Metal and metalloid content, bioavailability and sorption processes in glitter and raw glitter materials and associations with human and ecological risk concerns.

BACKGROUND: Microplastics comprise a significant group of emerging environmental contaminants with the capacity to adsorb several contaminants. These, in turn, undergo bioaccumulation and biomagnification processes throughout aquatic trophic chains. METHODS: Glitter, a microplastic powder composed of a combination of polymers, and raw glitter materials were investigated herein concerning metal and metalloid content, bioavailability, and sorption processes by inductively coupled plasma mass spectrometry (ICP-MS). RESULTS: Metal and metalloid concentrations were higher in glitter than in raw glitter materials, but all were below the limits established by the Brazilian National Health Surveillance Agency. Elements present in glitter originate mainly from pigments and, thus, depend on glitter color. The bioavailability of the determined elements concerning human skin was assessed. Low desorbed concentrations in solution indicate that glitter does not represent a health risk through dermal contact concerning metal and metalloid contamination. However, several elements were shown to undergo significant desorption and adsorption processes. CONCLUSION: The findings reported herein indicate seemingly low human health risks from dermal glitter contact but reinforce glitter risks as aquatic environment metal and metalloid transport vectors.

Before the Dam: A Fish-Mercury Contamination Baseline Survey at the Xingu River, Amazon Basin Before the Belo Monte Dam.

In the last decades, mega-diverse rivers worldwide (e.g., the Amazon and their tributaries) have experienced several human-driven transformations, although impact assessments oftentimes lack baseline data or reference values to better estimate observed disturbances. Herein, we assessed THg and δ15N in fish muscle tissue to determine biomagnification processes, also including seasonal assessments, concerning Hg in fish from the Belo Monte reservoir, located on the Xingu River, prior to its impoundment. A total of 312 samples from 19 fish species belonging to distinct feeding guilds were sampled. None presented THg concentrations above the threshold levels of 0.5 µg g-1 established by the World Health Organization. However, biomagnification was observed (R = 0.8; p < 0.001), also confirmed by a trophic magnification factor of 1.76 for the entire food web. The THg content in the water was less than 0.001 mg L-1 and only Hemiodus unimaculatus presented seasonal variations for THg. Therefore, at the time of this study, the Xingu River presented low Hg contamination levels in both water and target fish.

Cytosolic and Metallothionein-Bound Hepatic Metals and Detoxification in a Sentinel Teleost, Dules auriga, from Southern Rio de Janeiro, Brazil.

Dules auriga, a native Brazilian teleost, was applied as a sentinel species regarding metal contamination at Ilha Grande Bay, previously considered a reference site in Southeastern Brazil. Cytosolic (S50) and metallothionein-bound (HTS50) hepatic iron (Fe), zinc (Zn), copper (Cu), manganese (Mn), cadmium (Cd), and silver (Ag) were determined by inductively coupled plasma optical emission spectrometry (ICP-OES), while metallothionein (MT) concentrations were determined by polarography. Ag concentrations in both cytosolic fractions were below the limit of detection. All other HTS50 metal contents were significantly lower than S50 contents. No significant associations were found for MT. Fe and Mn S50 were positively and moderately correlated to total length, as well as HTS50 Mn, while total weight was correlated to both Mn fractions, suggesting that environmental Mn and Fe concentrations may influence fish growth. A moderate correlation between the condition factor and the S50 Cu fraction was observed, also indicating that Cu may affect fish growth. Inter-element correlations were observed, including between Cd, a toxic element, and Mn and Zn, both essential elements. Calculated molar ratios indicate that both Mn and Zn are in molar excesses compared with Cd, corroborating literature assessments regarding protective Mn and Zn effects against Cd. Lack of MT correlations suggests that metal concentrations may not be high enough to reach an MT induction threshold and that MT variability is probably linked to environmental metal concentrations. Therefore, the increased environmental contaminant levels observed in the study area indicate the need for biomonitoring efforts aiming at the application of efficient mitigation measures.

Metal-Associated Biomarker Responses in Crabs from a Marine Protected Area in Southeastern Brazil.

The environmental quality of a Ramsar wetland site located at the Cananéia-Iguape-Peruíbe Protected Area (CIP-PA), in São Paulo, Brazil, was assessed by geochemical analyses and biomarker assessments (GPx, GST, GSH, GST, MT, LPO, DNA damage) performed in swimming crab Callinectes danae Smith, 1869 organs (posterior and anterior gills and hepatopancreas) to estimate sediment contaminant bioavailability. The results indicated that two sampling stations, PT and PM, exhibited the worst environmental conditions, as sediments collected at both points contained metal contamination, while crabs exhibited significant responses for GPx, GST, and LPO (mostly during winter). Sediment contamination tended to be associated to fine sediments (both seasons) and organic matter (winter). During the summer survey, Pb concentrations in sediments of station PT exceeded the Brazilian Sediment Quality Guidelines (SQGs) and the Canadian Interim Marine Sediment Quality Guidelines. Metal concentrations in sediments sampled in winter were higher compared with summer, with Co, Ni, and Pb exceeding SQGs levels at PT, whereas Co, Ni, Hg, Zn, and Pb exceeded SQGs at PM. Biomarker induction during summer appeared to be caused by natural variables (water salinity and temperature, and molting cycle), whereas oxidative stress and tissue damage during winter appeared to be more clearly linked to metal contamination. Anterior gills presented the clearest signs of seasonal variability, being more responsive to sediment contamination. The results suggest that metals originated from the upper Ribeira de Iguape River are transported toward the estuarine system, causing effects on C. danae individuals. Additionally, seasonality is a strong factor concerning CIP-PA toxicity, since the rainfall regime significantly modifies the freshwater flow and, consequently, estuarine water salinity, suspended particle and metal inputs, as well as the location of depositional areas. Thus, efforts to mitigate CIP-PA contamination should be based on the control of upstream pollution sources.

Hepatic Metal and Metallothionein Levels in a Potential Sentinel Teleost, Dulesauriga, from a Southeastern Brazilian Estuary.

The identification of estuarine sentinel species is of paramount importance. The potential of the species Dules auriga sampled from Guanabara Bay, Brazil regarding metal contamination was assessed. Hepatic metallothionein (MT) and Fe, Zn, Cu, Mn, Cd and Ag concentrations were determined in cytosolic fractions (S50 and HT S50) by polarography and ICP-OES, respectively. HT S50 Fe, Mn and Zn were lower than in the S50 fraction, indicating MT-detoxification. MT was correlated to HT S50 Zn, indicating Zn homeostasis. Zn was negatively correlated to weight (TW) and length (TL), suggesting environmental Zn influence. A moderate negative correlation between HT S50 Cu and the condition factor (CF) was observed, indicating that increasing environmental Cu concentrations may decrease D. auriga CF. Several inter-elemental correlations were observed. No MT, TW and TL correlations were found. Thus, MT variability is probably linked to environmental metal concentrations. D. auriga fulfills most sentinel requisites, and MT may be an adequate metal contamination biomarker in this species.

Inihibition of mullet (M. liza) brain acetylcholinesterase activity by in vitro polycyclic aromatic hydrocarbon exposure.

Polycyclic aromatic hydrocarbons (PAH) have been reported as acetylcholinesterase (AChE) inibitors, although in vitro studies on PAH effects on AChE activity are scarce and have only been performed using electric eel brain extracts. Thus, this study investigated PAH effects on brain AChE activity in a tropical fish species in Southeastern Brazil, mullet (Mugil liza). Mullet specimens were obtained from Guanabara Bay (N = 20), Rio de Janeiro, Brazil. Brain AChE was extracted and exposed to an environmentally relevant concentration of Pyrene, Chrysene, Phenanthrene, and Naphthalene, and PAH metabolites, 2-Naphthol and 1-OH-Pyrene. AChE activity inhibition was observed, although no difference was observed between high- and low- molecular weight PAH. 2-Naphthol was a less potent AChE inhibitor than Naphthalene, albeit non-significantly. Further studies are required, since only one PAH concentration was used herein. Mullet brain extracts seem to be adequate to assess possible neurotoxic PAH effects on fish AChE.

Metal loads and biomarker suite responses in a tropical carnivorous fish indicative of anthropogenic impacts in a Southeastern Brazilian lagoon.

Tropical coastal lagoons are highly productive environments exhibiting high biodiversity. However, the use of these ecosystems by local communities is of concern, since this generally leads to environmental degradation. The Imboassica coastal lagoon, located in Macaé city, in Northern Rio de Janeiro, is an important ecosystem in the state, however, already displaying signs of anthropogenic impacts. Carnivorous fish Hoplias malabaricus specimens were sampled from this impacted site, as well as from a reference area. Fish from Imboassica Lagoon presented lower condition factor, lower cholinesterase activity, and higher percentage of erythrocyte micronuclei when compared to fish from the reference site. Metals in fish from Imboassica Lagoon were always higher than Encantada Lagoon, with some seasonal differences, where some metals were higher in the rainy season compared to the dry season in muscle tissue, with the exception of Cu, Fe, Sr, and Zn; and in the liver, except for Ba, Cd, Cr, Ni, and Sr. Cr and Mn in the edible muscle portion of the fish were higher than the limits established by Brazilian and International legislations as permissible for human consumption, thus leading to concerns regarding public health risks for the local population that use fish as their main protein source.

Glutathione peroxidase and glutathione S-transferase in blood and liver from a hypoxia-tolerant fish under oxygen deprivation.

Liver enzyme activities can be employed as biomarkers, but liver can only be obtained with death of the specimen. On the other hand, blood withdrawal is a non-lethal procedure. Accordingly, the hypothesis of this study is to verify if glutathione peroxidase (GPX) and glutathione S-transferase (GST) activities in blood parallel those in the liver of the hypoxia-tolerant fish, Piaractus mesopotamicus (pacu), submitted to hypoxia conditions. GPX was assayed with H2O2 in cytosols from both liver and erythrocytes and exhibited no significant variation, either in erythrocytes or in liver, when comparing pacus under normoxia with those under hypoxia (42 h). GST activity with chloro-dinitrobenzene (CDNB), an artificial substrate suitable for almost all GST isoenzymes, was compared to activity with 4-hydroxy-nonenal (4-HNE), a physiological endogenous substrate. GST activity with CDNB did not change in liver or in erythrocyte cytosols in pacus under hypoxia compared to those under normoxia. On the other hand, a significant decrease in erythrocyte activity with 4-HNE was observed after 42 h of hypoxia in both erythrocytes and liver, which may be a response to increased lipid oxidation in erythrocytes. Erythrocyte GST activity was 3-fold higher with 4-HNE than with CDNB, indicating that 4-HNE is a more appropriate substrate to determine GST activity in pacu erythrocytes.

Correction to: Cryptosporidium Spp. Contamination in Perna perna Mussels Destined for Human Consumption in Southeastern Rio de Janeiro, Brazil.

The original version of this article unfortunately contained a mistake.

Cryptosporidium spp. Contamination in Perna perna Mussels Destined for Human Consumption in Southeastern Rio de Janeiro, Brazil.

Cryptosporidium spp. has been recognized as an important pathogen. As bivalve mollusks are noted as potential sources of several pathogens due to their consumption as foodstuffs, the aim of this study was to investigate the occurrence of Cryptosporidium spp. oocysts in Perna perna mussels and in seawater samples from a mussel farm in Southeastern Brazil, where mussels are grown directly in the sea, attached to ropes. Oocysts were observed by microscopy and confirmed by an enzyme-linked immunosorbent assay. Oocysts were present in mussel gills and GI tracts, as well as in the seawater. Of the 100 females, 10% and 11% showed contaminated GI tracts and gills, respectively, while this rate was lower in males, at 5% and 8.9%. Oocysts were present in higher amounts in the GI tract compared to gills and water. Contamination of the study area is apparent, leading to public health risks. More in-depth studies are needed, including molecular investigations, to identify Cryptosporidium species in mussels, as well as the implementation of monitoring actions in animals destined for human consumption.

Evaluation of a Brain Acetylcholinesterase Extraction Method and Kinetic Constants after Methyl-Paraoxon Inhibition in Three Brazilian Fish Species.

Acetylcholinesterase (AChE) is an important enzyme in the control of the neuronal action potential and sensitive to organophosphate inhibition. Brain fish AChE is less sensitive to organophosphate inhibition than AChE from terrestrial animals, although this sensitivity is variable among species and has not yet been fully evaluated in fish species. In this setting, inhibition kinetic constants for progressive irreversible inhibition of brain acetylcholinesterase due to methyl-paraoxon exposure were determined in three fish species (Mugil liza, Genidens genidens and Lagocephalus laevigatus) and hen (Gallus domesticus). Enzyme extraction using a detergent was shown to be adequate, and samples presented activity inhibition in high substrate concentrations and suppression of inhibition by methyl-paraoxon in the presence of the substrate, similar to kinetic patterns from purified enzyme preparations. Catfish (G. genidens) AChE presented the highest sensitivity among the evaluated fish species (IC50 = 1031.20 nM ± 63.17) in comparison to M. liza and L. laevigatus (IC50: 2878.83 ± 421.94 and 2842.5 ± 144.63 nM respectively). The lower dissociation constant (Kd = 20.3 ± 2.95 µM) of catfish AChE showed greater enzyme affinity for methyl-paraoxon, explaining this species higher sensitivity to organophosphates. Hen AChE presented higher ki (900.57 ± 65.3 mM-1min-1) and, consequently, greater sensitivity to methyl-paraoxon, explained by a lower Kd (0.6 ± 0.13 µM). Furthermore, hen AChE did not differentiate between the propionylthiocholine and acetylthiocholine substrates, indicating easier access of methyl-paraoxon to the hen enzyme activity site. The results obtained herein indicate a suitable extraction of AChE and, despite different inhibition kinetic constants, demonstrate that fish AChE is less sensitive to methyl-paraoxon, probably due to reduced access to the catalytic center which provides greater enzyme substrate selectivity.

Acute selenium selenite exposure effects on oxidative stress biomarkers and essential metals and trace-elements in the model organism zebrafish (Danio rerio).

Selenium (Se) is an essential trace-element that becomes toxic when present at high concentrations. Little is known regarding Se effects on parameters such as oxidative stress biomarkers. The aim of the present study was to investigate the effects of acute selenium exposure on oxidative stress biomarkers in a model organism, zebrafish (Danio rerio). Fish were exposed to selenium selenite at 1mgL(-1). Reduced glutathione (GSH), and metallothionein (MT) concentrations were determined in liver, kidney and brain, with MT also being determined in bile. Essential metals and trace-elements were also determined by inductively coupled mass spectrometry (ICP-MS) in order to verify possible metal homeostasis alterations. GSH concentrations in liver, kidney and brain increased significantly (1.05±0.03µmolg(-1) ww, 1.42±0.03µmolg(-1) ww and 1.64±0.03µmolg(-1) ww, respectively) in the Se-exposed group when compared to the controls (0.88±0.05µmolg(-1) ww, 0.80±0.04µmolg(-1) ww and 0.89±0.03µmolg(-1) ww for liver, kidney and brain, respectively). MT levels in Se-exposed liver (0.52±0.03µmolg(-1) ww) decreased significantly in comparison to the control group (0.64±0.02µmolg(-1) ww), while levels in bile increased, albeit non-significantly. This is in accordance with previous studies that indicate efficient biliary MT action, leading to a rapid metabolism and elimination of contaminants from the body. Levels in the brain increased significantly after Se-exposure (0.57±0.01µmolg(-1) ww) when compared to the control group (0.35±0.03µmolg(-1) ww) since this organ does not present a detoxification route as quick as the liver-gallbladder route. Several metal and trace-elements were altered with Se-exposure, indicating that excess of selenium results in metal dyshomeostasis. This is the first report on metal dyshomeostasis due to Se-exposure, which may be the first step in the mechanism of action of selenium toxicity, as is postulated to occur in certain major human pathophysiologies.

Disruption of zinc and copper interactions with Aß(1-40) by a non-toxic, isoniazid-derived, hydrazone: a novel biometal homeostasis restoring agent in Alzheimer's disease therapy?

Disruptions of biometal-Aß(1-40) interactions by an isoniazid-derived hydrazone, INHHQ, were demonstrated via in vitro NMR titrations. The compound has adequate theoretical BBB absorption properties, assessed by in silico studies. In vivo acute toxicity assays indicate that INHHQ is innocuous up to 300 mg kg(-1), showing potential as an anti-Alzheimer's drug.

Polybrominated diphenyl ethers (PBDES) and polychlorinated biphenyls (PCBS) in mussels and two fish species from the estuary of the Guanabara Bay, Southeastern Brazil.

The present investigation aimed to analyze PBDE and PCB contamination in mussels (Perna perna) and two commercially important fish species, croaker (Micropogonias furnieri) and mullet (Mugil liza), in the Guanabara Bay, the most important Brazilian estuary, by gas chromatography coupled to mass spectrometry, in order to further knowledge regarding these compounds in the southern hemisphere. This is also the first report of PBDE in this mussel species in the Guanabara Bay. Fish were captured in September (dry season, winter) and March (wet season, summer) 2007 and September 2008. Mussels were collected in August (dry season, winter) 2006, in February (wet season, summer) 2007, and in August 2007 (winter). The results show that all samples showed higher PCB contamination when compared to other ecosystems around the world. On the other hand, PBDEs presented lower concentrations in 41 % of the samples. Croakers presented the highest PCB and PBDE levels, with mullet showing intermediary values and mussels, the lowest.

Enzymatic GST levels and overall health of mullets from contaminated Brazilian Lagoons.

Glutathione S-transferase (GST) assays in non-mammalian organisms are usually conducted inappropriately, since no previous standardization of the optimal concentrations of proteins and substrates and adequate pH is conducted. Standardization is a key task to adjust enzyme assays at their kinetically correct maximal initial velocities, if one wants these velocities to indicate the amount of enzyme in a sample. In this paper GST assays were standardized in liver cytosol to compare seasonal GST levels in liver of mullet from two contaminated lagoons in the Rio de Janeiro to those from a reference bay. GST potential as a biomarker of sublethal intoxication in this species was also evaluated. Mullet liver GST levels assayed with substrates that corresponded to three different GST isoenzymes varied throughout the year. The differences indicated that mullets are suffering from sublethal intoxication from contaminants in these lagoons. Seasonal variations of activity were relevant, since these could indicate differences in xenobiotic input into the areas. An analysis of overall mullet health condition using a morphological index (the Fulton Condition Factor) and macroscopic abnormalities corroborated the differences in GST levels, with fish from one of the sites in worse overall health condition showing lower and significantly different FCF when compared to the reference site. Therefore, GST standardized activity levels are useful biomarkers of environmental contamination for mullet.

Metal and physico-chemical variations at a hydroelectric reservoir analyzed by Multivariate Analyses and Artificial Neural Networks: environmental management and policy/decision-making tools.

This paper compared and evaluated seasonal variations in physico-chemical parameters and metals at a hydroelectric power station reservoir by applying Multivariate Analyses and Artificial Neural Networks (ANN) statistical techniques. A Factor Analysis was used to reduce the number of variables: the first factor was composed of elements Ca, K, Mg and Na, and the second by Chemical Oxygen Demand. The ANN showed 100% correct classifications in training and validation samples. Physico-chemical analyses showed that water pH values were not statistically different between the dry and rainy seasons, while temperature, conductivity, alkalinity, ammonia and DO were higher in the dry period. TSS, hardness and COD, on the other hand, were higher during the rainy season. The statistical analyses showed that Ca, K, Mg and Na are directly connected to the Chemical Oxygen Demand, which indicates a possibility of their input into the reservoir system by domestic sewage and agricultural run-offs. These statistical applications, thus, are also relevant in cases of environmental management and policy decision-making processes, to identify which factors should be further studied and/or modified to recover degraded or contaminated water bodies.

Alterations in morphometric and organosomatic indices and histopathological analyses indicative of environmental contamination in Mullet, Mugil liza, from Southeastern Brazil.

Mullet (Mugil liza) were sampled in five different areas along the Guanabara Bay, southeastern Brazil, classified as non-contaminated, moderately contaminated and contaminated. Morphometric (Fulton condition factor, relative condition factor and weight to length scaling coefficient) and organosomatic (hepatosomatic index) indices of environmental stress were analysed. Fish from the differentially contaminated areas show statistically different Fulton and relative condition factors and hepatosomatic indices, but not the weight to length scaling coefficient. The Kn and the FCF followed the same trend, with fish from São Gonçalo (1.07 ± 0.04 and 0.89 ± 0.03), Itaipu (0.84 ± 0.01 and 0.86 ± 0.01) and the Rodrigo de Freitas Lagoon (1.03 ± 0.01 and 0.87 ± 0.20) showing higher FCFs than fish from Magé (0.96 ± 0.01 and 0.81 ± 0.01). Fish from Itaipu showed significantly higher HSI values than the other sampling sites (1.68 ± 0.07), with fish from Olaria and Ipiranga showing the lowest (1.56 ± 0.12 and 1.60 ± 0.07, respectively).

First-time report of metalloproteinases in fish bile and their potential as bioindicators regarding environmental contamination.

Gallbladder bile from 2 fish species, mullet (Mugil liza) and tilapias (Tilapia rendalli), contain substantial matrix metalloproteinases (MMPs). Extensive purification studies were conducted in order to obtain workable samples for SDS-PAGE and zymography analysis. Proteinase activities were assayed by gelatin substrate zymography. Several protein bands were observed, corresponding to molecular weights of 200, 136, 43, 36, 34, 29, 23 and 14 kDa in mullet bile and 179, 97, 79, 61, 54, 45, 36, 33 and 21 kDa in tilapia bile. Specific inhibitor studies were conducted, in which MMPS were inhibited by EDTA and 1,10 phenanthroline, but not by serine and cysteine protease inhibitors, such as phenylmethylsulfonyl fluoride (PMSF) and transepoxysuccinyl-l-leucylamido-l-guanidino butane (E-64), confirming the proteinase identities as MMPs. Differences in proteinase expression were observed in fish from a contaminated and reference site. Some studies regarding MMPs in different fish tissues exist, however this is the first study conducted in fish bile, and their involvement in detoxification processes and organism protection against the effects of aquatic contaminants may be a possibility.

Mercury and methyl mercury ratios in caimans (Caiman crocodilus yacare) from the Pantanal area, Brazil.

The Pantanal region is the largest floodplain area in the world and of great biological importance due to its unique flora and fauna. This area is continuously undergoing increasing anthropogenic threats, and has also experienced mercury contamination associated with gold mining and other anthropogenic activities. Pantanal caimans are top-level predators, and, as such, show great potential to accumulate mercury (Hg) by biomagnification. In this study 79 specimens from four locations in the Pantanal were analyzed for total Hg and methyl mercury (MeHg) by cold vapor atomic absorption spectrometry. Total Hg contents ranged from 0.02 to 0.36 µg g(-1) (ww), and most specimens presented MeHg ratios above 70%. One of the sites, impacted by anthropogenic activities, presented significantly higher total Hg in comparison to three less impacted sites, supporting the hypothesis that caimans can, in fact, be considered effective bioindicators of ecosystem health.