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The occurrence of pharmaceutical compounds in wastewater treatment plants and surface waters has been detected worldwide, constituting a potential risk for aquatic ecosystems. Adult zebrafish, of both sexes, were exposed to three common pharmaceutical compounds (atenolol, ketoprofen and diclofenac) and their UV photolysis by-products over seven days. The results show that diclofenac was removed to concentrations < LOD after 5 min of UV irradiation. The oxidative stress response of zebrafish to pharmaceuticals and their photolysis by-products was evaluated through oxidative stress enzymes (glutathione-S-transferase, catalase, superoxide dismutase) and lipid peroxidation. Results suggest that the photolysis by-products of diclofenac were more toxic than those from the other compounds tested, showing an increase in GST and CAT levels, which are also supported by higher MDA levels. Overall, the toxicity of waters containing atenolol and ketoprofen was reduced after the parent compounds were transformed by photolysis, whereas the toxicity increased significantly from the by-products generated through diclofenac photolysis. Therefore, diclofenac photolysis would possibly necessitate higher irradiation time to ensure that the associated by-products are completely degraded to harmless form(s). [ABSTRACT FROM AUTHOR]

The aim of the present study was to assess the toxic effects of a secondary treated effluent from a modern pulp mill processing Eucalyptus globulus on both sexes of goldfish. The effects in the exposed fish were investigated by measuring biomarkers which have been used to assess the effects of BKME on aquatic biota, with particular emphasis in fish. However, few studies have been focused on effluents from pulp mill plants processing eucalyptus. The relative proportion of cytochrome P450 (CYP1A) and vitellogenin (VTG) induction were analyzed in the livers of males and females goldfish, exposed to different concentrations of a bleached Kraft pulp mill (BKPM) effluent. Somatic indices such as HSI (hepatosomatic index) and GSI (gonadosomatic index) were calculated and a significant reduction was found in males GSI. A significant increase of HSI was recorded for both sexes exposed to 50% BKPM effluent. Exposure to BKPM effluent induced CYP1A synthesis in both sexes and decreased VTG synthesis in females according to the different effluent concentrations suggesting that the secondary treated effluent contained compounds that affect fish negatively. The results suggest that BKPM effluent can cause sex-related biochemical changes in xenobiotic metabolism. [ABSTRACT FROM AUTHOR]

Abstract: To assess the estrogenicity of a municipal sewage treatment plant, sexually mature crucian carp of both sexes were exposed for 28 days to domestic treated sewage effluent running with a constant flow of water, with tap water and with different concentrations of the sewage effluent (25%, 50% and 100%). Vitellogenin (VTG), gonadosomatic index (GSI), hepatosomatic index (HSI), and histological abnormalities were used to assess the estrogenic potency of the effluent. Results show a significant () increase in VTG levels in all exposed male fish. A significant () reduction in the GSI was only observed in fish exposed to 100% effluent. Morphological changes were detected by histological evaluation, revealing severe effects on the testes. Spermatogenesis was progressively reduced to total inhibition in fish exposed to 100% effluent. One of the most pronounced effects detected was the presence of oocytes in male gonads, observed in 20% of males exposed to 100% sewage effluent. [Copyright &y& Elsevier]

Abstract: To assess the estrogenic potency of the treated domestic sewage effluent from a sewage treatment plant (STP) located in Lisbon (Chelas), 60 mirror carp (Cyprinus carpio) were exposed to different concentrations of the sewage effluent (0%, 25%, 50% and 100%) for two periods of 28 days in two different seasons (winter/spring). Vitellogenin induction in males was used as a biomarker of exposure to xenoestrogens. At the end of the experiment, blood samples were taken for vitellogenin analysis and the fish were sacrificed and dissected. Gonad samples were taken for histological evaluation of the sewage effects. The results showed an increase in vitellogenin induction in exposed fish, both males and females, depending on the different dilutions of the sewage effluent. In comparison with controls, the gonadosomatic index decreased significantly (P<0.05) in fish exposed to 100% treated effluent. Although statistically not significant, the hepatosomatic index (HSI) was high in all exposed fish. Histological abnormalities in fish gonads were evaluated and related to the different percentages of sewage effluent. Seasonal variations found in estrogenic responses were attributed to weather influences on sewage dilution. [Copyright &y& Elsevier]

Climate change has pervasive effects on marine ecosystems, altering biodiversity patterns, abundance and distribution of species, biological interactions, phenology, and organisms’ physiology, performance and fitness. Fish early life stages have narrow thermal windows and are thus more vulnerable to further changes in water temperature. The aim of this study was to address the sensitivity and underlying molecular changes of larvae of a key fisheries species, the sea bream Sparus aurata , towards ocean warming. Larvae were exposed to three temperatures: 18 °C (control), 24 °C (warm) and 30 °C (heat wave) for seven days. At the end of the assay, i) survival curves were plotted for each temperature treatment and ii) entire larvae were collected for proteomic analysis via 2D gel electrophoresis, image analysis and mass spectrometry. Survival decreased with increasing temperature, with no larvae surviving at 30 °C. Therefore, proteomic analysis was only carried out for 18 °C and 24 °C. Larvae up-regulated protein folding and degradation, cytoskeletal re-organization, transcriptional regulation and the growth hormone while mostly down-regulating cargo transporting and porphyrin metabolism upon exposure to heat stress. No changes were detected in proteins related to energetic metabolism suggesting that larval fish may not have the energetic plasticity needed to sustain cellular protection in the long-term. These results indicate that despite proteome modulation, S. aurata larvae do not seem able to fully acclimate to higher temperatures as shown by the low survival rates. Consequently, elevated temperatures seem to have bottleneck effects during fish early life stages, and future ocean warming can potentially compromise recruitment’s success of key fisheries species. [ABSTRACT FROM AUTHOR]

The influence of increasing temperatures in thermal and oxidative stress responses were studied in the muscle of several estuarine fish species (Diplodus vulgaris, Diplodus sargus, Dicentrarchus labrax, Gobius niger and Liza ramada). Selected fish were collected in July at the Tagus estuary (24±0.9°C; salinity of 30±4‰; pH=8). Fish were subjected to a temperature increase of 1°C.h−1 until they reached their Critical Thermal Maximum (CTMax), starting at 24°C (control temperature). Muscle samples were collected during the trial and results showed that oxidative stress biomarkers are highly sensitive to temperature. Results from stress oxidative enzymes show alterations with increasing temperature in all tested species. Catalase (CAT; EC 1.11.1.6) activity significantly increased in L. ramada, D. labrax and decreased in D. vulgaris. Glutathione S-transferase (GST; EC 2.5.1.18) activity increased in L. ramada, D. sargus, D. vulgaris, and D. labrax. In G. niger it showed a cycle of increase-decrease. Lipid peroxidation (LPO) increased in L. ramada, D. sargus and D. labrax. With respect to correlation analysis (Pearson; Spearman r), the results showed that oxidation products and antioxidant defenses were correlated in L. ramada (LPO-CAT and LPO-GST, D. sargus (LPO-CAT), and D. labrax (LPO-CAT). Oxidative biomarkers were correlated with thermal stress biomarker (Hsp70) in L. ramada (CAT-Hsp70), D. vulgaris (LPO-Hsp70), D. labrax (GST-Hsp70) and G. niger (LPO-Hsp70). In conclusion, oxidative stress does occur with increasing temperatures and there seems to be a relation between thermal stress response and oxidative stress response. The results suggest that oxidative stress biomarkers should be applied with caution, particularly in field multi-species/multi-environment studies. [ABSTRACT FROM AUTHOR]

Abstract: Three white-rot fungi (Pleurotus sajor caju, Trametes versicolor and Phanerochaete chrysosporium) and one soft-rot fungi (Rhizopus oryzae) species confirmed their potential for future applications in the biological treatment of effluents derived from the secondary treatment of a bleached kraft pulp mill processing Eucalyptus globulus. Among the four species P. sajor caju and R. oryzae were the most effective in the biodegradation of organic compounds present in the effluent, being responsible for the reduction of relative absorbance (25–46% at 250 nm and 72–74% at 465 nm) and of chemical oxygen demand levels (74 to 81%) after 10 days of incubation. Laccase (Lac), lignin (Lip) and manganese peroxidases (MnP) expression varied among fungal species, where Lac and LiP activities were correlated with the degradation of organic compounds in the effluent treated with P. sajor caju. The first two axes of a principal component analysis explained 88.9% of the total variation among sub-samples treated with the four fungus species, after different incubation periods. All the variables measured contributed positively to the first component except for the MnP enzyme activity which was the only variable contributing negatively to the first component. Absorbances at 465 nm, LiP and Lac enzyme activities were the variables with more weight on the second component. P. sajor caju revealed to be the only species able to perform the biological treatment without promoting an increment in the toxicity of the effluent to the Vibrio fischeri, as it was assessed by the Microtox® assay. The opposite was recorded for the treatments with the other three species of fungus. EC50–5 min values ranging between 28 and 57% (effluent concentrations) were recorded even after 10 to 13 days of treatment with P. chrysosporium, R. oryzae or with T. versicolor. [Copyright &y& Elsevier]

• Mussels exhibited intraspecific differences depending on each environmental scenario. • Harbour mussels exhibited differences in lethal and sublethal responses. • Female mussels showed more sensitivity after chemical and warming insults. • Lethal responses could determine the non-reversibility imprint of biomarker responses. The intertidal mussel B. rodriguezii is a representative species from hard bottom substrates where both anthropogenic and natural stressors are present. Pre-exposure to these different stressors can modify the tolerance to additional stressors such as warming. Moreover, this tolerance can vary depending on intraspecific variables such as the organism's sex. The effects of warming and its intraspecific variability in representative coastal species are crucial to understanding the tolerance to future environmental scenarios. The mussels were collected in different environmental scenarios, including low (Control), chemical (Harbour) and natural stressed (Estuary) sites, and then exposed to different water temperatures (10–30 °C) for 14 days. Lethal and sublethal responses were evaluated in different mussel populations. Thus, cumulative death rate, air survival time, heat shock proteins (HSC70/HSP70), total ubiquitin, catalase (CAT), glutathione-s-transferase (GST) and lipid peroxidation (TBARS) were assessed in mussels from different areas and different sexes. The results revealed diminished air survival time and high cumulative mortality rate in mussels collected at the harbour and those exposed to higher temperatures, respectively. The sublethal responses of the field animals showed different patterns according to the different areas investigated. Besides, the results revealed that these differences were also observed between sexes. Regarding the sublethal responses in mussels exposed to warming, the interactive effects of temperature and sites showed a strong influence on all biochemical parameters analyzed (p < 0.001). Therefore, harbour mussels showed a distinct pattern compared to other locations and reflecting the most damaging effects of warming. The influence of sex and its interactions with warming were also crucial in most of the sublethal responses (p < 0.05). Multivariate analysis was performed with all sublethal responses, and the different warming scenarios showed different groups according to the sites. In the predicted warming scenarios, males showed no differences between sites. In contrast to males, females showed differences between sites in the predicted and the worse-case warming scenarios. Our results highlight the importance of compensatory mechanisms in the mussel warming tolerance like HSP70. The influence of sex is also crucial in understanding warming tolerance in mussels chronically exposed to pollutants in their natural environment. Also, lethal endpoints are essential for understanding the non-reversibility signature of the observed biochemical responses. [ABSTRACT FROM AUTHOR]

• Seasonal proteome variation in intertidal shrimps was analyzed in situ. • Shrimps were collected during spring and summer low tides. • Cytoskeleton, energy metabolism and transcription were modulated by season. • Inter-individual variation in protein levels was low. • Molecular plasticity is crucial to cope with environmental fluctuations. The ability of intertidal organisms to maintain their performance via molecular and physiological adjustments under low tide, seasonal fluctuations and extreme events ultimately determines population viability. Analyzing this capacity in the wild is extremely relevant since intertidal communities are under increased climate variability owing to global changes. We addressed the seasonal proteome signatures of a key intertidal species, the shrimp Palaemon elegans , in a natural setting. Shrimps were collected during spring and summer seasons at low tides and were euthanized in situ. Environmental variability was also assessed using hand-held devices and data loggers. Muscle samples were taken for 2D gel electrophoresis and protein identification through mass spectrometry. Proteome data revealed that 55 proteins (10.6% of the proteome) significantly changed between spring and summer collected shrimps, 24 of which were identified. These proteins were mostly involved in cytoskeleton remodelling, energy metabolism and transcription regulation. Overall, shrimps modulate gene expression leading to metabolic and structural adjustments related to seasonal differences in the wild (i.e. abiotic variation and possibly intrinsic cycles of reproduction and growth). This potentially promotes performance and fitness as suggested by the higher condition index in summer-collected shrimps. However, inter-individual variation (% coefficient of variation) in protein levels was quite low (min-max ranges were 0.6–8.3% in spring and 1.2–4.8% in summer), possibly suggesting reduced genetic diversity or physiological canalization. Protein plasticity is relevant to cope with present and upcoming environmental variation related to anthropogenic forcing (e.g. global change, pollution) but low inter-individual variation may limit evolutionary potential of shrimp populations. [ABSTRACT FROM AUTHOR]