Non-destructive biomarkers can reveal effects of the association of microplastics and pharmaceuticals or personal care products.

Methods to assess the effects of contaminants on marine organisms typically involve euthanasia to obtain samples, but less invasive techniques may be more appropriate for working with threatened species. In this study, were assessed the biological responses of crabs exposed to microplastics and contaminants of emerging concern. Biochemical and cellular effects (lipid peroxidation, DNA damage, cholinesterase activity, and lysosomal membrane stability) in hemolymph were analyzed in a kinetic study, at 3 and 7 days, in U. cordatus exposed to microplastics spiked with Triclosan (TCS) or 17α-Ethynylestradiol (EE2). The results showed that the contaminants were produced toxic effects in the crabs exposed either to the microplastics alone (oxidative stress, genotoxicity, and neurotoxicity), or to microplastics with TCS or EE2 adsorbed (neurotoxic and cytotoxic). The present study showed the responsiveness of non-lethal analyzes to understanding the biological effects of combined exposure to microplastics and chemical pollution.

Evaluation of an alternative technique for preserving crustaceans in dry conditions with joint mobility: a proposal for didactic purposes

Although crustaceans are traditionally preserved in liquids (formaldehyde and/or ethyl alcohol), those substances tend to alter their morphological aspects. Glycerin, used in human anatomy, is considered a good substitute for formaldehyde, as it preserves animals in states similar to in vivo conditions. There are no records in the literature, however, concerning the use of glycerin for conserving invertebrates. The objective of this work was to elaborate and evaluate alternative techniques for conserving the crustacean Ucides cordatus (Linnaeus, 1763). Six fixatives (1, 3, 4 and 5% formaldehyde, 70% alcohol, and dietrich solution) and two controls (positive and negative) were tested, as well as the effects of freezing before fixation on the integrity of U. cordatus specimens. Our results were evaluated with respect to nine variables. The treatments that demonstrated the best aesthetic results were 4% formaldehyde and 70% ethyl alcohol. The freezing of the animals resulted in brittle organs in all treatments tested. The technique discussed here is extremely promising for the conservation of animals for educational purposes, as it produces preserved specimens that are aesthetically similar to their in vivo conditions.(AU)

Evaluation of an alternative technique for preserving crustaceans in dry conditions with joint mobility: a proposal for didactic purposes

Although crustaceans are traditionally preserved in liquids (formaldehyde and/or ethyl alcohol), those substances tend to alter their morphological aspects. Glycerin, used in human anatomy, is considered a good substitute for formaldehyde, as it preserves animals in states similar to in vivo conditions. There are no records in the literature, however, concerning the use of glycerin for conserving invertebrates. The objective of this work was to elaborate and evaluate alternative techniques for conserving the crustacean Ucides cordatus (Linnaeus, 1763). Six fixatives (1, 3, 4 and 5% formaldehyde, 70% alcohol, and dietrich solution) and two controls (positive and negative) were tested, as well as the effects of freezing before fixation on the integrity of U. cordatus specimens. Our results were evaluated with respect to nine variables. The treatments that demonstrated the best aesthetic results were 4% formaldehyde and 70% ethyl alcohol. The freezing of the animals resulted in brittle organs in all treatments tested. The technique discussed here is extremely promising for the conservation of animals for educational purposes, as it produces preserved specimens that are aesthetically similar to their in vivo conditions.

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.

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.

Magnetite nanoparticles coated with oleic acid: accumulation in hepatopancreatic cells of the mangrove crab Ucides cordatus.

The field of nanotechnology had enormous developments, resulting in new methods for the controlled synthesis of a wide variety of nanoscale materials with unique properties. Efficient methods such as thermal decomposition for efficient size control have been developed in recent years for the synthesis of oleic acid (OA)-coated magnetite (Fe3O4) nanoparticles (MNP-OA). These nanostructures can be a source of pollution when emitted in the aquatic environment and could be accumulated by vulnerable marine species such as crustaceans. In this work, we synthesized and characterized MNP-OA of three different diameters (5, 8, and 12 nm) by thermal decomposition. These nanoparticles were remarkably stable after treatment with high affinity iron chelators (calcein, fluorescent desferrioxamine, and fluorescent apotransferrin); however, they displayed pro-oxidant activity after being challenged with ascorbate under two physiological buffers. Free or nanoparticle iron displayed low toxicity to four types of hepatopancreatic cells (E, R, F, and B) of the mangrove crab Ucides cordatus; however, they were promptly bioavailable, posing the risk of ecosystem disruption due to the release of excess nutrients.

Evaluation of iron loading in four types of hepatopancreatic cells of the mangrove crab Ucides cordatus using ferrocene derivatives and iron supplements.

The mangrove crab Ucides cordatus is a bioindicator of aquatic contamination. In this work, the iron availability and redox activity of saccharide-coated mineral iron supplements (for both human and veterinary use) and ferrocene derivatives in Saline Ucides Buffer (SUB) medium were assessed. The transport of these metallodrugs by four different hepatopancreatic cell types (embryonic (E), resorptive (R), fibrillar (F), and blister (B)) of U. cordatus were measured. Organic coated iron minerals (iron supplements) were stable against strong chelators (calcein and transferrin). Ascorbic acid efficiently mediated the release of iron only from ferrocene compounds, leading to redox-active species. Ferrous iron and iron supplements were efficient in loading iron to all hepatopancreatic cell types. In contrast, ferrocene derivatives were loaded only in F and B cell types. Acute exposition to the iron compounds resulted in cell viability of 70-95%, and to intracellular iron levels as high as 0.40 µmol L-1 depending upon the compound and the cell line. The easiness that iron from iron metallodrugs was loaded/transported into U. cordatus hepatopancreatic cells reinforces a cautionary approach to the widespread disposal and use of highly bioavailable iron species as far as the long-term environmental welfare is concerned.

Metal toxicity assessment by sentinel species of mangroves: In situ case study integrating chemical and biomarkers analyses.

Globally, there is a lack of knowledge about tropical ecotoxicology dealing with the potential impact of metal contamination in mangrove ecosystem. This habitat is considered a nursery for several animal species, among them the "uçá"-crab (Ucides cordatus), known as a key species due to its biological and economical importance. This study evaluated the association involving metal contamination (Cd, Cu, Pb, Cr, Mn and Hg) in water, sediment, red-mangrove vegetation (Rhizophora mangle) and tissues of uçá crab, together with its geno-cytotoxic responses, based on micronucleated hemocytes frequency and the retention time of neutral red in lysosomes. We assessed six mangrove areas with distinct pollution levels in São Paulo State, Brazil, where the water and sediment contamination by metals were associated with accumulation of these pollutants in biotic compartments (mangrove leaves and crab). In U. cordatus, metal accumulation was best explained by metal concentration found in leaves of R. mangle than in the water or sediment, indicating that feeding drives metal exposure in this organism. Mercury (Hg) concentration in sediment, copper (Cu) concentration in hepatopancreas of U. cordatus and lead (Pb) in water and green leaves of R. mangle showed a significant correlation with genotoxic impact in U. cordatus. However, copper concentration (in green/senescent leaves and hepatopancreas) and lead (in sediment), were the major metals affecting lysosomal membrane integrity. Therefore, representatives of all compartments were associated with cyto and genotoxicity in this species, thus requiring a holistic approach to issues related to sublethal damage. Probability estimates of cytogenetic impacts related to metal concentration in abiotic compartments (significantly correlated with known biomarkers: Hg in sediment; and Pb in water and sediment) are also presented. Our results highlight the need for environmental restoration of mangroves areas contaminated with metals, responsible for cytogenetic injuries and revealing a pre-pathological condition in this sentinel species, in addition to ecological disturbances.

Habits and customs of crab catchers in southern Bahia, Brazil.

BACKGROUND: Brazilian mangrove forests are widely distributed along the coast and exploited by groups of people with customs and habits as diverse as the biology of the mangrove ecosystems. This study identifies different methods of extracting crabs that inhabit the mangrove belts; some of these activities, such as catching individual crabs by hand, are aimed at maintaining natural stocks of this species in Mucuri (south Bahia), Brazil. METHODS: In the studied community, illegal hunting activities that violate Brazilian legislation limiting the use of tangle-netting in mangrove ecosystem were observed. RESULTS: According to our observations, fishermen, to catch individual crabs, use the tangle-netting technique seeking to increase income and are from families that have no tradition of extraction. CONCLUSIONS: This analysis leads us to conclude that catchers from economically marginalised social groups enter mangroves for purposes of survival rather than for purposes of subsistence, because the catching by tangle-netting is a predatory technique. Tangle-netting  technique increase caught but also increases their mortality rate. We emphasise that traditional catching methods are unique to Brazil and that manual capturing of crab should be preserved through public policies aimed at maintaining the crab population.

Importância do rotífero como alimento no desenvolvimento larval do caranguejo-uçá / Rotifer importance as food on the larval rearing of the mud crab

O objetivo deste trabalho foi verificar os efeitos da exclusão de rotífero e sua substituição por ração no desenvolvimento, crescimento e sobrevivência de larvas do caranguejouçá (Ucides cordatus). As larvas foram cultivadas individualmente e submetidas a seis tratamentos (T1 a T5 e TR), com 50 réplicas, de zoea (Z1 a Z5) até o estágio de megalopa (M). A alimentação foi composta por náuplios de artêmia e microalgas em todos os tratamentos. O rotífero foi introduzido também como alimento de T1 a T5, exceto em TR, em que a ração comercial microparticulada foi administrada como alimento. O rotífero foi então fornecido nos tratamentos T1-Z1, T2-Z2, T3-Z3, T4-Z4 e T5-Z5 e posteriormente excluído. A baixa sobrevivência com reduzido número de megalopas, observada em TR, aponta que a substituição de rotífero por ração não é uma alternativa viável. A exclusão do rotífero pode ser realizada após a mudança para Z2, mantendo o fornecimento de náuplios de artêmia e microalgas, sem impactar no crescimento, sobrevivência e resistência ao estresse, assegurando megalopas com qualidade para repovoamentos e reduzindo, consequentemente, o manejo e custos envolvidos na produção.

The aim of this study was to assess the effects of the exclusion of rotifer or its replacement by commercial feed in the survival, larval development and growth during the larval rearing of mud crab (Ucides cordatus). Larvae were individually reared and subjected to 6 treatments (T1 to T5 and TR) with 50 replicates, from zoea (Z1 to Z5) to the megalopa stage (M). Feeding was composed of Artemia nauplii and microalgae in all treatments. Rotifer was also introduced as food from T1 to T5, except for TR, where microparticulate commercial feed was provided as food. Rotifer was then supplied in treatments T1-Z1, T2 –Z2, T3-Z3, T4-Z4 and T5-Z5, and subsequently excluded. The low survival with reduced number of megalopae observed when rotifer was replaced with the commercial feed, shows that this is not a viable alternative. The exclusion of rotifer can be performed after switching from Z1 to Z2, maintaining the supply of Artemia nauplii and microalgae, without impact on growth, survival and stress resistance, ensuring megalopae with quality for restoking and thereby reducing handling and the costs involved in the production.

Importância do rotífero como alimento no desenvolvimento larval do caranguejo-uçá / Rotifer importance as food on the larval rearing of the mud crab

O objetivo deste trabalho foi verificar os efeitos da exclusão de rotífero e sua substituição por ração no desenvolvimento, crescimento e sobrevivência de larvas do caranguejouçá (Ucides cordatus). As larvas foram cultivadas individualmente e submetidas a seis tratamentos (T1 a T5 e TR), com 50 réplicas, de zoea (Z1 a Z5) até o estágio de megalopa (M). A alimentação foi composta por náuplios de artêmia e microalgas em todos os tratamentos. O rotífero foi introduzido também como alimento de T1 a T5, exceto em TR, em que a ração comercial microparticulada foi administrada como alimento. O rotífero foi então fornecido nos tratamentos T1-Z1, T2-Z2, T3-Z3, T4-Z4 e T5-Z5 e posteriormente excluído. A baixa sobrevivência com reduzido número de megalopas, observada em TR, aponta que a substituição de rotífero por ração não é uma alternativa viável. A exclusão do rotífero pode ser realizada após a mudança para Z2, mantendo o fornecimento de náuplios de artêmia e microalgas, sem impactar no crescimento, sobrevivência e resistência ao estresse, assegurando megalopas com qualidade para repovoamentos e reduzindo, consequentemente, o manejo e custos envolvidos na produção.(AU)

The aim of this study was to assess the effects of the exclusion of rotifer or its replacement by commercial feed in the survival, larval development and growth during the larval rearing of mud crab (Ucides cordatus). Larvae were individually reared and subjected to 6 treatments (T1 to T5 and TR) with 50 replicates, from zoea (Z1 to Z5) to the megalopa stage (M). Feeding was composed of Artemia nauplii and microalgae in all treatments. Rotifer was also introduced as food from T1 to T5, except for TR, where microparticulate commercial feed was provided as food. Rotifer was then supplied in treatments T1-Z1, T2 –Z2, T3-Z3, T4-Z4 and T5-Z5, and subsequently excluded. The low survival with reduced number of megalopae observed when rotifer was replaced with the commercial feed, shows that this is not a viable alternative. The exclusion of rotifer can be performed after switching from Z1 to Z2, maintaining the supply of Artemia nauplii and microalgae, without impact on growth, survival and stress resistance, ensuring megalopae with quality for restoking and thereby reducing handling and the costs involved in the production.(AU)

Sewage input reduces the consumption of Rhizophora mangle propagules by crabs in a subtropical mangrove system.

Mangrove forests are highly productive and play a major role in global carbon cycling. Their carbon accumulation can be influenced through the consumption of nutrient-poor leaves and propagules by herbivore crabs. Anthropogenic nutrient input from sewage contamination is widespread in these often naturally nutrient-limited ecosystems. We hypothesised that sewage-mediated nutrient input to mangrove stands of Paranaguá Bay (southern Brazil), would alter the nutrient sources available for crabs, e.g. through microphytobenthos increase, and that this would reflect in their feeding behaviour. We predicted that propagules of Rhizophora mangle in contaminated stands would experience lower grazing pressure from their two main local consumers (Ucides cordatus and Goniopsis cruentata). We compared herbivory rates on R. mangle propagules in sewage contaminated and uncontaminated mangrove stands. We found that herbivory rates were significantly lower in contaminated than uncontaminated forests, but this pattern could not be clearly attributed to increased nutrient availability.

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.

First assessment of trace metal concentration in mangrove crab eggs and other tissues, SE Brazil.

The mangrove crab Ucides cordatus is widespread in the Brazilian coast, which has an important role in nutrient cycling. This species reproduces in summer and females carry eggs about a month, when they maintain contact with water and sediments. It remains unclear if trace metals can be absorbed or adsorbed by the eggs during development. The present study aims to investigate, for the first time, trace metal concentrations in ovigerous female tissues and eggs of U. cordatus in two areas with different metal pollution levels in the Southeastern Brazil. Samples were collected in two different mangroves, Guanabara Bay (GB) highly polluted environment and Paraíba do Sul River (PSR). In both populations, we observed significant increase of V, Cr, and Mn concentrations along eggs maturation. The higher metals averages were found in PSR population. This trend was reported since the 1990s and lower concentrations in GB marine organisms were attributed to reducing conditions, high organic load, and the presence of sulfide ions. These conditions restrict the bioavailability of metals in the bay, with exception of Mn. No significant differences were observed in gills and muscles. In both populations of the present study, V, Zn, As, and Pb were higher in eggs of initial stage, whereas Mn, Ni, Cu, and Cd were higher in hepatopancreas. Beside this, V, Cr, Mn, As, and Pb showed an increase concerning egg development. Thus, V, As, and Pb in eggs come from two sources previous discussed: females and environment. Zinc came mainly from females due to essential function. Those new information should be considered as one of the mechanisms of trace metal transfer to the trophic chain, between benthonic and pelagic environment.

Characterization of cadmium plasma membrane transport in gills of a mangrove crab Ucides cordatus.

Membrane pathway for intracellular cadmium (Cd(2+)) accumulation is not fully elucidated in many organisms and has not been studied in crab gill cells. To characterize membrane Cd(2+) transport of anterior and posterior gill cells of Ucides cordatus, a hypo-hyper-regulating crab, a change in intracellular Cd(2+) concentration under various experimental conditions was examined by using FluoZin, a fluorescent probe. The membrane Cd(2+) transport was estimated by the augmentation of FluoZin fluorescence induced by extracellular application of CdCl2 and different inhibitors. Addition of extracellular calcium (Ca(2+)) to the cells affected little the fluorescence of FluoZin, confirming that Cd(2+) was the main ion increasing intracellular fluorescence. Ca(2+) channels blockers (nimodipine and verapamil) decreased Cd(2+) influx as well as vanadate, a Ca(2+)-ATPase blocker. Chelating intracellular Ca(2+) (BAPTA) decreased Cd(2+) influx in gill cells, while increasing intracellular Ca(2+) (caffeine) augmented Cd influx. Cd(2+) and ATP added at different temporal conditions were not effective at increasing intracellular Cd(2+) accumulation. Ouabain (Na(+)/K(+)-ATPase inhibitor) increased Cd(2+) influx probably through a change in intracellular Na and/or a change in cell membrane potential. Routes of Cd(2+) influx, a non-essential metal, through the gill cell plasma membrane of crabs are suggested.

Cultivo de larvas de Ucides cordatus (LINNAEU, 1763) sob diferentes intensidades luminosas / Rearing of Ucides cordatus (LINNAEU, 1763) larvae under different light intensities

O caranguejo-uçá, Ucides cordatus, é uma espécie típica dos manguezais brasileiros e tem grande importância econômica para as populações litorâneas tradicionais. O presente trabalho investigou a influência da intensidade luminosa sobre a sobrevivência e a taxa de desenvolvimento larval de U. cordatus. Três intensidades luminosas foram avaliadas: claro - 710 lux, penumbra - 210 lux e escuro - 1 lux, em duas condições de cultivo, individual e coletivo. Houve diferenças significativas entre as taxas de sobrevivência das larvas zoea e as três intensidades luminosas avaliadas (p<0,05). As maiores taxas de ecdise para o estágio de megalopa foram obtidas no tratamento claro (42% nos cultivos coletivos e 30% nos cultivos individuais). No tratamento escuro, a metamorfose para megalopa foi de apenas 16% nos cultivos coletivos e de 7% nos cultivos individuais. Estes resultados indicam que a manutenção das larvas em baixas intensidades luminosas afeta negativamente a sobrevivência larval de U. cordatus...

Ucides cordatus is an edible crab species typical of Brazilian mangroves, and traditionally represents an important economic resource for many coastal populations. The present study investigated the influence of light intensity on the survival and rate of larval development of U. cordatus. Three different levels of luminosity were evaluated: 710 (Light), 210 Lux (Shaded) and 1 Lux (Dark), both in individual and collective cultivation conditions. Significant differences were found for survival of zoea larvae under the different light intensities (P<0.05). The greatest survival rates as well as rates of ecdysis to the megalopa stage were obtained under Light conditions (42% in collective cultures and 30% in individual cultures). In Dark conditions events of metamorphosis to megalopa stage was observed only in 16% of collective cultures and 7% of individual cultures. The result indicates that low light intensities may negatively affect larval survivorship during U. cordatus larval cultivations...

Cultivo de larvas de Ucides cordatus (LINNAEU, 1763) sob diferentes intensidades luminosas / Rearing of Ucides cordatus (LINNAEU, 1763) larvae under different light intensities

O caranguejo-uçá, Ucides cordatus, é uma espécie típica dos manguezais brasileiros e tem grande importância econômica para as populações litorâneas tradicionais. O presente trabalho investigou a influência da intensidade luminosa sobre a sobrevivência e a taxa de desenvolvimento larval de U. cordatus. Três intensidades luminosas foram avaliadas: claro - 710 lux, penumbra - 210 lux e escuro - 1 lux, em duas condições de cultivo, individual e coletivo. Houve diferenças significativas entre as taxas de sobrevivência das larvas zoea e as três intensidades luminosas avaliadas (p<0,05). As maiores taxas de ecdise para o estágio de megalopa foram obtidas no tratamento claro (42% nos cultivos coletivos e 30% nos cultivos individuais). No tratamento escuro, a metamorfose para megalopa foi de apenas 16% nos cultivos coletivos e de 7% nos cultivos individuais. Estes resultados indicam que a manutenção das larvas em baixas intensidades luminosas afeta negativamente a sobrevivência larval de U. cordatus.(AU)

Ucides cordatus is an edible crab species typical of Brazilian mangroves, and traditionally represents an important economic resource for many coastal populations. The present study investigated the influence of light intensity on the survival and rate of larval development of U. cordatus. Three different levels of luminosity were evaluated: 710 (Light), 210 Lux (Shaded) and 1 Lux (Dark), both in individual and collective cultivation conditions. Significant differences were found for survival of zoea larvae under the different light intensities (P<0.05). The greatest survival rates as well as rates of ecdysis to the megalopa stage were obtained under Light conditions (42% in collective cultures and 30% in individual cultures). In Dark conditions events of metamorphosis to megalopa stage was observed only in 16% of collective cultures and 7% of individual cultures. The result indicates that low light intensities may negatively affect larval survivorship during U. cordatus larval cultivations.(AU)

Characterization of copper transport in gill cells of a mangrove crab Ucides cordatus.

The branchial epithelium of crustaceans is exposed to the environment and is the first site affected by metal pollution. The aim of this work was to characterize copper (Cu) transport using a fluorescent dye, Phen Green, in gill cells of a hypo-hyper-regulator mangrove crab Ucides cordatus. The results showed that added extracellular CuCl2 (0, 0.025, 0.150, 0.275, 0.550 and 1.110 µM) showed typical Michaelis-Menten transport for Cu in anterior and posterior gill cells (Vmax for anterior and posterior gills: 0.41 ± 0.12 and 1.76 ± 0.27 intracellular Cu in µM × 22.10(4)cells(-1)× 300 s(-1) respectively and Km values: 0.44 ± 0.04 and 0.32 ± 0.13 µM, respectively). Intracellular Cu was significantly higher for posterior gill cells compared to anterior gill cells, suggesting differential accumulation for each gill type. Extracellular Ca at 20mM decreased cellular Cu transport for both anterior and posterior gill cells. Nifedipine and verapamil, calcium channel inhibitors from plasma membrane, decreased Cu transport and affected Km for both gills. These results could be due to a competition between Cu and Ca. Amiloride, a Na/Ca exchanger inhibitor, as well as bafilomycin, a proton pump inhibitor, caused a decrease of intracellular Cu compared to control. Ouabain and KB-R 7943, acting on Na homeostasis, similarly decreased intracellular Cu in both gill cells. Besides that, gill cells exposed to ATP and Cu simultaneously, showed an increase in intracellular copper, which was inhibited by vanadate, an inhibitor of P-type ATPase. These results suggest either the presence of a Cu-ATPase in crab gill cells, responsible for Cu influx, or the effect of a change in electrochemical membrane potential that could also drive Cu to the gill cell interior. Caffeine increased intracellular Cu, suggesting that intracellular Ca could be affecting Cu uptake. Overall the results show that copper uptake in gill cells of crabs is regulated by intracellular Ca, Ca channels and by Na exchangers. This is the first report of Cu transport characterization in whole gill cells of crabs.

Calcium transport in gill cells of Ucides cordatus, a mangrove crab living in variable salinity environments.

Crustaceans show discontinuous growth and have been used as a model system for studying cellular mechanisms of calcium transport, which is the main mineral found in their exoskeleton. Ucides cordatus, a mangrove crab, is naturally exposed to fluctuations in calcium and salinity. To study calcium transport in this species during isosmotic conditions, dissociated gill cells were marked with fluo-3 and intracellular Ca(2+) change was followed by adding extracellular Ca(2+) as CaCl2 (0, 0.1, 0.25, 0.50, 1.0 and 5mM), together with different inhibitors. For control gill cells, Ca(2+) transport followed Michaelis-Menten kinetics with Vmax=0.137±0.001 ∆Ca(2+)i (µM×22.10(4)cells(-1)×180s(-1); N=4; r(2)=0.99); Km=0.989±0.027mM. The use of different inhibitors for gill cells showed that amiloride (Na(+)/Ca(2+) exchange inhibitor) inhibited 80% of Ca(2+) transport in gill cells (Vmax). KB-R, an inhibitor of Ca influx in vertebrates, similarly caused a decrease in Ca(2+) transport and verapamil (Ca(2+) channel inhibitor) had no effect on Ca(2+) transport, while nifedipine (another Ca(2+) channel inhibitor) caused a 20% decrease in Ca(2+) affinity compared to control values. Ouabain, on the other hand, caused no change in Ca(2+) transport, while vanadate increased the concentration of intracellular calcium through inhibition of Ca(2+) efflux probably through the plasma membrane Ca(2+)-ATPase. Results show that transport kinetics for Ca(2+) in these crabs under isosmotic conditions is lower compared to a hyper-regulator freshwater crab Dilocarcinus pagei studied earlier using fluorescent Ca(2+) probes. These kinds of studies will help understanding the comparative mechanisms underlying the evolution of Ca transport in crabs living in different environments.

Comparison of three quick methods to estimate crab size in the land crabs Cardisoma guanhumi Latreille, 1825 and Ucides cordatus (Crustacea: Brachyura: Gecarcinidae and Ucididae) / Comparación de tres métodos rápidos para estimar el tamaño del cangrejo en los cangrejos terrestres Cardisoma guanhumi Latreille, 1825 y Ucides cordatus (Crustacea: Brachyura: Gecarcinidae y Ucididae)

Quick, reliable and non destructive methods are necessary to estimate size structure on commercial land crabs, in order to acquire relevant information concerning the health of their populations. Cardisoma guanhumi and Ucides cordatus are two land crabs that are exploited at a high scale and also in an artisan way in the Caribbean area and in the coasts of Brazil, which populations are endangered due to uncontrolled exploitation. The purpose of this work is to provide various methods to estimate indirectly crab body size. Sampling was carried out in Carenero (C. guanhumi) and Cumaná (U. cordatus) (Venezuela). For each species, three methods were used to measure burrow diameter (Vernier, internal spring caliper and photograph), and these were correlated with real body size of the crabs. Model II linear regression analyzes, i.e. Ordinary Least Squares and Mayor Axis, were used to build and test the performance of forecasting models. Cardisoma guanhumi showed a high bivariate data dispersion using Vernier and photo measuring methods, increasing these towards larger animals. Less dispersion was achieved with the spring caliper method; this resulted in the most accurate measurements of indirectly estimated body size in C. guanhumi (r²= 0.61), whereas Vernier measurements were the least precise. On the other hand, all three methods gave reliable estimates for U. cordatus, being the Vernier method the most accurate (r²= 0.71). However, in both species, all forecasting equations overestimated the size of smaller crabs (those below the mean) but underestimated the size of larger crabs. Nevertheless, all three methods were statistically significant for each of the species, and looking at the above mentioned underand overestimations, they can serve as reliable and fast non-destructive tools to be used by resource managers and field biologists to acquire size structure information concerning these two species. Vernier and internal spring caliper ...

Para la estimación de la estructura de tamaños en cangrejos terrestres comerciales y la obtención de información relevante para su manejo, es necesario utilizar métodos rápidos, confiables y no destructivos. Cardisoma guanhumi y Ucides cordatus son dos cangrejos terrestres que son explotados comercialmente en el Caribe y en Brasil. El propósito de este trabajo es suministrar métodos indirectos para la estimación del tamaño del caparazón de los cangrejos y por consiguiente, de la estructura de tallas. Los muestreos se llevaron a cabo en Carenero (C. guanhumi) y en Cumaná (U. cordatus) (Venezuela). Se utilizaron tres métodos para estimar el diámetro de sus madrigueras: Vernier, compás y fotografía. Estos se correlacionaron con el tamaño real del cangrejo. Se aplicó el análisis de regresión Ordinary Least Squares Model II y la capacidad de predicción se probó utilizando el modelo II Mayor Axis para las regresiones. Cardisoma guanhumi mostró una fuerte dispersión de sus datos en los métodos de Vernier y fotografía. Menos dispersión se obtuvo con el método del compás y fue el más preciso (r²= 0.61). Para U. cordatus las medidas con Vernier fueron la más adecuadas (r²= 0.71). Sin embargo los tres métodos fueron confiables. Los diferentes métodos mostraron ventajas y desventajas y dependerá del que aplique los métodos, decidir cuál será el más adecuado para sus propósitos.