A biomarker approach to study the effects of polluted Brazilian urban reservoirs in a native fish.

Two of the largest water reservoirs in the Metropolitan Region of São Paulo, Brazil (MRSP), named Billings and Guarapiranga, are facing high levels of anthropic impact. This is evidenced by the presence of contaminants and pollutants, which are deteriorating their water quality. Therefore, this study evaluated antioxidant defense enzymes, lipoperoxidation and genotoxicity, in adult females of a native species, Astyanax altiparanae from the Guarapiranga and Billings reservoirs. The study also aimed to evaluate these biomarkers during two different periods of the year, the rainy (summer) and dry (winter) seasons. The oxidative stress was evaluated by the activity of enzymes such as glutathione peroxidase, glutathione S-transferases, superoxide dismutase, and catalase in the gills and liver, and the occurrence of lipoperoxidation was also evaluated in both organs. The genotoxicity was assessed by performing comet assay, micronucleus, and nuclear abnormality tests on blood samples. The results showed that fish from both reservoirs are subjected to oxidative stress and genotoxic damage, mainly during winter, but fish living in Billings showed greater alterations than fish from Guarapiranga. Likewise, the results of the principal component analysis suggested that caffeine, nitrogenous compounds, and some metals might be triggering these toxic effects in fish.

Neurotoxic, biotransformation, oxidative stress and genotoxic effects in Astyanax altiparanae (Teleostei, Characiformes) males exposed to environmentally relevant concentrations of diclofenac and/or caffeine.

The present study evaluated neurotoxic, biotransformation, genotoxic and antioxidant responses to relevant environmental concentrations of diclofenac (0.4 µg L-1) and caffeine (27.5 µg L-1), separate and combined, in adult males of the freshwater fish Astyanax altiparanae after a subchronic exposure (14 days). Fish exposed to diclofenac and caffeine, both separate and combined, revealed a neurotoxic effect through the inhibition of acetylcholinesterase activity in the muscle, while diclofenac alone and in combination caused cyclooxygenase inhibition. Caffeine alone produces genotoxicity on this species but, when combined with diclofenac, it potentiates hepatic lipoperoxidation and the inhibition of oxidative stress enzymes, while diclofenac alone or in combination produces a general inhibition of important enzymes. This study suggests that aquatic contamination produced by these pharmaceuticals has the potential to affect homeostasis and locomotion in A. altiparanae and compromise their immune system and general health.

Nonsteroidal anti-inflammatory drugs act as endocrine disruptors in Astyanax lacustris (Teleostei: Characidae) reproduction: An ex vivo approach.

Pharmaceutical products can act as endocrine-disrupting compounds (EDCs), affecting the physiological processes of animals, such as development or reproduction. This study aimed to investigate the influence of different concentrations of nonsteroidal anti-inflammatory drugs (NSAIDs) diclofenac (DCF) and ibuprofen (IBU) alone and mixed (MIX) on gonadotropin gene expression and gonadal steroid release using Astyanax lacustris pituitary and testes explant systems, respectively. The explant organs were maintained for 12 h in Leibovitz (L-15) medium supplemented with 0, 0.1, 1, 10, 100, and 1000 ng L-1 of DCF, IBU, and MIX (ratio 1:1 of the same concentrations of DCF and IBU alone) and gonadotropin releasing-hormone (cGnRH2) stimulation in pituitary explants and human chorionic gonadotropin (hCG) stimulation in testes explants. The pituitary glands and the media from the testicular explants were collected for gene expression analysis including the ß subunit of the follicle-stimulating hormone (fshß) and luteinizing hormone (lhß) and secreted gonadal steroid concentration analysis, respectively. Both DCF and IBU (alone and mixed) decreased pituitary gene expression of fshß and lhß and this inhibitory effect was evident even at low concentrations. In the testes, DCF and IBU did not change the levels of estradiol, and both pharmaceuticals increased the release of 11-ketotestosterone at low doses, while only IBU decreased the levels of testosterone in all concentrations. IBU's inhibitory effect in the testes was not triggered by the mixture of the two drugs. These results suggest that NSAIDs, may interfere in fish reproduction by acting as EDCs, thereby negatively affecting A. lacustris spermatogenesis and maturation.

Paternal exposure to aluminum, acidity, and temperature affect fatty acid seminal profile, embryonic and larval development of Astyanax altiparanae.

We investigated the effects of water acidity, temperature, and aluminum (Al) on the fatty acid (FA) seminal profile, reproductive parameters (fertilization and hatching) and embryonic development of Astyanax altiparanae. We treated males with different experimental treatments, corresponding to the combination of water temperature (20 °C; 25 °C), pH (neutral - 7.0; acidic - 5.5), and the absence or presence of Al (0.5 mg L-1). After 96 h, we analyzed the FA profile of semen and performed artificial fertilization in activating medium with neutral pH or activating medium in the same experimental conditions of the males (neutral pH, acidic pH, and Al) to evaluate fertilization and hatching rates and to monitor embryonic development. Polyunsaturated FA percentage decreased in semen of fish from the neutral group, while monounsaturated FA increased in all groups maintained at 20 °C compared to 25 °C. Aluminum exposure decreased the percentage of C20:4n6 and increased the percentage of C22:5n3 at 20 °C. Males exposed to acidic pH and Al showed lower fertilization and hatching rates, as well as increased mortality of embryos and larvae. Moreover, Al favoured a higher percentage of abnormal larvae. Fertilization in Al activating medium harmed the embryos and larvae since fertilization and hatching rates decreased. Finally, temperature influenced fertilization time, hatching rate, and the morphology of embryos and larvae. Males exposed to Al had lower fertilizing capacity, which negatively affected the embryonic development of the species. Furthermore, Al activating medium reduced the number of fertilized oocytes, hatched embryos, and normal larvae. All events were temperature dependent.

The role of free fatty acids in the inflammatory and cardiometabolic profile in adolescents with metabolic syndrome engaged in interdisciplinary therapy.

The purpose of the present study was to evaluate if interdisciplinary therapy can influence the cardiometabolic and serum free fatty acid profile. The second aim was to evaluate if there is an association between serum free fatty acids, inflammation and cardiometabolic biomarkers in obese adolescents with and without metabolic syndrome submitted to a long-term interdisciplinary therapy. The study involved 108 postpuberty obese adolescents, who were divided according to metabolic syndrome (MetS) diagnosis: MetS (n=32) and Non-MetS (n=76). The interdisciplinary therapy consisted of a 1-year period of nutrition, psychology, physical exercise and clinical support. After therapy, both groups improved metabolic, inflammatory (leptin, adiponectin, leptin/adiponectin ratio, adiponectin/leptin ratio and C-reactive protein) and cardiometabolic profile (PAI-1 and ICAM). Metabolic syndrome prevalence reduced from 28.70% to 12.96%. Both groups reduced myristic acid (C14:0) and increased docosahexaenoic acid (DHA, C22:6n3), heneicosapentaenoic acid (HPA, C21:5n3) and arachidonic acid (C20:4n6). After adjustment for metabolic syndrome and the number of metabolic syndrome parameters, multiple regression analysis showed that changes in VCAM and PAI-1 were negatively associated with changes in cis-linoleic acid (C18:2n6c). Additionally, changes in trans-linoleic acid (C18:2n6t) were also positively associated with these biomarkers. Moreover, leptin and leptin/adiponectin ratio were negatively associated with changes in docosapentaenoic acid (DPA, C22:5n3) and stearidonic acid (SDA, C18:4n3). Adiponectin/leptin ratio was positively associated with docosapentaenoic acid (DPA, C22:5n3). Changes in adiponectin were positively correlated with changes in omega 3, such as heneicosapentaenoic acid (HPA, C21:5n3) and docosapentaenoic acid (DPA, C22:5n3). Results support that interdisciplinary therapy can control inflammatory and cardiometabolic profile in obese adolescents. Moreover, serum fatty acids can be influenced by lifestyle changes and are able to modulate these biomarkers.

The role of ovarian steroids in reproductive plasticity in Hoplias malabaricus (Teleostei: Characiformes: Erythrinidae) in tropical reservoirs with different degrees of pollution.

Chemicals in aquatic environments may lead to impairment of fish reproduction, but some species display plasticity levels to survive and reproduce in such localities. We investigated the reproductive cycle in a teleost species, Hoplias malabaricus, which inhabits a variety of environments with different degrees of pollution. Adult females were sampled at two locations in São Paulo State (Brazil), the reference Ponte Nova (PN) reservoir and the polluted Billings (BIL) reservoir. Metabolic and endocrine processes were analyzed throughout the annual cycle with special attention to ovarian histology, gonadosomatic index (GSI), fecundity, ovarian energetic substrates and plasma steroids analyses, in addition to physical and chemical analyses of the water. Pollutants were frequent in BIL but not in PN. The GSI, including the predominance of vitellogenic oocytes, was higher in spring and summer in both locations, but the oocyte recruitment dynamics was different. During winter, females from BIL presented vitellogenic oocytes and high levels of 11-ketotestosterone, which indicated precocity in the vitellogenic phase in relation to the females from PN. In animals from PN, high deposition of lipids occurred in the ovaries. However, plasma estradiol levels did not vary throughout the annual cycle. In animals from BIL, plasma estradiol levels peaked during the summer, but the ovarian lipid content remained unchanged throughout the year. The data suggest that the presence of pollutants cause various endocrine and metabolic responses and especially affects reproductive plasticity, which could explain why H. malabaricus survives under unfavorable environments.