Although it is fairly well known that eggshells are the major source of Ca and other micronutrients for developing avian embryos, potential anatomical and/or ecological correlates explaining the variation in embryo-induced depletion of micronutrients from eggshells remain to be addressed. Using unhatched eggs at various developmental stages and post-hatched eggshells of wild and captive-bred birds of a large precocial species, the Capercaillie Tetrao urogallus, we explored two major questions. (i) Do egg size, regional eggshell thickness as well as levels of two major micronutrients (Ca and Mg) and eight trace elements (Cr, Cu, Mn, Fe, Co, Cd, Pb and Zn) measured in egg shells and contents vary with embryonic age within and between wild and captive-bred bird eggs? (ii) How the proportions of eggshell elements become depleted during the entire process of embryogenesis? The eggshells of wild Capercaillies were thinner at the equator and the sharp pole, and contained more Mg and Ca, while in their contents there was more Cu, Cd and Pb compared with captive birds. All ten metal concentrations measured in the egg contents increased (from 15% to 285% more) with the age of embryos. The concentrations of six elements measured in eggshells (Cu, Mn, Fe, Co, Cd and Zn) decreased consistently (from 15 to 70% less) during embryogenesis. This suggests the presence of these elements in the innermost decalcified shell layer and their further mobilization into the embryos/egg contents. Our results strongly imply that most trace elements present in the egg contents/developing embryos are mobilized from the eggshells. We highlight the fact that knowledge of the magnitude of embryogenesis-related functional changes in the physical (such as shell thickness) and chemical traits of the eggs and eggshells of oviparous vertebrates is fundamental for drawing correct inferences regarding the response of an individual to variable environmental conditions.

Fipronil, a broad-spectrum chiral insecticide, has been documented to induce significant neurotoxicity to nontarget aquatic species; however, whether its neurotoxicity behaves enantioselectively and what molecular mechanisms correspond to the neurotoxicity remain unanswered. To date, few investigations have focused on the genomic mechanisms responsible for the enantioselective toxicity of chiral pesticides. The epigenetic modifications, especially DNA methylation, caused by the pesticides are also blind spot of the research works. Video tracking showed that R-fipronil exhibited more intense neurotoxicity, as well as the induction of more severe anxiety-like behavior, such as boosted swimming speed and dysregulated photoperiodic locomotion, to embryonic and larval zebrafish compared with S-fipronil. The MeDIP-Seq analysis, combined with Gene Ontology and KEGG, revealed that R-fipronil disrupted five signaling pathways (MAPK, Calcium signaling, Neuroactive ligand-receptor interaction, Purine metabolism, and Endocytosis) to a greater extent than S-fipronil through the hypermethylation of several important neuro-related genes, whereas no significant alterations of global DNA methylation were observed on the two enantiomers. To summarize, our data indicated that the fipronil-conducted enantioselective neurotoxicity likely applied its enantioselectivity by the dysregulation of DNA methylation. Our study also provided novel epigenetic insights into the study of enantioselective biological effects and the relevant underlying mechanisms of chiral insecticide.

In recent years, the impact of Plant Protection Products (PPPs) on insect pollinator decline has stimulated significant amounts of research, as well as political and public interest. PPP residues have been found in various bee-related matrices, resulting in governmental bodies worldwide releasing guidance documents on methods for the assessment of the overall risk of PPPs to different bee species. An essential part of these risk assessments are PPP residues found in pollen and nectar, as they represent a key route of exposure. However, PPP residue values in these matrices exhibit large variations and are not available for many PPPs and crop species combinations, which results in inaccurate estimations and uncertainties in risk evaluation. Additionally, residue studies on pollen and nectar are expensive and practically challenging. An extrapolation between different cropping scenarios and PPPs is not yet justified, as the behaviour of PPPs in pollen and nectar is poorly understood. Therefore, this review aims to contribute to a better knowledge and understanding of the fate of PPP residues in pollen and nectar and to outline knowledge gaps and future research needs. The literature suggests that four primary factors, the crop type, the application method, the physicochemical properties of a compound and the environmental conditions have the greatest influence on PPP residues in pollen and nectar. However, these factors consist of many sub-factors and initial effects may be disguised by different sampling methodologies, impeding their exact characterisation. Moreover, knowledge about these factors is ambiguous and restricted to a few compounds and plant species. We propose that future research should concentrate on identifying relationships and common features amongst various PPP applications and crops, as well as an overall quantification of the described parameters; in order to enable a reliable estimation of PPP residues in pollen, nectar and other bee matrices.

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.

The presence of PM₂.₅ may affect the photodegradation of benzene in the natural atmosphere. On one hand, the photodegradation of benzene may be promoted with the increase in PM₂.₅ concentrations, owing to adsorption and catalysis effect of PM₂.₅ surface; On the other hand, PM₂.₅ can scatter or block ultraviolet light and lead to weakening the photochemical reactions in the atmospheric system. It is very difficult to prove which process is dominant in the real atmosphere due to the complexity of the atmosphere. Based on coupling detrended fluctuation analysis, the goal of this work is to reveal the role of PM₂.₅ in the photodegradation of benzene in real atmosphere over long time scales. The 9 years regular monitoring data from 2007 to 2016 in Puzi of Taiwan are analyzed. A new nonlinear parameter (PDB) is established to characterize the photodegradation degree of atmospheric benzene. Based on sliding window technique, the correlations between the temporal variation of PDB and PM₂.₅ are analyzed. The results show that there is a positive correlation between PDB and PM₂.₅ in daytime and little correlation between them in nighttime. It indicates that PM₂.₅ mainly plays the promoting effect on the photodegradation of atmospheric benzene. This is the first study to directly determine the role of PM₂.₅ in the photochemical behavior of atmospheric benzene based on long term field observation data. Moreover, the results suggest that the regional transport of PM₂.₅ could seriously affect the geochemistry cycle of some VOCs. This research provides a new analysis method to directly quantify the effect of PM₂.₅ on the photodegradation of VOCs in the real atmosphere. It is helpful for evaluating the role of PM₂.₅ in the complex photochemical system.

This study characterized spatio-temporal variations in the concentration of benzene, toluene, ethylbenzene, and xylene (BTEX) compounds in the vicinity of gas and compressed natural gas (CNG) stations in Tehran, Iran. Health risk assessment (HRA) was computed using Monte Carlo simulations (MCS) for evaluating inhalation lifetime cancer risk (LTCR), the hazard quotient (HQ), and sensitivity analysis (SA) for BTEX exposure in different age groups (birth to <81) and as a function of distance (0–250 m) from the center of the stations. For all monitoring stations, the average values of benzene, toluene, ethylbenzene, and xylene in winter were 466.09 ± 132.25, 873.13 ± 233.51, 493.05 ± 141.22, and 910.57 ± 145.40 μg m⁻³, respectively. The mean wintertime ratios of T/B for the 12 stations ranged from 1.69 to 2.04. Furthermore, there was no significant relationship between the concentration of BTEX with either the specific month or distance from the center of stations (p > 0.05). Factors promoting BTEX formation in the study region were fuel evaporation and gas/CNG station emissions. The LTCRs for the target compounds in the winter for different age groups and distances from the center of stations was limited to 2.11 × 10⁻⁴ to 1.82 × 10⁻³ and 2.30 × 10⁻⁴ to 2.01 × 10⁻³, respectively, which exceeded proposed values by U.S. EPA. Moreover, the HQs for BTEX for three age groups and distances were limited to between 2.89 × 10⁻⁵ and 9.33 × 10⁻², which were lower than the acceptable limit (HQs < 1). The results of this work are applicable to similar areas that are heavily populated with vehicular traffic. This study motivates a closer look at mitigation strategies to limit the health effects of carcinogenic emissions such as benzene and ethylbenzene from gas/CNG stations.

Perfluorooctanoic acid (PFOA) toxicity is of considerable concern due to its wide application, environmental persistence, and bioaccumulation. In the current study, we used a scaffold-free three-dimensional (3D) spheroid model of mouse liver cells (AML12) to explore the toxicity of PFOA and emerging alternatives (HFPO-DA and PFO4DA). Comparing the short-term (24 and 72 h treatment) toxicity of PFOA between conventional 2D monolayer cells and 3D spheroids, we found that spheroids had higher EC₅₀ values and lower ROS levels after treatment, indicating their greater resistance to PFOA. Cell viability (i.e., adenosine triphosphate (ATP) content and lactate dehydrogenase (LDH) leakage) and liver-specific function (i.e., albumin secretion) were stable in spheroids through 28 day of culture. However, under 100 and 200 μM-PFOA treatment for 28 day, ROS levels, LDH leakage, and caspase3/7 activity all increased significantly. As a sensitive parameter, ROS showed a significant increase at 21 day, even in the 50 μM-PFOA group. Consistent with the elevation of ROS and caspase3/7, the expressions of oxidative stress- and apoptosis-related genes, including Gsta2, Nqo1, Ho-1, caspase3, p53, and p21, were induced in dose- and time-dependent manners after PFOA exposure. The peroxisome proliferator-activated receptor alpha (PPARα) pathway was also activated after treatment, with significant induction of its target genes, Fabp4 and Scd1. Similar to PFOA, both HFPO-DA and PFO4DA activated the PPARα pathway, induced ROS levels, and initiated cell damage, though at a relatively lower extent than that of PFOA. Our results imply that the 3D spheroid model is a valuable tool in chronic toxicological studies.

Environmental chemicals such as phthalate esters may have adverse effects on anogenital distance (AGD), but the evidence in both genders has not been reviewed systematically. The objective of the present study is to conduct a systematic review and meta-analysis of studies that analyzed the relationship between exposure to phthalates and AGD. English papers published up to March 2018 were searched in PubMed, Scopus, Clarivate-Web of Science, and Google scholar. We applied fixed-effects models to calculate pooled beta coefficient [β]. In the case of heterogeneity, random-effects models were used. Using the comprehensive search strategies, 313 papers were identified and after screening, 10 of them were included in this study. In primary analyses, we found that exposure to phthalates was not associated with short AGD (β = −0.11; 95% CI, −0.27, 0.06; I² = 0%). However, results of subgroup analyses indicated that in boys, the sum of di-2-ethylhexyl phthalate (∑DEHP) metabolites had significant association with the risk of shortened anopenile distance (AGDAP) (β = −0.915, 95% CI: 1.629, −0.2) and anoscrotal distance (AGDAS) (β = −0.857, 95% CI: 1.455, −0.26). In addition, urinary monobutyl phthalate (MBP), monoethyl phthalate (MEP), and monoisobutyl phthalate (MiBP) were associated with short AGDAP. We also observed significant association between monobenzylphthalate (MBzP) and anofourchette distance (AGDAF) in girls. Our study provided findings on significant association of exposure to ∑DEHP metabolites, MBP, MEP, and MiBP with shortened AGDAP in boys. The mechanisms of phthalates effect on AGD may involve receptors and enzymes involved in steroidgenesis, negative influence on Leydig cells, cell proliferation, gonocyte cell numbers, and testosterone production.

Biological tests with plant seeds have been adopted in many studies to investigate the phytotoxicity of pollutants to facilitate the control of risks and remain to be optimized. In this work, the experiment with a small sample size (Experiment 1) and the experiment with a large one (Experiment 2) were designed to study the effect of tetracycline (TC) on Chinese cabbage (Brassica rapa L.) at seed germination and radicle elongation stages. At the former stage, germination number data were obtained to analyze the germination energy (GE) and to judge the probability of the number of germinated seeds (Pₙ) by the binomial distribution model in Experiment 1. While germination time-to-number data were obtained to analyze the mean time to germination (MGT), the estimate of mean time to germination (EMGT) by survival analysis method, the time to germination for 50% of total seeds (T₅₀) and the germination rate (GR) besides GE in Experiment 2. At the latter stage, the data of radicle length (RL) were obtained in all the experiments and the influence from the former stage on this stage was excluded in Experiment 2 but not in Experiment 1. Results showed that TC had universal adverse effects on the latter stage but not on the former stage in the experiments. Considering the availability of germination data for statistical analysis and the robustness of RL data, the methods adopted in Experiment 2 were more feasible than those in Experiment 1. In addition, Chinese cabbage seeds with medium size have the character of rapid germination compared with the commonly used crop species and can be used to shorten the experimental cycle to study the responses of seeds to pollutants to evaluate the phytotoxicity.We introduced survival analysis method to analyze the germination time-to-number data obtained in seed germination test to evaluate the phytotoxicity of tetracycline.

Solid phase extraction has been increasingly applied for the detection of organic micropollutants (OMPs). However, time-consuming and high-cost disadvantages also limit the widespread use of this method, especially for the extraction of large-volume field water samples. In this study, a gas chromatography-mass spectrometry (GC-MS) method based on the magnetic microsphere (M150) solid-phase-extraction (MSPE) was established to investigate the OMPs in source water throughout the whole Huai River. In brief, the results demonstrated that the extraction efficiency of the M150 was superior to that of C₁₈ and HLB for the selected OMPs, including species of polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides (OCPs), phthalate esters (PAEs) and nitrobenzenes (NBs), and the method detection limits of M150 for these OMPs were comparable to those of C₁₈ and HLB. The optimized conditions of extraction and elution were the 100 mg/L dosages of microspheres, extraction time of 60 min and pH of 2, and the eluent with a similar polarity, hydrophobicity and molecular structure to the OMPs rendered higher elution efficiencies. A total of 21 types of OMPs affiliating to PAHs, OCPs, PAEs and NBs were detected by the established method, with the total concentrations of 505–2310 ng/L in source water of the Huai River. Spatial differences of the OMPs were also observed, demonstrating the link between pollutant profiles and geographical locations. This study provides an alternative to enrich OMPs in filed water samples, and it reveals pollutant profiles of source water throughout the whole Huai River.