Venlafaxine is an antidepressant and anti-anxiety drug that has been detected in municipal wastewater at low μg/L concentrations. In this study, the nest-defense behavior of adult male fathead minnows (Pimephales promelas) was observed in fish exposed for a full lifecycle to venlafaxine nominal concentrations of 0.88, 8.8, and 88 μg/L (i.e. 1, 9.3, 75 μg/L mean measured concentrations). Nest-defense behaviors quantified were the time taken to contact a dummy intruder fish (on a flexible stick, held near each nest) and the number of contacts made during a 1 min period. In male fathead minnows exposed to venlafaxine over a full lifecycle at environmentally relevant nominal concentrations (i.e. 0.88 and 8.8 μg/L) no significant effects were observed in behavior. However, in males exposed over a full lifecycle to the highest concentration of venlafaxine (i.e. 88 μg/L), nest-defense behaviors were increased in males with empty nests, as shown by the significantly elevated percentage of empty-nest males that made contact with the dummy intruder fish (89%) relative to the lower percentage of contacts (65%) among the Control males (p = 0.046). Lifecycle exposure to high venlafaxine (88 μg/L) caused males to over-protect their empty nests. Environmental venlafaxine concentrations are approximately 70 x lower than this, so it is unlikely that behavioral changes from venlafaxine exposure would occur in the environment. Normal nest defense behaviours in control males varied, depending on whether they were protecting empty nests or nests with eggs. Compared to Control males with empty nests, more Control males with eggs in their nests made contact with the dummy intruder fish (p = 0.014), contact was faster (i.e. <10 s, p = 0.011), and they hit the dummy intruder fish more times in 1 min (p = 0.031) This study is the first to assess reproductive behaviors in fish exposed to an antidepressant over a full lifecycle.

Atmospheric pollution by polycyclic aromatic hydrocarbons (PAHs) has become a serious problem, especially in Asia, as PAHs can severely affect ecologically important mountainous areas. Using pine needles and mosses as bio-indicators, this study examined PAH pollution in a mountainous study area and evaluated the influence of transboundary PAHs. PAHs in urban areas were also evaluated for comparison. The study sites were alpine areas and urban areas (inland or coastal cities) across central Japan, in the easternmost part of Asia where atmospheric pollutants are transported from mainland Asia. The mean PAH concentrations of pine needles and mosses were 198.9 ± 184.2 ng g⁻¹ dry weight (dw) and 131.8 ± 60.7 ng g⁻¹ dw (mean ± SD), respectively. Pine needles preferentially accumulated PAHs with low molecular weights (LMW PAHs) and exhibited large differences in both PAH concentration and isomer ratios between alpine and urban sites. These differences can be explained by the strong influence of LMW PAHs emitted from domestic sources, which decreased and changed during transport from urban to alpine sites due to dry/wet deposition and photodegradation. In contrast, mosses accumulated a higher ratio of PAHs with high molecular weight (HMW PAHs). A comparison of isomer ratios showed that the PAH source for alpine moss was similar to that for northern coastal cities, which are typically influenced by long-transported PAHs from East Asia. Thus, these results indicate that alpine moss can also be strongly affected by the transboundary PAHs. It is likely that the uptake characteristics of moss, alpine climate, and alpine locations far from urban areas can strengthen the influence of transboundary pollution. Based on these results, the limitations and most effective use of bioindicators of PAH pollution for preserving alpine ecosystems are discussed.

Microbial source tracking (MST) DNA-based assays have been used to successfully solve fecal pollution problems in many countries, particularly in developed nations. However, their application in developing countries has been limited but continues to increase. In this study, sixteen endpoint and quantitative PCR (qPCR) assays targeting universal and human-, swine-, and cattle-specific Bacteroidales gene markers were modified for endpoint PCR, evaluated for their performance with sewage and fecal samples from the Tha Chin watershed and subsequently validated with samples from the Chao Phraya watershed, Thailand. Sample sizes of 81 composite samples (from over 1620 individual samples) of farm animals of each type as well as 19 human sewage samples from the Tha Chin watershed were calculated using a stratified random sampling design to achieve a 90% confidence interval and an expected prevalence (i.e., desired assay's sensitivity) of 0.80. The best universal and human-, swine-, and cattle-specific fecal markers were BacUni EP, HF183/BFDrev EP, Pig-2-Bac EP, and Bac3 assays, respectively. The detection limits for these assays ranged from 30 to 3000 plasmid copies per PCR. The positive predictive values were high in universal and swine- and cattle-specific markers (85–100%), while the positive predictive value of the human-specific assay was 52.2%. The negative predictive values in all assays were relatively high (90.8–100%). A suite of PCR assays in Thailand was established for potential MST use in environmental waters, which supports the worldwide applicability of Bacteroidales gene markers. This study also emphasizes the importance of using a proper sample size in assessing the performance of MST markers in a new geographic region.

Urban development has led to an increase in urban runoff, accompanied with a decrease in water quality during rain events. One of the major causes of the decrease in water quality is the presence of trace organic contaminants in urban runoff. However, little is known about the sources of organic contaminants in urban runoff, especially related to land-use and temporal trends in those associated land uses. The objective of this study was to assess the occurrence and concentration trends of organic contaminants for a high-density residential site and commercial strip site in Madison, WI. Flow-weighted samples of urban stormwater runoff, collected with an auto-sampler, were composited and analyzed, producing mean organic contaminants concentrations for each storm event. The contaminants, which include pesticides, flame retardants, polycyclic aromatic hydrocarbons, corrosion inhibitors, among others, were extracted and analyzed by gas chromatography coupled with mass spectrometry or liquid chromatography coupled with tandem mass spectrometry. There were 30 organic contaminants that had greater than 50% detections in at least one of the sites, and those organic contaminants did provide information on similarities and differences of organic contaminants in urban runoff derived from different land uses. The sum of the total measured pesticides showed no significant difference between sites; this was likely due to the considerable green space and associated pesticide use in both sites. However, there were higher total concentrations of organophosphate flame retardants and corrosion inhibitors in the residential site. The reason for this is unknown and will require follow-up studies; however, several hypotheses are presented. Conversely, there were higher total concentrations of polycyclic aromatic hydrocarbons in the commercial site; this is most likely due to higher vehicle traffic in the commercial site. These data show that land-use may be important in determining the composition and concentrations of trace organic contaminants in urban stormwater runoff.

Trace metal risk assessment and environmental quality standard definition require realistic models that quantify metal exposure and accumulation by biota. In the present study we propose a novel trait-based approach to predict whole body concentrations of metals in aquatic invertebrates from concentrations measured in different environmental compartments. Field data from a large riverine floodplain was used to calibrate and test the model. The prediction performance of the trait-based model was unbiased and uncertainty was below the twofold of measured concentrations for the four studied metals (Ni, Cu, Cd, Pb). The relative contribution of feeding, respiration and locomotion patterns as well as metal concentrations in three environmental compartments provided insights into the importance of different uptake pathways. The relation with the sediment (i.e., to what degree taxa live in or directly on the sediment) was shown to be the most important trait to predict metal accumulation. Overall, this study demonstrated the potential use of bioecological traits for the modeling of whole body metal concentrations of entire aquatic invertebrate communities.

Carbamazepine (CBZ) is an antiepileptic drug that is frequently detected in wastewater treatment plants, soil and plants after irrigation with treated wastewater or application of biosolids. However, little information is available on the fate and uptake of CBZ in edible vegetables. In this study, radioautographic visualization of the ¹⁴C distribution revealed that ¹⁴C-CBZ can be taken up by all three ready-to-eat vegetables. Furthermore, a mass-balance study was conducted to evaluate the dynamic processes of the uptake and translocation of CBZ by ¹⁴C labeling. ¹⁴C-CBZ was gradually taken up with the growth of vegetables, with maximum uptake ratios of 2.19 ± 0.15, 2.86 ± 0.24 and 0.25 ± 0.05% of applied ¹⁴C in celery, carrot and pak choi, respectively. The bioconcentration factors (BCFs) based on ¹⁴C measurements ranged from 7.6 to 26.1 for celery, 3.6–12.9 for carrot, and 4.4–44 for pak choi. ¹⁴C-CBZ was easily translocated from the roots to the leaves and/or stems. The amendment of biosolids had a significant inhibitory effect on the uptake and translocation of ¹⁴C-CBZ from soil.

There is increasing evidence that toxicant exposure can alter DNA methylation profile, one of the main epigenetic mechanisms, particularly during embryogenesis when DNA methylation patterns are being established. In order to investigate the effects of the antibacterial agent Triclosan on DNA methylation and its correlation with gene expression, zebrafish embryos were exposed during 7 days post-fertilization (starting at maximum 8-cells stage) to 50 and 100 μg/l, two conditions for which increased sensitivity and acclimation have been respectively reported. Although global DNA methylation was not significantly affected, a total of 171 differentially methylated fragments were identified by Reduced Representation Bisulfite Sequencing. The majority of these fragments were found between the two exposed groups, reflecting dose-dependant specific responses. Gene ontology analysis revealed that pathways involved in TGF-β signaling were enriched in larvae exposed to 50 μg/l, while de novo pyrimidine biosynthesis functions were overrepresented in fish exposed to 100 μg/l. In addition, gene expression analysis revealed a positive correlation between mRNA levels and DNA methylation patterns in introns, together with significant alterations of the transcription of genes involved in nervous system development, transcriptional factors and histone methyltransferases. Overall this work provides evidence that Triclosan alters DNA methylation in zebrafish exposed during embryogenesis as well as related genes expression and proposes concentration specific modes of action. Further studies will investigate the possible long-term consequences of these alterations, i.e. latent defects associated with developmental exposure and transgenerational effects, and the possible implications in terms of fitness and adaptation to environmental pollutants.

IMO’s Anti-Fouling Systems convention banned the use of organotin-based antifouling systems in 2008 as the ultimate effort to stop tributyltin (TBT) inputs into the marine environment. One of the hazardous effects of TBT is imposex (the superimposition of male sexual characters onto gastropod females), a phenomenon that may cause female sterility and the gastropod populations decline. Despite previous European Union legislation had already been shown effective in reducing the imposex levels along the Portuguese coast, this study intends to confirm these decreasing trends after 2008 and describe the global evolution in the last 15 years. Imposex levels were assessed in two bioindicators – the dog-whelk Nucella lapillus and the netted-whelk Nassarius reticulatus (Gastropoda, Prosobranchia) – in 2011 and 2014, and the results were compared with previous years. Both species showed progressive decreasing trends in imposex levels over the last 15 years; median values of the vas deferens sequence index (VDSI) fell from 3.96 to 0.78 in N. lapillus and from 3.39 to 0.29 in N. reticulatus. The temporal/spatial evolution of imposex suggests an apparent shift of TBT hotspots, being now restricted to fishing ports and marinas in detriment of large commercial harbours where TBT levels fell rapidly. Butyltins were measured in the whole tissues of N. lapillus females collected in 2014: monobutyltin (MBT) varied from < DL (detection limit: 1 ng Sn/g) to 13 ng Sn/g dw, dibutyltin (DBT) from 2.2 to 27 ng Sn/g dw and TBT from 1.5 to 55 ng Sn/g dw. Although TBT body burden has declined over time, the butyltin degradation index ([MBT]+[DBT])/[TBT] exhibited values < 1 in c. a. 90% of the sites assessed, suggesting that recent TBT inputs are still widespread in the Portuguese coast eventually due to illegal use of TBT antifouling systems and TBT desorption from sediments.

Glyphosate (GLY) is one of the most used herbicide worldwide. Considering that information concerning the impact of GLY on bivalves is scarce, in this study we evaluated for the first time the effects of environmentally realistic concentrations of GLY (10, 100 and 1000 μg/L) to the mussel Mytilus galloprovincialis. Mussels were exposed for 7, 14 and 21 days and several biomarkers were measured in haemocytes/haemolymph (total haemocyte counts, haemocyte diameter and volume, haemolymph pH, haemolymph lactate dehydrogenase activity, haemocyte lysate lysozyme and acid phosphatase activities), as well as in gills and digestive gland (antioxidant enzyme and acetylcholinesterase activities). The concentrations of GLY and its main metabolite aminomethylphosphonic acid in the experimental tanks were also measured. The MANOVA analysis demonstrated that the experimental variables considered (exposure concentration, exposure duration, and their interaction) affected significantly biomarker responses. In addition, the two-way ANOVA analysis indicated that GLY was able to affect most of the cellular parameters measured, whereas antioxidant enzyme activities resulted to be influenced moderately. Interestingly, exposure to GLY reduced significantly acetylcholinesterase activity in gills. Although preliminary, the results of this study demonstrated that GLY can affect both cellular and biochemical parameters in mussels, highlighting a potential risk for aquatic invertebrates.

Excess exposure to fluoride causes substantive health burden in humans and livestock globally. However, few studies have assessed the distribution and controls of variability of ambient background concentrations of fluoride in soil. Ambient background concentrations of fluoride in soil were collated for Greater Melbourne, Greater Geelong, Ballarat and Mitchell in Victoria, Australia (n = 1005). Correlation analysis and machine learning techniques were used to identify environmental and anthropogenic influences of fluoride variability in soil. Sub-soils (>0.3 m deep), in some areas overlying siltstone and sandstone, and to a lesser extent, overlying basalt, were naturally enriched with fluoride at concentrations above ecological thresholds for grazing animals. Soil fluoride enrichment was predominantly influenced by parent material (mineralogy), precipitation (illuviation), leaching during palaeoclimates and marine inputs. Industrial air pollution did not significantly influence ambient background concentrations of fluoride at a regional scale. However, agricultural practices (potentially the use of phosphate fertilisers) were indicated to have resulted in added fluoride to surface soils overlying sediments. Geospatial variables alone were not sufficient to accurately model ambient background soil fluoride concentrations. A multiple regression model based on soil chemistry and parent material was shown to accurately predict ambient background fluoride concentrations in soils and support assessment of fluoride enrichment in the environment.