Low concentrations of complex mixtures of pesticides and metabolites are toxic to common Carp brain cells (Cyprinus carpio carpio).

Pesticide use increases annually, and Brazil is the world's largest consumer. However, unlike the European Union (EU), there is no established limit value for pesticide mixtures in drinking water, and therefore the concentration of pesticides can reach 3354 times the EU limit. Thus, determining the risk of exposure to pesticide mixtures and their main metabolites is challenging and requires the use of alternative methods. In the present study, the Common Carp Brain (CCB) cell line was used to evaluate the in vitro toxicity of relevant pesticide mixtures (glyphosate, 2,4-D, atrazine, and mancozeb) and their main metabolites after 72 h of exposure. The tested concentrations were based on the Acceptable Daily Intake (ADI) defined by Brazilian legislation. The results showed that cells exposed to lower concentrations of the pesticide mixtures and the pesticide + metabolite mixtures were affected by a decrease in cell confluence, resazurin metabolism, and wound healing capacity. The IBR index showed that lower concentrations had more severe effects, suggesting the absence of safe concentrations of these pesticide and metabolite mixtures for the CCB cell line within the tested concentration range. These findings raise concerns about the effects of exposure to these substances on animal and human health.

Multispecies and multibiomarker assessment of fish health from Iguaçu River reservoir, Southern Brazil.

This study investigated the impact of micropollutants on fish health from Segredo hydroelectric reservoir (HRS) along the Iguaçu River, Southern Brazil, contaminated by urban, industrial, and agricultural activities. This is the first comprehensive study assessment in the river after the severe drought in the 2020s in three fish species from different trophic levels Astyanax spp. (water column depth/omnivorous), Hypostomus commersoni (demersal/herbivorous), and Pimelodus maculatus (demersal/omnivorous). Animals, water, and sediment samples were collected from three distinct sites within the reservoir: Floresta (upstream), Iratim (middle), and Station (downstream). The chemical analysis revealed elevated concentrations of metals (Al, Cu, Fe) and the metalloid As in water, or Cu, Zn, and As in sediment, surpassing Brazilian regulatory limits, while the organic pollutants as DDT, PAHs, PCBs, and PBDEs were found under the Brazilian regulatory limits. The metal bioaccumulation was higher in gills with no significant differences among sites. The species Astyanax spp. and H. commersoni displayed variations in hepatosomatic index (HSI) and P. maculatus in the condition factor index (K) between sites, while adverse effects due to micropollutants bioaccumulation were observed by biochemical, genotoxic, and histopathological biomarkers. The principal component analysis and integrated biomarker response highlighted the upstream site Floresta as particularly inhospitable for biota, with distinctions based on trophic level. Consequently, this multifaceted approach, encompassing both fish biomarkers and chemical analyses, furnishes valuable insights into the potential toxic repercussions of micropollutant exposure. These findings offer crucial data for guiding management and conservation endeavors in the Iguaçu River.

Glyphosate and aminomethylphosphonic acid metabolite (AMPA) modulate the phenotype of murine melanoma B16-F1 cells.

Pesticides are contaminants run-offs from agricultural areas with a global concern due to their toxicity for non-target organisms. The Brazilian Health Surveillance Agency reported about 63% of the food contain pesticide residues. Glyphosate is a herbicide used worldwide but its toxicity is not a consensus among specialists around the world. AMPA (aminomethylphosphonic acid) is a glyphosate metabolite that can be more toxic than the parental molecule. Melanoma murine B16-F1 cells were exposed to glyphosate and AMPA to investigate the cell profile and possible induction to a more malignant phenotype. Glyphosate modulated the multi-drug resistance mechanisms by ABCB5 gene expression, decreasing cell attachment, increasing cell migration and inducing extracellular vesicles production, and the cells exposed to AMPA revealed potential damages to DNA. The present study observed that AMPA exhibits high cytotoxicity, which suggests a potential impact on non-tumor cells, which are, in general, more susceptible to chemical exposure. Conversely, glyphosate favored a more metastatic and chemoresistant behavior in cancer cells, highlighting the importance of additional research in this area.

Pollutant bioaccumulation in sentinel fish chronically exposed in Iguaçu river reservoirs (Southern Brazil) and human health risk of fish consumption.

Bioaccumulation studies in fish mark the initial phase of assessing the risk of chemical exposure to biota and human populations. The Iguaçu River boasting a diverse endemic ichthyofauna, is grappling with the repercussions of human activities. This study delved into the bioaccumulation of micropollutants, the early-warning effects on Rhamdia quelen and Oreochomis niloticus in the Segredo Reservoir (HRS) and the potential risk of human exposure. Two groups of caged fish in three sites of the reservoir were exposed during the autumn-winter and spring-summer, while a third group (O. niloticus) underwent a twelve-month exposure, and inorganic and organic chemicals analysis in water, sediment, and biota. Additionally, metallothionein expression and genotoxicity were employed as biomarkers. PAHs, PCBs, Al, Cu, Fe, and As in water and DDTs, Cu, Zn, and As in sediment surpassed the thresholds set by Brazilian regulations, where DDT exhibited bioaccumulation in muscle, alongside metals in liver, kidney, gills, and muscle tissues. R. quelen showed metallothionein expression whereas DNA damage and NMA frequencies were elevated in target organs and in brain and erythrocytes of O. niloticus during summer. In this species the DNA damage in liver was remarkable after twelve months. Target Hazard Quotients and Cancer Risk values shedding light on the vulnerability of both children and adults. The reservoir's conditions led to heightened sensitivity to micropollutants for R. quelen species. The data presented herein provides decision-makers with pertinent insights to facilitate effective management and conservation initiatives within the Iguaçu Basin.

BDE-209 exposure in murine melanoma (B16-F1) cells modulates tumor malignancy and progression in vivo.

Melanoma is a type of skin cancer considered aggressive due to its high metastatic ability and rapid progression to other tissues and organs. BDE-209 (2,2',3,3',4,4',5,5',6,6'-decabromodiphenyl ether) is an additive used as a flame retardant and classified as a persistent organic pollutant that has a high bioaccumulation capacity due to its lipophilic nature. This substance has already been detected in rivers, air, soil, plants and even in different human biological samples, such as plasma, umbilical cord blood and breast milk, revealing a great concern to human populations. Thus, in the current study we investigated whether prior exposure of murine melanoma B16-F1 cells to BDE-209 modulates in vivo progression and malignancy of melanoma. B16-F1 cells were cultured and exposed in vitro to BDE-209 (0.01, 0.1 e 1 nM) for 15 days and then inoculated, via caudal vein, in C57BL/6 mice for experimental metastasis analysis after 20 days. Inoculation of BDE-209-exposed cells resulted in 82% increase of metastasis colonized area in the lungs of mice, downregulation of tumor suppressors genes, such as Timp3 and Reck, decrease of lipid peroxidation and increase of systemic and local inflammatory response. These findings are related to melanoma progression. Additionally, the histopathological analysis revealed greater number of focal points of metastases in the lungs and invasiveness of metastases to the mice brain (89%). The results showed that exposure to BDE-209 may alter the phenotype of B16-F1 cells, worsening their metastatic profile. Current data showed that BDE-209 may interfere with the prognosis of melanoma by modulating cells with less invasiveness capacity to a more aggressive profile.

Immunotoxicity of relevant mixtures of pesticides and metabolites on THP-1 cells.

Pesticides are used to combat agricultural pests but also trigger side effects on non-target organisms. Particularly, immune system dysregulation is a major concern due to the organism's increased vulnerability to diseases, including cancer development. Macrophages play essential roles in innate and adaptive immunity and can undergo classical (M1) or alternative (M2) activation. The M1 pro-inflammatory phenotype has an antitumor role, while M2 favors tumor promotion. Although previous studies have linked pesticide exposure to immune compromise, macrophage polarization is still poorly studied. Here, we investigated the effects of 72 h-long exposure to the mixture of four pesticides widely used in Brazil (glyphosate, 2,4-D, mancozeb, and atrazine), and their main metabolites (aminomethylphosphonic acid, 2,4-diclorophenol, ethylenethiourea, and desethylatrazine) on human leukemia monocytic THP-1 cell line at concentrations based on the Acceptable Daily Intake (ADI) value established in the country. The data revealed immunotoxicity related to impaired cell metabolism in all exposed groups, decreased cell attachment (Pes: 10-1; Met: 10-1; Mix: all concentrations), and disturbance in nitric oxide (NO) levels (Met: 10-1, 101; Mix: all concentrations). The polarization of macrophages towards a more pro-tumor M2-like phenotype was also supported by decreased secretion of the pro-inflammatory cytokine TNF-α (Pes 100, 101) and increased IL-8 (Pes 101). These outcomes alert about the risk of pesticide exposure in the Brazilian population.

BDE-209 and TCDD enhance metastatic characteristics of melanoma cells after chronic exposure.

TCDD (2,3,7,8-tetrachlorodibenzo-p-dioxin) and BDE-209 (decabromodiphenyl ether) are persistent organic pollutants (POPs) produced by industrial activities and associated with several diseases. TCDD is a known human carcinogen, but few studies investigated about the effects of exposure to both compounds, i.e., whether BDE-209 and TCDD can render tumor cells more aggressive and metastatic. In the current study we investigated if the exposure of B16-F1 and B16-F10 melanoma murine cells to environmental relevant concentrations of TCDD and BDE-209 at 24 h and 15-day exposure modulates the expression of genes related to metastasis, making the cells more aggressive. Both pollutants did not affect cell viability but lead to increase of cell proliferation, including the upregulation of vimentin, MMP2, MMP9, MMP14 and PGK1 gene expression and downregulation of E-cadherin, TIMP2, TIMP3 and RECK, strongly suggesting changes in cell phenotypes defined as epithelial to mesenchymal transition (EMT) in BDE-209 and TCDD-exposed cells. Foremost, increased expression of metalloproteinases and decreased expression of their inhibitors made B16-F1 cells similar the more aggressive B16-F10 cell line. Also, the higher secretion of extracellular vesicles by cells after acute exposure to BDE-209 could be related with the phenotype changes. These results are a strong indication of the potential of BDE-209 and TCDD to modulate cell phenotype, leading to a more aggressive profile.

Polybrominated diphenyl ethers BDE-47 and BDE-99 modulate murine melanoma cell phenotype in vitro.

Cancer is one of the leading causes of mortality worldwide. Even with the advances of pharmaceutical industry and treatments, the mortality rate for various types of cancer remains high. In particular, phenotypic alterations of tumor cells concerning drug efflux, migratory and invasive capabilities may represent a hurdle for cancer treatment and contribute to poor prognosis. In the present study, we investigated the effects of polybrominated diphenyl ethers (PBDEs) used as flame retardants on phenotypic features of melanoma cells that are important for cancer. Murine melanoma B16-F1 (less metastatic) and B16-F10 (more metastatic) cells were exposed to 0.01-1.0 nM of BDE-47 (2,2',4,4'-tetrabromodiphenyl ether), BDE-99 (2,2',4,4',5-pentabromodiphenyl ether), and the mixture of both (at 0.01 nM) for 24 h (acute exposure) and 15 days (chronic exposure). The polybrominated diphenyl ethers (PBDEs) did not affect cell viability but led to increased drug efflux transporter activity, cell migration, and colony formation, as well as overexpression of Abcc2 (ATP-binding cassette subfamily C member 2), Mmp-2 (matrix metalloproteinase-2), Mmp-9 (matrix metalloproteinase-9), and Tp53 (tumor protein p53) genes and downregulation of Timp-3 (tissue inhibitor of metalloproteinase 3) gene in B16-F10 cells. These effects are consistent with increased aggressiveness and malignancy of tumors due to exposure to the flame retardants and raise some concerns on the effects such chemicals may have on melanoma treatment and cancer prognosis.

Toxicological effects of silver nanoparticles and cadmium chloride in macrophage cell line (RAW 264.7): An in vitro approach.

BACKGROUND: Silver nanoparticles (AgNP) are largely used in nanotechnological products, but the real risks for human and environment are still poorly understood if we consider the effects of mixtures of AgNP and environmental contaminants, such as non-essential metals. METHODS: The aim of the present study was to investigate the cytotoxicity and toxicological interaction of AgNP (1-4 nm, 0.36 and 3.6 µg mL-1) and cadmium (Cd, 1 and 10 µM) mixtures. The murine macrophage cell line RAW 264.7 was used as a model. RESULTS: Effects were observed after a few hours (4 h) on reactive oxygen species (ROS) and became more pronounced after 24 h-exposure. Cell death occurred by apoptosis, and loss of cell viability (24 h-exposure) was preceded by increases of ROS levels and DNA repair foci, but not of NO levels. Co-exposure potentiated some effects (decrease of cell viability and increase of ROS and NO levels), indicating toxicological interaction. CONCLUSION: These effects are important findings that must be better investigated, since the interaction of Cd with AgNP from nanoproducts may impair the function of macrophages and represent a health risk for humans.

Effects of trophic 2,2', 4,4'-tetrabromodiphenyl ether (BDE-47) exposure in Oreochromis niloticus: A multiple biomarkers analysis.

Polybrominated diphenyl esters are emerging environmental contaminants with few toxicological data, being a concern for the scientific community. This study evaluated the effects of BDE-47 on the health of Oreochromis niloticus fish. The animals were exposed to three doses of BDE-47 (0, 0.253, 2.53, 25.3 ng g-1) every 10 days, for 80 days. The BDE-47 affected the hepatosomatic and gonadosomatic index in female and the condition factor by intermediate dose in both sexes. The levels of estradiol decreased and the T4 are increased, but the vitellogenin production was not modulated in male individuals. Changes in AChE, GST, LPO and histopathology were observed while the integrated biomarker response index suggests that the lowest dose of BDE-47 compromised the activity of antioxidant enzymes. The oral exposure to BDE-47 in environmental concentrations is toxic to O. niloticus and the use of multiple biomarkers is an attribution in ecotoxicology studies and biomonitoring programs.

2,4,6-Tribromophenol is toxic to Oreochromis niloticus (Linnaeus, 1758) after trophic and subchronic exposure.

The presence of 2,4,6-Tribromophenol (TBP) in the environment increased the risk of exposure to aquatic organisms affecting the animal development or metabolism. The current study investigated the low, subchronic and trophic effect of TBP in both, male and female adult of Oreochromis niloticus. The fish were exposed to 0.5 or 50 ng g-1 of TBP every ten days for 70 days. Then, hepatosomatic (HSI) and gonadosomatic (GSI) indexes, erythrocyte parameters (hemoglobin content, nuclear morphology and morphometrical abnormalities), biochemical endpoints (glutathione S-Transferase and catalase activities, non-protein thiols, lipid peroxidation and protein carbonylation levels in the liver; and acetylcholinesterase activity in the brain and muscle), histopathological analysis (liver) and vitellogenin levels (plasma) were considered. TBP affected the HSI in male and female fish, but not the GSI. Principal Component Analysis revealed that erythrocytes from males are more sensitive to TBP exposure. Likewise, TBP induced the expression of vitellogenin, CAT activity and liver lesion in male fish comparatively with control group, but GST and NPT were influenced only by sex. Finally, the results showed that the antioxidant mechanism and cholinesterase activity effects were more pronounced in male than in female. The current data shows evidences of estrogenic endocrine disruption and toxicity in O. niloticus exposed to TBP, revealing the risk of exposure to biota.

Stereological study and analysis of oxidative stress during renal aging in rats.

PURPOSE: To evaluate renal histological changes by stereology and morphometry and analyze the main markers of oxidative stress in rats undergoing natural aging. METHODS: Seventy two Wistar rats were divided into six groups of 12 rats each, which were euthanized at 3, 6, 9, 12, 18, and 24 months of age. Right kidney was stereologically and morphometrically analyzed to calculate the volumetric density (Vv[glom]), numerical density (Nv[glom]) and glomerular volume (Vol[glom]). Left kidney was used to determine the levels of nonprotein thiols, lipid peroxidation, and protein carbonylation, as well as the activities of superoxide-dismutase and catalase enzymes. RESULTS: Both Vv[glom] and Nv[glom] values showed gradual decreases between groups. Activity of superoxide-dismutase was elevated at 24 months of age, and the levels of nonprotein thiols were higher in older animals. Greater catalase activity and protein carbonylation were observed in animals between 6 and 12 months of age but lessened in older rats. Lipid peroxidation decreased in the older groups. CONCLUSIONS: Morphometric and stereological analyses revealed a gradual decrease in the volume and density of renal glomeruli during aging, as well as kidney atrophy. These findings related to oxidative stress clarify many changes occurring in kidney tissues during senescence in rats.

Malignancy and tumorigenicity of melanoma B16 cells are not affected by silver and gold nanoparticles.

Gold (AuNP) and silver (AgNP) nanoparticles have been incorporated into many therapeutic and diagnostic applications. However, previous studies revealed toxic properties as well as the hormesis phenomenon of many nanoparticles in different biological models. To evaluate the effects of low concentrations of AuNP and AgNP on murine melanoma cells B16F1 and B16F10 and relate them with phenotype changes, cells were exposed for 24 and 48 h. No cytotoxicity was observed for B16 cells through neutral red, MTT, trypan blue, and crystal violet assays at concentrations from 0.01 to 10 ng mL-1. Likewise, the nanoparticles did not interfere with drug-efflux activity, cell migration, cell cycle, and colony formation. Slight toxicity was observed for B16F10 exposed to 100 ng mL-1, with a decreased number of viable and attached cells, indicating differential sensitivity of B16F1 and B16F10 cells to the nanoparticles. Furthermore, colony size dispersion decreased for both B16 cell sub-lines. Therefore, there is no evidence that the tested concentrations of AuNP and AgNP can render B16 cells more aggressive and malignant, which is important since both nanoparticles are already largely used in nanotechnological products. Considering studies that have showed the hormesis effect of nanoparticles at low concentrations, which could protect cancer cells against chemotherapy, further investigation is advised.

Oral exposure to BDE-209 modulates metastatic spread of melanoma in C57BL/6 mice inoculated with B16-F10 cells.

Polybrominated diphenyl ethers (PBDEs) are brominated, persistent and bioaccumulative flame retardants widely used in the manufacture of plastic products. Decabromodiphenyl ether (BDE-209) is the most prevalent PBDE in the atmosphere and found in human blood, breast milk and umbilical cord. In vitro studies showed that BDE-209 interferes with murine melanoma cells (B16F10), modulating cell death rates, proliferation and migration, important events for cancer progression. In order to evaluate if BDE-209 modulates metastasis formation in murine models, C57BL/6 mice were exposed to BDE-209 (0.08, 0.8 and 8 µg/kg) via gavage (5-day intervals for 45 days) (9 doses in total). Then, mice were inoculated with melanoma cells (B16-F10) at caudal vein receiving 4 additional doses of BDE-209. At 20th day post-cell inoculation, blood, lung, liver, kidney and brain were sampled for hematological, biochemical and morphological analyses. The slightly higher levels of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in the blood and pro-oxidant state in the liver of BDE-exposed mice indicated liver damage. Although the in vivo approach is for metastasis formation in the lung, they were unexpectedly observed in non-target organs (liver, brain, kidney and gonads). The similarity test showed high proximity among individuals from the control and a dissimilarity index between the control and exposed groups. The present data corroborate the known hepatotoxicity of BDE-209 to mice (C57BL/6) and demonstrate for the first time the increase of metastatic dissemination of B16F10 cells in vivo due to previous and continuous BDE-209 exposure, revealing possible implications of this organic compound with melanoma malignancy related traits.

Low concentration of 2,4,6-tribromophenol (TBP) represents a risk to South American silver catfish Ramdia quelen (Quoy and Gaimard, 1824) population.

The 2,4,6-tribromophenol (TBP) is an environmental persistent pollutant widely used as flame retardant, antimicrobial and insecticide agent in wood preservation and plastic production. Currently, TBP is found in environmental compartments such as soil, freshwater, groundwater, sewage sludge and domestic dust, but the effects to biota and the risk of exposure to aquatic vertebrates are still scarce. In the present study, Rhamdia quelen fish embryos (8 h post-fertilization - hpf) were exposed to 0.3 and 3.0 µg L-1 of TBP until 96 hpf. Biochemical biomarkers, hatching, survival and larvae/embryo malformations were evaluated after exposure. Additionally, a mathematical model was proposed to evaluate the effects along further generations. The results showed that TBP decreased the survival level but did not cause significant difference in the hatching rates. After 72 and 96 hpf, individuals from the highest tested concentration group showed more severe malformations than individuals from control and the lower concentrations groups. The deformities were concentrated on the embryos facial region where the sensorial structures related to fish behavior are present. The biochemical biomarkers revealed both oxidative stress and neurotoxicity signs after exposure to the contaminant, while the application of the mathematical model showed a decrease of population in both tested TBP concentrations. In conclusion, the current results demonstrated that TBP is toxic to R. quelen embryos and represents a risk to population after early life stage exposure.

Stereological study and analysis of oxidative stress during renal aging in rats

Purpose To evaluate renal histological changes by stereology and morphometry and analyze the main markers of oxidative stress in rats undergoing natural aging. Methods Seventy two Wistar rats were divided into six groups of 12 rats each, which were euthanized at 3, 6, 9, 12, 18, and 24 months of age. Right kidney was stereologically and morphometrically analyzed to calculate the volumetric density (Vv[glom]), numerical density (Nv[glom]) and glomerular volume (Vol[glom]). Left kidney was used to determine the levels of nonprotein thiols, lipid peroxidation, and protein carbonylation, as well as the activities of superoxide-dismutase and catalase enzymes. Results Both Vv[glom] and Nv[glom] values showed gradual decreases between groups. Activity of superoxide-dismutase was elevated at 24 months of age, and the levels of nonprotein thiols were higher in older animals. Greater catalase activity and protein carbonylation were observed in animals between 6 and 12 months of age but lessened in older rats. Lipid peroxidation decreased in the older groups. Conclusions Morphometric and stereological analyses revealed a gradual decrease in the volume and density of renal glomeruli during aging, as well as kidney atrophy. These findings related to oxidative stress clarify many changes occurring in kidney tissues during senescence in rats.

Biomarker responses in fish exposed to polycyclic aromatic hydrocarbons (PAHs): Systematic review and meta-analysis.

Biomarkers of antioxidant and biotransformation systems are commonly used to assess polycyclic aromatic hydrocarbons (PAHs) pollution in fish. Despite their extensive application of biomarkers, contradictory results are vastly reported in the literature, even for the same species in similar contamination scenarios. This study aims to verify response patterns of biomarkers in fish exposed to PAHs. Through systematic reviews and meta-analyses, we were able to evaluate: (i) overall magnitude of PAHs effects on biotransformation and oxidative stress biomarkers; (ii) patterns of response among experimental approaches (laboratory, field and active biomonitoring), environment (marine and freshwater) and fish habitat (pelagic, demersal, etc.); (iii) effects of exposure route, time and concentration of PAHs; and (iv) which biomarkers respond best to PAHs exposure. Overall, biomarker responses were significantly affected by PAHs exposure. The activities of ethoxyresorufin-O-deethylase (EROD), glutathione S-transferase (GST), superoxide dismutase (SOD), glutathione peroxidase (GPx) and levels of oxidized glutathione (GSSG) and lipid peroxide (LPO) significantly increased in fish exposed to PAHs, whereas catalase (CAT) and glutathione reductase (GR) activities and levels of reduced glutathione (GSH) were not affected. Amongst responsive biomarkers, EROD and GST activities significantly differed among approaches and between marine and freshwater environments, but were not affected by fish habitat. GSSG levels were higher in fish from laboratory bioassays compared to the field, but did not differ between environments nor habitats. Exposure route played a major role only for GST and GPx responses. Finally, increasing PAHs concentration and exposure time had a significant effect on all assessed biomarkers, except for CAT. We conclude that EROD and GST are robust biomarkers to assess PAHs effects in fish. Contrarily, CAT is an inadequate biomarker of PAHs exposure since no significant response was observed. Our study also highlighted some research gaps in PAHs contamination studies, such as a clear lack of active biomonitoring experiments.

Comparative bioaccumulation and effects of purified and cellular extract of cylindrospermopsin to freshwater fish Hoplias malabaricus.

Many tropical freshwater ecosystems are impacted by cyanobacteria blooms increasing the risk of cyanotoxins exposure to aquatic organisms while human populations may be exposed by eating fish, drinking water, or dermal swimming. However, few toxicological data are available on the influence of cyanobacteria blooms in particular, cylindrospermopsin (CYN) on Brazilian neotropical fish. A number of studies demonstrated the ability of CYN to bioaccumulate in freshwater organisms and consequently enter the human food chain. The aim of the current study was to examine the effects of CYN following single intraperitoneal injection (50 µg/kg) of purified CYN (CYNp) or aqueous extract of CYN-producing cyanobacteria extract (CYNex) after 7 or 14 days. Biomarkers such as histopathology (liver), oxidative stress (liver and brain), and acetylcholinesterase (AChE) activity (muscle and brain) were utilized in order to assess the influence of CYN on Hoplias malabaricus. In terms of AChE activity, administration of CYNex and CYNp both muscle and brains were used as target tissues. In brain an increase of glutathione S-transferase (GST) activity and lipid peroxidation (LPO) levels was noted suggesting an imbalance in redox cycling. The majority of biomarkers did not present significant alterations in liver, but an elevation in superoxide dismutase (SOD) and glucose 6 phosphate dehydrogenase (G6PDH) activities was found. Different profiles of GST activity were observed in both studied groups (CYNex and CYNp) while LPO (CYNex and CYNp) and protein carbonylation (PCO) (CYNp) levels increased after exposure to CYN. The incidence of necrosis, melanomacrophages centers, and free melanomacrophages were detected as evidence of cell death and immune responses. Nonprotein thiols (NPT) levels were not markedly affected in both exposed groups. Data demonstrated that in vivo exposure to CYN produced biochemical and morphological disturbances in liver and brain of H. malabaricus.

Tribromophenol affects the metabolism, proliferation, migration and multidrug resistance transporters activity of murine melanoma cells B16F1.

Murine melanoma cells B16F1 were exposed to the flame retardant and wood preservative chemical 2,4,6-tribromophenol (TBP) during 24 and 48 h, at the concentrations found in human diet. TBP-exposed cells had increased MTT and Alamar blue® metabolism and ABCB5 mRNA levels (qPCR), but the cells had decreased proliferation (crystal violet assay), migration (scratch assay), and drug-effux transporters activity (rhodamine B efflux assay). Exposure to TBP did not affect the cell viability (neutral red and annexin V-PI assays), colony formation (colony number, clonogenic assay), and the levels of reactive oxygen species (DCF probe) or P53 mRNA (qPCR). The tested TBP concentrations had low toxicity to melanoma cells B16F1. However, dual effect on metastatic profile and chemoresistance suggests that the increase of ABCB5 positively modulates the cell chemoresistance, but decreases cell migration and proliferation. These findings may be explored in cancer therapy.

Diffuse sources of contamination in freshwater fish: Detecting effects through active biomonitoring and multi-biomarker approaches.

Aquatic organisms are usually exposed to a mixture of xenobiotics that may exert a large effect even in low concentrations, and when information is obtained exclusively from chemical analyses the prediction of the deleterious effects is potentially hindered. Therefore, the application of complementary monitoring methods is a priority. Here, in addition to chemical analyses, an active biomonitoring study using multiple biomarker responses in Nile tilapia Oreochromis niloticus was conducted to assess the effects of a contamination gradient along four reservoirs in Iguaçu River. Chemical analysis in the muscle showed high levels of metals in fish from the reservoir closest to an industrialized and environmentally degraded area, however fish exposed to all studied reservoirs showed hepatic alterations (necrosis and inflammatory processes). Also, significant variations of biochemical biomarkers were observed with no clear indication of contamination gradient, since an indicative of higher impact was found in an intermediary reservoir, including high concentrations of biliary polycyclic aromatic hydrocarbons (PAHs). However, nuclear morphological alterations (NMA) were less frequent at the same reservoir. Thus, the multi-biomarker approach allied to active biomonitoring is a practical and important tool to assess deleterious effects of contamination in freshwater, providing data for monitoring and conservation protocols.