Ivermectin toxicokinetics in rainbow trout (Oncorhynchus mykiss) following P-glycoprotein inhibition.

Changes to ivermectin (IVM [22,23-dihydro avermectin B1a + 22,23-dihydro avermectin B1b]) toxicokinetics (TK) with and without P-glycoprotein (P-gp) inhibition by cyclosporin A (CsA) were examined in rainbow trout (Oncorhynchus mykiss). Rainbow trout were injected with 175 µg/kg 3H-IVM (8.6 µCi/mg IVM) with or without co-administration of 480 µg/kg CsA into the caudal vasculature. Fish were sacrificed at various time points (0.25, 0.5, 1, 3, 24, 48, 96, and 168 h) for organ and tissue sampling (blood, liver, kidney, gill, intestines, brain [5 regions], eye, gonad, and fat) which were analyzed for IVM-derived radioactivity. The IVM concentration decreased over time in blood, liver, kidney, and gill, while concentrations in other tissues remained constant. The highest maximum IVM concentration (Cmax) was found in kidney, followed by liver; the lowest Cmax was found in eye, followed by brain and adipose tissue. The highest % of the administered dose was found in the blood 15 min post-IVM administration, followed by the intestine at 60 min post-IVM administration. P-gp inhibition by CsA did not significantly affect calculated TK parameters (AUC [7.33 ± 0.73 - 11.5 ± 2.5 mg•h/kg], mean residence time [84.7 ± 21 - 125 ± 55 h], T1/2 [58.7 ± 15 - 86.8 ± 38 h], clearance rate [0.0152 ± 0.0033 - 0.0239 ± 0.0024 L/kg•h], or volume of distribution [1.91 ± 0.47 - 2.02 ± 0.33 L/kg]), but resulted in small but significant changes in the % administered dose found in blood and medulla. These results suggest that P-gp plays a limited role in overall IVM TK, and that its role in xenobiotic protection may be much less robust in fish than it is in mammals.

The effects of P-glycoprotein induction on ivermectin-induced behavioural alterations in zebrafish (Danio rerio) under varying diets.

The neuroprotective effects of inducing the blood-brain barrier ATP-binding cassette protein transporter P-glycoprotein (P-gp) with clotrimazole (CTZ) in both fed and fasted zebrafish (Danio rerio) against the CNS-toxicant ivermectin (IVM, 22,23-dihydro avermectin B1a + 22,23-dihydro avermectin B1b) were examined. Zebrafish were administered 2 µmol/kg IVM intraperitoneally, and various behavioural assays (swimming performance, exploratory behaviour, olfactory responses, motor coordination, and escape responses) were used to measure neurological dysfunction. IVM administration alone caused a decrease in mean swim speed (91 % of controls), maximal speed (71 %), passage rate (81 %), 90° turns (81 %), and response to food stimulus (39 %). IVM exposure also increased the percent time that fish spent immobile (45 % increase over controls) and the percent of lethargic fish (40 % increase). Fish administered 30 µmol/kg of the P-gp inducer CTZ intraperitoneally 3 d prior to IVM exposure exhibited a change in only the % time spent immobile. These data indicate that P-gp induction may be limited in protecting the zebrafish CNS from IVM over baseline. Fasted fish did not differ from fed fish in the effects of IVM on behaviour, and no differences were seen following P-gp induction with CTZ. These results suggest that this chemical defence system is not downregulated when fish are challenged with limited energy availability.

rGO outperforms GO in generating oxidative stress and DNA strand breaks in zebrafish liver cells.

Graphene oxide (GO) and reduced graphene oxide (rGO) are both widely applicable and there is a massive production throughout the world which imply in inevitable contamination in the aquatic environment by their wastes. Nevertheless, information about their interaction at the cellular level in fish is still scarce. We investigated the metabolic activity, reactive oxygen species (ROS) production, responses of antioxidant defenses, and total antioxidant capacity (TAC) as well as oxidative stress and DNA integrity in zebrafish liver cells (ZFL) exposed to (0.001, 0.01, 0.1 and 1 µg mL-1) of GO and rGO after two exposure period (24 and 72 h). Higher ROS production and no significant changes in the antioxidant defenses resulted in lipid peroxidation in cells exposed to rGO. Cells exposed to GO increased the activity of antioxidant defenses sustaining the TAC and avoiding lipid peroxidation. Comet assay showed that both, GO and rGO, caused DNA strand breaks after 24 h of exposure; however, only rGO caused DNA damage after 72 h of exposure. The exposure to rGO was significantly more harmful to ZFL cells than GO, even at very low concentrations. The cells showed a high capacity to neutralize ROS induced by GO preventing genotoxic effects and metabolic activity, thus sustaining cell viability. The time of exposure had different impacts for both nanomaterials, GO caused more changes in 24 h showing recovery after 72 h, while cells exposed to rGO were jeopardized at both exposure times. These results indicate that the reduction of GO by removal of the oxygen functional groups (rGO) increased toxicity leading to adverse effects in the cells, even at very low concentrations.

Metal/metalloid bioconcentration dynamics in fish and the risk to human health due to water contamination with atmospheric particulate matter from a metallurgical industrial area.

Settleable atmospheric particulate matter (SeAPM) containing a mixture of metals, including metallic nanoparticles, has increased throughout the world, and caused environmental and biota contamination. The metal bioconcentration pattern in Nile tilapia (Oreochromis niloticus) was evaluated during a 30-day exposure to 1 g L-1 SeAPM and assessed the human health risk from consuming fish fillets (muscle) based on the estimated daily intake (EDI). SeAPM was collected surrounding an iron ore processing and steel industrial complex in Vitória city (Espírito Santo, Brazil) area. Water samples were collected daily for physicochemical analyses, and every 3 days for multi-elemental analyses. Metal bioconcentrations were determined in the viscera and fillet of fish every 3 days. The elements B, Al, V, Cr, Mn, Fe, Ni, Cu, Zn, As, Se, Rb, Sr, Ag, Cd, Pb, Hg, Ba, Bi, W, Ti, Zr, Y, La, Nb, and Ce were analyzed in SeAPM, water, and fish using inductively coupled plasma mass spectrometry. The metal concentration in SeAPM-contaminated water was higher than in control water. Most metals bioconcentrated preferentially in the fish viscera, except for the Hg and Rb, which bioconcentrated mostly in the fillet. The bioconcentration pattern was Fe > Al > Mn > Pb > V > La > Ce > Y > Ni > Se > As > W > Bi in the viscera; it was higher than the controls throughout the 30-day exposure. Ti, Zr, Nb, Rb, Cd, Hg, B, and Cr showed different bioconcentration patterns. The Zn, Cu, Sr, Sn, Ag, and Ta did not differ from controls. The differences in metal bioconcentration were attributed to diverse metal bioavailability in water and the dissimilar ways fish can cope with each metal, including inefficient excretion mechanisms. The EDI calculation indicated that the consumption of the studied fish is not safe for children, because the concentrations of As, La, Zr, and Hg exceed the World Health Organization's acceptable daily intake for these elements.

Reactive oxygen species and other biochemical and morphological biomarkers in the gills and kidneys of the Neotropical freshwater fish, Prochilodus lineatus, exposed to titanium dioxide (TiO2) nanoparticles.

This study investigated the action of titanium dioxide nanoparticles (TiO2-NPs), on the gills and kidneys of Neotropical freshwater fish, Prochilodus lineatus, with emphasis on reactive oxygen species (ROS) production, antioxidant responses, and morphological changes. Fish were exposed to 1, 5, 10, and 50 mg L-1 nominal TiO2-NPs suspended into water for 2 or 14 days. In gills, ROS decreased and glutathione (GSH) increased after 2 days, while ROS and GSH increased and superoxide dismutase activity decreased after 14 days. In kidneys, GSH and lipoperoxidation increased after 2 days and catalase activity decreased after 14 days. Common histopathologies in gills were epithelium hyperplasia, cellular hypertrophy, proliferation of mitochondria-rich cells (MRC), and lamellar stasis; in kidneys, there were cellular and nuclear hypertrophy, focal tubule degeneration, necrosis, and melanomacrophage (MM) proliferation. Although environmentally unlikely, high-dose exposures clarified biological effects of TiO2-NPs, such as ROS formation and MRC responses in the gills, which may impair ionic balance. It was also found that MM are likely responsible for eliminating NPs in the kidney. These findings will help to regulate TiO2-NP disposal, but longer-term studies are still needed.