Impact of aged and virgin polyethylene microplastics on multi end-points effects of freshwater fish tissues.

The buildup of plastic waste in aquatic environments presents serious threats to the environment, wildlife, and ultimately to humans. Specifically, microplastics (MPs) ingestion by aquatic animals leads to adverse physiological and toxicological effects. In addition, discarded MPs undergo aging and degradation processes which affect their morphological properties and chemical composition, enhancing the absorption of environmental pollutants. Under this prism, the present research was conducted to investigate and compare the impact of 'aged' versus pristine low-density polyethylene microplastics (PE-MPs) on various toxicity endpoints as biochemical and molecular parameters in the muscle tissue and liver of the freshwater fish species Perca fluviatilis. In parallel, the morphological, physicochemical, and structural changes occurred in "aged" PE-MPs, (after being exposed to UV radiation for 120 days) were studied, significantly illustrating signs of oxidation and crack propagation at the surface of the studied MPs. Fish were exposed to artificial diet reached with virgin and "aged" PE-MPs, sized 100-180 µm, at concentrations of 1 mg/g of dry food for a period of 15-days. Thereafter, liver and muscle tissues were analyzed in relation to multi oxidative parameters. Compared to the control group, the observed changes in the examined fish included increased activities of antioxidant enzymes, as superoxide dismutase, catalase and glutathione reductase, enhanced concentrations of malondialdehyde, protein carbonylation, HSP70 levels, elevated MAPK phosphorylation, induction of ubiquitin-proteins, as well as heightened levels of Bax/Bcl-2 proteins, caspases and differentiated levels of LC3 II/I, SQSTM1/p62. From the studied biomarkers, apoptosis, ubiquitin and hsp70 levels, showed a more sensitive response against the ingested MPs, followed by autophagy, p38MAPK levels, antioxidant enzymes, MDA and carbonylation levels. The effect of "aged" PE-MPs was more pronounced compared to that of the virgin ones. When evaluating the response of all oxidative stress biomarkers across the studied tissues, the liver demonstrates the highest response for the majority of the biomarkers against both virgin and "aged" PE-MPs. These findings strongly indicate that "aged" MPs activate the antioxidant defence mechanisms and impact the cellular well-being of the examined fish species.

Genotoxicity and metabolic changes induced via ingestion of virgin and UV-aged polyethylene microplastics by the freshwater fish Perca fluviatilis.

The present study aims to compare and assess the toxicity induced by aged (irradiated with ultraviolet radiation for 120 days) polyethylene microplastics (PE-MPs) in comparison to virgin (non-irradiated) ones, after feeding the freshwater fish Perca fluviatilis. To this end, MPs mediated genotoxicity was assessed by the investigation of micronucleus nuclear abnormalities frequency in fish blood, and the degree of DNA damage in the liver and muscle tissues, while metabolic alterations were also recorded in both tissues. Results showed that both virgin and aged PE-MPs induced signaling pathways leading to DNA damage and nuclear abnormalities, as well as metabolites changes in all tissues studied. Metabolic changes revealed that the metabolism of nucleic acids, energy, amino acids, and neurotransmitters was more disrupted in the liver and by aged PE-MPs compared to muscles. Fish fed with aged PE-MPs exhibited greater DNA damage, while blood cells of fish fed with virgin PE-MPs seemed to be more vulnerable to nuclear abnormalities in relation to those fed with aged PE-MPs. Moreover, aged PE-MPs induced more acute overall effects on the metabolic profiles of fish tissues, and initiated stronger stress responses, inflammation, and cellular damages in fish tissues in relation to virgin ones. Characterization of both virgin and aged MPs revealed that the latter exhibited lower crystallinity and melting point, more irregular shapes and higher moiety of oxygen and carbonyl groups, which could be attributed for their observed higher toxicity. The research outcomes provide significant insights for advancing toxicological investigations in this field.

Dose-Dependent Cytotoxicity of Polypropylene Microplastics (PP-MPs) in Two Freshwater Fishes.

The massive accumulation of plastics over the decades in the aquatic environment has led to the dispersion of plastic components in aquatic ecosystems, invading the food webs. Plastics fragmented into microplastics can be bioaccumulated by fishes via different exposure routes, causing several adverse effects. In the present study, the dose-dependent cytotoxicity of 8−10 µm polypropylene microplastics (PP-MPs), at concentrations of 1 mg/g (low dose) and 10 mg/g dry food (high dose), was evaluated in the liver and gill tissues of two fish species, the zebrafish (Danio rerio) and the freshwater perch (Perca fluviatilis). According to our results, the inclusion of PP-MPs in the feed of D. rerio and P. fluviatilis hampered the cellular function of the gills and hepatic cells by lipid peroxidation, DNA damage, protein ubiquitination, apoptosis, autophagy, and changes in metabolite concentration, providing evidence that the toxicity of PP-MPs is dose dependent. With regard to the individual assays tested in the present study, the biggest impact was observed in DNA damage, which exhibited a maximum increase of 18.34-fold in the liver of D. rerio. The sensitivity of the two fish species studied differed, while no clear tissue specificity in both fish species was observed. The metabolome of both tissues was altered in both treatments, while tryptophan and nicotinic acid exhibited the greatest decrease among all metabolites in all treatments in comparison to the control. The battery of biomarkers used in the present study as well as metabolomic changes could be suggested as early-warning signals for the assessment of the aquatic environment quality against MPs. In addition, our results contribute to the elucidation of the mechanism induced by nanomaterials on tissues of aquatic organisms, since comprehending the magnitude of their impact on aquatic ecosystems is of great importance.

Do poly(lactic acid) microplastics instigate a threat? A perception for their dynamic towards environmental pollution and toxicity.

Fears concerning microplastics (MPs) environmental fate and persistence are progressively expanding on a global basis, with the emphasis given to manufacturing bioplastics for substituting petro-derived plastics extensively growing. Among them, poly(lactic acid) (PLA) holds a pioneering role towards the replacement of conventional polymeric materials, owing to its multifunctional properties, enclosing superior mechanical properties, low cost, renewability, great biocompatibility, transparency, and thermoplasticity launching many fields of application. Due to the wide applicability of PLA in several sectors of everyday life, its waste to be released into the environment is expected to follow a growing tendency during the upcoming years. Even though PLA is a biodegradable polyester, it actually degrades under specific composting environments, including a rich oxygen environment with high temperatures (58-80 °C), high humidity (>60% moisture) as well as the presence of micro-organisms (thermophilic bacteria). Additionally, in various studies it has been implied that PLA displays slower degradation performance when found in blends with other conventional polymers, underlining the unspecified effects on PLA degradation profile, keeping thus the information about PLA degradation from a blur standpoint. Therefore, a deepened understanding of the fate and dynamic effects of PLA MPs is of primary importance. Nevertheless, the current examination of the effects of PLA MPs in terms of sorption capacities and toxicity is so far limited and broadly unexplored since the current scientific emphasis has been merely centered on the conventional MPs' behavior. In this light, the present review provides an inclusive overview of the ongoing research of poly(lactic acid) in the framework of microplastics' pollution, while the future trends and missing points in this context are highlighted.

Differentiation in the expression of toxic effects of polyethylene-microplastics on two freshwater fish species: Size matters.

The built up of microplastic (MPs) remains is shaping a new aquatic habitat and imposes the necessity for research of the effects that these relatively new pollutants exert on organisms, environment, and human health. The purpose of the present study was to verify if there is a particle-size dependence of fish response to MPs. Thus, we exposed two freshwater fish species, the zebrafish (Danio rerio) and perch (Perca fluviatilis) for 21 days to polyethylene microplastics (PE-MPs) sized 10-45 µm and 106-125 µm. Thereafter, in the liver and gills tissues, biochemical and molecular parameters and the metabolic profile were examined. Ex-vivo characterization by ATR-FTIR spectroscopy exhibited increased concentration of 10-45 µm PE-MPs in the liver of the two fish species while 106-125 µm PE-MPs mostly concentrated in fish gills. The penetration of PE-MPs to fish and the induced oxidative stress triggered changes in lipid peroxidation, DNA damage and ubiquitination and furthermore stimulated signal transduction pathways leading to autophagy and apoptosis. The smaller PE-MPs were more potent in inducing alterations to all the latter parameters measured than the larger ones. Tissue response in both fish seems to depend on the parameter measured and does not seem to follow a specific pattern. Our results showed that there is no clear sensitivity of one fish species versus the other, against both sizes of PE-MPs they were exposed. In perch the metabolic changes in gills were distinct to the ones observed in liver, following a size dependent pattern, indicating that stress conditions are generated through different mechanisms. All the parameters employed can be suggested further as biomarkers in biomonitoring studies against PE-MPs.

Toxicity and Functional Tissue Responses of Two Freshwater Fish after Exposure to Polystyrene Microplastics.

Microplastics (MPs)' ingestion has been demonstrated in several aquatic organisms. This process may facilitate the hydrophobic waterborne pollutants or chemical additives transfer to biota. In the present study the suitability of a battery of biomarkers on oxidative stress, physiology, tissue function and metabolic profile was investigated for the early detection of adverse effects of 21-day exposure to polystyrene microplastics (PS-MPs, sized 5-12 µm) in the liver and gills of zebrafish Danio rerio and perch, Perca fluviatilis, both of which are freshwater fish species. An optical volume map representation of the zebrafish gill by Raman spectroscopy depicted 5 µm diameter PS-MP dispersed in the gill tissue. Concentrations of PS-MPs close to the EC50 of each fish affected fish physiology in all tissues studied. Increased levels of biomarkers of oxidative damage in exposed fish in relation to controls were observed, as well as activation of apoptosis and autophagy processes. Malondialdehyde (MDA), protein carbonyls and DNA damage responses differed with regard to the sensitivity of each tissue of each fish. In the toxicity cascade gills seemed to be more liable to respond to PS-MPs than liver for the majority of the parameters measured. DNA damage was the most susceptible biomarker exhibiting greater response in the liver of both species. The interaction between MPs and cellular components provoked metabolic alterations in the tissues studied, affecting mainly amino acids, nitrogen and energy metabolism. Toxicity was species and tissue specific, with specific biomarkers responding differently in gills and in liver. The fish species that seemed to be more susceptible to MPs at the conditions studied, was P. fluviatilis compared to D. rerio. The current findings add to a holistic approach for the identification of small sized PS-MPs' biological effects in fish, thus aiming to provide evidence regarding PS-MPs' environmental impact on wild fish populations and food safety and adequacy.

Adverse effects polystyrene microplastics exert on zebrafish heart - Molecular to individual level.

In the present study the effects of sublethal concentrations of polystyrene microplastics (PS-MPs) on zebrafish were evaluated at multiple levels, related to fish activity and oxidative stress, metabolic changes and contraction parameters in the heart tissue. Zebrafish were fed for 21 days food enriched with PS-MPs (particle sizes 3-12 µm) and a battery of stress indices like DNA damage, lipid peroxidation, autophagy, ubiquitin levels, caspases activation, metabolite adjustments, frequency and force of ventricular contraction were measured in fish heart, parallel to fish swimming velocity. In particular, exposure to PS-MPs caused significant decrease in heart function and swimming competence, while enhanced levels of oxidative stress indices and metabolic adjustments were observed in the heart of challenged species. Among stress indices, DNA damage was more vulnerable to the effect of PS-MPs. Our results provide evidence on the multiplicity of the PS-MPs effects on cellular function, physiology and metabolic pathways and heart rate of adult fish and subsequent effects on fish activity and fish fitness thus enlightening MPs characterization as a potent environmental pollutant.

Assessing ecological water quality with macroinvertebrates and fish: a case study from a small Mediterranean river.

Biological elements, such as benthic macroinvertebrates and fish, have been used in assessing the ecological quality of rivers according to the requirements of the Water Framework Directive. However, the concurrent use of multiple organism groups provides a broader perspective for such evaluations, since each biological element may respond differently to certain environmental variables. In the present study, we assessed the ecological quality of a Greek river (RM4 type), during autumn 2003 and spring 2004 at 10 sites, with benthic macroinvertebrates and fish. Hydromorphological and physicochemical parameters, habitat structure, and riparian vegetation were also considered. Pollution sensitive macroinvertebrate taxa were more abundant at headwaters, which had good/excellent water quality according to the Hellenic Evaluation System (HES). The main river reaches possessed moderate water quality, while downstream sites were mainly characterised as having bad or poor water quality, dominated by pollution-tolerant macroinvertebrate taxa. Macroinvertebrates related strongly to local stressors as chemical degradation (ordination analysis CCA) and riparian quality impairment (bivariate analysis) while fish did not. Fish were absent from the severely impacted lower river reaches. Furthermore, external pathological signs were observed in fish caught at certain sites. A combined use of both macroinvertebrates and fish in biomonitoring programs is proposed for providing a safer assessment of local and regional habitat impairment.

Seasonal variation of fish abundance and biomass in gillnet catches of an East Mediterranean lake: Lake Doirani.

The seasonal variation of fish species composition and abundance in gillnet catches (14-90 mm knot-to-knot) from the Greek part of the transboundary Lake Doirani was studied during the period 2006-2007. A total of 8,419 specimens weighing 182.3 kg and belonging to 9 species were caught. Catch composition differed with season. Thus, Rhodeus meridionalis dominated in terms of NPUE the spring, Perca fluviatilis the summer and Albumus macedonicus the autumn and winter catches. Cyprinids were generally the most abundant, with the cyprinids:percids biomass ratio ranging from 1.7 in summer to 14.8 in winter, supporting the eutrophic character of the lake. Richness and Shannon-Wiener diversity and evenness indices differed seasonally (ANOVA; p < 0.05). The abundance-biomass comparative (ABC) curves showed that fish communities were dominated by one or a few opportunistic species (e.g. Rhodeus meridionalis, Albumus macedonicus), which while dominated in number did not dominate in biomass, being small bodied.