is a fungicide widely used in agriculture as well as an active ingredient in antifouling paints. Although it causes toxic effects on non-target organisms and can accumulate in fish tissues, little is known about its sublethal effects. Thus, genotoxic and mutagenic effects of intraperitoneal injected chlorothalonil in Micropogonias furnieri, an estuarine fish of frequent human consumption and a promising test-organism for ecotoxicological assays, were assessed. Chlorothalonil showed to be genotoxic (DNA damage by comet assay) and mutagenic (micronuclei, nuclear buds, apoptotic fragments, and bilobed cells) even at the lowest dose tested (0.35μgg-1) and in a dose-dependent manner (0.35 and 3.5μgg-1) for micronuclei, apoptotic fragments, and bilobed cells. As genomic instability may lead to carcinogenesis, the present evidence can assist decision-makers in banning this compound since any benefit toward food production is outweighed by the hazard to aquatic ecosystems and human health.

Single toxicity of arsenic and combined trace metal exposure to a microalga of ecological and commercial interest: Diacronema lutheri

Eco-toxicological assays with species of economic interest such as Diacronema lutheri are essential for industries that produce aquaculture feed, natural food additives and also in drug developing industries. Our study involved the exposure of a single and combined toxicity of arsenic (As V) to D. lutheri for the entire algal growth phase and highlighted that a combined exposure of As V with other essential (Copper, Cu; Nickel, Ni) and non-essential (Cadmium, Cd; Lead, Pb) trace metals reduced significantly the cell number, chlorophyll a content, and also significantly increased the de-epoxidation ratio (DR) as a stress response when compared to the single toxicity of As V. Arsenic, as one of the ubiquitous trace metal and an active industrial effluent is reported to have an increased bio-concentration factor when in mixture with other trace metals in this study. In the combined exposure, the concentration of total As bio-accumulated by D. lutheri was higher than in the single exposure. Hence, polluted areas with the prevalence of multiple contaminants along with the highly toxic trace metals like As can impose a greater risk to the exposed organisms that may get further bio-magnified in the food chain. Our study highlights the consequences and the response of D. lutheri in terms of contamination from single and multiple trace metals in order to obtain a safer biomass production for the growing need of natural derivatives.

Implications of kinetically-hindered metals in ecotoxicological studies: Effect of platinum spike aging on its toxicity to Dunaliella salina

Platinum (Pt) is considered an emerging environmental micro-contaminant due to its increasing use in anthropogenic activities during the past decades. However, there are still important gaps in the understanding of its biogeochemical behavior in the aquatic environment – e.g. its speciation, reactivity and fate – mainly as a result of the analytical challenge of the determination of its typical ultra-trace environmental concentrations. Also, Pt is a kinetically-hindered metal displaying slow reaction kinetics, which has important implications regarding eco-toxicological studies. That is, investigation of its toxicity under laboratory-controlled conditions may therefore require ensuring that equilibrium speciation conditions are reached before starting the experiments. In order to shed further light on this issue, in this study we have monitored the speciation changes during aging of the Pt(IV) spikes in controlled media (seawater) using an UV-Vis spectrophotometry. Platinum toxicity to the green microalgae Dunaliella salina was then compared, using standardized tests, with fresh and aged Pt(IV) spikes at the mg L−1 concentration range. Following 96-hour exposure, ecotoxicological assays consisting in spectrometric measurements of chlorophyll-a concentrations and Effective Concentrations (EC) of Pt resulting in the inhibition of 10% and 50% of algae growth rate were calculated (EC10 and EC50, respectively). Daily monitoring of Pt speciation reflected the transition from PtCl62- (spike) to hydrolyzed species, probably in the form [PtCl3−n(OH)3+n]2-, n = 0–3. Exposure experiments showed that after a short period of aging (10 days), Pt(IV) toxicity increased one order of magnitude compared to freshly spiked media. These results confirm the relevance of considering spike aging to ensure that speciation equilibrium conditions are attained in order to produce environmental realistic eco-toxicological data.

Effects of the endocrine disruptor 4-nonylphenol on the demography of rotifers Plationus patulus and Brachionus havanaensis: a multigenerational study

The emerging contaminant 4-nonylphenol is an active ingredient in many surfactants. Though 4-nonylphenol has been reported in Mexican waterbodies, no stringent law is available to establish its permissible limits. Most ecotoxicological assays using zooplankton species are based on previously unexposed populations, and multigenerational experiments are rare. Plationus patulus and Brachionus havanaensis are widely distributed species in Mexico and have been used in ecotoxicological assessments. In this work, the median lethal concentration of 4-nonylphenol (LC50, 24 h) for both rotifer species was derived. Based on our acute toxicity data (24 h LC50 of 4-nonylphenol for P. patulus and B. havanaensis were 500 and 250 µg L−1, respectively), three sublethal concentrations (µg L−1) (7.81, 15.82 and 31.25 for B. havanaensis and 15.62, 31.25 and 62.5 for P. patulus) were used in the life table demography tests for two successive generations (F0 and F1) of either rotifer species. Regardless of the concentrations used, 4-nonylphenol had an adverse effect on both rotifer species over two successive generations. P. patulus was more sensitive than B. havanaensis in chronic toxicity tests. F1 generation of both rotifer species was far more adversely affected than the F0 in all toxicant treatments.

A Multidisciplinary Approach to Evaluate the Effects of Contaminants of Emerging Concern on Natural Freshwater and Brackish Water Phytoplankton Communities.

Simple SummaryZinc oxide nanoparticles (ZnO NPs) and potassium dichromate (K2Cr2O7) are two contaminants of emerging concern (CECs; chemicals not commonly monitored in the environment but has the potential to enter the environment and cause known or suspected adverse ecological and/or human health effects). On this path, phytoplankton species, because of their photosynthetic activity, are vital for providing oxygen, and environmental impacts on such organisms can disrupt an entire ecosystem’s integrity. Generally, ecotoxicological assays on monospecific phytoplankton cultures provide useful information about the cellular effects of toxic compounds; however, they have limited application for detecting the effects of environmental pollutants on multiple species communities like in nature. For this reason, in this study, we took for the first time an ecotoxicological (growth rate and inhibition growth rate), ecological (taxonomic identification of species and diversity in communities), and biochemical (photosynthetic pigments) approach to evaluate the effects of ZnO NPs and K2Cr2O7 on natural freshwater and brackish water phytoplankton communities. Results show that both chemicals have negative effects on natural phytoplankton communities with an alteration of the growth rate, species composition, and photosynthetic activity. However, the exposure to ZnO 10 mg/L acts as a growth stimulant for phytoplanktonic communities. Our findings provide evidence for alterations in natural phytoplankton after exposure to such CECs.Ecotoxicological assays on monospecific phytoplankton have limited application for detecting the effects of environmental pollutants on multiple species communities. With this study, we took an ecotoxicological, ecological, and biochemical approach to evaluate the effects of two contaminants of emerging concern (zinc oxide nanoparticles, ZnO NPs, and potassium dichromate, K2Cr2O7) at different concentrations (K2Cr2O7 5.6–18–50 mg/L; ZnO NPs 10–100–300 mg/L) on natural freshwater and brackish water phytoplankton communities. Cell density and absorbance values decreased in freshwater and brackish water phytoplankton communities after exposure to ZnO NPs (100 mg/L and 300 mg/L only for freshwater), whereas growth rate was increased in both freshwater and brackish water phytoplankton communities after exposure to ZnO NPs 10 mg/L. Differently, there was no clear relationship between concentration and inhibition growth after exposure to K2Cr2O7: the lowest cell density was recorded after exposure to 18 mg/L. Moreover, the evenness index value was lower compared to the other concentrations, indicating the growth of a few, albeit resistant species to higher K2Cr2O7 concentrations. Generally, Bacillariophyceae and Dinoficee were prevalent in phytoplankton cultures after exposure to ZnO NPs and K2Cr2O7. The Shannon-Wiener index was slightly higher in the negative than the positive controls, but diversity was low after all treatments in both ecotoxicological assays. The evenness index was always very close to zero, indicating the numerical predominance of one or very few species. Finally, the decrease in chlorophyll-a and pheophytin-a in both ecotoxicological assays indicated a change in photosynthetic activity. Our findings provide evidence for alterations in natural phytoplankton after exposure to emerging contaminants that can disrupt an entire ecosystem’s integrity.

Unlocking Kautsky’s dark box: Development of an optical toxicity classification tool (OPTOX index) with marine diatoms exposed to emerging contaminants

Chlorophyll a induction curves, or Kautsky curves, have been extensively used to study physiological stress conditions in phototrophic organisms, with the analysis of several derived parameters. Nevertheless, these variables use only about 10 % of the information comprised in the complete Kautsky curve dataset, leaving 90 % of the photochemical data within an underutilized dark box, that is not translated into photochemically relevant variables. By observing the variable fluorescence profiles from marine diatoms exposed to a myriad of emerging and classical contaminants, several fluorescence profile alterations were detected, with significant deviations from the control conditions concomitant with the degree of growth inhibition imposed by the chemical stressor. The Linear Discriminant Analysis (LDA) analysis based on the normalized variable chlorophyll a fluorescence profiles revealed a high discriminatory efficiency of the type of contaminant to which the cultures were exposed, indicating that the exposure to different chemical stressors (contaminants) results in specific fluorescence profiles that can be used as descriptors of these exposure conditions. Analysing the individual contaminant LDA analysis, a very low overlap between samples exposed to different concentrations was observed, indicating a high discriminatory power of the variable fluorescence profiles. When evaluating the blind-test classification efficiencies, provided by this contaminant-specific LDA approach, it was possible to observe a high degree of efficiency in almost all contaminants tested, and for most of the concentrations applied. With this in mind, the produced linear discriminants and proportion of traces was used to compute an optical toxicity classification tool - the OPTOX index. The index revealed a high degree of correlation with the growth inhibition observed and/or with the exogenous dose of contaminant applied. The developed OPTOX index, a unifying tool enclosing all the fluorescence data provided by the chlorophyll a induction curve, proved to be an efficient tool to apply in ecotoxicological assays using marine model diatoms with a high degree of reliability for classifying the exposure of the cells to emerging contaminants. Additionally, the data analysis pipeline, as well as the index development methodology here proposed, can be easily transposed to other autotrophic organisms subjected to different ecotoxicological test conditions calibrated and validated against known biochemical or morphological descriptors of stress, integrating this way a large amount of data that was until know completely overlooked and left within an underutilized and undervalued dark box.

Gut and faecal bacterial community of the terrestrial isopod Porcellionides pruinosus: potential use for monitoring exposure scenarios.

This work aimed to characterize the gut and faeces bacterial communities (BC) of Porcellionides pruinosus using high-throughput sequencing. Isopods were collected from the field and kept in laboratory conditions similar to those normally applied in ecotoxicology tests. Faeces and purged guts of isopods (n = 3 × 30) were analysed by pyrosequencing the V3-V4 region of 16 S rRNA encoding gene. Results showed that gut and faecal BCs were dominated by Proteobacteria, particularly by an OTU (Operational Taxonomic Unit) affiliated to genus Coxiella. Diversity and richness values were statistically higher for faecal BC, mainly due to the occurrence of several low-abundance phylotypes. These results may reflect faecal carriage of bacterial groups that cannot settle in the gut. BCs of P. pruinosus comprised: (1) common members of the soil microbiota, (2) bacterial symbionts, (3) bacteria related to host metabolic/ecological features, and (4) bacterial etiological agents. Comparison of BC of this isopod species with the BC from other invertebrates revealed common bacterial groups across taxa. The baseline information provided by this work will assist the design and data interpretation of future ecotoxicological or biomonitoring assays where the analysis of P. pruinosus BC should be included as an additional indicator. CAPSULE: Terrestrial isopods bacterial communities might support ecotoxicological assays and biomonitoring processes as a valuable tool.

Assessment of effluents quality through ecotoxicological assays: evaluation of three wastewater treatment plants with different technologies.

The water quality of the effluents is mainly focused on physicochemical and microbiological parameters. However, the ecotoxicological assessments are crucial to ensure an effective water quality of the effluents. This work aims to assess the ecotoxicity of effluents originated from WWTPs with different wastewater treatment technologies. For that, effluent samples from three WWTPs with different treatment processes were seasonally collected. Physicochemical parameters were determined, the toxicity towards daphnia, protozoan, and microalgae organisms was evaluated, and data correlated. Toxicity assays showed different susceptibility of the organisms to the effluents and that toxicity is dependent on the season and wastewater treatment technology. No toxicity was observed to daphnia in winter and spring, but ~100% of mortality was observed in effluent from WWTP A in summer. Growth inhibition was observed for both protozoan and microalgae for all effluents and in all seasons with highest values in spring in WWTP C (~80%) for the protozoan while the highest microalgae growth inhibition percentage was observed for WWTP B in both spring (~80%) and summer (~80%). These results show that effluents might have negative impacts into their receiving water systems and highlight that a global assessment of effluent quality should include ecotoxicological assays to complement physicochemical and microbiological data for an operative environmental management of wastewater treatment plants.

A Comparison of Short-Term and Continuous Exposures in Toxicity Tests of Produced Waters, Condensate, and Crude Oil to Marine Invertebrates and Fish.

Petroleum hydrocarbons can be discharged into the marine environment during offshore oil and gas production or as a result of oil spills, with potential impacts on marine organisms. Ecotoxicological assay durations (typically 24–96 h) used to characterize risks to exposed organisms may not always reflect realistic environmental exposure durations in a high‐energy offshore environment where hydrocarbons are mixed and diluted rapidly in the water column. To investigate this, we adapted 3 sensitive toxicity tests to incorporate a short‐term pulse exposure to 3 petroleum‐based products: a produced water, the water‐accommodated fraction (WAF) of a condensate, and a crude oil WAF. We measured 48‐h mobility of the copepod Acartia sinjiensis, 72‐h larval development of the sea urchin Heliocidaris tuberculata, and 48‐h embryo survival and deformities of yellowtail kingfish Seriola lalandi, after exposure to a dilution series of each of the 3 products for 2, 4 to 12, and 24 h and for the standard duration of each toxicity test (continuous exposure). Effects on copepod survival and sea urchin larval development were significantly reduced in short‐term exposures to produced water and WAFs compared to continuous exposures. Fish embryos, however, showed an increased frequency of deformities at elevated concentrations regardless of exposure duration, although there was a trend toward increased severity of deformities with continuous exposure. The results demonstrate how exposure duration alters toxic response and how incorporating relevant exposure duration to contaminants into toxicity testing may aid interpretation of more realistic effects (and hence an additional line of evidence in risk assessment) in the receiving environment. Environ Toxicol Chem 2021;40:2587–2600. © 2021 CSIRO. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Insights into the Restoration of Tributyltin Contaminated Environments Using Marine Bacteria from Portuguese Fishing Ports

Tributyltin (TBT) is an organotin chemical mainly used as biocide in marine antifouling paints. Despite the restrictions and prohibitions on its use, TBT is still an environmental problem due to its extensive application and subsequent release into the environment, being regarded as one of the most toxic chemicals released into the marine ecosystems. Microorganisms inhabiting impacted sites are crucial for their restoration since they have developed mechanisms to tolerate and break down pollutants. Nonetheless, transformation products resulting from the degradation process may still be toxic or, sometimes, even more toxic than the parent compound. The determination of the parent and degradation products by analytical methods, although necessary, may not be ecologically relevant since no information is provided regarding their ecotoxicity. In this study, marine bacteria collected from seven Portuguese fishing ports were isolated and grown in the presence of TBT. Bacteria that exhibited higher growth were used to bioremediate TBT-contaminated waters. The potential of these bacteria as bioremediation agents was evaluated through ecotoxicological assays using the sea snail Gibbula umbilicalis as model organism. Data suggested that some TBT-tolerant bacteria, such as Pseudomonas putida, can reduce the toxicity of TBT contaminated environments. This work contributes to the knowledge of TBT-degrading bacteria.

Polypyrrole-Based Nanorobots Powered by Light and Glucose for Pollutant Degradation in Water.

Novel photoactive and enzymatically active nanomotors were developed for efficient organic pollutant degradation. The developed preparation route is simple and scalable. Light-absorbing polypyrrole nanoparticles were equipped with a bi-enzyme [glucose oxidase/catalase (GOx/Cat)] system enabling the simultaneous utilization of light and glucose as energy sources for jet-induced nanoparticle movement and active radical production. The GOx utilizes glucose to produce hydrogen peroxide, which is subsequently degraded by Cat, resulting in the generation of active radicals and/or oxygen bubbles that propel the particles. Uneven grafting of GOx/Cat molecules on the nanoparticle surface ensures inhomogeneity of peroxide creation/degradation, providing the nanomotor random propelling. The nanomotors were tested for their ability to degrade chlorophenol, under various experimental conditions, that is, with and without simulated sunlight illumination or glucose addition. In all cases, degradation was accelerated by the presence of the self-propelled nanoparticles or light illumination. Light-induced heating also positively affects enzymatic activity, further accelerating nanomotor diffusion and pollutant degradation. In fact, the chemical and photoactivities of the nanoparticles led to more than 95% removal of chlorophenol in 1 h, without any external stirring. Finally, the quality of the purified water and the extent of pollutant removal were checked using an eco-toxicological assay, with demonstrated significant synergy between glucose pumping and sunlight illumination.

Effects of Land Use and Pollution Loadings on Ecotoxicological Assays and Bacterial Taxonomical Diversity in Constructed Wetlands

Freshwater ecosystems are affected by anthropogenic alterations. Different studies have extensively studied the concentrations of metals, nutrients, and water quality as measurements of pollution in freshwater ecosystems. However, few studies have been able to link these pollutants to bioindicators as a risk assessment tool. This study aimed to examine the potential of two bioindicators, plant ecotoxicological assays and sediment bacterial taxonomic diversity, in ecological risk assessment for six freshwater constructed wetlands in a rapidly urbanizing watershed with diverse land uses. Sediment samples were collected summer, 2015 and 2017, and late summer and early fall in 2016 to conduct plant ecotoxicological assays based on plant (Lepidium, Sinapis and Sorghum) growth inhibition and identify bacterial taxonomical diversity by the 16S rRNA gene sequences. Concentrations of metals such as lead (Pb) and mercury (Hg) (using XRF), and nutrients such as nitrate and phosphate (using HACH DR 2800TM spectrophotometer) were measured in sediment and water samples respectively. Analyses of response patterns revealed that plant and bacterial bioindicators were highly responsive to variation in the concentrations of these pollutants. Hence, this opens up the scope of using these bioindicators for ecological risk assessment in constructed freshwater wetland ecosystems within urbanizing watersheds.

Acartia tonsa Dana 1849 as a Model Organism: Considerations on Acclimation in Ecotoxicological Assays.

The copepod Acartia tonsa was standardized as model organism in acute toxicity bioassays due to its key position in coastal food chains and high sensitivity. Once bioassays are performed according to a protocol their results may become tools for the protection of aquatic ecosystems. However, there are divergences in bioassays methods using A. tonsa. This study aimed to investigate: (i) the need for acclimation of A. tonsa collected from the environment for use in acute toxicological bioassays; and (ii) differences in sensitivity between copepods collected from the environment and laboratory-grown copepods. Laboratory-grown copepods are more sensitive to SDS thanA. tonsa from theenvironment. The acclimation time of 30h helped organisms to recover from stress of collection/handling and changing environment/conditions. Therefore, laboratory-grown copepods showed to be more sensitive than organisms from environment; and for ecotoxicological bioassays acclimating A. tonsa collected from the environment for 30h can be adopted.

Toxicity of metal cations and phenolic compounds to the bioluminescent fungus Neonothopanus gardneri

Fungi play a key role in the soil ecosystem, where they occupy the first level of the food chain. Hence, they are considered suitable model organisms to conduct ecotoxicological assays for the evaluation of soil condition after proper soil extraction. Fungi bioluminescence-based bioassays are useful considering that their luminescence is an early toxicity endpoint and light emission a promptly detectable signal. In this paper, we describe a toxicological bioassay that relies on a 24-h variation of total light emitted by the mycelium of the bioluminescent fungus Neonothopanus gardneri when exposed to a toxicant. The current bioluminescent assay, which uses a fungus of the Omphalotus lineage, fills a gap covering all the representative species of bioluminescent fungi. Among the compounds tested here, Cd(II) showed the highest toxicity, followed by 4-nitrophenol, phenol and Cu(II), respectively. We also found that N. gardneri presents a predictable bioluminescence and growth pattern, and is highly sensitive to these compounds. The aforementioned characteristics offer valuable advantages and make N. gardneri the ideal candidate for toxicological studies with basidiomycetes.

The sensitivity of multiple ecotoxicological assays for evaluating Microcystis aeruginosa cellular algal organic matter and contribution of cyanotoxins to the toxicity

Secondary metabolites of cyanobacteria and algae released during algal blooms often exhibit toxic effects, but only a small number of the metabolites are the subject of routine analytical screenings. Alternatively, ecotoxicological assays offer a better representation of the overall negative effects. The aim of this work was to compare multiple assays in their sensitivity towards cellular algal organic matter (COM) of the toxin-producing cyanobacterium Microcystis aeruginosa. Multiple endpoints were investigated: mortality, growth inhibition, bioluminescence inhibition, genotoxicity, endocrine-disrupting effects, oxidative stress, and the induction of ethoxyresorufin-O-deethylase (EROD). Three rainbow trout (Oncorhynchus mykiss) cell lines as well as representatives of bacteria, yeasts, algae, vascular plants, and crustaceans were employed, and the results were expressed per mg of dissolved organic carbon (DOC) in the COM. M. aeruginosa COM was toxic to the RTgill-W1, RTG-2, and RTL-W1 cell lines (EC50 values ranging from 0.48 ± 0.02 to 1.9 ± 0.1 mgDOC/L), to the crustacean Thamnocephalus platyurus (LC50 = 20 ± 1 mgDOC/L), and to Lepidium sativum (IC50 = 241 ± 13 mgDOC/L). In contrast, no effect was observed for bacteria and yeasts, and the growth of the alga Desmodesmus subspicatus was even stimulated. No genotoxicity, endocrine-disrupting effects or increase in oxidative stress or EROD activity was detected. The content of six microcystins (MC-LR, MC-RR, MC-YR, MC-LY, MC-LW, and MC-LF), anatoxin-a, cylindrospermopsin, and nodularin in the M. aeruginosa COM was determined by liquid chromatography–tandem mass spectrometry. An artificially prepared mixture of the detected cyanotoxins in the corresponding concentrations did not induce response in the O. mykiss cell lines and T. platyurus, suggesting that other cyanobacterial metabolites are responsible for the toxicity of M. aeruginosa.

Organic solvents alter photophysiological and oxidative stress profiles of the coral Zoanthus sp. - Towards an optimization of ecotoxicological protocols.

Coral reefs are declining, affected by climate change and escalating anthropogenic pressures, such as pollution or habitat alteration. Consequently, ecotoxicological assays with tropical corals have increased, specifically towards the study of emergent or persistent pollutants. However, standardized methodology to test for corals is non-existent, and their response to organic solvents, recurrently required in ecotoxicological appraisals, remains unknown. Therefore, we aimed to establish a threshold for the safe use of the selected solvents in ecotoxicological studies with these organisms. We assessed the oxidative stress response (antioxidant response and oxidative damage), cellular energy allocation and photophysiology of the photosynthetic coral Zoanthus sp. (Anthozoa, Hexacorallia) exposed to six doses of three different organic solvents (ethanol, methanol and dimethyl sulfoxide - DMSO). Our results suggest that the coral is more sensitive to methanol and DMSO than to ethanol. Methanol and DMSO LOEC were 0.01mLL-1 affecting maximum quantum yield (Fv/Fm) and glutathione S-transferase (GST) activity, respectively, while for ethanol was 0.03mLL-1, influencing Fv/Fm. Despite the higher tolerance of Zoanthus sp. to ethanol, 2.9mLL-1 of this organic solvent was the only treatment causing mortality. Based on these findings, thresholds for the use of organic solvents with tropical corals can now be adopted. Nevertheless, species specificities should not be overlooked.