Large-scale ecotoxicological studies have technical and ethical limitations, both related to the need to expose large numbers of individuals to potentially harmful compounds. The computational modeling is a complementary useful and predictive tool that overcomes these limitations. Considering the increasing interest in the effects of pesticides on behavioral traits, the aim of this study was to evaluate the effects of chlorpyrifos (CPF) on intra- and inter-specific interactions of anuran tadpoles, complementing traditional ecotoxicological tools with a theoretical analysis verified by computational simulations. Experiments were developed under two consecutive phases: a first phase of exposure (treated and control group), and a second phase of interactions. The second phase consisted of evaluating the effects of CPF on intra- and inter-specific interactions of exposed C. ornata (Co) tadpoles acting as predators and unexposed Rhinella fernandezae (Rf) tadpoles acting as prey (Experiment I), under different predator-prey proportions (0/10 = 0Co–10Rf, 2/8, 4/6, 6/4, 8/2, 10/0). Also, intraspecific interactions of three Co tadpoles under different conditions of exposure were evaluated (Experiment II: 3 exposed Co, 2 exposed Co/1 non-exposed, 1 exposed Co/2 non-exposed). During the exposure phase, chlorpyrifos induced significant mortality from 48 h (48 h: p < 0.05, 72 h–96 h: p < 0.001), irregular swimming, tail flexure, and the presence of subcutaneous air. Also, it induced effects on the sounds emitted after 96 h of exposure, registering a smaller number of pulses and higher dominant frequencies, and altered intra- and inter-specific interactions. During the interaction phase, the larvae continued to show sound effects, however, the antipredator mechanism continued to be operating and efficient. Finally, it was possible to model the behavior of the larvae under the effects of chlorpyrifos. We conclude that experimental data and computational modeling matched. Therefore, computational simulation is a valuable ecotoxicological tool that provides new information and allows prediction of natural processes.

Microplastics are highly bioavailable to marine organisms, either through direct ingestion, or indirectly by trophic transfer from contaminated prey. The latter has been observed for low-trophic level organisms in laboratory conditions, yet empirical evidence in high trophic-level taxa is lacking. In natura studies face difficulties when dealing with contamination and differentiating between directly and indirectly ingested microplastics. The ethical constraints of subjecting large organisms, such as marine mammals, to laboratory investigations hinder the resolution of these limitations. Here, these issues were resolved by analysing sub-samples of scat from captive grey seals (Halichoerus grypus) and whole digestive tracts of the wild-caught Atlantic mackerel (Scomber scombrus) they are fed upon. An enzymatic digestion protocol was employed to remove excess organic material and facilitate visual detection of synthetic particles without damaging them. Polymer type was confirmed using Fourier-Transform Infrared (FTIR) spectroscopy. Extensive contamination control measures were implemented throughout. Approximately half of scat subsamples (48%; n = 15) and a third of fish (32%; n = 10) contained 1–4 microplastics. Particles were mainly black, clear, red and blue in colour. Mean lengths were 1.5 mm and 2 mm in scats and fish respectively. Ethylene propylene was the most frequently detected polymer type in both. Our findings suggest trophic transfer represents an indirect, yet potentially major, pathway of microplastic ingestion for any species whose feeding ecology involves the consumption of whole prey, including humans.

As a result of the efforts to introduce the principle of the 3Rs (replacement, reduction, and refinement) into animal testing, alternative in vitro skin corrosion test methods have been developed and standardized globally. However, alternative in vitro skin corrosion test methods have some limitations in terms of the use of humanely killed rats or commercial models and kits. The present study focused on the applicability of invertebrates as alternative in vivo skin models. Even though earthworm skin comprises the same biomolecules as human skin, the possibility of using earthworm skin as an alternative for skin testing remains unexplored. In this study, we developed a half-pipe tool for earthworm skin corrosion testing and optimized the test protocol. Subsequently, the applicability of the earthworm half-pipe assay for corrosion testing with six chemicals, including inorganic acids, organic acids, and alkalis, was investigated using stereomicroscopy and electron microscopy. It was observed that the specific concentrations for earthworm skin corrosion were lower than those for animal or in vitro tests. Therefore, the sensitivity of the earthworm half-pipe assay indicates that it could be useful as a screening tool before conducting in vivo animal tests or in vitro skin tests. This new method can contribute to research on alternative skin corrosion tests by reducing ethical issues, time, and cost while achieving effective results.

The world's oceans are under increasing pressure from anthropogenic activities, including significant and rapidly increasing inputs of plastic pollution. Seabirds have long been considered sentinels of ocean health, providing data on physical and chemical pollutants in their marine habitats. However, long-term data that can elucidate important patterns and changes in seabird exposure to marine pollutants are relatively limited but are urgently needed to identify and support effective policy measures to reduce plastic waste. Using up to 12 years of data, we examined the benefits and challenges of different approaches to monitoring plastic in seabirds, and the relationship between plastic and body size parameters. We found the mass and number of ingested plastics per bird varied by sample type, with lavage and road-kill birds containing less plastic (9.17–9.33 pieces/bird) than beach-washed or otherwise dead birds (27.62–32.22 pieces/bird). Beached birds therefore provide data for only a particular subset of the population, mostly individuals in poorer body condition, including those severely impacted by plastics. In addition, the mass and number of plastics in beached birds were more variable, therefore the sample sizes required to detect a change in plastic over time were significantly larger than for lavaged birds. The use of lavaged birds is rare in studies of plastic ingestion due to ethical and methodological implications, and we recommend future work on ingested plastics should focus on sampling this group to ensure data are more representative of a population's overall exposure to plastics.

Criteria continuous concentrations (CCCs) are useful for describing chronic exposure to pollutants and setting water quality standards to protect aquatic life. However, because of financial, practical, or ethical restrictions on toxicity testing, few data are available to derive CCCs. In this study, CCCs for 34 metals or metalloids were derived using quantitative ion character-activity relationships–species sensitivity distributions (QICAR–SSD) and the final acute-chronic ratio (FACR) method. The results showed that chronic toxic potencies were correlated with several physico-chemical properties among eight species chosen, where the softness index was the most predictive characteristic. Predicted CCCs for most of the metals, except for Lead and Iron, were within a range of 10-fold of values recommended by the U.S. EPA. The QICAR–SSD model was superior to the FACR method for prediction of data-poor metals. This would have significance for predicting toxic potencies and criteria thresholds of more metals or metalloids.

Estrogenic endocrine disrupting chemicals (EDCs) are natural hormones, synthetic compounds or industrial chemicals that mimic estrogens due to their structural similarity with estrogen's functional moieties. They typically enter aquatic environments through wastewater treatment plant effluents or runoff from intensive livestock operations. Globally, most natural and synthetic estrogens in receiving aquatic environments are in the low ng/L range, while industrial chemicals (such as bisphenol A, nonylphenol and octylphenol) are present in the μg to low mg/L range. These environmental concentrations often exceed laboratory-based predicted no effect concentrations (PNECs) and have been evidenced to cause negative reproductive impacts on resident aquatic biota. In vertebrates, such as fish, a well-established indicator of estrogen-mediated endocrine disruption is overexpression of the egg yolk protein precursor vitellogenin (Vtg) in males. Although the vertebrate Vtg has high sensitivity and specificity to estrogens, and the molecular basis of its estrogen inducibility has been well studied, there is growing ethical concern over the use of vertebrate animals for contaminant monitoring. The potential utility of the invertebrate Vtg as a biomonitor for environmental estrogens has therefore gained increasing attention. Here we review evidence providing support that the molluscan Vtg holds promise as an invertebrate biomarker for exposure to estrogens. Unlike vertebrates, estrogen signalling in invertebrates remains largely unclarified and the classical genomic pathway only partially explains estrogen-mediated activation of Vtg. In light of this, in the latter part of this review, we summarise recent progress towards understanding the molecular mechanisms underlying the activation of the molluscan Vtg gene by estrogens and present a hypothetical model of the interplay between genomic and non-genomic pathways in the transcriptional regulation of the gene.

In vitro methods have gained rising importance in ecotoxicology due to ethical concerns. The aim of this study was to assess the single and combined in vitro effects of gold, as nanoparticle (AuNPs) and ionic (Au⁺) form, and the pharmaceutical gemfibrozil (GEM). Sparus aurata liver organ culture was exposed to gold (4 to 7200 μg·L⁻¹), GEM (1.5 to 15,000 μg·L⁻¹) and combination 80 μg·L⁻¹ gold +150 μg·L⁻¹ GEM for 24 h. Endpoints related with antioxidant status, peroxidative/genetic damage were assessed. AuNPs caused more effects than Au⁺, increasing catalase and glutathione reductase activities and damaging DNA and cellular membranes. Effects were dependent on AuNPs size, coating and concentration. GEM damaged DNA at an environmentally relevant concentration, 1.5 μg·L⁻¹. Overall, the effects of the combined exposures were higher than the predicted, based on single exposures. This study showed that liver culture can be a useful model to study contaminants effects.

Exposure to complex organic substances present in textile wastewater has been considered a threat to human health and aquatic organisms. Development of appropriate treatment mechanisms, as well as sensitive monitoring assays, is considered important in order to safeguard and protect the delicate natural equilibrium in the environment. In this study, combined coagulation/flocculation and electrohydraulic discharge (EHD) system were explored for treatment of textile wastewater. Pre- and post-treatment samples were used to evaluate process efficiencies. Process efficiencies were evaluated using physicochemical characteristics, and cytotoxicity and inflammatory activities induced in macrophage RAW264.7 cell line. The RAW264.7 cell line was evaluated as an alternative to animals and human blood culture models, whose routine applications are hindered by stern ethical requirements. The toxicity of effluent was evaluated using WST-1 assay. The inflammatory activities were evaluated in RAW264.7 cell culture supernatant using nitric oxide (NO) and interleukin 6 (IL-6) as biomarkers of inflammation. The levels of NO and IL-6 were determined using the Griess reaction assay and double-antibody sandwich enzyme-linked immunoassay (DAS ELISA), respectively. Overall, the results of this study show that combined approaches and not the single EHD system are sufficient for complete removal of chemical oxygen demand (COD) and total organic carbon (TOC), toxicity and inflammatory activities in textile wastewater. The study shows that induction of NO and IL-6 secretions in macrophage RAW264.7 cells is a very sensitive model system to monitor the efficiency of textile effluent treatment processes.

To address the frequent emergence of ecological problems, ecology has intersected with various disciplines. From the perspective of linguistic ecology, ecological literacy is an important concept that combines the subjects of ecology and linguistics. It not only discusses ecological issues, but also establishes a linguistic framework. Here, we constructed a quantitative method of assessing ecological literacy from the perspective of linguistic ecology. Ecological literacy was divided into five parts: ecological knowledge literacy, ecological awareness literacy, ecological ethics literacy, ecological emotional literacy, and ecological behavioral literacy. Each of these was set with four quantitative indicators that were evaluated through eight questions. A case study was conducted to investigate the ecological literacy of the inhabitants of Guiyang City, one of China’s top ten ecologically advanced cities. The results showed that the proposed assessment method was an effective way to evaluate the level of ecological literacy comprehensively. In the case analysis, the overall ecological literacy level of Guiyang inhabitants was relatively good, and the levels of the five specific dimensions of them in descending order were as follows: ecological ethics literacy, ecological emotional literacy, ecological awareness literacy, ecological knowledge literacy, and ecological behavioral literacy. The results of this study are conducive to the production of targeted ways to improve the level of ecological literacy for sustainable development.

The integration of Industry 4.0 (I4.0) has emerged as an innovative paradigm for industrial firms contemplating environmental and economic issues. This study explicates the role of I4.0 technologies (I4.0TEC) in reinforcing the management of environmental assets (ENVASS) as well as optimizing financial performance (FP). The data in this research was collected from 738 industrial firms in Malaysia between 2009 and 2018. The analyses of ordinary least square statistics (OLS) and structural equation modeling (SEM) delineated three major findings. The individual effect of ENVASS, robotization, and flexibility in production technologies has a marginal impact on sales, exports, and labor productivity indicators. The complementarities of these variables represent a similar effect on the performance indicators. The findings related to gross operating margin elucidate that ENVASS and I4.0TEC have neither individual nor complementarity effects. This was explained by developing a robust model by integrating ENVASS, I4.0TEC, spending and investing in R&D, flexibility in production, and human capital management. Our findings have confirmed that the proposed model offers a functional toolkit for the firms considering optimizing their profitability by leveraging ENVASS and I4.0TEC. This research also contributes to developing an ethical business model for the circular economy.