Histopathology of the mangrove land crab Ucides cordatus (Ocypodidae) affected by lethargic crab disease

Lethargic crab disease (LCD) has caused extensive epizootic mortality of the mangrove land crab Ucides cordatus (Linnaeus 1763) (Brachyura: Ocypodidae) along the Brazilian coast. Direct culture of tissue samples from sick crabs and subsequent isolation and purification identified the causative agent as an Exophiala species of fungus. The histopathology of crabs with variable signs of LCD indicates that the most affected tissues are the epidermis, connective tissue, heart, hepatopancreas, nervous system, and gills. Gonads, somatic muscles, and digestive system are less affected by the fungus. The observed pathology is compatible with the clinical signs of LCD. Necrosis, tissue degeneration, and congestion of hemal sinuses and vessels are present in heavily infected organs. Nerve fibers may be compressed by accumulations of yeast-like cells. In heavy infections the tissue of gill lamellae is destroyed with subsequent dilation or compression. Cellular immune responses include hemocytic infiltration, agglutination and encapsulation, and phagocytosis. Phagocytosis of yeast-like cells is abundant in the connective tissue associated with the exoskeleton. These results indicate that LCD is the result of a systemic phaeohyphomycosis caused by a species of Exophiala. The present study also suggests that dispersal of the fungus within the crab occurs through the hemal system.     

Drinking and osmoregulation in the mangrove crab Ucides cordatus following exposure to benzene

In benzene-exposed Ucides cordatus acclimated for 96 h to 9 and 34 per thousand SW, haemolymph, urine and gastric juice are isosmotic with each other, but differ significantly in osmolality from external media. In both salinities, under benzene action, urine K+ excretion and calcium absorption are increased significantly, whereas Na+ absorption and Mg2+ excretion show U/B ratios similar to control values. In 9 per thousand SW, some ionic exchanges via benzene-exposed gills are possibly hastened. Benzene exposure decreases significantly branchial chamber water osmolality, [Na+] and [K+], whereas [Ca2+] and [Mg2+] are unaffected. However, faster medium exchange presumably occurs in 34 per thousand SW, both crab groups show branchial chamber water osmotic and ionic concentrations similar to surrounding medium. Benzene exposure unaffected gastric juice composition. In both media, [Ca2+] and [Mg2] accumulate several times higher than surrounding media, and [Na+] and [K+] are significantly hypo-ionic to haemolymph. Na+ and K+ G/H ratios are lower in crabs acclimated to 34 per thousand SW than in crabs acclimated to 9 per thousand SW. Drinking rates are enhanced by benzene exposure and are higher at 34 per thousand SW than in seawater isosmotic with the haemolymph (26 per thousand SW). Benzene exposure affects significantly osmoregulatory capability, slowing haemolymph dilution after transfer to clean 9 per thousand SW. Lower haemolymph dilution rate accounts for higher osmolality, but 48 h after transfer there is no recovery like in control crabs. Haemolymph transfusion experiments suggest an interaction among effects of benzene and hormonal factors, possibly on water influx.     

Separation and viability of gill and hepatopancreatic cells of a mangrove crab Ucides cordatus

The gills contain essential cells for respiration and osmoregulation, whereas the hepatopancreas is the site of digestion, absorption, and nutrients storage. The aim of this work was to separate and characterize gill and hepatopancreatic cells of the mangrove crab, Ucides cordatus. For gills, the methodology consisted of an enzymatic cellular dissociation using Trypsin at 0.5%, observation of cellular viability with Tripan Blue, and separation of cells using discontinuous sucrose gradient at concentrations of 10%, 20%, 30%, and 40%. The hepatopancreatic cells were dissociated by magnetic stirring, with posterior separation by sucrose gradient at the same concentrations above. For gills, a high cellular viability was observed (92.5±2.1%), with hemocyte cells in 10% sucrose layer (57.99?±?0.17%, *P?相似文献   

Polymorphic microsatellite DNA markers in the mangrove crab Ucides cordatus (Brachyura: Ocypodidae)

The isolation and characterization of the first polymorphic microsatellite markers for the mangrove crab Ucides cordatus are described. The number of alleles at each locus ranged from three to 25, mean of nine alleles, in 46 crabs captured in two Brazilian mangroves. The markers averaged high levels of observed (0.709 ± 0.183) and expected (0.716 ± 0.170) heterozygosities. Departures from Hardy-Weinberg equilibrium were observed at two loci. Linkage disequilibrium tests were not significant and no evidence of null alleles was detected. All these microsatellite loci are expected to be useful in estimating fine-scale population processes of this valuable mangrove species currently subjected to excessive fishing efforts.     

Isolation and characterization of 11 microsatellite loci in the mangrove crab, Ucides cordatus (Ocypodidae: Ucidinae)

Eleven polymorphic microsatellite loci were described for the mangrove crab, Ucides cordatus, an important fishery resource on the Brazilian coast. The number of alleles observed at each locus varied between eight and 23. Observed and expected mean heterozygosities were 0.791 and 0.893 respectively. Amplification of all loci was highly successful, under the same polymerase chain reaction conditions. With the exception of P2D3, all loci adhered to the assumptions of the Hardy-Weinberg equilibrium and did not present deviations reflecting linkage disequilibrium. Given this, these markers will be extremely useful in future management programmes for U. cordatus stocks.     

Phenotypical traits and gonadal development in mangrove land crab,Ucides cordatus (Decapoda: Ocypodidae)

Castilho, G.G., Ostrensky, A., Pie, M.R. and Boeger, W.A. 2010. Phenotypical traits and gonadal development in mangrove land crab, Ucides cordatus (Decapoda: Ocypodidae). —Acta Zoologica (Stockholm) 92 : 393–397. The association between phenotypical and reproductive traits has been the focus of considerable research in decapod crustaceans, particularly with the goal of facilitating the management and sustainable use of natural stocks of commercial interest. The goal of the present study is to evaluate the existence of a relationship between ovarian maturation and the coloration of the ovary and carapace of Ucides cordatus using histological methods and macroscopic observation. Monthly samples of adult females were obtained between October 2002 and March 2005 in mangroves of the Baía de Antonina, Southern Brazil (25º25′S; 48º42′W). A total of 367 specimens were collected throughout the study. Each specimen was classified subjectively into five ovarian colorations (whitish, yellow, orange, brown, and purple) and five carapace colorations (yellow, blue, dark gray, brown, and purple). Although substantial differences in ovarian development were found among females with different carapace and ovarian colorations, the variability within each class precluded their use as reliable indicators of ovarian development.     

A morphological study on gills of a crab acclimated to fresh water

The gills of the fully euryhaline Chinese crab Eriocheir sinensis were studied by light and electron microscopy. In these Phyllobranchiates, the gills consist of a double row of lamellae extending laterally from a central shaft. Haemolymph flow pattern inside the gill is described and the existence of a complex secondary vascularization inside the platelets is reported. It is shown that important differences exist between the ultrastructure of the three anterior and the three posterior pairs of large gills. The epithelium of the posterior gills is much thicker and possesses an extensive elaboration of the plasma membranes in the form of infoldings, crypts and interdigitations, along which are packed numerous mitochondria. The presence of such a complex membrane system opening to the extracellular space and closely associated with mitochondria is common to all salt-transporting tissues. This study corroborates the idea that the posterior pairs of gills of Eriocheir sinensis are the only ones implicated in active Na+ uptake when the crab lives in dilute aquatic environment. The epithelium of anterior gills is much thinner and the cells poor in intracellular organelles. It seems to be involved essentially in respiration. Thus this work clearly corroborates the existence already suggested by physiological approach of a functional difference between the different pairs of E. sinensis branchiae with respect to their participation in the respiration and in the regulation of the blood ions content. Common to both types of gills is the presence of a lamellar septum separating the haemolymph space into two compartments. The part played by that structure in determining the pattern of haemolymph flow, together with periodic bridges forming pillars across the haemolymph space, is emphasized.     

Disposition of trace elements in the mangrove ecosystem and their effects on Ucides cordatus (Linnaeus, 1763) (Crustacea,Decapoda)

BioMetals - The tropical mangrove at the Estação Ecológica do Lameirão (Southeastern Brazil), an ecosystem of high socio-economic value, receives a large load of pollutants from...     

Net uptake of chloride across the posterior gills of the Chinese crab (Eriocheir sinensis)

Two methods are commonly used for the determination of transbranchial net fluxes of Na+ and Cl-: direct analysis of changes in ion concentrations in the external medium using flame spectrophotometry or titration (net flux method), and measurement of unidirectional ion fluxes by means of radioactive tracers (tracer method). When we applied both methods in the same preparation, the isolated perfused posterior gill of freshwater-acclimated Eriocheir sinensis, to determine net fluxes of Cl-, the results differed substantially. In artificial fresh water (AFW) containing NaCl, the net flux method yielded a net uptake, but the tracer method showed a net efflux of Cl-. The net uptake of Cl- was abolished in Na(+)-free AFW indicating that Cl- uptake is coupled with the uptake of Na+. Applying the tracer method, net efflux of Cl- remained almost unchanged in Na(+)-free AFW. This suggests the opposite mechanism, i.e. uncoupled uptake of Na+ and Cl-. The discrepancy in the results obviously depends on the method employed. Since the data obtained with the net flux method explain the osmoregulatory performance of crabs living in fresh water, we consider this method as appropriate for determining net transbranchial ion fluxes.     

Modulation of ion uptake across posterior gills of the crab Chasmagnathus granulatus by dopamine and cAMP

Cyclic AMP (cAMP) and dopamine modulate ion uptake across isolated and perfused posterior gills of Chasmagnathus granulatus acclimated to 10 per thousand salinity. Addition of cAMP agonists, such as cp-cAMP, forskolin, and IBMX, produced a significant increase in the transepithelial potential difference (Vte), which reflects ion transport activity. Dopamine (DA) also had a stimulatory effect on ion uptake, increasing Vte and Na(+) influx, although this effect was transient, since both variables remained elevated for less than 30 min. In addition, the dose-response curve for DA concentration-Vte was biphasic, and the maximum stimulation was obtained with 10 micromol l(-1). When the effects of forskolin and DA on the Na(+)/K(+)-ATPase activity were tested, they correlated well with the Vte and Na(+) influx experiments; the enzyme activity increased significantly after preincubation of gill fragments for 10 min with forskolin or DA (51 and 64%, respectively), but there was no effect after pre-incubation with DA for 20 min. Finally, KT5720, a specific inhibitor of cAMP-dependent protein kinase (PKA), completely abolished the stimulatory effect of DA on Vte, suggesting the involvement of PKA in this mechanism.     

Burrows of the Semi-Terrestrial Crab Ucides cordatus Enhance CO2 Release in a North Brazilian Mangrove Forest

Ucides cordatus is an abundant mangrove crab in Brazil constructing burrows of up to 2 m depth. Sediment around burrows may oxidize during low tides. This increase in sediment-air contact area may enhance carbon degradation processes. We hypothesized that 1) the sediment CO₂ efflux rate is greater with burrows than without and 2) the reduction potential in radial profiles in the sediment surrounding the burrows decreases gradually, until approximating non-bioturbated conditions. Sampling was conducted during the North Brazilian wet season at neap tides. CO₂ efflux rates of inhabited burrows and plain sediment were measured with a CO₂/H₂O gas analyzer connected to a respiration chamber. Sediment redox potential, pH and temperature were measured in the sediment surrounding the burrows at horizontal distances of 2, 5, 8 and 15 cm at four sediment depths (1, 10, 30 and 50 cm) and rH values were calculated. Sediment cores (50 cm length) were taken to measure the same parameters for plain sediment. CO₂ efflux rates of plain sediment and individual crab burrows with entrance diameters of 7 cm were 0.7–1.3 µmol m⁻² s⁻¹ and 0.2–0.4 µmol burrows⁻¹ s⁻¹, respectively. CO₂ released from a Rhizophora mangle dominated forest with an average of 1.7 U. cordatus burrows⁻¹ m⁻² yielded 1.0–1.7 µmol m⁻² s⁻¹, depending on the month and burrow entrance diameter. Laboratory experiments revealed that 20–60% of the CO₂ released by burrows originated from crab respiration. Temporal changes in the reduction potential in the sediment surrounding the burrows did not influence the CO₂ release from burrows. More oxidized conditions of plain sediment over time may explain the increase in CO₂ release until the end of the wet season. CO₂ released by U. cordatus and their burrows may be a significant pathway of CO₂ export from mangrove sediments and should be considered in mangrove carbon budget estimates.     

A membrane-bound hemoglobin from gills of the green shore crab Carcinus maenas

Most hemoglobins serve for the transport or storage of O(2). Although hemoglobins are widespread in "entomostracan" Crustacea, malacostracans harbor the copper-containing hemocyanin in their hemolymph. Usually, only one type of respiratory protein occurs within a single species. Here, we report the identification of a hemoglobin of the shore crab Carcinus maenas (Malacostraca, Brachyura). In contrast to the dodecameric hemocyanin of this species, C. maenas hemoglobin does not reside in the hemolymph but is restricted to the gills. Immunofluorescence studies and cell fractioning showed that C. maenas hemoglobin resides in the membrane of the chief cells of the gill. To the best of our knowledge, this is the first time that a membrane-bound hemoglobin has been identified in eukaryotes. Bioinformatic evaluation suggests that C. maenas hemoglobin is anchored in the membrane by N-myristoylation. Recombinant C. maenas hemoglobin has a hexacoordinate binding scheme at the Fe(2+) and an oxygen affinity of P(50) = 0.5 Torr. A rapid autoxidation rate precludes a function as oxygen carrier. We rather speculate that, analogous to prokaryotic membrane-globins, C. maenas hemoglobin carries out enzymatic functions to protect the lipids in cell membrane from reactive oxygen species. Sequence comparisons and phylogenetic studies suggested that the ancestral arthropod hemoglobin was most likely an N-myristoylated protein that did not have an O(2) supply function. True respiratory hemoglobins of arthropods, however, evolved independently in chironomid midges and branchiopod crustaceans.     

Phosphofructokinase from the posterior gills of the euryhaline crab,Eriocheir sinensis: evidence for its regulation by phosphorylation

Phosphofructokinase from the posterior gills of the euryhaline crab Eriocheir sinensis acclimated to freshwater is likely regulated in part via phosphorylation induced by endogenous cyclic nucleotide-dependent protein kinases. Phosphofructokinase from gill extracts devoid of low molecular weight compounds by chromatography through a PD10 Sephadex column, incubated in the presence of cAMP or cGMP protein kinases activators (cAMP or cGMP, Mg-ATP and Mg²⁺), shows an increased catalytic activity. This treatment is accompanied by ³²P incorporation into the proteins immunoprecipitated with anti-mammalian phosphofructokinase polyclonal antibodies cross-reacting with the analog crustacean enzyme. Our results indicate that the covalent modification induced by these nucleotide-dependent protein kinases activates the glycolytic enzyme by increasing its affinity for its substrate and, when the activation is specifically due to cAMP-dependent protein kinases, by also reducing the homotropic cooperativity between its multiple substrate binding sites.     

Mangrove habitat partitioning by Ucides cordatus (Ucididae): effects of the degree of tidal flooding and tree-species composition during its life cycle

Environmental factors strongly affect mangrove crabs, and some factors modulate population structure and habitat partitioning during the crabs’ life cycle. However, the effect of these environmental factors on habitat selection by mangrove crabs is still unknown. We evaluated habitat selection by the mangrove crab Ucides cordatus in mangrove forests with different degrees of predominance of Rhizophora mangle, Laguncularia racemosa or Avicennia schaueriana, two tidal flooding levels (less- and more-flooded), and two biological periods (breeding and non-breeding seasons). Sampling was conducted in four mangrove forests with different influences of these biotic and abiotic parameters. We used the data for sex ratio to explain environmental partitioning by this species. Females predominated in R. mangle mangroves, independently of the biological period (breeding or non-breeding seasons), and males predominated only in the less-flooded L. racemosa mangroves. The flooding level affected the sex ratio of U. cordatus, with a predominance of males in less-flooded mangroves, independently of the biological period; and a gender balance in the more-flooded mangroves only during the breeding season. Outside the breeding season, the largest specimens were recorded in the R. mangle mangroves, but in the breeding season, the largest crabs were recorded in the L. racemosa mangroves with a higher level of flooding. These results suggest that tree-species composition and tidal flooding level can have a significant effect on the habitat partitioning of sexes and sizes of the mangrove crab U. cordatus both during and outside the breeding season.     

Some functional and morphological adaptations in the gills of dipnoi

Summary The gill epithelium of the Neoceratodus forsteri contains ce0lls which produce generations of dark vesicles. These vesicles move in waves towards the surface of the cells, break the cellular membranes and pour their content onto the cellular surfaces. This surface coat consists of threads and granules. The breaking of cell membranes by the densely situated vesicles leads to formation of processes, which join frequently to form a surface net: conversely the cytoplasm of the epithelial cells forms large invaginations penetrated by fluid. The reduction of cytoplasm and the mechanical loss of the cytoplasmic processes leads finally to the destruction of the cell.A small number of granulated cells with long processes extending between other cells have also been observed. The border between the gill epithelium and the connective tissue is characterised by the presence of bundles of thin fibers penetrating the epithelium.The above findings are discussed in relation to the differentiation of the epithelial cells, the facilitation of respiration in poorly oxygenated waters, the protection of gill surfaces against abrasion and with regard to the survival of the lung fish in different conditions.     

The life history of the mangrove tree crab, Aratus pisoni

Aratus pisoni (Milne Edwards) breeds throughout the year according to a lunar rhythm; the hatching of the eggs occurs at both full and new moon. Females become mature at about six months old or 12 mm carapace width and from then on, ovulate, on the average, once every intermoult; the number of eggs laid is directly proportional to the volume of the crab. By the time a female is eighteen months old at a size of 18 mm carapace width it will have laid 27,500 eggs and passed through seven moults. Mortality is greatest during the larval life and is caused mainly by predation.     

The organization of the mitochondrial control region in 2 Brachyuran Crustaceans: Ucides cordatus (Ocypodidae) and Cardisoma guanhumi (Gecarcinidae)

The control region (CR) is the largest noncoding segment of the mitochondrial DNA and includes the major regulatory elements for its replication and expression. In addition, the high level of intraspecific genetic variability found in the CR favors its use in phylogeographical and population genetic studies of a variety of organisms. However, most of the work on the structure of the CR has focused on vertebrates and insects, and little is known about the evolution of the CR in other taxa. In this study, we sequenced the entire CR of several individuals of 2 crab species: Ucides cordatus (Ocypodidae) and Cardisoma guanhumi (Gecarcinidae). There were neither large conserved regions in the CR of either species nor any similarity among species at the nucleotide level. However, the spatial pattern of genetic variability on the CR was similar among species. In addition, interesting similarities were found in the formation of stable secondary structures and in the position of regulatory elements. These results indicate that the evolution of CR in crustaceans is a remarkably dynamic process, with most homology among species being found at the secondary level.     

Effect of salinity on osmoregulatory patch epithelia in gills of the blue crab Callinectes sapidus

In euryhaline crabs, ion-transporting cells are clustered into osmoregulatory patches on the lamellae of the posterior gills. To examine changes in the branchial osmoregulatory patch in the blue crab Callinectes sapidus in response to change in salinity and to correlate these changes with other osmoregulatory responses, crabs were acclimated to a range of salinities between 10 and 35 ppt. When crabs that had been acclimated to 35 ppt were subsequently transferred to 10 ppt, both the size of the osmoregulatory patch on individual gill lamellae and the specific activity of Na+, K+-ATPase in whole-gill homogenates increased only after the first 24 h of exposure to dilute seawater. Enzyme activity and size of patch area increased gradually and reached their maxima (increasing by 200% and 60%, respectively) 6 days following transfer to 10 ppt seawater and then remained at these levels. Patch size at acclimation varied inversely with the salinity for seawater dilutions below 26 ppt (the isosmotic point of the crab), although it did not vary in salinities at or above 26 ppt. Thus, the size of the patch clearly is modulated with acclimation salinity, but it increases only in those salinities in which the crab hyperosmoregulates. An increase in the total RNA/DNA ratio in gill homogenates, the lack of mitotic figures in the lamellae, and the lack of incorporation of bromodeoxyuridine into nuclei of lamellar epithelial cells during acclimation to dilute seawater were interpreted as evidence that no cell proliferation had occurred and that increases in the size of the osmoregulatory patch occurred through differentiation of existing gas exchange cells or of undifferentiated epithelial cells into ion-transporting cells.