Bethanie Carney Almroth

Protein carbonyls and antioxidant defenses in corkwing wrasse (Symphodus melops) from a heavy metal polluted and a PAH polluted site.

The use of fish in environmental monitoring has become increasingly important in recent years as anthropogenic substances, many of which function as prooxidants, are accumulating in aquatic environments. We have measured a battery of antioxidant defenses as a measure of oxidative status, as well as protein carbonylation as a measure of oxidative damage, in corkwing wrasse (Symphodus melops) captured near a disused copper mine, where water and sediment are contaminated with heavy metals, and an aluminum smelter, a site contaminated with PAHs. Results were compared to two different reference sites. Fish at the heavy metal site had lower glucose-6-phosphate dehydrogenase activity and elevated protein carbonyls (1.8 times) compared to fish from the reference site. At the PAH site, EROD was increased 2-fold, while total glutathione and methemoglobin reductase concentration, were decreased. No differences were seen in protein carbonyl levels at the PAH site. Measures of both antioxidant defenses and oxidative damage should be used when assessing effects of xenobiotics on oxidative stress in fish species.

Oxidative stress, evident in antioxidant defences and damage products, in rainbow trout caged outside a sewage treatment plant

Sewage treatment plants (STPs) are common sources of a wide variety of chemicals entering into the aquatic environment. We have investigated the effects of STP effluent on oxidative stress parameters in rainbow trout via measurements of several enzymatic antioxidants as well as the molecular antioxidant glutathione (GSH). In addition, we have measured levels of oxidative damage, i.e. protein carbonyls and lipid peroxides. Our findings indicate that STP effluent does contain prooxidants that affect fish. Oxidative damage was a more consistent indicator of exposure to prooxidants than antioxidant enzymes. Protein carbonyls and lipid peroxides were both elevated (1.3- and 1.8-fold increases, respectively) in fish caged at the STP site compared with reference site as was the amount of oxidized GSH (GSSG), which showed a 3-fold increase. Antioxidant enzymes showed no clear-cut response. Additionally, older pollutants that have been present in down stream sediment for decades had no effects on the parameters measured here.

The impact of urban environment on oxidative damage (TBARS) and antioxidant systems in lungs and liver of great tits, Parus major.

A direct negative link between human health and urban pollution levels generated by increased internal levels of oxyradicals is well established. The impact of urban environment on the physiology of wild birds is however, poorly investigated. Here we compare oxidative damage (i.e., lipid peroxidation, measured as TBARS) and different antioxidant enzymes (glutathione reductase (GR), glutathione-S-transferase (GST), and catalase (CAT)) in lungs of urban and rural great tits, Parus major. In addition, we investigated enzymatic (i.e., CAT) and non-enzymatic (i.e., carotenoids) antioxidant levels in liver tissue. There was no significant difference in lipid peroxidation in lungs between the environments. Among the antioxidant enzymes measured in lungs, only CAT showed a tendency towards increased activity in the urban environment. In contrast, CAT in livers was highly non-significant. However, there was a significantly higher concentration of dietary carotenoids (i.e., lutein (Lut) and zeaxanthin (Zx)) in urban males, along with a sex-specific difference in composition (Lut:Zx ratio) between the environments. Taken together, these results suggest that great tit lungs and livers do not seem to be negatively affected, regarding oxidative stress, by living in an urban environment.

Screening for microplastics in sediment, water, marine invertebrates and fish: Method development and microplastic accumulation

Measurements of microplastics in biota and abiotic matrices are key elements of exposure and risk assessments for this emerging environmental pollutant. We investigated the abundance of microplastics in field-collected biota, sediment and water. An improved sediment extraction method, based on density separation was developed. For analysis of microplastics in biota we found that an adapted enzymatic digestion protocol using proteinase K performed best, with a 97% recovery of spiked plastic particles and no observed degradation effects on the plastics in subsequent Raman analysis. Field analysis revealed that 8 of 9 tested invertebrate species from the North Sea and 68% of analyzed individuals of brown trout (Salmo trutta) from the Swedish West Coast had microplastics in them. Based on the number of plastic particles per kg d.w. the microplastic concentrations found in mussels were approximately a thousand-fold higher compared to those in sediment and surface water samples from the same location.

Early-age changes in oxidative stress in brown trout, Salmo trutta.

Fish are often used as models for studies investigating the ability of xenobiotics to induce oxidative stress, though age or developmental stage of the individuals studied has been given little attention. Oxidative stress in other organisms is associated with aging as well as with periods of rapid growth, which occurs in young brown trout. We measured protein carbonyls, 20S proteosome activity and glutathione (GSH) levels in farmed Salmo trutta in four different age groups from 5 months to 3 years. We found an increase in protein carbonyls and a decrease in 20S proteosome activity in both brain and liver tissues of the fish with increasing size and age. Total GSH levels in liver tissue declined as fish aged and the GSSG:GSH ratio increased. Five month and 1 year old trout were treated with paraquat (PQ) to induce oxidative stress. Five month old fish showed no changes in the measured parameters while 1 year old fish had both an increase in protein carbonylation in liver tissue and a decrease in 20S proteosome activity in brain tissue. These results indicate that oxidative stress biomarkers are affected by age or rapid growth in brown trout, and that individuals of different ages respond differently to oxidative stress induced by PQ.

Oxidative stress in growth hormone transgenic coho salmon with compressed lifespan--a model for addressing aging.

Abstract Growth hormone (GH) transgenic fish have dramatically enhanced growth rates, increased oxygen demands and reactive oxygen species production. GH-transgenic coho salmon provide an opportunity to address effects of increased metabolism on physiological aging. The objective of this study was to compare oxidative stress in wild-type (WT) and GH-transgenic (T) coho salmon (Oncorhynchus kisutch) of different ages (1 and 2 years). Antioxidant enzyme activity, protein carbonyls (PC) and glutathione (GSH, GSSG) were measured. PC correlated to growth rates in individual fish. T fish exhibited lower antioxidant enzyme activities and GSH levels compared to the WT, while levels of PC and GSSG were higher. Age affects were observed in both WT and T fish; enzyme activities and GSH decreased while PC and GSSG increased. Our results support the metabolic rate theory of aging. This study aims to be a platform for continued studies of the theories of aging using fish as model organisms.

A critical perspective on early communications concerning human health aspects of microplastics

Microplastic research in recent years has shown that small plastic particles are found almost everywhere we look. Besides aquatic and terrestrial environments, this also includes aquatic species intended for human consumption and several studies have reported their prevalence in other food products and beverages. The scientific as well as public debate has therefore increasingly focused on human health implications of microplastic exposure. However, there is a big discrepancy between the magnitude of this debate and actual scientific findings, which have merely shown the presence of microplastics in certain products. While plastics can undoubtedly be hazardous to human health due to toxicity of associated chemicals or as a consequence of particle toxicity, the extent to which microplastics in individual food products and beverages contribute to this is debatable. Considering the enormous use of plastic materials in our everyday lives, microplastics from food products and beverages likely only constitute a minor exposure pathway for plastic particles and associated chemicals to humans. But as this is rarely put into perspective, the recent debate has created a skewed picture of human plastic exposure. We risk pulling the focus away from the root of the problem: the way in which we consume, use and dispose of plastics leading to their widespread presence in our everyday life and in the environment. Therefore we urge for a more careful and balanced discussion which includes these aspects.

Gender differences in health and aging of Atlantic cod subject to size selective fishery

Summary We have analyzed health and physiological aging parameters in male and female Atlantic cod, Gadus morhua, captured in Kattegat, Skagerrak and in Öresund. Gender differences were clearly evident in a number of variables. Males had longer liver telomeres and higher catalase activities than females, while females had higher superoxide dismutase activity, liver somatic index and condition factor. Effects of age were found for males where levels of the antioxidant glutathione and telomere length declined with age, indicating physiological aging. Liver somatic index increased and percentage oxidized glutathione decreased with age. Between-site comparisons of males show that percentage oxidized glutathione and catalase were lowest in Kattegat, whereas protein carbonyls and condition factor were higher in Skagerrak. Females, on the other hand, showed no differences between sites or indications of somatic aging or age-related effects in egg quality, indicating that older and larger female cod are healthy and show no changes in eggs with age. In contrast, males showed indications of physiological aging and lower condition than females. The results emphasize the importance of conserving old mature fish, in particular high egg-productive females, when managing fisheries.

Waterborne beclomethasone dipropionate affects the physiology of fish while its metabolite beclomethasone is not taken up

Asthma is commonly treated with inhalable glucocorticosteroids, including beclomethasone dipropionate (BDP). This is a synthetic prodrug which is metabolized to the more active monopropionate (BMP) and free beclomethasone in humans. To evaluate potential effects of residual drugs on fish, we conducted a 14 day flow-through exposure experiment with BDP and beclomethasone using rainbow trout, and analyzed effects on plasma glucose, hepatic glutathione and catalase activity together with water and body concentrations of the BDP, BMP and beclomethasone. We also analyzed hepatic gene expression in BDP-exposed fish by microarray and quantitative PCR. Beclomethasone (up to 0.65 μg/L) was not taken up in the fish while BDP (0.65 and 0.07 μg/L) resulted in accumulation of both beclomethasone, BMP and BDP in plasma, reaching levels up to those found in humans during therapy. Accordingly, exposure to 0.65 μg/L of BDP significantly increased blood glucose as well as oxidized glutathione levels and catalase activity in the liver. Exposure to beclomethasone or the low concentration of BDP had no effect on these endpoints. Both exposure concentrations of BDP resulted in significantly higher transcript abundance of phosphoenolpyruvate carboxykinase involved in gluconeogenesis, and of genes involved in immune responses. As only the rapidly metabolized prodrug was potent in fish, the environmental risks associated with the use of BDP are probably small. However, the observed physiological effects in fish of BDP at plasma concentrations known to affect human physiology provides valuable input to the development of read-across approaches in the identification of pharmaceuticals of environmental concern.

Oxidative damage in rainbow trout caged in a polluted river.

Sewage treatment works (STWs) are a common source of chemicals entering into the aquatic environment. In order to assess effects of these effluents on oxidative stress parameters in aquatic organisms, we caged rainbow trout at five sites: upstream, near an STW effluent, and three sites downstream in the river Viskan in western Sweden for 14 days during autumn, 2006. We then measured protein carbonyls in plasma as well as 20S proteosome activity and lipid peroxidation products, i.e. MDA and 4-HNE, in liver samples. Levels of both lipid and protein oxidative damage products were elevated in fish caged near the STW effluent while 20S activity showed no differences. This argues that complex mixtures of chemicals entering into the aquatic environment do have deleterious effects on fish. Additionally, oxidative stress parameters can serve as a biomarker in aquatic organisms.