The potential risks of phthalates affecting human and animal health as well as the environment are emerging as serious concerns worldwide. However, the mechanism by which phthalates induce developmental effects is under debate. Herein, we found that embryonic exposure of zebrafish to di-(2-ethylhexyl) phthalate (DEHP) and di-butyl phthalate (DBP) increased the rate of heart defects including abnormal heart rate and pericardial edema. Changes in the transcriptional profile demonstrated that genes involved in the development of the heart, such as tbx5b, nppa, ctnt, my17, cmlc1, were significantly altered by DEHP and DBP at 50 μg/L, which agreed with the abnormal cardiac outcomes. Methylated DNA immunoprecipitation sequencing (MeDIP-Seq) further showed that significant hypomethylation of nppa and ctnt was identified after DEHP and DBP exposure, which was consistent with the up-regulation of these genes. Notably, hypermethylation on the promoter region (<1 kb) of tbx5b was found after DEHP and DBP exposure, which might be responsible for its decrease in transcription. In conclusion, phthalates have the potential to induce cardiac birth defects, which might be associated with the transcriptional regulation of the involved developmental factors such as tbx5b. These findings would contribute to understand the molecular pathways that mediated the cardiac defects caused by phthalates.

Water pollution by pesticides used in agriculture is currently a major concern both in Spain and in Europe as a whole, prompting the need to evaluate water quality and ecological risk in areas of intensive agriculture. This study involved monitoring pesticide residues and certain degradation products in surface and ground waters of the Denomination of Origin (DO) Jumilla vineyard area in Spain. Sixty-nine pesticides were selected and evaluated at twenty-one sampling points using a multi-residue analytical method, based on solid-phase extraction (SPE) and analysis by liquid chromatography coupled with mass spectrometry (LC-MS), providing reliable results. Twenty-six compounds from those selected were detected in the samples analyzed (eleven insecticides including one degradation product, nine herbicides, and six fungicides) and fifteen of them were found in concentrations over 0.1 μg L⁻¹ (upper threshold established by the EU for pesticides detected in waters for human consumption). Indoxacarb was present in more than 70% of the samples, being the most frequently detected compound in water samples. Some pesticides were ubiquitous in all the water samples. Ecotoxicological risk indicators, toxic units (TUs) and risk quotients (RQs), for algae, Daphnia magna and fish were calculated to estimate the environmental risk of the presence of pesticides in waters. The compounds with the highest risk were the herbicides pendimethalin, with RQ values > 1 for the three aquatic organisms, and diflufenican, posing a high risk for algae and fish, and the insecticide chlorpyrifos, with a high risk for Daphnia magna and fish. The ∑TUi determined for water at each sampling point posed only a high risk for the three aquatic organisms in a sample. These results are important for considering the selection of pesticides with less environmental risk in intensive agricultural areas.

Plastic debris has become a major threat to the marine environment and wildlife. Sea turtles are particularly vulnerable, and are known to ingest plastic debris globally; however, information from Greek waters is still absent. In this study, 36 stranded dead loggerhead turtles (Caretta caretta) were collected from the Greek coastline area, and their gastrointestinal content was analysed for ingested plastic debris. Twenty-six individuals (72%) were found to have ingested plastic, with an average of 7.94 ± 3.85 (SE) plastic items per turtle. In total, 286 plastic items were counted and categorised by size, shape, colour, and polymer type. Fourier Transform Infrared Spectrometry revealed that polypropylene and polyethylene were the dominant polymer plastic types found. Results indicated a variation in plastic ingestion amongst life stages of the loggerhead specimens. This study provides evidence of plastic ingestion by loggerhead turtles in Greek waters.

Shellfish constitute an important component of human diet, especially for those living in coastal regions. Shellfish have attracted extensive attention due to high enrichment of heavy metals. The aims of this study were to investigate the levels of trace elements in shellfish from coastal waters of Shenzhen, China and to assess human intake risks. Nine elements, including chromium (Cr), copper (Cu), iron (Fe), zinc (Zn), manganese (Mn), selenium (Se), cadmium (Cd), arsenic (As) and lead (Pb) were measured in 216 shellfish samples from eight species. Their concentrations (based on wet weight) were: Cr (0.28–21.4 mg kg⁻¹), Cu (1.40–158 mg kg⁻¹), Fe (16.5–5387 mg kg⁻¹), Zn (11.1–847 mg kg⁻¹), Mn (1.33–422 mg kg⁻¹), Se (0.15–11.8 mg kg⁻¹), Cd (0.02–18.4 mg kg⁻¹), Pb (<LOQ-10.9 mg kg⁻¹) and As (2.24–95.5 mg kg⁻¹), relatively greater than those reported in shellfish from other locations of China. Crassostrea ariakensis and Babylonia areolata were found to enrich As and Cd, respectively. The target hazard quotient (THQ) values of Cd and As were more than 1, suggesting considerable health risks from the consumption of shellfish of this zone. To our knowledge, this is the first study to assess the human risk exposure to trace elements via shellfish consumption in South China.

Environmental distribution and concentration of tetrabromobisphenol A bis- (2-hydroxyethyl) ether (TBBPA-DHEE) and tetrabromobisphenol A mono- (hydroxyethyl) ether (TBBPA-MHEE), are obscure due to the lack of available analytical methods. Here two novel immunoassays were established to systematically investigate their distributions in Taizhou, Eastern China. Five monoclonal antibodies against pollutants were generated with two designed haptens through animal immunization. After matched with different coating antigens/antibodies, ELISAs were established (LOD for TBBPA-DHEE, 0.12 ng/mL, based on OVA-M3/mAb-D4G6; LOD for TBBPA-MHEE, 0.79 ng/mL, based on OVA-M3/mAb-D2G6) and applied for investigation of their occurrences at a typical e-waste recycling area after 2-year samples collection, where the total 33 water, 32 soil and 16 biological samples were collected with the highest concentrations of 3.46 ng/mL, 2.76 ng/g (dry weight, dw) and 5.01 ng/g (dw), respectively. Meanwhile, our study also indicated that at the centralizing e-waste recycling sites the serious pollution for both chemicals still existed despite of various efforts. Besides, obvious improvements were observed at an abandoned e-waste recycling region treated and remedied for many years by the local Chinese government. These findings highlight the importance of policy decisions in treatment of pollutants to reduce organic pollutant-related health risks.

Cylindrospermopsin (CYN) is an emerging cyanotoxin increasingly being found in freshwater cyanobacterial blooms worldwide. Humans and animals are exposed to CYN through the consumption of contaminated water and food as well as occupational and recreational water activities; therefore, it represents a potential health threat. It exhibits genotoxic effects in metabolically active test systems, thus it is considered as pro-genotoxic. In the present study, the advanced 3D cell model developed from human hepatocellular carcinoma (HepG2) cells was used for the evaluation of CYN cyto-/genotoxic activity. Spheroids were formed by forced floating method and were cultured for three days under static conditions prior to exposure to CYN (0.125, 0.25 and 0.5 μg/mL) for 72 h. CYN influence on spheroid growth was measured daily and cell survival was determined by MTS assay and live/dead staining. The influence on cell proliferation, cell cycle alterations and induction of DNA damage (γH2AX) was determined using flow cytometry. Further, the expression of selected genes (qPCR) involved in the metabolism of xenobiotics, proliferation, DNA damage response, apoptosis and oxidative stress was studied. Results revealed that CYN dose-dependently reduced the size of spheroids and affected cell division by arresting HepG2 cells in G1 phase of the cell cycle. No induction of DNA double strand breaks compared to control was determined at applied conditions. The analysis of gene expression revealed that CYN significantly deregulated genes encoding phase I (CYP1A1, CYP1A2, CYP3A4, ALDH3A) and II (NAT1, NAT2, SULT1B1, SULT1C2, UGT1A1, UGT2B7) enzymes as well as genes involved in cell proliferation (PCNA, TOP2α), apoptosis (BBC3) and DNA damage response (GADD45a, CDKN1A, ERCC4). The advanced 3D HepG2 cell model due to its more complex structure and improved cellular interactions provides more physiologically relevant information and more predictive data for human exposure, and can thus contribute to more reliable genotoxicity assessment of chemicals including cyanotoxins.

Because oil spills frequently occur in coastal regions that serve as spawning habitat, characterizing the effects of oil in estuarine fish carries both economic and environmental importance. There is a breadth of research investigating the effects of crude oil on fish, however few studies have addressed how transcriptional responses to oil change throughout development or how these responses might be conserved across taxa. To investigate these effects, we performed RNA-seq and pathway analysis following oil exposure 1) in a single estuarine species (Cyprinodon variegatus) at three developmental time points (embryos, yolk-sack larvae, free-feeding larvae), and 2) in two ecologically similar species (C. variegatus and Fundulus grandis), immediately post-hatch (yolk-sack stage). Our results indicate that C. variegatus embryos mount a diminished transcriptional response to oil compared to later stages, and that few transcriptional responses are conserved throughout development. Pathway analysis of larval C. variegatus revealed dysregulation of similar biological processes at later larval stages, including alteration of cholesterol biosynthesis pathways, cardiac development processes, and immune functions. Our cross-species comparison showed that F. grandis exhibited a reduced transcriptional response compared to C. variegatus. Pathway analysis revealed that the two species shared similar immune and cardiac responses, however pathways related to cholesterol biosynthesis exhibited a divergent response as they were activated in C. variegatus but inhibited in F. grandis. Our results suggest that examination of larval stages may provide a more sensitive estimate of oil-impacts than examination of embryos, and challenge assumptions that ecologically comparable species respond to oil similarly.

Marine oil spill often causes contamination of drinking water sources in coastal areas. As the use of oil dispersants has become one of the main practices in remediation of oil spill, the effect of oil dispersants on the treatment effectiveness remains unexplored. Specifically, little is known on the removal of dispersed oil from contaminated water using conventional adsorbents. This study investigated sorption behavior of three prototype activated charcoals (ACs) of different particle sizes (4–12, 12–20 and 100 mesh) for removal of dispersed oil hydrocarbons, and effects of two model oil dispersants (Corexit EC9500A and Corexit EC9527A). The oil content was measured as n-alkanes, polycyclic aromatic hydrocarbons (PAHs), and total petroleum hydrocarbons (TPHs). Characterization results showed that the smallest AC (PAC100) offered the highest BET surface area of 889 m2/g and pore volume of 0.95 cm3/g (pHPZC = 6.1). Sorption kinetic data revealed that all three ACs can efficiently adsorb Corexit EC9500A and oil dispersed by the two dispersants (DWAO-I and DWAO-II), and the adsorption capacity followed the trend: PAC100 > GAC12 × 20 > GAC4 × 12. Sorption isotherms confirmed PAC100 showed the highest adsorption capacity for dispersed oil in DWAO-I with a Freundlich KF value of 10.90 mg/g∙(L/mg)1/n (n = 1.38). Furthermore, the presence of Corexit EC9500A showed two contrasting effects on the oil sorption, i.e., adsolubilization and solubilization depending on the dispersant concentration. Increasing solution pH from 6.0 to 9.0 and salinity from 2 to 8 wt% showed only modest effect on the sorption. The results are useful for effective treatment of dispersed oil in contaminated water and for understanding roles of oil dispersants.

The increasing availability of water quality datasets has led to a greater focus on hydrologic and water quality analysis, thus requiring more efficient and accurate modelling methods. Data mining techniques have been increasingly used for water quality analysis and prediction of the concentration and load of nitrogen pollutants instead of more traditional simulation methods. In this study, we tested the multilayer perceptron (MLP), k-nearest neighbor (k-NN), random forest, and reduced error pruning tree (REPTree) methods, along with the traditional linear regression, to predict nitrate levels based on long-term data from six watersheds with different land-use practices in the midwestern United States. Both the concentration and load results indicated that REPTree had the best performance, with an R² of 0.61–0.85 and a relative absolute error of <75.8%. The different watershed types, however, influenced the performance of the data mining methods, where all four methods showed a higher accuracy for urban dominant watershed and lower accuracy for agricultural and forest watersheds. Out of these four methods, classification tree methods (REPTree and RF) performed better than cluster methods (MLP and k-NN) for agricultural and forested watersheds. Our results indicated that both the data structure based on the dominant land use and type of algorithmic method should be carefully considered for selecting a data mining method to predict nitrate concentration and load for a watershed.

The presence of microplastics in aquatic ecosystems has recently received increased attention. Small plastic particles may resemble natural food items of larval fish and other aquatic organisms, and create strong selective pressures on the feeding traits in exposed populations. Here, we examined if larval ingestion of 90 μm polystyrene microspheres, in the presence of zooplankton (Artemia nauplii, mean length = 433 μm), shows adaptive variation in the European whitefish (Coregonus lavaretus). A full-factorial experimental breeding design allowed us to estimate the relative contributions of male (sire) and female (dam) parents and full-sib family variance in early feeding traits, and also genetic (co)variation between these traits. We also monitored the magnitude of intake and elimination of microplastics from the alimentary tracts of the larvae. In general, larval whitefish ingested small numbers of microplastics (mean = 1.8, range = 0–26 particles per larva), but ingestion was marginally affected by the dam, and more strongly by the full-sib family variation. Microsphere ingestion showed no statistically significant additive genetic variation, and thus, no heritability. Moreover, microsphere ingestion rate covaried positively with the ingestion of Artemia, further suggesting that larvae cannot adaptively avoid microsphere ingestion. Together with the detected strong genetic correlation between food intake and microplastic intake, the results suggest that larval fish do not readily possess additive genetic variation that would help them to adapt to the increasing pollution by microplastics. The conflict between feeding on natural food and avoiding microplastics deserves further attention.