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.

Metallic nanoparticle contamination from environmental atmospheric particulate matter in the last slab of the trophic chain: Nanocrystallography, subcellular localization and toxicity effects.

Atmospheric particulate material (PM) from mining and steel industries comprises several metallic contaminants. PM10 samples collected in a Brazilian region with a recognized influence of the steel and iron pelletizing industries were used to investigate metallic nanoparticle incorporation into human fibroblast cells (MRC-5). MRC-5 cells were exposed to 0 (control, ultrapure water), 2.5, 5, 10, 20 and 40 µg PM10 mL-1, for 24 h. Cytotoxic and genotoxic dose-response effects were observed on lysosome and DNA structure, and concentrations high as 20 and 40 µg PM10 mL-1 induced elevated cell death. Ultrastructure analyses showed aluminosilicate, iron, and the emerging metallic contaminants titanium, bismuth, and cerium nanoparticles were incorporated into lung cells, in which the nanocrystallography analysis indicated the bismuth as Bi2O3. All internalized metallic nanoparticles were free and unbound in the cytoplasm and nucleus thereby indicating bioavailability and potential interaction to biological processes and cellular structures. Pearson's correlation analysis showed Fe, Ni, Al, Cr, Pb and Hg as the main cytotoxic elements which are associated with the stainless steel production. The presence of internalized nanoparticles in human lung cells exposed to environmental atmospheric matter highlights the need for a greater effort by regulatory agencies to understand their potential damage and hence the need for future regulation, especially of emerging metallic contaminants.

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.

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.

Interrogating pollution sources in a mangrove food web using multiple stable isotopes.

Anthropogenic activities including metal contamination create well-known problems in coastal mangrove ecosystems but understanding and linking specific pollution sources to distinct trophic levels within these environments is challenging. This study evaluated anthropogenic impacts on two contrasting mangrove food webs, by using stable isotopes (δ13C, δ15N, 87Sr/86Sr, 206Pb/207Pb and 208Pb/207Pb) measured in sediments, mangrove trees (Rhizophora mangle, Laguncularia racemosa, Avicennia schaueriana), plankton, shrimps (Macrobranchium sp.), crabs (Aratus sp.), oysters (Crassostrea rhizophorae) and fish (Centropomus parallelus) from both areas. Strontium and Pb isotopes were also analysed in water and atmospheric particulate matter (PM). δ15N indicated that crab, shrimp and oyster are at intermediate levels within the local food web and fish, in this case C. parallelus, was confirmed at the highest trophic level. δ15N also indicates different anthropogenic pressures between both estuaries; Vitória Bay, close to intensive human activities, showed higher δ15N across the food web, apparently influenced by sewage. The ratio87Sr/86Sr showed the primary influence of marine water throughout the entire food web. Pb isotope ratios suggest that PM is primarily influenced by metallurgical activities, with some secondary influence on mangrove plants and crabs sampled in the area adjacent to the smelting works. To our knowledge, this is the first demonstration of the effect of anthropogenic pollution (probable sewage pollution) on the isotopic fingerprint of estuarine-mangrove systems located close to a city compared to less impacted estuarine mangroves. The influence of industrial metallurgical activity detected using Pb isotopic analysis of PM and mangrove plants close to such an impacted area is also notable and illustrates the value of isotopic analysis in tracing the impact and species affected by atmospheric pollution.

Accumulation of organotins in seafood leads to reproductive tract abnormalities in female rats.

Organotins (OTs) are environmental contaminants used as biocides in antifouling paints that have been shown to be endocrine disrupters. However, studies evaluating the effects of OTs accumulated in seafood (LNI) on reproductive health are particularly sparse. This study demonstrates that LNI leads to impairment in the reproductive tract of female rats, as the estrous cycle development, as well as for ovary and uterus morphology. Rats were treated with LNI, and their reproductive morphophysiology was assessed. Morphophysiological abnormalities, such as irregular estrous cycles, abnormal ovarian follicular development and ovarian collagen deposition, were observed in LNI rats. An increase in luminal epithelia and ERα expression was observed in the LNI uteri. Together, these data provide in vivo evidence that LNI are toxic for reproductive morphophysiology, which may be associated with risks to reproductive function.

Changes in bioaccumulation and translocation patterns between root and leafs of Avicennia schaueriana as adaptive response to different levels of metals in mangrove system.

Espírito Santo estuaries (Brazil) are impacted by industrial activities, resulting in contamination of water and sediments. This raise questions on biological uptake, storage and consequences of metal contamination to mangrove plants. The goal of this work was evaluating accumulation and translocation of metals from sediment to roots and leaves of Avicennia schaueriana, growing in areas with different degrees of contamination, correlating bioaccumulation with changes in its root anatomy. Highest bioconcentration factors (BCFs) were observed in plants growing in less polluted areas. Conversely, highest translocation factors were found in plants from highest polluted area, evidencing an adaptive response of A. schaueriana to less favourable conditions. Namely, the absorption of metals by roots is diminished when facing highest levels of metals in the environment; alternatively, plants seem to enhance the translocation to diminish the concentration of toxic metals in roots. Root also responded to highly polluted scenarios with modifications of its anatomy.

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.