Interspecific competitive interactions affect body size and oxidative status of two nonnative salmonid species.

In fish, interspecific interactions between nonnative and other sympatric species are considered determinants in shaping species assemblages. Such interactions can also arise between nonnative fish species only, including salmonids such as the brown trout (Salmo trutta, Linnaeus, 1758) and the rainbow trout (Oncorhynchus mykiss, Walbaum, 1792), returning contrasting outcomes. The present manipulative experiment was aimed at exploring the effect of interspecific competition on the body growth and the oxidative status of parr (2 + -year-old individuals) of the brown trout and the rainbow trout. Allopatric (intraspecific competition) and sympatric (interspecific competition) populations of these species were experimentally recreated in two wild streams. At the end of a 2-month-long experiment, changes in specific growth rate (SGR), oxidative status (i.e., levels of reactive oxygen species and activity of antioxidant enzymes such as superoxide dismutase - SOD, catalase - CAT and glutathione peroxidase - GPx) and oxidative damage (i.e., lipid peroxidation) were investigated in brown and rainbow trout individuals maintained in allopatric or sympatric populations. Sympatric interactions between rainbow and brown trout parr resulted in a significant decrease in SGR of brown trout individuals only. Moreover, an overall modulation of the oxidative status, in terms of an increase in ROS levels coupled with the activation of SOD and CAT activity, occurred in brown trout individuals under sympatric conditions. These findings might suggest that, under sympatric conditions, parr of the rainbow trout are more competitive than brown trout for food acquisition. However, this competition affected the antioxidant defenses of the brown trout only, probably because of reduced ingestion of dietary antioxidants or increased physical activity and aggressive behavior. Thus, interspecific interactions can induce physiological and phenotypic effects on parr of nonnative salmonids, with potential consequences on the establishment of populations of these species in freshwater ecosystems.

Trends and potential human health risk of trace elements accumulated in transplanted blue mussels during restoration activities of Flekkefjord fjord (Southern Norway).

The monitoring of contaminants represents a priority to preserve the integrity of marine ecosystems, as well as to plan and to manage restoration activities in order to protect environmental and human health. In the present study, a 6-months active biomonitoring was performed to explore the levels of eighteen trace and toxic elements, including heavy metals (TEs; i.e. Al, As, Ca, Cd, Cr, Cu, Fe, Hg, K, Mg, Mn, Na, Ni, P, Pb, Sr, Ti, and Zn), accumulated in soft tissues of blue mussel (Mytilus edulis Linnaeus, 1758) individuals transplanted at different depths (5- and 15-m depth) in five locations within the Flekkefjord fjord (Southern Norway). As this area suffered a long-lasting contamination due to both organic and inorganic contaminants, a series of restoration activities were activated to tackle and to prevent potential risks for ecosystem and local population. Our results demonstrated that the levels of TEs accumulated in edible tissues of transplanted mussels in the Flekkefjord fjord were generally low before the beginning of the restoration activities. However, location- and time-specific differences in the accumulation of TEs were noted after the implementation of such activities. Interestingly, the levels of Fe and Mn significantly increased after the beginning of the restoration activities, likely because the release of these TEs from the slag used in such operations and/or resuspension of contaminated sediments. However, assuming that native mussels can accumulate the same TEs at levels measured in transplanted individuals, our results suggest a substantial safety for human consumption of native mussels from the Flekkefjord fjord, regardless of restoration activities.

Prenatal yolk corticosterone exposure promotes skeletal growth and induces oxidative imbalance in yellow-legged gull embryos.

Maternally derived hormones induce variation in offspring phenotype, with consequences that can carry over into post-natal life and even into adulthood. In birds, maternal egg corticosterone (CORT) is known to exert contrasting effects on offspring morphology, physiology and behaviour after hatching. However, information on the effects of CORT exposure on pre-hatching embryonic development is limited. We experimentally increased yolk CORT levels in yellow-legged gull (Larus michahellis) eggs, and assessed the effects on embryo pre-hatching development and oxidative status of brain and liver. CORT-supplemented embryos reached a larger skeletal size and liver mass compared with controls. Embryos from CORT-injected last-laid eggs showed decreased activity of the hepatic antioxidant enzymes superoxide dismutase and catalase, while intermediate-laid eggs showed increased levels of lipid peroxidation. However, elevated yolk CORT did not affect oxidative stress endpoints in the brain. Our results indicate that elevated yolk CORT levels affect prenatal embryo development by promoting skeletal growth, and induce laying sequence- and organ-specific oxidative imbalance, with potential adverse consequences during postnatal life, especially for late-hatched offspring.

Oxidative stress-related effects induced by micronized polyethylene terephthalate microparticles in the Manila clam.

Microplastic (MP) contamination represents a serious threat for marine organisms. Several lab studies demonstrated adverse effects induced by exposure to different MP polymers toward diverse marine species. However, the information regarding toxicity of polyethylene terephthalate (PET) MPs is largely unknown. The present study was aimed at investigating the adverse effects induced by 7-day exposure to two concentrations (0.125 or 12.5 µg/ml) micronized, irregular shaped and variable size PET microparticles (PET-MPs) toward Manila clam (Ruditapes philippinarum). Histological analyses were performed to assess tissue damage on digestive glands, gonads, gut and gills, whereas oxidative stress-related effects, including the concentration of pro-oxidant molecules, activity of antioxidant (superoxide dismutase - SOD, catalase - CAT and glutathione peroxidase - GPx) and detoxifying (glutathione S-transferase - GST) enzymes, as well as levels of lipid peroxidation, were determined in gills and digestive gland. Our results showed that clams ingest and egest micronized PET-MPs, but no marked histological alterations to bivalve tissues occurred. Although PET-MPs did not produce oxidative stress in the digestive gland, these materials significantly altered oxidative status of gills, leading to lipid peroxidation. No apparent clear indication of a weakness of bivalve health status was obtained in this study.

Prenatal independent and combined effects of yolk vitamin E and corticosterone on embryo growth and oxidative status in the yellow-legged gull.

Variation in the concentration of antioxidants and hormones of maternal origin in the eggs of birds can have a profound influence on offspring phenotype both prenatally and postnatally. Egg maternal substances can have interacting effects, but experimental studies of the consequences of the combined variation in the egg concentration of such molecules are extremely rare, particularly as far as prenatal stages are concerned. We manipulated the yolk concentration of vitamin E and corticosterone, which are, respectively, the main antioxidant and the main glucocorticoid hormone in bird eggs, both independently and simultaneously, and we tested their separate and combined effects on growth and oxidative status in the liver and in the brain of yellow-legged gull (Larus michahellis) embryos. Egg supplementation of relatively large physiological doses of corticosterone depressed embryo growth (total body mass, tarsus length and liver mass), whereas administration of vitamin E in association with corticosterone restored normal growth. Vitamin E did not affect embryo growth when administered alone. We further analysed the independent and combined effects of vitamin E and corticosterone on liver and brain total antioxidant capacity, the concentration of reactive oxygen molecules and lipid peroxidation. Vitamin E significantly reduced liver total antioxidant capacity, while corticosterone depressed brain lipid peroxidation. Prenatal exposure to vitamin E and corticosterone appears to have antagonistic effects on body growth, although vitamin E is not limiting in yellow-legged gull eggs. In combination with the results of previous experiments on the same species applying smaller experimental doses or focusing on the postnatal rather than prenatal life stages, our findings indicate that the effects of a physiological increase in the egg concentration of these substances can be life stage and dose specific, implying that generalizing prenatal effects of egg compounds may not be feasible.

Age- and sex-dependent variation in the activity of antioxidant enzymes in the brown trout (Salmo trutta).

Oxidative stress is defined as the imbalance between pro-oxidant and antioxidant molecules in favor of the former and it represents one of the main driving forces of aging. To counteract the harmful effects of oxidative stress, organisms evolved a complex antioxidant system. According to the free radical theory of aging, while the production of reactive oxygen species (ROS) increases with age, the antioxidant defenses decline. Although this relationship has been investigated in diverse vertebrate taxa, the information in fish is scant and inconsistent, particularly for wild populations. Thus, the aim of the present study was the investigation of age- and sex-related changes of the antioxidant enzymes activity in free-living individuals of the brown trout (Salmo trutta). We measured the activity of the main enzymes involved in antioxidant protection, namely superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione S-transferase (GST), as well as lipid peroxidation, in the gills and the liver dissected from brown trout (1+- to 5+-year-old). A significant age-dependent variation in the activity of antioxidant enzymes was noted, with the exception of CAT. GPx activity followed a significant increasing trend with age in both the organs, while SOD decreased in the liver. Increased GST activity was found in the gills only. Lipid peroxidation levels significantly decreased with age in both the organs. SOD and CAT showed sex-dependent differences in the liver of brown trout, with males showing lower enzymatic activity than females. Our data contribute to improve the knowledge on the relationship between antioxidant enzyme activity, aging, and sex in fish.

Effect of yolk corticosterone on begging in the yellow-legged gull.

Behavioral lateralization is widespread across vertebrates. The development of lateralization is affected by both genetic and environmental factors. In birds, maternal substances in the egg can affect offspring lateralization via activational and/or organizational effects. Corticosterone affects the development of brain asymmetry, suggesting that variation in yolk corticosterone concentration may also influence post-natal behavioral lateralization, a hypothesis that has never been tested so far. In the yellow-legged gull (Larus michahellis), we increased yolk corticosterone concentration within physiological limits and analyzed the direction of lateralization of hatchlings in reverting from supine to prone position ('RTP' response) and in pecking at dummy parental bills to solicit food provisioning ('begging' response). We found that corticosterone treatment negatively affected the frequency of begging and it may cause a slight leftward lateralization. However, the direction of lateralization of the RTP response was not affected by corticosterone administration. Thus, our study shows a maternal effect mediated by corticosterone on a behavioral trait involved in parent-offspring communication during food provisioning events. The findings on lateralization are not conclusive due to the weak effect size but provide information for further ecological and evolutionary studies, investigating mechanisms underlying the development of lateralization.

Independent and combined effects of egg pro- and anti-oxidants on gull chick phenotype.

Oviparous mothers transfer to their eggs components that have both independent and combined effects on offspring phenotype. The functional interaction between egg components, such as antioxidants and hormones, suggests that a change in the concentration of one component will have effects on offspring traits that depend on the concentration of other interacting components. However, the combined effects of variation in different egg components are virtually unknown. Bird eggs contain vitamin E, a major antioxidant, and also maternal corticosterone. The independent consequences of variation in the egg concentrations of these compounds for offspring phenotype are largely unknown and no study has investigated their combined effects. We manipulated the concentration of vitamin E and corticosterone in the eggs of the yellow-legged gull (Larus michahellis) by administering a physiological (2 s.d.) dose both independently and in combination. We tested for an effect on chick post-natal growth, plasma antioxidant capacity (TAC) and oxidative compounds (TOS). Separate administration of vitamin E or corticosterone caused a reduction in body mass relative to controls, whereas the combined administration of the two compounds reversed their negative effects. These results suggest that maternal egg components, such as antioxidants and steroid hormones, interact and mothers must balance their concentrations in order to achieve optimal offspring phenotype. The functional relationship between vitamin E and corticosterone is corroborated by the observation of positive covariation between these compounds.

Telomere length is reflected by plumage coloration and predicts seasonal reproductive success in the barn swallow.

Individuals differ in realized fitness but the genetic/phenotypic traits that underpin such variation are often unknown. Telomere dynamics may be a major source of variation in fitness traits because physiological telomere shortening depends on environmental and genetic factors and may impair individual performance. Here, we showed that, in a population of a socially monogamous, biparental passerine bird, the barn swallow (Hirundo rustica), breeding in northern Italy, telomere length (TL) of both adult males and females positively correlated with seasonal reproductive and fledging success, as expected because long telomeres are supposed to boost performance. Telomere length was correlated with sexually dimorphic coloration in both sexes, showing for the first time in any species that coloration reliably reflects TL and may mediate mutual mate choice, leading to the observed positive assortative mating for TL in the barn swallow. Thus, TL appears to be associated with variation in a major fitness trait and may be an ultimate target of mate choice, as individuals of both sexes can use coloration to adaptively choose high-quality mates that possess long telomeres.

Contrasting effects of increased yolk testosterone content on development and oxidative status in gull embryos.

Hormone-mediated maternal effects generate variation in offspring phenotype. In birds, maternal egg testosterone (T) exerts differential effects on offspring traits after hatching, suggesting that mothers experience a trade-off between contrasting T effects. However, there is very little information on T pre-natal effects. In the yellow-legged gull (Larus michahellis), we increased yolk T concentration within physiological limits and measured the effects on development and oxidative status of late-stage embryos. T-treated embryos had a larger body size but a smaller brain than controls. Males had a larger brain than females, controlling for overall size. T treatment differentially affected brain mass and total amount of pro-oxidants in the brain depending on laying order. T-treatment effects were not sex dependent. For the first time in the wild, we show contrasting T pre-natal effects on body mass and brain size. Hence, T may enforce trade-offs between different embryonic traits, but also within the same trait during different developmental periods.

Antioxidants and embryo phenotype: is there experimental evidence for strong integration of the antioxidant system?

Organisms have evolved complex defense systems against oxidative stress. Bird eggs contain maternally derived antioxidants that protect embryos from oxidative damage. The antioxidant system components are thought to be integrated, but few studies have analyzed the covariation between antioxidant concentrations, embryo 'oxidative status' and morphology. In addition, no study has tested the effects of experimental change in yolk antioxidant concentration on other antioxidants, on their reciprocal relationships and on their relationships with embryo oxidative status or growth, which are expected if antioxidants defenses are integrated. In yellow-legged gull (Larus michahellis) embryos, we analyzed the covariation between several antioxidants, markers of 'oxidative status' [total antioxidant capacity (TAC), concentration of pro-oxidants (TOS), lipid peroxidation (LPO) and protein carbonylation (PC)] in the yolk, liver and brain, and morphology. Yolk and liver antioxidant concentrations were positively correlated reciprocally and with embryo size, and positively predicted TAC but not oxidative status. TOS and LPO were positively correlated in the liver, while TAC and LPO were negatively correlated in the brain. Weak relationships existed between antioxidants and TOS, PC and LPO. The effects of antioxidants on oxidative status and morphology were non-synergistic. An experimental physiological increase in yolk vitamin E had very weak effects on the relationships between other antioxidants or oxidative status and vitamin E concentration, the concentration of other antioxidants or oxidative status; the covariation between other antioxidants and oxidative status, and relationships between morphology or oxidative status and other antioxidants, challenging the common wisdom of strong functional relationships among antioxidants, at least for embryos in the wild.

Yolk testosterone affects growth and promotes individual-level consistency in behavioral lateralization of yellow-legged gull chicks.

Behavioral lateralization is common in animals and may be expressed at the individual- and at the population-level. The ontogenetic processes that control lateralization, however, are largely unknown. Well-established sex-dependence in androgen physiology and sex-dependent variation in lateralization have led to the hypothesis that testosterone (T) has organizational effects on lateralization. The effects of T exposure in early life on lateralization can be efficiently investigated by manipulating T levels in the cleidoic eggs of birds, because the embryo is isolated from maternal and sibling physiological interference, but this approach has been adopted very rarely. In the yellow-legged gull (Larus michahellis) we increased yolk T concentration within the physiological limits and tested the effects on the direction of lateralization in two functionally fundamental behaviors (begging for parental care and escape to cover) of molecularly sexed hatchlings. We also speculated that T may intervene in regulating consistency, rather than direction of lateralization, and therefore tested if T affected the 'repeatability' of lateral preference in consecutive behavioral trials. T treatment had no effect on the direction of lateralization, but enhanced the consistency of lateral preference in escape responses. Sex did not predict lateralization. Neither behavior was lateralized at the population-level. We therefore showed for the first time in any species an effect of egg T on consistency in lateralization. The implications of the effect of T for the evolution of trade-offs in maternal allocation of egg hormones, and the evolutionary interpretations of findings from our studies on lateralization among unmanipulated birds are discussed.

Dietary flavonoids advance timing of moult but do not affect redox status of juvenile blackbirds (Turdus merula).

Flavonoids are the most abundant plant polyphenols, widely occurring in fruits and berries, and show a strong antioxidant activity in vitro Studies of avian species feeding on berries suggest that dietary flavonoids have health-promoting effects and may enhance the expression of melanin-based plumage traits. These effects are probably mediated by the antioxidant activity of flavonoids. However, the effect of dietary flavonoids on oxidative status has never been investigated in any bird species. We analysed the effects of dietary flavonoids on blood non-enzymatic antioxidants and protein oxidative damage of juvenile European blackbirds (Turdus merula). In addition, we analysed the effects of flavonoid-enriched diet on body condition and on the timing of moult from juvenile to adult plumage. Dietary flavonoids did not significantly affect redox status but significantly advanced the onset of moult, hastening plumage development. Moulting birds showed higher protein oxidative damage compared with those that had not yet started moulting. The probability of initiating moult after 40 days of dietary treatment was higher for birds with low circulating levels of oxidizing agents and high glutathione concentration. The metabolization of flavonoids could have altered their redox potential, resulting in no net effects on redox status. However, flavonoid consumption before and during moult may contribute to enhance plumage development. Moreover, our findings suggest that moulting feathers may result in redox imbalance. Given their effect on moult and growth of melanin-rich feathers, fruit flavonoids may have contributed to shape plant fruiting time in relation to fruit consumption preferences by birds.

Genotoxic effects induced by the exposure to an environmental mixture of illicit drugs to the zebra mussel.

Despite the growing interest on the presence of illicit drugs in freshwater ecosystems, just recently the attention has been focused on their potential toxicity towards non-target aquatic species. However, these studies largely neglected the effects induced by exposure to complex mixtures of illicit drugs, which could be different compared to those caused by single psychoactive molecules. This study was aimed at investigating the genetic damage induced by a 14-day exposure to a realistic mixture of the most common illicit drugs found in surface waters worldwide (cocaine, benzoylecgonine, amphetamine, morphine and 3,4-methylenedioxymethamphetamine) on the zebra mussel (Dreissena polymorpha). The mixture caused a significant increase of DNA fragmentation and triggered the apoptotic process and micronuclei formation in zebra mussel hemocytes, pointing out its potential genotoxicity towards this bivalve species.

Sublethal effects induced by morphine to the freshwater biological model Dreissena polymorpha.

Opioids are considered as emerging contaminants in aquatic ecosystems, mainly due to their large illicit consume worldwide. Morphine (MOR) is the main opiate and it was commonly found at measurable concentrations in freshwaters. Even though its occurrence is well documented, just limited information is available regarding its hazard to nontarget organisms. The aim of this study was of the evaluation of sublethal effects induced by MOR to the freshwater bivalve Dreissena polymorpha. We exposed mussels to two MOR concentrations (0.05 µg/L and 0.5 µg/L) for 14 days and we investigated the sublethal effects by a suite of biomarkers. The Neutral Red Retention Assay (NRRA) was used as a test of cytotoxicity, while the oxidative stress was evaluated by the activity of antioxidant and detoxifying enzymes, namely catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) and glutathione-S-transferase (GST), and by measuring the levels of lipid peroxidation (LPO) and protein carbonylation (PCC). The genetic damage was assessed by the Single Cell Gel Electrophoresis (SCGE) assay, the DNA diffusion assay and the micronucleus test (MN test). Finally, the filtration rate of D. polymorpha was evaluated in order to investigate possible physiological effects. Both tested concentrations reduced the lysosome membrane stability of bivalves, but only the highest MOR concentration induced significant changes in the activity of antioxidant enzymes (SOD, CAT, and GPx) and increase in lipid peroxidation levels. Slight increase in primary DNA fragmentation was noticed, while no fixed genetic damage and alterations of the filtering rate were found.

Geographical and ecological factors affect microplastic body burden in marine fish at global scale.

Microplastic (MP) contamination has been identified as a worrisome environmental issue at the global level. Fish are the taxonomic group more extensively investigated to assess MP contamination in marine environment. A large variability in MP bioaccumulation (i.e., body burden) was reported in fish but to date there is a dearth of information concerning the drivers underlying this process. The present systematic review aimed at summarizing the results of the scientific literature on MP body burden in the digestive tract of marine fish to quantitatively shed light on the contribution of different geographical (i.e., latitudinal origin of the sample, distance from the coastline and field- or marked-collected) and ecological (i.e., trophic strategy, milieu, and body size) factors driving bioaccumulation. The mean (±SE) MPs/individual was 4.13 ± 2.87, and the mean MPs/ww (i.e., MPs/g) was 5.92 ± 0.94. Overall, MP abundance expressed as MPs/individual of fish from tropical areas was significantly higher compared to the other latitudinal bands, with species sampled close to the coastline that accumulated a larger number of MPs compared to those collected offshore. Neither the trophic strategy, nor the milieu and the market or field origin of fish explained the MP body burden. However, fish body size resulted as a determinant of MP body burden (as MPs/individual), with small fish accumulating a lower amount of MPs compared to larger ones. Qualitatively, but not statistically significant, similar results were generally obtained for MPs/ww, except for an opposite, and significant, variation according to species body size. Our findings showed that geographical, rather than ecological factors represent the main drivers of MP body burden in marine fish, suggesting that environmental variables and/or local pollution sources mainly contribute to explaining the large variability underlying the ingestion and bioaccumulation processes of these contaminants.

Multi-level toxicity assessment of polylactic acid (PLA) microplastics on the cladoceran Daphnia magna.

The accumulation of plastics waste in the environment has raised a worrisome concern, moving the society to seek out for sustainable solutions, such as the transition from the use of fossil-based, conventional plastics to bioplastics (BPs). However, once in the environment bioplastics have the same probability to accumulate and experience weathering processes than conventional plastics, leading to the formation of microplastics (MPs). However, to date the information on the potential toxicity of MPs originated from the weathering of bioplastics is limited. Thus, this study aimed at investigating the adverse effects induced by the exposure to MPs made of a bioplastic polymer, the polylactic acid (PLA), towards the freshwater cladoceran Daphnia magna. Organisms were exposed for 21 days to three concentrations (0.125 µg/mL, 1.25 µg/mL and 12.5 µg/mL) of PLA microplastics (hereafter PLA-MPs). A multi-level approach was performed to investigate the potential effects through the biological hierarchy, starting from the sub-individual up to the individual level. At the sub-individual level, changes in the oxidative status (i.e., the amount of reactive oxygen species and the activity of antioxidant and detoxifying enzymes) and oxidative damage (i.e., lipid peroxidation) were explored. Moreover, the total caloric content as well as the content of protein, carbohydrate and lipid content assess were used to investigate the effects on energy reserves. At individual level the changes in swimming activity (i.e., distance moved and swimming speed) were assessed. Our results showed that the exposure to PLA-MPs induced a slight modulation in the oxidative status and energy reserves, leading to an increase in swimming behavior of treated individuals compared to control conspecifics. These results suggest that the exposure to MPs made of a bioplastic polymer can induce adverse effects similar to those caused by conventional polymers.

Potential Effects of Environmental and Occupational Exposure to Microplastics: An Overview of Air Contamination.

Microplastics (MPs) are now ubiquitous environmental contaminants that lead to unavoidable human exposure; they have received increasing attention in recent years and have become an emerging area of research. The greatest concern is the negative impacts of MPs on marine, fresh-water, and terrestrial ecosystems, as well as human health, to the extent that the World Health Organization (WHO) calls for increased research and standardized methods to assess exposure to MPs. Many countries and international organizations are implementing or proposing legislation in this regard. This review aims to summarize the current state of legislation, indoor and outdoor contamination, and potential human health risk due to exposure to airborne MPs, considering that occupational exposure to MPs is also becoming a growing area of concern. Even though research regarding MPs has continuously increased in the last twenty years, the effects of MPs on human health have been scarcely investigated, and toxicity studies are still limited and not directly comparable, due to the lack of standardized studies in this field.

Comparison of the potential toxicity induced by microplastics made of polyethylene terephthalate (PET) and polylactic acid (PLA) on the earthworm Eiseniafoetida.

A growing number of studies have demonstrated that microplastic (MP) contamination is widespread in terrestrial ecosystems. A wide array of MPs made of conventional, fossil-based polymers differing in size and shape has been detected in soils worldwide. Recently, also MPs made of bioplastics have been found in soils, but there is a dearth of information concerning their toxicity on soil organisms. This study aimed at exploring the potential toxicity induced by the exposure for 28 days to irregular shaped and differently sized MPs made of a fossil-based (polyethylene terephthalate - PET) and a bioplastic (polylactic acid - PLA) polymer on the earthworm Eisenia foetida. Two amounts (1 g and 10 g/kg of soil, corresponding to 0.1% and 1% of soil weight) of both MP types were administered to the earthworms. A multi-level approach was used to investigate the MP-induced effects at sub-individual and individual level. Changes in the activity of antioxidant and detoxifying enzymes, as well as in lipid peroxidation levels, were investigated at specific time-points (i.e., 7, 14, 21 and 28 days) as sub-individual responses. Histological analyses were performed to assess effects at tissue level, while the change in digging activity was considered as a proxy of behavioral effects. Earthworms ingested MPs made of both the polymers. MPs made of PET did not induce any adverse effect at none of the biological levels. In contrast, MPs made of PLA caused the modulation of earthworms' oxidative status as showed by a bell-shaped activity of superoxide dismutase coupled with an increase in glutathione peroxidase activity. However, neither oxidative and tissue damage, nor behavioral alteration occurred. These findings suggest that the exposure to bio-based MPs can cause higher toxicity compared to fossil-based MPs.

Exposure to Microplastics Made of Plasmix-Based Materials at Low Amounts Did Not Induce Adverse Effects on the Earthworm Eisenia foetida.

The implementation of recycling techniques represents a potential solution to the plastic pollution issue. To date, only a limited number of plastic polymers can be efficiently recycled. In the Italian plastic waste stream, the residual, non-homogeneous fraction is called 'Plasmix' and is intended for low-value uses. However, Plasmix can be used to create new materials through mechanical recycling, which need to be tested for their eco-safety. This study aimed to investigate the potential toxicity of two amounts (0.1% and 1% MPs in soil weight) of microplastics (MPs) made of naïve and additivated Plasmix-based materials (Px and APx, respectively) on the earthworm Eisenia foetida. Changes in oxidative status and oxidative damage, survival, gross growth rate and reproductive output were considered as endpoints. Although earthworms ingested both MP types, earthworms did not suffer an oxidative stress condition or growth and reproductive impairments. The results suggested that exposure to low amounts of both MPs can be considered as safe for earthworms. However, further studies testing a higher amount or longer exposure time on different model species are necessary to complete the environmental risk assessment of these new materials.