Identification of Additives in Disposable Face Masks and Evaluation of Their Toxicity Using Marine Medaka (Oryzias melastigma).

The COVID-19 pandemic has resulted in huge amounts of face masks worldwide. However, there is a lack of awareness on the additives and their potential risk to aquatic ecosystems of face masks. To address this issue, the additives and their toxicity in 13 face masks (e.g., polypropylene, polyethylene, and polylactic acid) were determined using nontarget analysis and bioassays. A total of 826 organic additives including intermediates (14.8%), surfactants (9.3%), plasticizers (8.2%), and antioxidants (6.1%) were tentatively identified, with 213 compounds being assigned confidence levels of 1 and 2. Interestingly, polylactic acid masks contained more additives than most polypropylene or polyethylene masks. Among these additives, the concentration of tris(2-ethylhexyl) phosphate in masks was 9.4-978.2 ng/g with a 100% detection frequency. Furthermore, 13 metals such as zinc (up to 202.0 µg/g), copper (32.5 µg/g), and chromium (up to 5.7 µg/g) were detected in the face masks. The methanol extracts of the masks showed the developmental toxicity, swimming behavior, and/or endocrine disruption in embryos/larvae of Oryzias melastigma. The findings demonstrate that face masks contain various toxic additives to marine medaka, which deserves close attention to pollution by face masks.

Plastic pollution from takeaway food industry in China.

China's takeaway food industry is growing rapidly, and bringing unprecedented demand for plastic packaging, which results in serious plastic pollution and increasing emissions of plasticizers of phthalate esters (PAEs) and greenhouse gases (GHGs). This study assesses the current and future situation of plastic usage for takeaway food packaging in China, and also analyzes the PAEs and GHG emissions brought by these plastics under different scenarios. From 2010 to 2020, the plastic usage grew from 2.92 to 101 × 104 tons, and brought 112-3845 kg PAEs and 43.6-1438 kt CO2e GHG emissions. Their distribution exhibited a clear 'two-line' pattern: higher features mostly located in Beijing-Guangzhou and Beijing-Shanghai railways. The socio-economic factors model performed better than the growth rate model for plastic usage prediction from 2021 to 2060. It is predicted that 40.6 Mt. plastic would be consumed in 2060, and they will bring 155 tons PAEs and 37.0 Mt. CO2e GHGs. At that time, biodegradable plastic replaced or plastic cycling cannot significantly contribute to national carbon reduction, unless using a temperature change of 2 °C scenario. Our work improves the understanding of PAEs and GHG emission from plastic pollution, and provides insight into long-term dynamics in the plastics management of takeaway food industry.

Endocrine disruptor responses in the embryos of marine medaka (Oryzias melastigma) after exposure to aged plastic leachates.

The issue of the additives leached from plastics has attracted widespread attention. More crucially, endocrine disruptor status for several leached additives has been established. However, little is known about the overall endocrine disrupting effects of aged plastic leachates. Therefore, the transcriptional responses of endocrine-related genes were assessed in the embryos of marine medaka (Oryzias melastigma), which were exposed to the leachates from aged plastics that were immersed into the simulated seawater (SW) or fish digest (FD). The results revealed that there was a great difference between the SW and FD leachates in the transcripts of endocrine-related genes. With the exception of cyp1a, all target genes had their transcripts potentially down-regulated by the FD leachates. Chgl (a biomarker for estrogens), pparß (related to lipid metabolism), and cyp19a (related to sexual differentiation and reproduction) transcripts tended to be repressed by the SW leachates, while pparα, pparγ and cyp1a (mediating metabolism of xenobiotics) transcripts were stimulated. In addition, a redundancy analysis was carried out to determine the relationship between the leached additives and the transcriptional changes. However, the additives only partially explained the variation in the transcripts of endocrine-related genes (24.8%), indicating that other leached additives may have an impact on target gene transcription. This study provided molecular evidence of the aged plastic leachates' endocrine disrupting effects. Exploring the primary factors that affect the transcriptional alterations would require more research.

Exposure to estrone disrupts the endocrine system of western mosquitofish (Gambusia affinis).

Estrone (E1) is one of the predominant natural estrogens detected in aquatic environments, yet little is known about its effects on the endocrine system in fish. In this study, the sex ratio, secondary sexual characteristics, gonadal histology, and transcriptional levels of genes closely related to sex differentiation and hypothalamic-pituitary-gonadal-liver (HPGL) axis were assessed in western mosquitofish (Gambusia affinis) after a full life-cycle exposure to E1 (0, 25.4, 143, 740, and 4300 ng/L) for 119 days. The results showed that exposure to 4300 ng/L of E1 resulted in 100% female and inhibited the growth of females. Exposure to environmentally relevant concentrations of E1 (143 and 740 ng/L) led to obvious feminization of skeletons and anal fins in males. Exposure to 740 and 4300 ng/L of E1 increased the proportion of mature spermatocytes in females, and exposure to 143 and 740 ng/L decreased the proportion of mature spermatocytes in males. Moreover, the transcripts of genes related to sex differentiation and HPGL axis were changed in the E1-exposed adult fish and embryos inside females. This study has provided valuable data on the endocrine disruption effects of E1 at environmentally relevant concentrations in G. affinis.

Toxicity and Estrogenicity of Bisphenol TMC in Oryzias melastigma via In Vivo and In Silico Studies.

Bisphenol 4-[1-(4-hydroxyphenyl)-3,3,5-trimethylcyclohexyl] phenol (BPTMC), as a substitute for bisphenol A, has been detected in environments. However, the ecotoxicological data of BPTMC are extremely scarce. Here, the lethality, developmental toxicity, locomotor behavior, and estrogenic activity of BPTMC at different concentrations (0.25-2000 µg/L) in marine medaka (Oryzias melastigma) embryos were examined. In addition, the in silico binding potentials of O. melastigma estrogen receptors (omEsrs) with BPTMC were assessed by docking study. Low-concentration BPTMC exposure (including an environmentally relevant concentration, 0.25 µg/L) resulted in stimulating effects, including hatching rate, heart rate, malformation rate, and swimming velocity. However, elevated concentrations of BPTMC led to an inflammatory response, changed heart rate and swimming velocity in the embryos and larvae. In the meantime, BPTMC (including 0.25 µg/L) altered the concentrations of estrogen receptor, vitellogenin, and endogenous 17 ß-estradiol as well as the transcriptional levels of estrogen-responsive genes in the embryos or/and larvae. Furthermore, elaborate tertiary structures of omEsrs were built by ab initio modeling, and BPTMC exerted potent binding potential with three omEsrs with -47.23, -49.23, and -50.30 kJ/mol for Esr1, Esr2a, and Esr2b, respectively. This work suggests that BPTMC has potent toxicity and estrogenic effects in O. melastigma.

A comparative study on endocrine disrupting effects of leachates from virgin and aged plastics under simulated media in marine medaka larvae (Oryzias melastigma).

Marine plastic pollution has garnered substantial attention, but the potential endocrine disrupting effects of plastic leachates in marine organisms remain unclear. In this study, the larvae of marine medaka (Oryzias melastigma) were exposed to the leachates from virgin and aged plastics soaked in simulated seawater and fish digest for 3 days. The concentrations of vitellogenin (VTG), estradiol (E2), and 11-ketotestosterone (11-KT), as well as the transcripts of endocrine-related genes were measured in the larvae. The results revealed that endogenous E2 was more sensitive to plastic leachates than VTG and 11-KT, which was significantly affected by 26.7 % of all plastic leachates. Among all genes, estrogen receptor α was impacted mostly, being up-regulated by 53.3 % of leachates from aged plastics. The comparative results demonstrated that the leachates from plastics with different statuses caused a greater difference than those from plastics in different simulated media, and the leachates from aged plastics resulted in higher endocrine disrupting effects than those from virgin plastics. In addition, seven leached additives (plasticizers and flame retardants) could explain 25.6 % of the hormonal effects using redundancy analysis, indicating that other additives in the plastic leachates can also play important roles in regulating the endocrine system of O. melastigma larvae.

Chemical characteristics and toxicological effects of leachates from plastics under simulated seawater and fish digest.

In recent years, the ecological risks of plastics to marine environments and organisms have attracted increasing attention, especially the leachates from plastics. However, a comprehensive knowledge about the leaching characteristics and subsequent toxicological effects of leachates is still sparse. In this study, 15 different plastic products were immersed in simulated seawater and fish digest for 16 h. The leachates were analyzed through non-target and target analyses and their toxicological signatures were assessed by bioassays. In total, 240 additives were identified from the plastic leachates, among which plasticizers represented the most (16.7%), followed by antioxidants (8.7%) and flame retardants (7.1%). Approximately 40% of plastic leachates exhibited significant inhibitory effects on the bioluminescence using a recombinant luminescent assay. In addition, both the hyperactive and hypoactive behaviors were displayed in the larvae of marine medaka (Oryzias melastigma) exposed to some plastic leachates. In general, the number and amount of identified compounds under simulated fish digest were less than those under simulated seawater. However, the simulated fish digest leachates triggered higher toxicity. Redundancy analysis demonstrated that identified additives did not adequately explain the toxicological effects. Future research should focus on the identification of more additives in the plastic leachates and their potential ecological risks.

Imidacloprid and thiamethoxam affect synaptic transmission in zebrafish.

Imidacloprid (IMI) and thiamethoxam (THM) are two commonly applied neonicotinoid insecticides. IMI and THM could cause negative impacts on non-target organisms like bees. However, the information about neurotoxicity of IMI and THM in fish is still scarce. Here we investigated the effects of IMI and THM on locomotor behavior, AChE activity, and transcription of genes related to synaptic transmission in zebrafish exposed to IMI and THM with concentrations of 50 ng L-1 to 50,000 ng L-1 at 14 day post fertilization (dpf), 21 dpf, 28 dpf and 35 dpf. Our results showed that IMI and THM significantly influenced the locomotor activity in larvae at 28 dpf and 35 dpf. THM elevated AChE activity at 28 dpf. The qPCR data revealed that IMI and THM affected the transcription of marker genes belonging to the synapse from 14 dpf to 35 dpf. Furthermore, IMI and THM mainly affected transcription of key genes in γ-aminobutyric acid, dopamine and serotonin pathways in larvae at 28 dpf and 35 dpf. These results demonstrated the neurotoxicity of IMI and THM in zebrafish. The findings from this study suggested that IMI and THM in the aquatic environment may pose potential risks to fish fitness and survival.

Endocrine disrupting effects of binary mixtures of 17ß-estradiol and testosterone in adult female western mosquitofish (Gambusia affinis).

Androgens and estrogens often co-exist in aquatic environments and pose potential risks to fish populations. However, little is known about the endocrine disrupting effects of the mixture of androgens and estrogens in fish. In this study, transcriptional level of target genes related to the hypothalamic-pituitary-gonadal-liver (HPGL) axis, sex hormone level, VTG protein concentration, histology and secondary sex characteristic were assessed in the ovaries and livers of adult female western mosquitofish (Gambusia affinis) exposed to 17ß-estradiol (E2), testosterone (T), and mixtures of E2 and T for 91 days. The results showed that the transcriptional expression of cytochrome P450, family 19, subfamily A, polypeptide 1a (Cyp19a1a) was suppressed in the 200 ng/L T treatment and the 50 ng/L E2 + 200 ng/L T treatment in the ovaries. Steroidogenic acute regulatory protein (Star) and Cyp11a1 showed a similar expression pattern in the T treatment to its corresponding T + E2 mixtures. In the ovaries, the concentrations of 17ß-estradiol and testosterone were decreased in most treatments compared with the solvent control. VTG protein was induced in all steroid treatment. However, exposure to T or E2 + T mixture did not cause the abnormal cells of the ovaries and livers and an extension of the anal fins in female G. affinis. This study demonstrates that chronic exposure to E2, T and their mixtures affects the transcripts of genes in the HPGL axis, steroid hormone level and VTG protein concentration in the ovaries and livers, but fails to cause the histopathological effect of the ovaries and livers and alter the morphology of the anal fins in G. affinis.