Impacts of in vivo and in vitro exposures to tamoxifen: comparative effects on human cells and marine organisms

Tamoxifen (TAM) is a first generation-SERM administered for hormone receptor-positive (HER+) breast cancer in both pre- and post-menopausal patients and may undergo metabolic activation in organisms that share similar receptors and thus face comparable mechanisms of response. The present study aimed to assess whether environmental trace concentrations of TAM are bioavailable to the filter feeder M. galloprovincialis (100 ng L-1) and to the deposit feeder N. diversicolor (0.5, 10, 25 and 100 ng L-1) after 14 days of exposure. Behavioural impairment (burrowing kinetic), neurotoxicity (AChE activity), endocrine disruption by alkali-labile phosphate (ALP) content, oxidative stress (SOD, CAT, GPXs activities), biotransformation (GST activity), oxidative damage (LPO) and genotoxicity (DNA damage) were assessed. Moreover, this study also pertained to compare TAM cytotoxicity effects to mussels and targeted human (i.e. immortalized retinal pigment epithelium - RPE; and human transformed endothelial cells - HeLa) cell lines, in a range of concentrations from 0.5 ng L-1 to 50 μg L-1. In polychaetes N. diversicolor, TAM exerted remarkable oxidative stress and damage at the lowest concentration (0.5 ng L-1), whereas significant genotoxicity was reported at the highest exposure level (100 ng L-1). In mussels M. galloprovincialis, 100 ng L-1 TAM caused endocrine disruption in males, neurotoxicity, and an induction in GST activity and LPO byproducts in gills, corroborating in genotoxicity over the exposure days. Although cytotoxicity assays conducted with mussel haemocytes following in vivo exposure was not effective, in vitro exposure showed to be a feasible alternative, with comparable sensitivity to human cell line (HeLa).info:eu-repo/semantics/publishedVersio

Multi-generational effects under single and pulse exposure scenarios in two monophyletic Daphnia species

Anthropogenic activities commonly relate to a set of diffuse and point contamination sources, from industrial, domestic or agricultural outputs, characterized by a chemical cocktail exposure and consequent disturbances of natural ecosystems. Different species may present different sensitivities to contaminants, even when phylogenetically close. This study used two monophyletic Daphnia species from tropical and temperate environments, Daphnia similis and Daphnia magna respectively, to evaluate the variation of their sensitivity to Pb (if any) and fitness during a multi-generational exposure and recovery. To accomplish that, standard acute immobilization tests were done on specific generations. Tests were carried out with exposures to 1) potassium dichromate (K2Cr2O7) to evaluate organisms' sensitivity/fitness, 2) Pb, to monitor variation on Pb sensitivity and 3) the fungicide mancozeb, providing a pulse toxicity approach on generational Pb acclimated daphnids. Since growth is an important trait related to organisms' fitness, organisms' size measurements were also monitored. In addition, organisms were maintained under two different dietary regimes. Our results indicate no variation on daphnids sensitivity to K2Cr2O7, except for D. similis from a recovery period under food restriction. However, a lower Pb sensitivity was seen for both species throughout generations. Both species also showed that under food restriction neonates' sizes were larger than those kept under regular food, while reproduction was considerably reduced. Food restriction also generated opposite outcomes on both species, such as D. magna epigenetic changes and D. similis phenotypic acclimation to Pb. Besides, D. magna pre-exposed to Pb presented lower sensitivity to mancozeb, while the contrary was shown by D. similis. This study indicates that daphnids are capable of acquiring a lower sensitivity to Pb across a long-term exposure, and that Pb pre-exposure can affect the sensitivity to other chemicals. Also, different patterns in multi-generational responses from monophyletic species (especially under oligotrophic media, common on natural habitats) acknowledge the use of representative or native species to assess the effect of contaminants, since monophyletic species can provide different toxicity outputs.publishe

Influence of seasonality and pollution on the hematological parameters of the estuarine fish Centropomus parallelus

This study investigated the hematological parameters of the tropical estuarine fish Centropomus parallelus and their use as a non-destructive biomarker for aquatic pollution. Individuals were collected, in summer and winter, at two estuaries, Cananéia (CAN) and São Vicente (SVE), and blood was extracted by caudal puncture. The evaluated parameters were hematocrit (Ht), red blood cells (RBC), Mean Corpuscular Volume (MCV), and the leukocyte (WBC) and thrombocyte counts. Fishes from CAN showed higher values of absolute number of thrombocytes in the summer. The fishes from SVE presented lower values of Ht and MCV in winter. Comparing the hematological parameters of fishes from these two sites, Ht, MCV, WBC and RBC were higher in fishes from SVE in the summer, whereas during the winter, Ht and thrombocytes were higher in animals from SVE. The results allow attributing the changes in the blood of fishes to seasonality and the presence of contaminants.No presente estudo, foram utilizados parâmetros hematológicos do peixe tropical estuarino Centropomus parallelus como biomarcador não destrutivo. Os peixes foram coletados, no verão e no inverno, em dois estuários: Cananéia (CAN) e São Vicente (SVE). Foram analisados: hematócrito (Ht), contagem de eritrócitos (RBC), volume corpuscular médio (VCM), leucograma (WBC) e trombograma. Os peixes de ambos locais apresentaram variações no quadro hematológico devido à sazonalidade e aos níveis de degradação ambiental. Os peixes de Cananéia apresentaram aumento no número absoluto de trombócitos no verão, enquanto aqueles coletados em São Vicente apresentaram hematócrito (Ht) e Volume Corpuscular Médio (MCV) menor no inverno. Comparando o quadro hematológico dos peixes dos dois estuários, observamos maiores valores de hematócrito (Ht), Volume Corpuscular Médio (MCV), leucócitos (WBC), e número de eritrócitos (RBC) nos peixes de São Vicente no verão, enquanto no inverno, Ht e trombócitos foram também maiores nos peixes desse local. Os resultados sugerem que as alterações observadas no quadro hematológico se deveram à qualidade ambiental dos estuários e à sazonalidade

Water toxicity and cyto-genotoxicity biomarkers in the fish Oreochromis niloticus (Cichlidae)

Micronucleus (MN) and nuclear abnormalities (NA) tests were used as biomarkers in the fish Oreochromis niloticus from a polluted pond in winter, spring and summer. Samples of water column were taken aiming to evaluate physical-chemical parameters and acute toxicity to the microcrustacean Daphnia similis. The pond waters were toxic to D. similis in spring and winter, but this toxicity was not detected in summer. The pond waters presented low levels of dissolved oxygen in spring and summer; higher ammonia concentrations in winter, and high hardness and conductivity. Fish collected at the polluted pond exhibited higher rates of DNA damage, as indicated by increased rates of MN and NA in erythrocytes when compared to the negative control, which were associated to water toxicity and physical-chemical variable. Thus, this study shows that results of water toxicity tests and genotoxic biomarkers in fish may be associated and theses approaches may be used together for the water quality assessment.Keywords: fish, cyto-genotoxicity, pollution, ecotoxicology, water quality

Cytogenotoxicity biomarkers in fat snook Centropomus parallelus from Cananéia and São Vicente estuaries, SP, Brazil

The aquatic environment receives many contaminants that can induce damages at the molecular, biochemical, cellular and physiological levels. Centropomus parallelus, an important food resource for local populations, is a predator fish that feeds on small fishes and benthic invertebrates, thus being vulnerable to the bioconcentration and biomagnification processes. This study aimed to evaluate cytogenotoxic responses in erythrocytes from C. parallelus juveniles collected in the Cananéia and São Vicente estuaries, both in winter and in summer. After anesthesia, blood samples were collected by caudal puncture. Blood smears were prepared on glass slides and stained with May-Grünwald-Giemsa dye. Two thousand cells were analyzed per slide (1000x), and nuclear abnormalities (NA) and micronuclei (MN) were scored. The São Vicente sample showed MN and NA frequencies (%/1000 cells) of 0.325 and 3.575, in winter, and of 0.125 and 2.935 in summer respectively; the Cananéia sample showed frequencies of 0.0325 and 0.03, in winter, and of 0.065 and 0.355 in summer, respectively. The rates found in São Vicente were significantly higher than those found in Cananéia, evidencing that the levels of pollution in that estuary were high enough to induce genetic damages

Hematological analysis of Micropogonias Furnieri, Desmarest, 1823, Scianidae, from two estuaries of Baixada Santista, São paulo Brazil

Alterações hematológicas em peixes são consideradas uma importante ferramenta para avaliar processos patológicos decorrentes da exposição a poluentes ambientais. Micropogonias furnieri (Desmarest, 1823) (corvina) é comumente encontrada em regiões estuarinas e eventualmente está exposta a inúmeros contaminantes. No presente estudo foi avaliado o quadro hematológico de indivíduos de M. furnieri coletados na Baixada Santista: o Sistema Estuarino de Santos, considerado poluído, e o estuário do Rio Itanhaém (controle). Foram avaliados o número de Eritrócitos (Er), o Hematócrito (Ht), a taxa de Hemoglobina (Hb), o Volume Corpuscular Médio (VCM) e a Concentração de Hemoglobina Corpuscular Média (CHCM). Nos peixes coletados no Sistema Estuarino de Santos, os níveis de Ht foram significativamente menores, enquanto os níveis de CHCM e Hb foram significativamente mais altos, indicando que os prováveis efeitos estejam atribuídos aos diferentes níveis de contaminação encontrados nos estuários.Hematological alterations in fish are considered a useful tool to evaluate pathological processes resulting from the exposure to environmental pollutants. The whitemouth croaker Micropogonias furnieri is a common species in estuarine areas and potentially exposed to many contaminants. In the present study, the hematological characteristics of fish collected at two sites in Baixada Santista (Santos Estuarine System - SES, a polluted site; and the Estuary of Itanhaém River - EIR, unpolluted site) del was analysed. The following blood descriptors were analyzed: number of Erythrocytes (Er), Hematocrit (Ht), Hemoglobin (Hb), Mean Corpuscular Volume (MCV) and Mean Corpuscular Hemoglobin concentration (MCHC). Fish from SES exhibited significant lower levels of Ht and increase on MCHC and Hb. Such differences are likely related to the different contamination levels found in these estuaries