Microplastics and 17α Ethinylestradiol: How Do Different Aquatic Invertebrates Respond to This Combination of Contaminants?

The synthetic hormone 17α ethinyl estradiol (EE2) is a molecule widely used in female contraceptives and recognized as a contaminant of attention (Watch List) in the European Union due to its high consumption, endocrine effects and occurrence in aquatic environments. Its main source of introduction is domestic sewage where it can be associated with other contaminants such as microplastics (MPs). Due to their characteristics, they can combine with each other and exacerbate their isolated effects on biota. This study evaluated the combined effects of microplastics (MPs) and 17α ethinylestradiol (EE2) on two tropical estuarine invertebrate species: Crassostrea gasar and Ucides cordatus. Polyethylene particles were spiked with EE2 and organisms were exposed to three treatments, categorized into three groups: control group (C), virgin microplastics (MPs), and spiked microplastics with EE2 (MPEs). All treatments were evaluated after 3 and 7 days of exposure. Oysters exhibited changes in phase 2 enzymes and the antioxidant system, oxidative stress in the gills, and reduced lysosomal membrane stability after exposure to MPs and MPEs. Crabs exposed to MPs and MPEs after seven days showed changes in phase 1 enzymes in the gills and changes in phases 1 and 2 enzymes in the hepatopancreas, such as disturbed cellular health. The combined effects of microplastics and EE2 increased the toxicity experienced by organisms, which may trigger effects at higher levels of biological organization, leading to ecological disturbances in tropical coastal ecosystems.

Settleable atmospheric particulate matter harms a marine invertebrate: Integrating chemical and biological damage in a bivalve model.

Some atmospheric pollutants may affect aquatic ecosystems after settling, generating contamination, bioaccumulation, and threats to aquatic species. Metallurgical processes result in the emission of settleable atmospheric particulate matter (SePM), including metals and metalloids, along with rare earth elements (REE) that are considered emerging contaminants. We report the 30-day exposure of brown mussels (Perna perna) to SePM collected in a metallurgical area of southeast Brazil close to estuarine ecosystems, followed by a 30-day clearance period, to evaluate the toxic potential of SePM to this model mollusk. The bioaccumulation of 28 elements identified in SePM and the sublethal effects were evaluated. REEs were found in SePM (Ce, Y, and La). Significant bioaccumulation of eight metals (Fe, Ni, Cu, Zn, Rb, Sr, Cd, and Ba) was found in the bivalves and correlates with the cytotoxicity and genotoxicity, showing a dose-dependent mode and suggesting a pre-pathological condition that could lead to ecological disturbances over time. Conversely, the unchanged lipid-peroxidation level after SePM exposure could indicate the effectiveness of the antioxidant system in protecting gills and digestive glands. The clearance period was not enough to successfully reverse the negative effects observed. So far, the current results enhance the comprehension of the negative role of SePM on metal bioaccumulation and metal-induced toxicity to aquatic biota. Thus, this report adds innovative findings on the role of SePM in aquatic pollution in coastal areas affected by atmospheric pollution, which should be relevant for future public policies to verify and control the environmental pollution.

Lead toxicity on a sentinel species subpopulation inhabiting mangroves with different status conservation.

Lead is a priority pollutant introduced in the aquatic environment by different sources commonly located in estuarine regions, such as ports, marinas and industries. Environmental agencies around the world set the maximum allowable concentration of lead in effluents, surface water and sediment, but few studies reported its accumulation and chronic toxicity in mangrove benthic invertebrates using concentrations believed to be safe. In the case of Brazilian mangrove environments, Ucides cordatus is a crab species of choice to be used in bioaccumulation studies. We have assessed biomarkers' responses (DNA strand breaks, micronucleated cells, metallothioneins, enzymatic activity of aminolevulinic acid dehydratase and neutral red retention time) and the total bioaccumulation in six tissues of U. cordatus crabs resident to mangrove areas under different conservation status during a 28-day period bioassay. We also investigated Pb subcellular partition and biomarkers' responses using a supposedly safe concentration (10 µg L-1). During the Pb exposure, the highest concentration of Pb was observed in crab gills. Crabs also showed a high ability to allocate Pb in detoxified forms. Multivariate analysis pointed out that bioaccumulation (total, active and detoxified) is linked to biomarkers. Even in supposedly safe dosage, U. cordatus triggered its defense mechanisms expressing more metallothioneins and presented relevant cyto-genotoxic damage. Our data suggest the development of biological tolerance to Pb in crabs from polluted areas. Our results provided a new insight about lead toxicity even at concentrations considered environmentally safe, which could support new strategies to manage estuarine areas considering their respective conservation status.

Population structure of Callichirus major (Say 1818) (Crustacea: Callianassidae) and conservation considerations at Southeast coast of São Paulo, Brazil.

Globally, there is limited knowledge about management and conservation of Callianassidae family. Their catches seem to be recently increasing in several parts of the world and evidences raise concern on the need of specialized literature focused on populations monitoring. At Southeast of Brazil, the species Callichirus major (Say 1818), known as the ghost shrimp, is frequently captured for use as bait in sport fishing and, according to previous studies, the overexploitation could threaten the population. The aim of this study was evaluated over a year this harmful human pressure on the species C. major in two beaches contiguous but susceptible to different anthropogenic impacts due to the existence of legislation on one beach that prohibits the capture of this crustacean. The population of C. major presented where the prohibition regulation does not exist a density almost three times lower when compared to the beach that prohibits the capture, with individuals with smaller carapace oval area and a shorter reproductive period. This paper alert to some evidence of C. major structure population disturbance resulting from its exploitation at Southeast of Brazil. Recommendations were also addressed to conservation management considering the species' ecological importance.

Mangrove metal pollution induces biological tolerance to Cd on a crab sentinel species subpopulation.

Metals are persistent pollutants, able to accumulate in the biota and magnify in trophic web. In the specific case of cadmium contamination, it has been the subject of considerable interest in recent years because of its biological effects and it is one of major pollutant in estuarine areas. Ucides cordatus is considered a mangrove local sentinel crab species in Brazil and there are previous studies reporting crab subpopulations living from pristine to heavily metal impacted areas in São Paulo coast (Southeastern Brazil). Taking into account the background knowledge about these subpopulations, we proposed the hypothesis that crabs from a highly polluted mangrove (Cubatão - CUB) have developed biological tolerance to cadmium compared to animals from an Environmental Protected Area (Jureia - JUR). Aiming to verify this hypothesis, we have investigated total bioaccumulation and subcellular partition of Cd, besides biomarkers' responses during a long-term exposure bioassay (28 days, with weekly sampling) using a supposedly safe Cd concentration (0.0022 mg L-1). Specimens from the pristine area (JUR) accumulated higher total Cd, as such as in its biologically active form in gills. Animals living in the polluted site (CUB) presented higher amounts of Cd in the mainly detoxifying tissue (hepatopancreas), which could be considered a pathway leading to tolerance for this metal. Multivariate analysis indicated that bioaccumulation (active, detoxified and total Cd) is linked to geno-cytotoxic damages. CUB subpopulation was considered more tolerant since it presented proportionally less damage and more capacity to allocate Cd in the main detoxifying forms and tissues.

Multi-level biological responses in Ucides cordatus (Linnaeus, 1763) (Brachyura, Ucididae) as indicators of conservation status in mangrove areas from the western atlantic.

There is a global lack of knowledge on tropical ecotoxicology, particularly in terms of mangrove areas. These areas often serve as nurseries or homes for several animal species, including Ucides cordatus (the uçá crab). This species is widely distributed, is part of the diet of human coastal communities, and is considered to be a sentinel species due to its sensitivity to toxic xenobiotics in natural environments. Sublethal damages to benthic populations reveal pre-pathological conditions, but discussions of the implications are scarce in the literature. In Brazil, the state of São Paulo offers an interesting scenario for ecotoxicology and population studies: it is easy to distinguish between mangroves that are well preserved and those which are significantly impacted by human activity. The objectives of this study were to provide the normal baseline values for the frequency of Micronucleated cells (MN‰) and for neutral red retention time (NRRT) in U. cordatus at pristine locations, as well to indicate the conservation status of different mangrove areas using a multi-level biological response approach in which these biomarkers and population indicators (condition factor and crab density) are applied in relation to environmental quality indicators (determined via information in the literature and solid waste volume). A mangrove area with no effects of impact (areas of reference or pristine areas) presented a mean value of MN‰<3 and NRRT>120min, values which were assumed as baseline values representing genetic and physiological normality. A significant correlation was found between NRRT and MN, with both showing similar and effective results for distinguishing between different mangrove areas according to conservation status. Furthermore, crab density was lower in more impacted mangrove areas, a finding which also reflects the effects of sublethal damage; this finding was not determined by condition factor measurements. Multi-level biological responses were able to reflect the conservation status of the mangrove areas studied using information on guideline values of MN‰, NRRT, and density of the uçá crab in order to categorize three levels of human impacts in mangrove areas: PNI (probable null impact); PLI (probable low impact); and PHI (probable high impact). Results confirm the success of U. cordatus species' multi-level biological responses in diagnosing threats to mangrove areas. Therefore, this species represents an effective tool in studies on mangrove conservation statuses in the Western Atlantic.

Accumulation of six metals in the mangrove crab Ucides cordatus (Crustacea: Ucididae) and its food source, the red mangrove Rhizophora mangle (Angiosperma: Rhizophoraceae).

The crab Ucides cordatus and the red mangrove Rhizophora mangle are endemic mangrove species and potential bio-accumulators of metals. This study quantified the accumulation of six metals (Cd, Cr, Cu, Hg, Mn and Pb) in different organs (claw muscle, hepatopancreas and gills) of U. cordatus, as well as in different maturation stages of the leaves (buds, green mature, and pre-abscission senescent) of R. mangle. Samples were collected from mangrove areas in Cubatão, state of São Paulo, a heavily polluted region in Brazil. Data for metal contents in leaves were evaluated by one-way ANOVA; while for crabs a factorial ANOVA was used to investigate the effect of different tissues, animal size and the interactions between them. Means were compared by Tukey test at five percent, and the association between the metal concentrations in each crab organ, depending on the size, was evaluated by Pearson's linear correlation coefficient (r). Concentrations of Pb and Hg were undetectable for the different leaf stages and crab tissues, while Cd concentrations were undetectable in the leaf stages. In general, the highest accumulation of metals in R. mangle leaves occurred in pre-abscission senescent and green mature leaves, except for Cu, which was found in the highest concentrations in buds and green mature leaves. For the crab, Cd, Cu, Cr and Mn were present in concentrations above the detection limit, with the highest accumulation in the hepatopancreas, followed by the gills. Cu was accumulated mostly in the gills. Patterns of bioaccumulation between the crab and the mangrove tree differed for each metal, probably due to the specific requirements of each organism for essential metals. However, there was a close and direct relationship between metal accumulation in the mangrove trees and in the crabs feeding on them. Tissues of R. mangle leaves and U. cordatus proved effective for monitoring metals, acting as important bioindicators of mangrove areas contaminated by various metals.