Training the next generation of plastics pollution researchers: tools, skills and career perspectives in an interdisciplinary and transdisciplinary field.

Plastics pollution research attracts scientists from diverse disciplines. Many Early Career Researchers (ECRs) are drawn to this field to investigate and subsequently mitigate the negative impacts of plastics. Solving the multi-faceted plastic problem will always require breakthroughs across all levels of science disciplinarity, which supports interdisciplinary discoveries and underpins transdisciplinary solutions. In this context, ECRs have the opportunity to work across scientific discipline boundaries and connect with different stakeholders, including industry, policymakers and the public. To fully realize their potential, ECRs need to develop strong communication and project management skills to be able to effectively interface with academic peers and non-academic stakeholders. At the end of their formal education, many ECRs will choose to leave academia and pursue a career in private industry, government, research institutes or non-governmental organizations (NGOs). Here we give perspectives on how ECRs can develop the skills to tackle the challenges and opportunities of this transdisciplinary research field and how these skills can be transferred to different working sectors. We also explore how advisors can support an ECRs' growth through inclusive leadership and coaching. We further consider the roles each party may play in developing ECRs into mature scientists by helping them build a strong foundation, while also critically assessing problems in an interdisciplinary and transdisciplinary context. We hope these concepts can be useful in fostering the development of the next generation of plastics pollution researchers so they can address this global challenge more effectively.

Histological, enzymatic and chemical analyses of the potential effects of differently sized microplastic particles upon long-term ingestion in zebrafish (Danio rerio).

In microplastics (MPs) research, there is an urgent need to critically reconsider methodological approaches and results published, since public opinion and political decisions might be based on studies using debatable methods and reporting questionable results. For instance, recent studies claim that MPs induce intestinal damage and that relatively large MPs are transferred to, e.g., livers in fish. However, there is methodological criticism and considerable concern whether MP transfer to surrounding tissues is plausible. Likewise, there is an ongoing discussion in MP research if MPs act as vectors for adsorbed hazardous chemicals. In this study, effects of very small (4-6 µm) and very large (125-500 µm) benzo(a) pyrene (BaP)-spiked polyethylene (PE) particles administered via different uptake routes (food chain vs. direct uptake) were compared in a 21-day zebrafish (Danio rerio) feeding experiment. Particular care was taken to prevent cross-contamination of MPs during dissection and histological sample preparation. In contrast to numerous reports in literature describing similar approaches, independent of exposure route and MP size, no adverse effects could be detected. Likewise, no BaP accumulation could be documented, and MPs were exclusively seen in the lumen of the intestinal tract, which, however, did not induce any histopathological effects. Results indicate that in fish MPs are taken up, pass along the intestinal lumen and are excreted without any symptoms of adverse effects.

Novel aspects of uptake patterns, metabolite formation and toxicological responses in Salmon exposed to the organophosphate esters-Tris(2-butoxyethyl)- and tris(2-chloroethyl) phosphate.

Given the compound differences between tris(2-butoxyethyl)- and tris(2-cloroethyl) phosphate (TBOEP and TCEP, respectively), we hypothesized that exposure of juvenile salmon to TBOEP and TCEP will produce compound-specific differences in uptake and bioaccumulation patterns, resulting in potential formation of OH-metabolites. Juvenile salmon were exposed to waterborne TCEP or TBOEP (0.04, 0.2 and 1 mg/L) for 7 days. The muscle accumulation was measured and bioconcentration factor (BCF) was calculated, showing that TCEP was less accumulative and resistant to metabolism in salmon than TBOEP. Metabolite formations were only detected in TBOEP-exposed fish, showing seven phase I biotransformation metabolites with hydroxylation, ether cleavage or combination of both reactions as important metabolic pathways. In vitro incubation of trout S9 liver fraction with TBOEP was performed showing that the generated metabolite patterns were similar to those found in muscle tissue exposed in vivo. However, another OH-TBOEP isomer and an unidentified metabolite not present in in vivo exposure were observed with the trout S9 incubation. Overall, some of the observed metabolic products were similar to those in a previous in vitro report using human liver microsomes and some metabolites were identified for the first time in the present study. Toxicological analysis indicated that TBOEP produced less effect, although it was taken up faster and accumulated more in fish muscle than TCEP. TCEP produced more severe toxicological responses in multiple fish organs. However, liver biotransformation responses did not parallel the metabolite formation observed in TBOEP-exposed fish.

Lipid peroxidation and oxidative stress responses in juvenile salmon exposed to waterborne levels of the organophosphate compounds tris(2-butoxyethyl)- and tris(2-chloroethyl) phosphates.

There is limited knowledge on the toxicological, physiological, and molecular effects attributed to organophosphate (OP) compounds currently used as flame retardants or additives in consumer products. This study investigated the effects on oxidative stress and lipid peroxidation in juvenile Atlantic salmon liver and brain samples after exposure to two OP compounds, tris(2-butoxyethyl) phosphate (TBOEP) and tris(2-chloroethyl) phosphate (TCEP). In this study, groups of juvenile Atlantic salmon were exposed using a semistatic experimental protocol over a 7-d period to 3 different concentrations (0.04, 0.2, or 1 mg/L) of TBOEP and TCEP. When toxicological factors such as bioaccumulation and bioconcentration, and chemical structural characteristics and behavior, including absorption to solid materials, are considered, these concentrations represent environmentally relevant concentrations. The concentrations of the contaminants were derived from levels of their environmental occurrence. The expression of genes related to oxidative stress-glutathione peroxidase (GPx), glutathione reductase (GR), glutathione S-transferase (GST)-and to lipid peroxidation-peroxisome proliferator-activated receptors (PPAR)-were determined using quantitative (real-time) polymerase chain reaction (PCR). The presence of PPAR proteins was also investigated using immunochemical methods. Levels of thiobarbituric acid-reactive substances (TBARS) in liver were used as a measure of lipid peroxidation. Overall, our data show an increase in lipid peroxidation, and this was associated with an augmented expression of genes from the glutathione family of responses. Interestingly, PPAR expression in liver after exposure to TBOEP and TCEP was consistently decreased compared to controls, while expression in brain did not show a similar trend. The results suggest that OP contaminants may induce oxidative stress and thus production of reactive oxygen substances (ROS), and modulate lipid peroxidation processes in organisms.

Differential modulation of neuro- and interrenal steroidogenesis of juvenile salmon by the organophosphates - tris(2-butoxyethyl)- and tris(2-cloroethyl) phosphate.

Following the ban of polybrominated diphenyl ether (PBDEs) flame retardants under well-documented toxicity issues, organophosphate such as tris(2-butoxyethyl) phosphate (TBOEP) and tris(2-cloroethyl) phosphate (TCEP) were considered as potential substitutes. Although TBOEP and TCEP are consistently detected in the aquatic environment, there are few data about the possible toxicological effects of these compounds on aquatic organisms, including fish. In the present study, we have investigated the influence of TBOEP and TCEP on neuro- and interrenal steroidogenesis of juvenile Atlantic salmon (Salmo salar), after a seven-day exposure to four different concentrations (0 (control), 0.04, 0.2 and 1mg/L) of each compound. TBOEP and TCEP were diluted in Milli-Q water. The expression of genes involved in steroidogenesis (StAR, cyp19a, cyp19b, cholesterol side-chain cleavage enzyme (P450scc), 3ß-hydroxysteroid dehydrogenase (3ß-hsd), and 11ß-hydroxylase (cyp11ß)), were analyzed in the brain and head kidney using real-time PCR. Plasma 11-ketotestosterone (11-KT) analysis was performed using enzyme immunoassay (EIA). Our results showed that TBOEP accumulated more rapidly than TCEP in fish muscle tissue. Surprisingly, TBOEP produced less pronounced effects than TCEP on neural and interrenal steroidogenic responses, despite the observed rapid uptake and bioaccumulation pattern. Specifically, TBOEP produced significant and consistent concentration-specific alterations on neural- and interrenal steroidogenesis. Plasma levels of 11-KT were not significantly altered by any of the exposures. The increased expression of steroidogenic genes demonstrated in the present study could produce time-specific alterations in the production of glucocorticoids and steroid hormones that play integral roles in fish metabolism, stress responses and adaptation, sexual maturation, reproduction and migration with overt consequences on reproductive success and survival.