Ovaries of guppies (Poecilia reticulata) investigated in pre-embryonic, embryonic and post-embryonic stages after exposure to maghemite nanoparticles (y-Fe2O3) associated with Roundup® and glyphosate, followed by recovery period evaluation.

Iron oxide nanoparticles (IONP) are promising alternatives to environmental remediation, so this study investigates IONP single and associated to contaminants, in this case, glyphosate (GLY) and Roundup® (GBH) in Poecilia reticulata (guppy). The guppies have internal development, therefore this study analyzed female gonads to establish the developmental stages of P. reticulata and evaluate effects of exposure (7, 14 and 21 days) and post-exposure (same period) to the treatments with Iron ions 0.3 mg Fe/L (IFe); IONP 0.3 mg Fe/L; IONP 0.3 mgFe/L + GBH 0,65 mgGLY/L (IONP+GBH1); IONP 0.3 mgFe/L + GBH 1.30 mgGLY/L (IONP+GBH2); and IONP 0.3 mgFe/L + GLY 0.65 mg/L (IONP+GLY). The development was organized in immature, development, and gestation phases. The damage in all treatments after 21 days of exposure was evident in reaction patterns regressive inflammatory, and circulatory including total histopathologic index of liver, nevertheless there was a damage recovery trend during post-exposure period.

Co-exposure of iron oxide nanoparticles with glyphosate herbicides in Poecilia reticulata: Fish liver damages is reversible during iron accumulation and elimination period.

Iron oxide nanoparticles (IONPs) are advanced materials for water remediation technologies. It is therefore relevant to evaluate the cellular and tissue behavior of fishes in response to IONPs and their associations with agrochemicals such as glyphosate (GLY) and glyphosate-based herbicides (GBHs). Iron accumulation, tissue integrity and lipid distribution in the hepatocytes of Poecilia reticulata (guppy) were investigated in a control group and in groups exposed to soluble iron ions, namely IFe (0.3 mgFe/L), IONPs (0.3 mgFe/L), and IONPs, associated with GLY (0.65 mg/L), GBHs 0.65 mgGLY/L (IONPs + GBH1), and 1.30 mgGLY/L (IONPs + GBH2), for 7, 14, and 21 days, followed by an equal period of postexposure in clean reconstituted water. The results showed that the accumulation of iron was greater in the subjects in the IONP treatment group when compared to that in the Ife group. In addition, the subjects in the mixtures with GBHs had a greater accumulation of iron than those in the IONP + GLY treatment group. Tissue integrity assessments demonstrated an intense accumulation of lipids, formation of necrotic zones and leukocyte infiltrates in all the treated groups, with a greater quantity of lipids in the animals treated with IONP + GLY and IFe. During postexposure, the results indicated an elimination of iron in all treated groups, reaching the same level as the control group, throughout the 21 days postexposure. Thus, the damage caused to animal livers by IONP mixtures is reversible, providing promising results for the development of safe environmental remediation practices using nanoparticles.

Distribution and behavior of lipid droplets in hepatic cells analyzed by variations of citochemical technique and scanning electron microscopy.

Toxicity evaluations involve the analysis of multiple biomarkers. In this study, the liver, target organ analyzed by treatments with iron concentrations, indicated the accumulation of lipids as a response. Considering that the distribution of lipids in an organ is directly related to the induction of inflammatory processes by aquatic contaminants, this study proposes to carry out an integrative investigation of the behavior and the distribution of lipids in the liver tissue. Techniques of light and electron microscopy were performed in order to propose a new way of assessing and quantifying the distribution of lipid droplets, also presenting methodological alternatives that can be chosen by the reader according to the interests and resources available. Thus, it is assumed that the method begins with the fixation of the liver with Glutaraldehyde 2,5% in PBS 0,1 M and continues with post fixation with osmium tretoxide 1%, which marks lipids. For this proposition, two inclusion methodologies were performed to histological analyses in Historesin and ultrastructural analyses in EMBeed 812. For light microscopy (LM) analyses, cuts were obtained with 2,5 micrometers thickness, which were stained with (1) Mayers hematoxylin and (2) toluidine blue. The images obtained were processed in software Image J Fiji to evidence the lipid distribution in liver.•Cytological reactions with osmium tetroxide constitute low complexity methods that allow the optimization of the localization, identification and quantification of lipid droplets in the liver tissue when analyzed under the conventional light microscope.•Samples included in EMBeed 812 resin commonly used in Transmission Electron Microscopy can be analyzed by SEM-BEC, as complementary analyses for the detection of lipids.•Using SEM-BEC and conventional light microscopy, it is possible to quantify the area occupied by lipid droplets using Image J Fiji software, as these are contrasted due to the reaction with osmium tetroxide.

Co-exposure of iron oxide nanoparticles and glyphosate-based herbicide promote liver toxicity in guppy (Poecilia reticulata): A histochemical and ultrastructural approach.

Citrate functionalized iron oxide nanoparticles (IONPs) are employed for various purposes-including environmental remediation but the interaction of IONPs with aquatic contaminants is poorly understood. Among those, glyphosate-based herbicides are toxic and affect target organs such as the liver. Evaluations of livers of female Poecilia reticulata by exposures to IONPs at a concentration of 0.3 mg/L were performed with association to: (1) 0.65 mg of glyphosate per litter and (2) 1.3 mg of glyphosate per litter of Roundup Original, and (3) glyphosate P.A at 0.65 mg/L. These associations were carried out progressively, after 7, 14, and 21 days. We detected circulatory disturbances, inflammatory responses, activation of the immune system, regressive changes, and progressive responses with changes in the connective tissue and decreased glycogen reserve from days 14 to 21. Ultrastructural changes in the Disse space and microvilli of hepatocytes indicated decreased contact surface area. In general, the damage was time and concentration dependent, increasing from 7 to 14 days and tending to stabilize from 14 to 21 days. Therefore, herbicide-associated IONPs functioned as xenobiotics inducing intense cellular detoxification processes and activation of hepatic immune responses.

Recovery trend to co-exposure of iron oxide nanoparticles (γ-Fe2O3) and glyphosate in liver tissue of the fish Poecilia reticulata.

Citrate-coated iron oxide nanoparticles (IONPs) have potential use in environmental remediation, with possibilities in decontaminating aquatic environments exposed to toxic substances. This study analyzed IONPs associated to Roundup Original, a glyphosate-based herbicide (GBH), and pure glyphosate (GLY), through ultrastructural and histopathological biomarkers in liver tissue, from females of Poecilia reticulata exposed to: iron ions (0.3 mg/L) (IFe) and IONPs (0.3 mgFe/L) associated with GLY (0.65 mg/L) and GBH (0.65 mgGLY/L (IONP + GBH1) and 1.30 mgGLY/L (IONP + GBH2)) for a period of 7, 14 and 21 days, followed by an equal post-exposure period only in reconstituted water. For the assays, the synthetized IONPs had crystalline and rounded shape with an average diameter of 2,90 nm, hydrodynamic diameter 66,6 mV, zeta potential -55,4 and diffraction profile of maghemite (γ-Fe2O3). The data obtained by biomarkers indicated a high inflammatory response in all treatments. These same parameters, considered during the post-exposure period indicated recovery in reaction patterns of circulatory disturbances and regressive changes, resulting in average reductions of 37,53 points in IFe, 21 points in IONP + GBH1, 15 points in IONP + GBH2 and 11 points in IONP + GLY in total histopathological index of liver after 21 days post-exposure. However, although the cellular and tissue responses were significant, there was no change in the condition factor and hepatosomatic index, denoting resilience of the experimental model.