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.

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.