As agriculture expands to provide food and wellbeing to the world’s growing population, there is a simultaneous increasing concern about the use of agrochemicals, which can harm non-target organisms, mainly in the aquatic environment. The fungicide Mancozeb (MZ) has been used on a large-scale and is a potent inducer of oxidative stress. Therefore, there is an urgent need for the development of more sensitive biomarkers designed to earlier biomonitoring of this compound. Here we tested the hypothesis that behavioral changes induced by sublethal MZ concentrations would occur first as compared to biochemical oxidative stress markers. Embryos at 4 h post-fertilization (hpf) were exposed to Mancozeb at 5, 10 and 20 μg/L. Controls were kept in embryo water only. Behavioral and biochemical parameters were evaluated at 24, 28, 72, and 168 hpf after MZ exposure. The results showed that MZ significantly altered spontaneous movement, escape responses, swimming capacity, and exploratory behavior at all exposure times. However, changes in ROS steady-stead levels and the activity of antioxidant enzymes were observable only at 72 and 168 hpf. In conclusion, behavioral changes occurred earlier than biochemical alterations in zebrafish embryos exposed to MZ, highlighting the potential of behavioral biomarkers as sensitive tools for biomonitoring programs.

The immune organs, like thymus, are one of the targets of hydrogen sulfide (H₂S). Previously we reported that H₂S induced the differential expression of mRNAs that implicating apoptosis in thymus, however, the roles of noncoding RNAs (ncRNAs) in H₂S-induced thymus injury are still unknown. Pollution gases could alter the expression of ncRNAs, which have been shown to play important roles in many physiological and pathophysiological processes, including immune activity. This study revealed that H₂S exposure induced 9 differentially expressed circRNAs and 15 differentially expressed miRNAs in chicken thymus. Furthermore, the circRNA - miRNA - mRNA network was constructed. We discovered that circR-PTPN23 - miR-15a - E2F3 was involved in the cell cycle and apoptosis. Further, an in vitro H₂S exposure model was established using HD11 cell line and demonstrated that H₂S suppressed cell proliferation and induced apoptosis. Moreover, ciR-PTPN23 and E2F3 were downregulated, but miR-15a was upregulated in both the thymus and HD11 cell line after H₂S exposure. Bioinformatics analysis revealed that ciR-PTPN23 directly bound to miR-15a and that E2F3 was the target gene of miR-15a. Knocking down ciR-PTPN23 suppressed HD11 proliferation and caused G1 arrest and apoptosis, however, this phenomenon could be partially reversed by ciR-PTPN23 overexpression or miR-15a silencing. In summary, the ciR-PTPN23 - miR-15a - E2F3 axis was involved in H₂S-induced cell proliferation suppression and apoptosis.

The long term vertical and horizontal mobility of mercury (Hg) in soils of agricultural areas of a historically contaminated Italian National Relevance Site (SIN Brescia-Caffaro) was investigated. The contamination resulted from the continuous discharge of Hg in irrigation waters by an industrial plant (Caffaro S.p.A), equipped with a mercury-cell chlor-alkali process. The contamination levels with depth ranged from about 20 mg/kg dry weight (d.w.) of soil in the top (plow) layer to less than 0.1 mg/kg d.w. at 1 m depth. The concentrations varied also spatially, up to one order of magnitude within the same field and showing a decreasing trend from the Hg source (i.e., irrigation ditches). The concentration profiles and gradients measured were explained considering Hg loading, soil properties, such as the texture, organic carbon content, pH and cation exchange capacity. A Selective Sequential Extraction (SSE) was also applied on soil samples from an ad hoc greenhouse experiment to investigate the role of different plant species in influencing Hg speciation in soils. Although most of the extracted Hg was included in scarcely mobile or immobile forms, some plant species (i.e., alfalfa) showed to importantly increase the soluble and exchangeable fractions with respect to the unplanted control soils, thus affecting mobility and potential bioavailability of Hg.

1,2-dibromo-4-(1,2-dibromoethyl)-cyclohexane (DBE-DBCH) is a brominated flame retardant used in commercial and industrial applications. The use of DBE-DBCH containing products has resulted in an increased release into the environment. However, limited information is available on the long-term effects of DBE-DBCH and its effects in aquatic invertebrates. Thus, the present study was aimed at determining how DBE-DBCH diastereomers (αβ and γδ) affects aquatic invertebrates using Daphnia magna as a model organism. Survival, reproduction, feeding, swimming behavior and toxicogenomic responses to environmental relevant concentrations of DBE-DBCH were analyzed. Chronic exposure to DBE-DBCH resulted in decreased lifespan, and reduced fecundity. Expression of genes involved in reproductive processes, vtg1 and jhe, were also inhibited. DBE-DBCH also induced hypoxia by inhibiting the transcription of genes involved in heme biosynthesis and oxygen transport. Furthermore, DBE-DBCH also inhibited feeding resulting in emptiness of the alimentary canal. Increased expression of the stress response biomarkers was observed following DBE-DBCH exposure. In addition, DBE-DBCH diastereomers also altered the swimming behavior of Daphnia magna. The present study demonstrates that DBE-DBCH cause multiple deleterious effects on Daphnia magna, including effects on reproduction and hormonal systems. These endocrine disrupting effects are in agreement with effects observed on vertebrates. Furthermore, as is the case in vertebrates, DBE-DBCH γδ exerted stronger effects than DBE-DBCH αβ on Daphnia magna. This indicate that DBE-DBCH γδ has properties making it more toxic to all so far studied animals than DBE-DBCH αβ.

Cadmium (Cd) contamination in soil and crops caused by mining activities has become a prevalent concern in the world. Given that different crops have varying Cd bioaccumulation factors, crops with low Cd bioaccumulation abilities can be selected for the safe usage of Cd -contaminated lands. This study aimed to investigate Cd contamination in soil and crops and the influencing factors of soil Cd activity in a tin mining area (TMA) and control area (CA) and to put forward suggestions for the safe usage of farmlands by developing prediction models of Cd content in different crop grains. We collected 72 and 40 pairs of rice and maize grain samples, respectively, along with their rhizosphere soil samples and 6176 topsoil samples. The results showed that compared with the CA, the Cd pollution was more severe in the cultivated soil and crop grains around TMA. Furthermore, rice has a strong ability to transport Cd from soil to grains, whereas maize has a poor Cd uptake ability. The total organic carbon, CaO, pH, and Mn in soil play key roles in the transfer of Cd from soil to crop grains. Using these parameters and Cd concentration in soil, two sets of accurate Cd prediction models were developed for maize and rice. Based on the Cd concentration in the topsoil and predicted Cd concentration in crop grains, the safe utilization scheme of farmland was proposed. The proportions of priority protection, safe exploitation, planting adjustment, and strict control were 72.59%, 22.77%, 3.16%, and 1.48% in the TMA, respectively. The values reached 80.51% (priority protection), 19.12% (safe exploitation), 0.37% (planting adjustment), and 0% (strict control) in the CA. Thus, given the difference between Cd accumulation in rice and maize, adjustment of planting crops in contaminated farmlands can be applied to maximize the use of farmland resources.

All European bats are protected by the EU and Associated Members legal regulations. Being insectivorous and top predators, they can be particularly exposed to persistent organic and inorganic pollutants. It is surprising how little is known about the impact of environmental pollutants on bats from physiological to populational levels. In this study we focused on contamination with trace metals of first-year bats from Kharkiv city, NE Ukraine. Tissues from the carcasses of two species, Nyctalus noctula (n = 20) and Eptesicus serotinus (n = 20), were used for metal analysis. The samples of external (wing membrane, fur) and internal (liver, lung, kidney, bones) tissues were analysed for contents of Pb, Cu, Zn, and Cd to see whether fur or wing membrane can be used as proxies for metal contamination of the vital internal tissues. In E. serotinus, significant positive correlations in Pb concentrations were found between all external and internal tissues. For Cd only, correlation between the fur and lung was found, for Cu between the fur and liver, and for Zn between the fur and kidney. In contrast, for N. noctula, only one such correlation was found – between Zn concentrations in the fur and kidney. The tissues differed significantly in concentrations of all studied metals, with no difference between the species. The results showed that the fur and wing membrane can be used as good proxies for Pb concentrations in internal organs of E. serotinus, but not necessarily for other metals or for N. noctula. The results for Pb are, however, encouraging enough to conclude that the topic is worth further studies, covering more species, a wider age range and more diverse environments.

Arsenic (As) is a ubiquitous environmental carcinogen that enters the human food chain mainly through rice grains. In the present study, we evaluated the potential of thiourea (TU; non-physiological reactive oxygen species scavenger) in mitigating the negative effects of arsenic (As) stress in indica rice variety IR64, with the overall aim to reduce grain As accumulation. At seedling stage, As + TU treatment induced the formation of more numerous and longer crown roots compared with As alone. The As accumulation in main root, crown root, lower leaf and upper leaf was significantly reduced to 0.1-, 0.14-, 0.16-, 0.14-fold, respectively in As + TU treated seedlings compared with those of As alone. This reduced As accumulation was also coincided with light-dependent suppression in the expression levels of aquaporins and photosynthesis-related genes in As + TU treated roots. In addition, the foliar-supplemented TU under As-stress maintained reducing redox conditions which decreased the rate of As accumulation in flag leaves and, eventually grain As by 0.53-fold compared with those of As treatment. The agronomic feasibility of TU was validated under naturally As contaminated sites of Nadia (West Bengal, India). The tiller numbers and crop productivity (kg seed/ha) of TU-sprayed plants were increased by 1.5- and 1.18-fold, respectively; while, grain As accumulation was reduced by 0.36-fold compared with those of water-sprayed control. Thus, this study established TU application as a sustainable solution for cultivating rice in As-contaminated field conditions.

Mercury (Hg) is an environmental contaminant that poses a threat to aquatic systems globally. Temporal evaluations of Hg contamination have increased in recent years, with studies focusing on how anthropogenic activities impact Hg bioavailability in a variety of aquatic systems. While it is common for these studies and ecological risk assessments to evaluate Hg bioaccumulation and effects in wildlife, there is a paucity of information regarding Hg dynamics in reptiles. The goal of this study was to investigate temporal patterns in total mercury (THg) and methylmercury (MeHg) concentrations across a 36-year period, as well as evaluate relationships among and between destructive (kidney, liver, muscle) and non-destructive (blood, tail) tissue types in a common watersnake species. To accomplish this, we measured THg and MeHg concentrations in multiple tissues from brown watersnakes (Nerodia taxispilota) collected from Steel Creek on the Savannah River Site (SRS; Aiken, SC, USA) from two time periods (1983–1986 and 2019). We found significant and positive relationships between tail tips and destructive tissues. In both time periods, THg concentrations varied significantly by tissue type, and destructive tissues exhibited higher but predictable THg values relative to tail tissue. Methylmercury concentrations did not differ among tissues from the 1980s but was significantly higher in muscle compared to other tissues from snakes collected in 2019. Percent MeHg of THg in N. taxispilota tissues mirrored patterns reported in other reptiles, although the range of % MeHg in liver and kidney differed between time periods. Both THg and MeHg concentrations in N. taxispilota declined significantly from the 1980s to 2019, with average values 1.6 to 4-fold lower in contemporary samples. Overall, our data add further evidence to the utility of watersnakes to monitor Hg pollution in aquatic environments and suggest attenuation of this contaminant in watersnakes in our study system.

Inorganic arsenic (iAs) is a naturally occurring metalloid present in drinking water and polluted air exposing millions of people globally. Epidemiological studies have linked iAs exposure to the development of numerous diseases including cognitive impairment, cardiovascular failure and cancer. Despite intense research, an effective therapy for chronic arsenicosis has yet to be developed. Laboratory studies have been of great benefit in establishing the pathways involved in iAs toxicity and providing insights into its mechanism of action. However, the in vivo analysis of arsenic toxicity mechanisms has been difficult by the lack of reliable in vivo biomarkers of iAs’s effects. To address this issue we have applied the use of our recently developed stress reporter models to study iAs toxicity. The reporter mice Hmox1 (oxidative stress/inflammation; HOTT) and p21 (DNA damage) were exposed to iAs at acute and chronic, environmentally relevant, doses. We observed induction of the oxidative stress reporters in several cell types and tissues, which was largely dependent on the activation of transcription factor NRF2. We propose that our HOTT reporter model can be used as a surrogate biomarker of iAs-induced oxidative stress, and it constitutes a first-in-class platform to develop treatments aimed to counteract the role of oxidative stress in arsenicosis. Indeed, in a proof of concept experiment, the HOTT reporter mice were able to predict the therapeutic utility of the antioxidant N-acetyl cysteine in the prevention of iAs associated toxicity.

Acoustic pollution in aquatic environments has increased with adverse effects on many aquatic organisms. However, little work has been done considering the effects of the vibratory component of acoustic stimuli, which can be transmitted in the substrate and propagated into the aquatic medium. Benthic marine organisms, including many invertebrates, are capable of sensing seabed vibration, yet the responses they trigger on organism have received little attention. This study investigates the impact of underwater vibration on the physiology and behaviour of a ubiquitous inhabitant of coastal areas of the northern hemisphere, the shore crab Carcinus maenas. We developed a novel vibratory apparatus with geophones supported on a softly sprung frame to induce a seabed vibration of 20 Hz frequency, as observed during dredging, piling and other anthropogenic activities. The geophone internal mass caused the frame to vibrate in a controlled manner. Our results show that transition from ambient to anthropogenic vibrations induced an increase in activity and antennae beats in shore crabs, indicating perception of the vibratory stimulus and a higher stress level. There was also a trend on sex-specific responses to anthropogenic vibration, with males showing a higher activity level than females. However, no effect of anthropogenic vibrations was found upon oxygen consumption. These results show that anthropogenic underwater vibration induces behavioural responses in Carcinus maenas. This highlights the importance of evaluating man-made vibratory activities on coastal invertebrates and the necessity of evaluating anthropogenic effects on both sexes.