Integrating chemical and biological data by chemometrics to evaluate detoxification responses of a neotropical bivalve to metal and metalloid contamination.

Mangroves represent a challenge in monitoring studies due to their physical and chemical conditions under constant marine and anthropogenic influences. This study investigated metals/metalloids whole-body bioaccumulation (soft tissues) and the risk associated with their uptake, biochemical and morphological detoxification processes in gills and metals/metalloids immobilisation in shells of the neotropical sentinel oyster Crassostrea rhizophorae from two Brazilian estuarine sites. Biochemical and morphological responses indicated three main mechanisms: (1) catalase, superoxide dismutase and glutathione played important roles as the first defence against reactive oxygen species; (2) antioxidant capacity against peroxyl radicals, glutathione S-transferase, metallothionein prevent protein damage and (3) metals/metalloids sequestration into oyster shells as a mechanism of oyster detoxification. However, the estimated daily intake, target hazard quotient, and hazard index showed that the human consumption of oysters would not represent a human health risk. Among 14 analysed metals/metalloids, chemometrics indicate that Mn, As, Pb, Zn and Fe overload the antioxidant system leading to morphological alterations in gills. Overall, results indicated cellular vacuolization and increases in mucous cell density as defence mechanisms to prevent metals/metalloids accumulation and the reduction in gill cilia; these have long-term implications in respiration and feeding and, consequently, for growth and development. The integration of data from different sites and environmental conditions using chemometrics highlights the main biological patterns of detoxification from a neotropical estuarine bivalve, indicating the way in which species can cope with metals/metalloids contamination and its ecological consequences.

P-glycoprotein inhibition affects ivermectin-induced behavioural alterations in fed and fasted zebrafish (Danio rerio).

The role of the blood-brain barrier ATP-binding cassette protein transporter P-glycoprotein (P-gp) in protecting zebrafish (Danio rerio) from the central nervous system neurotoxicant ivermectin (IVM, 22,23-dihydroavermectin B1a + 22,23-dihydroavermectin B1b) was examined in the absence and presence of the competitive inhibitor cyclosporin A (CsA). Zebrafish injected intraperitoneally with 1, 2, 5, or 10 µmol/kg IVM exhibited mortality 30 min following administration at the highest dose. At sublethal doses > 1 µmol/kg, IVM altered the swimming performance, exploratory behaviour, motor coordination, escape response and olfactory response in exposed fish. When fish were exposed to IVM in the presence of CsA, alterations in swimming and behaviours increased significantly and at the highest IVM/CsA ratio resulted in a complete lack of exploratory and olfactory behaviours. In separate experiments, fish were either fed or fasted, and the effects of IVM and CsA administration were examined. The effects of IVM administration and the exacerbated effects seen with CsA co-administration were not affected by fasting. This study provides evidence that P-gp provides a protective role in the BBB of fish against environmental neurotoxicants. The results also show that P-gp activity is maintained even under conditions of food deprivation, suggesting that this chemical defence system is prioritized over other energy expenditures during diet limitation.

Different trophodynamics between two proximate estuaries with differing degrees of pollution.

Mangroves are complex ecosystems with widely varying abiotic factors such as salinity, pH, redox potential, substratum particle size, dissolved organic matter and xenobiotic concentrations, and a high biodiversity. This paper presents the trophodynamic pathways of accumulation and transfer of metals and metalloids (B, Al, V, Cr, Mn, Fe, Ni, Cu, Zn, Ag, As, Se, Rb, Sr, Pb and Hg), in three trophic chains (plant-crab-fish, plankton-shrimp-fish and plankton-oyster) of similar food webs, corresponding to two mangrove estuaries (Santa Cruz and Vitória Bay, separated by 70 km) in the Espírito Santo State (Brazil). Although the trophic transfer patterns are affected by physical variables, metal and metalloids were found in all trophic levels. We observed similar trophodynamics between both estuaries with some elements, but unequal transfer patterns in other cases, thus questioning the effectiveness of 15N to determine the food chain when the aquatic biota is affected by anthropogenic contaminants. Thus, in the Santa Cruz estuary, most metals were biomagnified through the food web. Conversely, Vitória Bay presented mostly biodilution, suggesting that metal/metalloid transference patterns in mangrove ecosystems may be affected by different anthropogenic contamination inputs. These results indicate the importance of knowing the complete food web when evaluating the trophic transfer of elements, including an evaluation of the differential impact of pollution on diverse components of the food chain.

Atmospheric particulate matter from an industrial area as a source of metal nanoparticle contamination in aquatic ecosystems.

Air pollution legislation and control worldwide is based on the size of particulate matter (PM) to evaluate the effects on environmental and human health, in which the small diameter particles are considered more dangerous than larger sizes. This study investigates the composition, stability, size and dispersion of atmospheric settleable particulate matter (SePM) in an aqueous system. We aimed to interrogate the changes in the physical properties and characteristics that can contribute to increased metal uptake by aquatic biota. Samples collected in an area influenced by the steel and iron industry were separated into 8 fractions (425 to ≤10 µm) and analysed physically and chemically. Results from ICP-MS and X-ray showed that the PM composition was mainly hematite with 80% of Fe, followed by Al, Mn and Ti. Among 27 elements analysed we found 19 metals, showing emerging metallic contaminants such as Y, Zr, Sn, La, Ba and Bi. Scanning electron microscopy (SEM) showed that SePM fractions are formed by an agglomeration of nanoparticles. Furthermore, dynamic light scattering (DLS), zeta potential and nanoparticle tracking analysis (NTA) demonstrated that SPM were dissociated in water, forming nanoparticles smaller than 200 nm, which can also contribute to water pollution. This study highlights that SePM contamination may be substantially higher than expected under that allowed in atmospheric regulatory frameworks, thereby extending their negative effect to water bodies upon settling, which is an underexplored area of our knowledge. We therefore provide important insights for future investigations on safety regulations involving SePM in the environment, indicating the need to revise the role of SePM, not solely associated with air pollution but also considering their deleterious effects on water resources.

Trophic transfer of emerging metallic contaminants in a neotropical mangrove ecosystem food web.

Emerging metallic contaminants (EMCs) are of concern due their presence in aquatic ecosystems and the lack of environmental regulations in several countries. This study verifies the presence of EMCs in two neotropical mangrove estuarine ecosystems (Espírito Santo Brazil) by evaluating abiotic and biotic matrices across six trophic levels (plankton, oyster, shrimp, mangrove trees, crabs and fish) and hence interrogates the trophic transfer of these elements and their possible input sources. Using the oyster Crassostrea rhizophorae as a biomonitor, ten EMCs (Bi, Ce, La, Nb, Sn, Ta, Ti, W, Y and Zr) were determined. Bi input was from iron export and pelletizing industries; Ce, La and Y inputs were mainly associated with solid waste from steel production, while Zr, Nb and Ti were related to atmospheric particulate matter emissions. EMCs were detected at various trophic levels, showing biomagnification for most of them in the Santa Cruz estuary but biodilution in Vitória Bay. These contrasting results between the estuaries could be attributed to different pollution degrees, needing further research to be fully understood. This is the first report demonstrating EMCs trophic pathways in situ, constituting an essential baseline for future research and safety regulations involving EMCs in the environment.

Overview of the toxic effects of titanium dioxide nanoparticles in blood, liver, muscles, and brain of a Neotropical detritivorous fish.

The toxicity of titanium dioxide nanoparticles (TiO2 -NP) in the blood, liver, muscle, and brain of a Neotropical detritivorous fish, Prochilodus lineatus, was tested. Juvenile fish were exposed to 0, 1, 5, 10, and 50 mg L-1 of TiO2 -NP for 48 hours (acute exposure) or 14 days (subchronic exposure) to evaluate changes in hematology, red blood cell (RBC) genotoxicity/mutagenicity, liver function (reactive oxygen species (ROS) production, antioxidant responses, detoxification, and histopathology), acetylcholinesterase (AChE) activity in muscles and brain, and Ti bioaccumulation. TiO2 -NP did not cause genetic damage to RBC, but acutely decreased white blood cells (WBC) and increased monocytes. Subchronically, RBC decreased, mean cell volume and hemoglobin increased, and WBC and lymphocytes decreased. Therefore, NP has the potential to affect immune system and increase energy expenditure, reducing the fish's ability to avoid predator and to resist pathogens. In the liver, acute exposure decreased ROS and increased glutathione (GSH) content, while subchronic exposure decreased superoxide dismutase activity and increased glutathione-S-transferase (GST) activity and GSH content. GSH and GST seem to play an essential role in metabolizing NP and ROS, likely increasing hepatocytes' metabolic rate, which may be the cause of observed cell hypertrophy, disarrangement of hepatic cords and degenerative morphological alterations. Although most studies indicate that the kidney is responsible for metabolizing and/or eliminating TiO2 -NP, this study shows that the liver also has a main role in these processes. Nevertheless, Ti still accumulated in the liver, muscle, and brain and decreased muscular AChE activity after acute exposure, showing neurotoxic potential. More studies are needed to better understand the biochemical pathways TiO2 -NP are metabolized and how its bioaccumulation may affect fish homeostasis and survival in the environment.

Nanoparticle transport and sequestration: Intracellular titanium dioxide nanoparticles in a neotropical fish.

Intracellular titanium dioxide nanoparticles (TiO2-NP) with rutile crystalline form and dimensions varying from 43 to 67 nm × 64 to 93 nm are reported for the first time as being sequestered from the environment. TiO2-NP were identified inside all organs/tissues (muscle, kidney, gonad, hepatopancreas and gill) in both the cytoplasm and nucleus of the neotropical fish Centropomus parallelus, captured in an area affected by metallurgical activity. Atmospheric particulate matter (PM) sampled in the same area showed the presence of TiO2-NP with the same rutile crystalline form and dimensions varying from 16 to 93 nm × 45 to 193 nm, thus indicating the smelting and iron processing industries as the most probable source of TiO2-NP. In any sample, chemical analyses identify and quantify Ti concentration and nanocrystallography identified the structure of TiO2-NP. The Ti concentration in the sediment and atmospheric PM varied between years and it was mirrored by the Ti concentration in the fish organs. The gill has a higher Ti concentration varying from 5.50 to 14.57 µg g-1 dry weight and the gonad was the organ with lowest Ti level, 0.25 to 0.87 µg g-1 dry weight. In the muscles, Ti concentration varied from 0.85 to 3.34 µg g-1 dry weight. This contamination may be likely to affect the surrounding biota and food uptake, including the humans living in the city close to the metallurgical complex. These findings emphasised the needs to improve methods to reduce PM (including nanoparticles) arising from human activities and to evaluate the toxicokinetic and effects of TiO2-NP in the biota and human health.

Differential biochemical responses to metal/metalloid accumulation in organs of an edible fish (Centropomus parallelus) from Neotropical estuaries.

Metal/metalloid accumulation in fish organs elicits biochemical responses indicating the overall fish and environmental health status. This study evaluated the bioaccumulation of metals and metalloid in relation to a suite of biochemical biomarkers (superoxide dismutase, catalase, glutathione-S-transferase, Na+/K+-ATPase, H+-ATPase, acetylcholinesterase activities and the levels of glutathione, metallothionein, lipid peroxidation and oxidized protein) in different organs of fish, Centropomus parallelus, in Vitória Bay and Santa Cruz estuaries (State of Espírito Santo, Brazil) with distinct contamination levels. Metal and metalloid concentrations differ in each organ and were significantly higher in winter than in summer. Chemometric evaluation performed between metal/metalloid accumulation and the biomarkers revealed a complex scenario in which the biomarker responses depend on both metal accumulation and organ/tissue sensitivity. The metal levels in gills indicate fish contamination mainly via water and the low sensitivity of this organ to most metals. Biomarker responses suggested that the metal elimination pathway is through the gills and kidney. The hepatopancreas and kidneys were the most important detoxification organs while muscle was the less reactive tissue. In general, the finding suggested that, C. parallelus is partly able to tolerate such metal contamination. However, it is emphasized that the biomarker responses imply an energetic cost and may affect the growth rate and reproduction. Given the ecological and economic importance of C. parallelus, the level of toxic metals/metalloids in juvenile fish is an important early-warning for the maintenance, conservation and commercial use of this species.

Mitochondria-rich cells adjustments and ionic balance in the Neotropical fish Prochilodus lineatus exposed to titanium dioxide nanoparticles.

Manufactured titanium dioxide nanoparticles (TiO2-NP) have been intensely applied in numerous industrial products and may be a risk for aquatic systems as they are not completely removed from domestic and industrial wastes after water treatment. This study evaluated the osmo- and ionic balance, Na+/K+-ATPase, H+-ATPase and carbonic anhydrase activities and the mitochondria-rich cells (MRC) in the gills and kidney of the Neotropical fish Prochilodus lineatus after 2 (acute) and 14 (subchronic) days of exposure to nominal 0, 1, 5, 10 and 50 mg L-1 TiO2-NP. The nominal concentrations corresponded to 0.0, 0.6, 1.6, 2.7 and 18.1 mg L-1 suspended TiO2-NP, respectively, in the water column one hour after NP introduction and were maintained for at least 24 h. Acute exposure to TiO2-NP decreased plasma osmolality and Ca2+ levels. Na+/K+-ATPase, H+-ATPase and carbonic anhydrase activities were inhibited in the gills, but not in the kidney. Total MRC density did not change in gills and kidneys. At gill surface, total MRC density decreased in fish exposed to 50 mg L-1 TiO2-NP and the total MRC fractional surface area unchanged although, there were some changes in the fractional area of MRC with apical microvilli (MRCm) and MRC with apical sponge-like structure (MRCs). MRCm was more abundant than MRCs. After subchronic exposure, there was no change in plasma osmolality, ionic balance and enzyme activities. Total gill MRC density increased in the filament epithelium and renal tubules. In the gills, MRC contacting water exhibited some adjustments. Total MRC and fractional surface area unchanged, but there was an increase of MRCs contacting water at gill surface after exposure to10 and 50 mg L-1 TiO2-NP. MRC proliferation in filament epithelium and in renal tubules as well as the increasing MRCs at gill surface may have contributed to avoid change in plasma osmolality, ionic balance and enzyme activities and suggested a cellular physiological and morphological response to restore and maintain osmotic and ionic homeostasis after subchronic exposure.

Matching metal pollution with bioavailability, bioaccumulation and biomarkers response in fish (Centropomus parallelus) resident in neotropical estuaries.

Two neotropical estuaries affected by different anthropogenic factors were studied. We report levels of metals and metalloids in water and sediment as well as their influence on genetic, biochemical and morphological biomarkers in the native fish Centropomus parallelus. Biomarkers reflected the fish health status. Multivariate statistics indicated both spatial and temporal changes in both water and sediment, which are linked to the elemental composition and health status of inhabitant fish, showing the biggest influence of surface water, followed by sediments and interstitial water. Bioaccumulation in fish muscle was useful to identify elements that were below detection limits in water, pointing out the risk of consuming fish exceeding allowance limits for some elements (As and Hg in this case). Multivariate statistics, including physical, chemical and biological issues, presents a suitable tool, integrating data from different origin allocated in the same estuary, which could be useful for future studies on estuarine systems.