Both Ca deficiency and metal exposure may affect physiological and nutritional condition of breeding females altering their ability to deposit essential resources (e.g. Ca, antioxidants) into the eggs. This effect of the maternal investment into egg quality is not strictly limited to the embryonic period, but may persist after hatching, since nutrient levels in yolks can compromise nestling antioxidant status, growth and fledging success. The goal of this study was to investigate how metal pollution and Ca availability during the breeding season affect oxidative stress biomarkers and plasma biochemistry in adult female pied flycatchers (Ficedula hypoleuca). In addition, we aim to evaluate how maternal antioxidant status and body condition relate to breeding parameters and offspring oxidative balance. Females breeding in a metal-polluted area in SW Finland showed higher metal concentrations compared to the control area, although current levels were below the toxic level able to affect female physiology. In addition, Ca availability was not constraining female oxidative status and general health in the study area. Interestingly, our results suggested that antioxidant response to metals was better when Ca concentrations were high enough to cover the physiological Ca requirements in breeding females. There seems to be a subtle balance between the concentrations of Ca in the organism and the tolerance to metal-related effects that requires further research. This study supports that offspring oxidative balance and nestling development are affected by maternal body condition and antioxidant status.

The growing use of octocrylene (OC) in sunscreens has posed a great threat to aquatic organisms. In the present study, to assess its reproductive toxicity and mechanism, paired Japanese medaka (Oryzias latipes) (F0) were exposed to OC at nominal concentrations of 5, 50, and 500 μg/L for 28 d. Significant increases were observed in the gonadosomatic index (GSI) and hepatosomatic index (HSI) of F0 medaka at 500 μg/L OC (p < 0.05) without significant differences in fecundity. The fertility was significantly decreased at all treatments (p < 0.05). Significant increases in the percent of mature oocytes were observed at 5 and 500 μg/L OC, in which contrary to the percent of spermatozoa (p < 0.05). The plasma sex hormones and vitellogenin levels significantly increased in males at all treatments and in females at 50 and 500 μg/L OC (p < 0.05). In addition, the levels of fshβ and lhβ in the brains and the levels of fshr, lhr and cyp17α in the gonads were significantly upregulated in males at all treatments (p < 0.05), in line with those of ar, erα, erβ and cyp19β in the brains of male and female. The upregulation of vtg in male and female livers was observed only at 500 μg/L OC and upregulation of star and hsd3β was observed in testis at all treatments (p < 0.05). Continued exposure to OC significantly induced increases in the time to hatching, morphological abnormality rates, and cumulative death rates of F1 embryos, inconsistent with body length of F1 larvae (p < 0.05). Therefore, the responses of the exposed fish at the biochemical and molecular levels indicated reproductive toxicity and estrogenic activity of OC, providing insights into the mechanism of OC.

Research using various species of wild and cultured fish has identified negative effects of short-term exposure to microbeads. Although wild animals might be contaminated with microbeads and/or other pharmaceuticals, data regarding the long-term effects remain limited. To clearly elucidate the effects of microbeads, studies of long-term exposure using animal models are necessary. Our aim was to elucidate the effects of microbeads alone on the growth and fecundity of medaka following long-term exposure (12 weeks). In experiment 1, fish groups (except controls) were temporarily exposed to polyethylene microbeads (10–63 μm diameter) a low dose of 0.065 microbeads-mg/L and high dose of 0.65 microbeads-mg/L. In experiment 2, see-through medaka and fluorescent polyethylene microbeads (10–45 μm diameter) were used to estimate the retention time of ingested microbeads in the digestive tract, which was 4–9 days. The low dose of microbeads did not affect growth but did decrease the number of eggs and the hatching rate. The high dose decreased growth, the number of eggs, and hatching rate. Growth differences were recognized for the first time at 7 weeks, and differences in the number of eggs at 12 weeks. Thus, long-term tests using medaka indicated that microbeads per se exhibit growth inhibition and reproductive toxicity. These effects could be associated with nutritional factors resulting from the long retention time of microbeads in the digestive tract. We also determined the dose that affects only fecundity. This suggests that normal growth of medaka in the wild does not mean the environment is free from microbead contamination. We are thus attempting to identify new biological indexes for monitoring the status of microbead contamination using our system.

The ubiquity of microplastics in the world's ocean has aroused great concern. However, the ecological effects of microplastics at environmentally realistic concentrations are unclear. Here we showed that exposure of marine medaka (Oryzias melastigma) to environmentally relevant concentrations of 10 μm polystyrene microplastics for 60 days not only led to microplastic accumulation in the gill, intestine, and liver, but also caused oxidative stress and histological changes. Moreover, 2, 20, and 200 μg/L microplastics delayed gonad maturation and decreased the fecundity of female fish. Alterations of the hypothalamus-pituitary-gonadal (HPG) axis were investigated to reveal the underlying mechanisms, and gene transcription analysis showed that microplastic exposure had significantly negative regulatory effects in female HPG axis. Transcription of genes involved in the steroidogenesis pathway in females were also downregulated. This disruption resulted in decreased concentrations of 17β-estradiol (E₂) and testosterone (T) in female plasma. Furthermore, parental exposure to 20 μg/L microplastics postponed the incubation time and decreased the hatching rate, heart rate, and body length of the offspring. Overall, the present study demonstrated for the first time that environmentally relevant concentrations of microplastics had adverse effects on the reproduction of marine medaka and might pose a potential threat to marine fish populations.

Bisphenol A (BPA) is an endocrine disruptor whose ubiquitous presence in the environment has been related with impairment of male reproduction. BPA can cause both transcriptomic and epigenetic changes during spermatogenesis. To evaluate the potential effects of male exposure to BPA, adult zebrafish males were exposed during spermatogenesis to doses of 100 and 2000 μg/L, which were reported in contaminated water bodies and higher than those allowed for human consumption. Fertilization capacity and survival at hatching were analysed after mating with untreated females. Spermatogenic progress was analysed through a morphometrical study of testes and apoptosis was evaluated by TUNEL assay. Testicular gene expression was evaluated by RT-qPCR and epigenetics by using ELISA and immunocytochemistry. In vitro studies were performed to investigate the role of Gper. Chromatin fragmentation and the presence of transcripts were also evaluated in ejaculated sperm. Results on testes from males treated with the highest dose showed a significant decrease in spermatocytes, an increase in apoptosis, a downregulation of ccnb1 and sycp3, all of which point to an alteration of spermatogenesis and to meiotic arrest and an upregulation of gper1 and esrrga receptors. Additionally, BPA at 2000 μg/L caused missregulation of epigenetic remodelling enzymes transcripts in testes and promoted DNA hypermethylation and H3K27me3 demethylation. BPA also triggered an increase in histone acetyltransferase activity, which led to hyperacetylation of histones (H3K9ac, H3K14ac, H4K12ac). In vitro reversion of histone acetylation changes using a specific GPER antagonist, G-36, suggested this receptor as mediator of histone hyperacetylation. Males treated with the lower dose only showed an increase in some histone acetylation marks (H3K14ac, H4K12ac) but their progeny displayed very limited survival at hatching, revealing the deleterious effects of unbalanced paternal epigenetic information. Furthermore, the highest dose of BPA led to chromatin fragmentation, promoting direct reproductive effects, which are incompatible with embryo development.

Stability of silver sulfide nanoparticle (Ag₂S-NP) in the environment has recently drawn considerable attention since it is associated with environmental risk. Although the overestimated stability of Ag₂S-NP in aqueous solution has already been recognized, studies on transformation of Ag₂S-NP in environmental water are still very scarce. Here we reported that Ag₂S-NP could undergo dissolution by manganese(IV) oxide (MnO₂), an important naturally occurring oxidant in the environment, even in environmental water, although the dissolved silver would probably be adsorbed onto the particles (>0.45 μm) in environmental water, mitigating the measurable levels of dissolved silver. The extent and rate of Ag₂S-NP dissolution rose with the increasing concentration of MnO₂. In addition, environmental factors including natural organic matter, inorganic salts and organic acids could accelerate the Ag₂S-NP dissolution by MnO₂, wherein an increase in dissolution extent was also observed. We further documented that Ag₂S-NP dissolution by MnO₂ was highly dependent on O₂ and it was an oxidative dissolution, with the production of SO₄²⁻. Finally, dissolution of Ag₂S-NP by MnO₂ affected zebra fish (Danio rerio) embryo viability, showing significant reduction in embryo survival and hatching rates, compared to embryos exposed to Ag₂S-NP, MnO₂ or dissolved manganese alone. These findings would further shed light on the stability of Ag₂S-NP in the natural environment - essential for comprehensive nano risk assessment.

Mycotoxins are secondary metabolites produced by varieties of fungi that contaminate food and feed resources and are capable of inducing a wide range of toxicity. In the current study, we investigated developmental and behavioural toxicity in zebrafish larvae after exposure to six different mycotoxins; ochratoxin A (OTA), type A trichothecenes mycotoxin (T-2 toxin), type B trichothecenes mycotoxin (deoxynivalenol - DON), and zearalenone (ZEN) and its metabolites alpha-zearalenol (α-ZOL) and beta-zearalenol (β-ZOL). Developmental defects, hatching time, and survival were monitored until 96 h post fertilisation (hpf). The EC₅₀, LC₅₀, and IC₅₀ values were calculated. Subsequently, to assess behavioural toxicity, new sets of embryos were exposed to a series of non-lethal doses within the range of environmental and/or developmental concern. Results indicated that all the tested mycotoxins were toxic, they all induced developmental defects, and with the exception of OTA, all affected hatching time. Behavioural effects were only observed following exposure to OTA and ZEN and its metabolites, α ZOL and β ZOL. These results demonstrate that mycotoxins are teratogenic and can influence behaviour in a vertebrate model.

Mining in the Athabasca Oil Sands Region (AOSR) has contributed extensively to increased exposure of wildlife to naturally occurring polycyclic aromatic compounds (PACs), yet little is known about the toxicity of PACs to wildlife in this region. We identified reproductive and developmental changes in tree swallows (Tachycineta bicolor) breeding in close proximity to mining-related activities in the AOSR, and determined these changes in relation to the birds' exposure and accumulation of 41 PACs (parent-, alkylated-PAHs), dibenzothiophenes (DBTs; previously published), diet (carbon (δ13C), nitrogen (δ15N)), volatile organic compounds, and weather variables. Tree swallow pairs (N = 43) were compared among mining-related (OS1, OS2) and reference (REF1, REF2) sites. At OS2, clutch initiation was slightly advanced (2012) but reproductive success (65%) was much lower than at the other sites (≥ 79%). Fledgling production by each pair was influenced by the timing of clutch initiation (years combined); in a highly inclement brood rearing period (2013), additional influences included the nestlings' exposure to ΣDBTs, accumulation of C1-naphthalene, the trophic position of the prey in their diet (δ15N), and record-breaking rainfall. Nestlings at OS2 were significantly lighter at day (d) 9 and d14, and in poorer body condition (d9). Nestling body mass was influenced by multiple stressors that varied by site: mass of younger nestlings (d9) was related to dietary source (δ13C; e.g., wetlands, terrestrial fields), exposure and/or accumulation of C1-phenanthrenes, C2-fluorenes, Σalkyl-PAHs and ΣDBTs, while for older nestlings (d14), body mass was related to sex, hatch date and/or rainfall during brood rearing. The swallows' exposure and accumulation of parent-PACs, alkyl-PACs and DBTs, the timing of hatching, their diet and exposure to highly inclement rains, contributed to their reproductive and developmental changes.

Microcystin-LR (MCLR) has been reported to cause developmental neurotoxicity in zebrafish, but there are few studies on the mechanisms of MCLR-induced transgenerational effects of developmental neurotoxicity. In this study, zebrafish were exposed to 0, 1, 5, and 25 μg/L MCLR for 60 days. The F1 zebrafish embryos from the above-mentioned parents were collected and incubated in clean water for 120 h for hatching. After examining the parental zebrafish and F1 embryos, MCLR was detected in the gonad of adults and F1 embryos, indicating MCLR could potentially be transferred from parents to offspring. The larvae also showed a serious hypoactivity. The contents of dopamine, dihydroxyphenylacetic acid (DOPAC), serotonin, gamma-aminobutyric acid (GABA) and acetylcholine (ACh) were further detected, but only the first three neurotransmitters showed significant reduction in the 5 and 25 μg/L MCLR parental exposure groups. In addition, the acetylcholinesterase (AChE) activity was remarkably decreased in MCLR parental exposure groups, while the expression levels of manf, bdnf, ache, htr1ab, htr1b, htr2a, htr1aa, htr5a, DAT, TH1 and TH2 genes coincided with the decreased content of neurotransmitters (dopamine, DOPAC and serotonin) and the activity of AChE. Neuronal development related genes, α1-tubulin, syn2a, mbp, gfap, elavl3, shha and gap43 were also measured, but gap43 was the gene only up-regulated. Our results demonstrated MCLR could be transferred to offspring, and subsequently induce developmental neurotoxicity in F1 zebrafish larvae by disturbing the neurotransmitter systems and neuronal development.

Exposure experiments were conducted to evaluate the influence of dissolved organic matter (DOM) on the toxicity of ZnO-NPs (10–30 nm) and dissolved Zn at sub-lethal doses (50 and 5 ppm, respectively) to zebrafish (Danio rerio). Humic acid, alginic acid, bovine serum albumin and various natural DOM isolated from rivers as the Milwaukee River-WI (NOMW), Yukon River-AK (NOMA) and Suwannee River-GA DOM (NOMS) were used to represent humic substances (HA), carbohydrates (CHO), proteins (PTN), and natural organic matter (NOM), respectively. Initial experiments were carried out to confirm the toxic effect of ZnO-NPs at 50 ppm, followed by mitigation experiments with different types and concentrations of DOM (0.4–40 mg-C/L). Compared to 0% hatch of 50 ppm ZnO-NPs exposed embryos at 72 h post fertilization (hpf), NOMS, NOMW and HA had the best mitigative effects on hatching (53–65%), followed by NOMA, CHO and PTN (19–35%); demonstrating that the mitigation effects on ZnO-NPs toxicity were related to DOM's quantity and composition. At 96 hpf, 20% of embryos exposed to 50 ppm ZnO-NPs hatched, 100% of embryos reared in embryo medium hatched, and close to 100% of the embryos hatched upon mitigation, except for those mitigated with PTN which had less effect. Dissolved Zn (5 ppm) also exhibited the same toxicity on embryos as ZnO-NPs (50 ppm). However, in the presence of HA, NOM and CHO, the hatching rates at 72 and 96 hpf increased significantly compared to 5% hatch without DOM. The overall mitigation effects produced by DOM followed the order of HA ≥ NOMS > NOM (A&W) > CHO >> PTN, although specific mitigation effects varied with DOM concentration and functionalities. Our results also indicate that the toxicity of ZnO-NPs to embryos was mostly derived from NPs although dissolved Zn released from ZnO-NPs also interacted with embryos, affecting hatching, but to a less extent.