Differential adjustment in gill Na(+)/K(+)- and V-ATPase activities and transporter mRNA expression during osmoregulatory acclimation in the cinnamon shrimp Macrobrachium amazonicum (Decapoda, Palaemonidae)

We evaluate osmotic and chloride (Cl(-)) regulatory capability in the diadromous shrimp Macrobrachium amazonicum, and the accompanying alterations in hemolymph osmolality and [Cl(-)], gill Na(+)/K(+)-ATPase activity, and expression of gill Na(+)/K(+)-ATPase alpha-subunit and V-ATPase B subunit mRNA during salinity (S) acclimation. We also characterize V-ATPase kinetics and the organization of transport-related membrane systems in the gill epithelium. Macrobrachium amazonicum strongly hyper-regulates hemolymph osmolality and [Cl(-)] in freshwater and in salinities up to 25 parts per thousand S. During a 10-day acclimation period to 25 parts per thousand S, hemolymph became isosmotic and hypo-chloremic after 5 days, [Cl(-)] alone remaining hyporegulated thereafter. Gill Na(+)/K(+)-ATPase alpha-subunit mRNA expression increased 6.5 times initial values after 1 h, then decreased to 3 to 4 times initial values by 24 h and to 1.5 times initial values after 10 days at 25 parts per thousand S. This increased expression was accompanied by a sharp decrease at 5 h then recovery of initial Na(+)/K(+)-ATPase activity within 24 h, declining again after 5 days, which suggests transient Cl(-) secretion. V-ATPase B-subunit mRNA expression increased 1.5-fold within 1 h, then reduced sharply to 0.3 times initial values by 5 h, and remained unchanged for the remainder of the 10-day period. V-ATPase activity dropped sharply and was negligible after a 10-day acclimation period to 21 parts per thousand S, revealing a marked downregulation of ion uptake mechanisms. The gill epithelium consists of thick, apical pillar cell flanges, the perikarya of which are coupled to an intralamellar septum. These two cell types respectively exhibit extensive apical evaginations and deep membrane invaginations, both of which are associated with numerous mitochondria, characterizing an ion transporting epithelium. These changes in Na(+)/K(+)- and V-ATPase activities and in mRNA expression during salinity acclimation appear to underpin ion uptake and Cl(-) secretion by the palaemonid shrimp gill.

Structural and Biochemical Correlates of Na(+), K(+)-ATPase Driven Ion Uptake Across the Posterior Gill Epithelium of the True Freshwater Crab, Dilocarcinus pagei (Brachyura, Trichodactylidae)

To better comprehend the structural and biochemical underpinnings of ion uptake across the gills of true freshwater crabs, we performed an ultrastructural, ultracytochemical and morphometric investigation, and kinetically characterized the Na(+), K(+)-ATPase, in posterior gill lamellae of Dilocarcinus pagei. Ultrastructurally, the lamellar epithelia are markedly asymmetrical: the thick, mushroom-shaped, proximal ionocytes contain elongate mitochondria (41% cell volume) associated with numerous (approximate to 14 mu m(2) membrane per mu m(3) cytoplasm), deep invaginations that house the Na(+), K(+)-ATPase, revealed ultracytochemically. Their apical surface is amplified (7.5 mu m(2) mu m(-2)) by stubby evaginations whose bases adjoin mitochondria below the subcuticular space. The apical membrane of the thin, distal ionocytes shows few evaginations (1.6 mu m(2) mu m(-2)), each surrounding a mitochondrion, abundant in the cytoplasm below the subcuticular space; basolateral invaginations and mitochondria are few. Fine basal cytoplasmic bridges project across the hemolymph space, penetrating into the thick ionocytes, suggesting ion movement between the epithelia. Microsomal Na(+), K(+)-ATPase specific activity resembles marine crabs but is approximate to 5-fold less than in species from fluctuating salinities, and freshwater shrimps, suggesting ion loss compensation by strategies other than Na(+) uptake. Enzyme apparent K(+) affinity attains 14-fold that of marine crabs, emphasizing the relevance of elevated K(+) affinity to the conquest of fresh water. Western blotting and biphasic ouabain inhibition disclose two alpha-subunit isoforms comprising distinct functional isoenzymes. While enzyme activity is not synergistically stimulated by NH(4)(+) and K(+), each increases affinity for the other, possibly assuring appropriate intracellular K(+) concentrations. These findings reveal specific structural and biochemical adaptations that may have allowed the establishment of the Brachyura in fresh water. J. Exp. Zool. 313A:508-523, 2010. (C) 2010 Wiley-Liss, Inc.

Selection of shrimp breeders free of white spot syndrome and infectious hypodermal and hematopoietic necrosis

The objective of this work was to select surviving breeders of Litopenaeus vannamei from white spot syndrome virus (WSSV) outbreak, adapted to local climatic conditions and negatively diagnosed for WSSV and infectious hypodermal and hematopoietic necrosis virus (IHHNV), and to evaluate if this strategy is a viable alternative for production in Santa Catarina, Brazil. A total of 800 males and 800 females were phenotypically selected in a farm pond. Nested-PCR analyses of 487 sexually mature females and 231 sexually mature males showed that 63% of the females and 55% of the males were infected with IHHNV. Animals free of IHHNV were tested for WSSV, and those considered double negative were used for breeding. The post-larvae produced were stocked in nine nursery tanks for analysis. From the 45 samples, with 50 post-larvae each, only two were positive for IHHNV and none for WSSV. Batches of larvae diagnosed free of virus by nested-PCR were sent to six farms. A comparative analysis was carried out in growth ponds, between local post-larvae and post-larvae from Northeast Brazil. Crabs (Chasmagnathus granulata), blue crabs (Callinectes sapidus), and sea hares (Aplysia brasiliana), which are possible vectors of these viruses, were also evaluated. The mean survival was 55% for local post-larvae against 23.4% for post-larvae from the Northeast. Sea hares showed prevalence of 50% and crabs of 67% of WSSV.

Avaliação de contaminação por metais pesados em amostras de siris azuis

Metais pesados por não serem biodegradáveis podem se acumular nos tecidos vivos ao longo da cadeia alimentar chegando ao ser humano principalmente por meio da alimentação. Siris e outros organismos que se alimentam de matéria orgânica existente em sistemas aquáticos, podem absorver uma carga maior destes elementos tóxicos e assim, representar um risco potencial à saúde da população da região. Neste trabalho, foram realizados estudos quantitativos sobre o teor de Pb, Cd, Cr, Zn e Cu em amostras (n = 144) de siris azuis do gênero Callinectes sp., coletados ao longo do rio Cubatão, utilizando um Espectrofotômetro de Absorção Atômica por Chama (FSAA). Exceto para o Cr, onde o valor médio obtido neste trabalho ficou acima do estabelecido para qualquer tipo de alimento pela legislação brasileira, todos os outros resultados estiveram abaixo dos limites máximos recomendados por agências de saúde. Uma discussão é apresentada sobre a contribuição ao valor provisório de ingestão diária tolerável (PTDI), sugerido pela Organização Mundial de Saúde, devido ao consumo de carne de siri pela população.

Brachyuran soft-bottom assemblage from marine shallow waters in the southeastern Brazilian littoral.

The present investigation examined the assemblages of brachyuran crabs in three bays of southeastern So Paulo State, analyzing the numbers of species and individuals, biomass, and diversity. The crabs were collected monthly in Ubatumirim, Ubatuba, and Mar Virado bays in the Ubatuba region over 2 years (1998 and 1999). In each bay, six tows were made from a fishing boat. A total of 39,820 specimens, representing 11 super families, 39 genera, and 65 species of brachyurans, were obtained. The mean number of specimens and biomass varied seasonally, which can be mainly related with the population tendencies of two dominant species in the area (Callinectes ornatus and Hepatus pudibundus). The highest numbers of individuals and species were found in Ubatuba Bay, followed by Ubatumirim and Mar Virado. The highest diversity index was estimated for Ubatumirim (H' = 2.44), followed by Ubatuba (H' = 2.37) and Mar Virado (H' = 1.86). In general, the coastal region supported high abundance and species richness concerning brachyurans, with high diversity indices, probably because of its more-heterogeneous substrate, which may afford more refuges for certain species. These results suggested that environmental conditions in this area, although they showed certain differences, are favorable for the establishment and development of a diverse brachyuran guild.

Dimensionamento e sobreposição de nichos dos portunídeos (Decapoda, Brachyura), na Enseada da Fortaleza, Ubatuba, São Paulo, Brasil

The niche breadth of the Portunidae and their overlap on the subtidal sediments of Fortaleza bay, Ubatuba (São Paulo) was analyzed. Samples were made monthly from November/1988 to October/1989, inseven areas of the bay using a shrimp fishery boat equipped with two otter-trawls. Each area was characterized based on environmental factors such as depth, temperature, salinity, dissolved oxygen, organic matter and granulometric composition of the sediment. The number of individuais of each species was registered to each area (resource). Levins's standardized measure (BA) and niche percentage were calculated. Five species of swimming crabs were recorded in this study: Callinectes ornatus Ordway, 1863, Arenaeus cribrarius (Lamarck, 1818), Portunus spinimanus Latreille, 1819, Callinectes danae Smith, 1869 and Portunus spinicarpus (Stimpson, 1871). The widest ecological niche occurred to C. ornatus, present in all sampled areas (generalist species). Such fact can be related to high sediment tolerance when it is compared to P. spinimanus (especialist species) which was limited to the areas with coarse granulometric fractions. Highest niche overlap was verified between C. danae and A. cribrarius may be due to greater salinity tolerance of these species. The low occurrence of P. spinicarpus and its reduced niche size in Fortaleza Bay are due to association of this species to cold water currents (ACAS) more evidente in smaler depths during summer months. One future evaluation of the portunid diet can be useful to complement informations about this important aspect of the marine ecology.

Differential benthic occupation by crabs in the Ubatuba bay, São Paulo, Brazil

The goal of this work was to examine the occurrence of brachyurans in soft bottom sublittoral habitats and their association with some environmental factors. The relative abundance of crabs in two depth strata (from 4.5 to 9 m and from 9 to 18 m) was quantified. Crabs were collected with an otter-trawl with 3.7 m of mouth opening and 12-mm mesh in the cod end. Monthly sampling, consisted of a single trawl in each stratum during a 1-yr period, were carried out. Fifteen brachyuran and six anomuran species were found, including Callinectes ornatus Ordway, 1863; Callinectes danae Smith, 1869; Hepatus pudibundus (Herbst, 1785); Libinia spinosa H. Milne-Edwards, 1834; Persephona punctata (Linnaeus, 1758), and P. mediterranea (Herbst, 1794), which were the most abundant and frequent in the area. The most abundant swimming crabs in both strata were C. ornatus and C, dan(re. Size differences in C. ornatus were observed between strata, suggesting a spatial separation of juveniles and adult crabs.

Diel variations in decapod catch rate and size of captured individuals in a subtropical area of Brazil

Diel variations in decapod crustaceans catch rate, as well as variations in size of sampled individuals, were investigated in a sublittoral portion of Ubatuba Bay (23 degrees 20', 23 degrees 35'S and 44 degrees 50', 45 degrees 14'W) in order to detect differential patterns of occurrence. Three replicate trawls, each enclosing a 2,500 m(2) area, were performed at a median depth of 3.5 m during the waning moon period, in 3 consecutive summer and winter months. Trawls were conducted at dawn, noon, dusk and midnight. Hydrological and substratum features were monitored. Penaeoideans did not show a significant diel catch rate variation during the sampling periods, but the catch rate of brachyurans was highest at dusk and midnight during winter (p < 0.01). Fixing diel variation, catch rates of both brachyurans and penaeoideans are subjected to significant seasonal differences (p < 0.05). The largest specimens of Callinectes ornatus, Xiphopenaeus kroyeri, Rimapenaeus constrictus and Farfantepenaeus spp were found at twilight during summer. Differences on size of captured individuals mainly in samples of portunids and penaeids taken during the course of the day evidences that significant daily movements take place. This confirms that activity alterations depend on characteristics of daily schedules and on environmental demands of studied species.

Brachyuran community in Ubatuba Bay, northern coast of São Paulo State, Brazil

A bruchyuran crab assemblage from eight transacts at a non-consolidated sublittoral site in Ubatuba Bay was studied on a monthly basis from September 1995 to August 1996. Data about number of individuals of 50 species found and other information such as distribution of the dominant crabs are reported. The family Majidae was represented by 13 species, followed by Xanthidae (13), Portunidae (10), Leucosiidae (5), Calaphidae (2), Dromiidae (2), Parthenopidae (2), Goneplacidae (1), Pinnotharidne (1), and Ocvpodidae (1). The brachyuran taxocoenosis was dominated by Callinectes ornatus (60.4%), Callinectes danae (18.8%), and Hepatus pudibundus (7.7%). representing together 86.9% of the total number of collected brachyurans. The Shannon-Weaver diversity index ranged from 1.10 to 2.06 between transects, and from 1.33 to 2.22 between months, depending more on equitability than on richness.

Composição e abundância dos caranguejos (Decapoda, Brachyura) nas regiões de Ubatuba e Caraguatatuba, litoral norte paulista, Brasil

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Composição e distribuição dos Crustácea (Decapoda) na Lagoa do Peixe, Rio Grande do Sul, Brasil

The Lagoa do Peixe (Peixe's Lagoon) is located between Atlantic Ocean and Lagoa dos Patos and is 30 Km long and 0,5-2,0 Km wide (31º13'S, 50º55'W -31º26'S, 51º09'). For one year, the composition and distribution of Crustácea Decapoda were analyzed monthly (from July/1994 to June/1995), in seven subareas (terrestrial border and aquatic area) and physical and chemical parameters (depth, salinity, sediment texture, oxygen in water, organic material of sediment and water temperature) were registered. Decapoda from the families Grapsidae, Ocypodidae, Portunidae (Brachyura), Diogenidae (Anomura) and Penaeidae (Penaeidea) were captured. Chasmagnathus granulata Dana 1851 was found ali months near the canal connecting Atlantic Ocean and Lagoa do Peixe, but inside the lagoon it was collected only in October/94 and June/95, and in extreme subareas it was sampled in February/95 when the salinity was 31 at these sites. Callinectes sapidus Rathbun, 1896 and Cyrtograpsus angulatus Dana, 1851 were captured throughout the year, except October/94 and April/95, respectively. Both species were found in ali áreas of sample, indicating a high plasticity in relation to the factors analyzed in this study. Another species captured probably come in the lagoon in specific months, when the environment provides ideal conditions for their develo-pments.

SPERMATOGENESIS, SPERMATOPHORE, and SEMINAL FLUID PRODUCTION IN THE ADULT BLUE CRAB CALLINESTES DANAE (PORTUNIDAE)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Ocorrência de anormalidades morfológicas externas em caranguejos marinhos (Decapoda, Brachyura) no litoral norte do estado de são paulo

Deformities and abnormalities in crustaceans have been associated to genetic problem, which occurred during molt process, damage caused by ectobionts, predators or environmental stress caused by chemical wastes. Some crab specimens collected in the São Paulo littoral were found having body abnormalities. They belong to the following crab species: Callinectes ornatus (Ordway, 1863), Arenaeus cribrarius (Lamarck, 1818) and Leurocyclus tuberculosus (H. Milne Edwards; Lucas, 1843). Samplings were performed by trawling during July 2008, August and October 2009 at the Ubatuba region, São Paulo State, Brazil. Body abnormalities were verified in the cheliped dactyl (C. ornatus an adult male), carapace deformities (A. cribrarius an adult male) and abdominal alterations (C. ornatus an adult female; L. tuberculosus an adult male and an ovigerous female). The record and analysis of such occurrences can help in the distinction of natural or human impact caused alterations. In this way, the occurrence study of this kind of body alterations could provide tools in order to control unprotected environmental areas, as well as bring subsides to understand the unusual variations during the ontogeny of important species in the benthic community.

Epibiosis in decapod crustaceans by stalked barnacle Octolasmis lowei (Cirripedia: Poecilasmatidae)

Stalked barnacles Octolasmis lowei Darwin, 1851 are frequently found attached to decapod crustaceans. Their epibiotic association depends on many factors, which are mainly related to characteristics of the host's biology. This study evaluated the infestation and distribution of stalked barnacles in the branchial chambers of crabs, and analyzed the data with respect to the host's sex, maturity stage, molt cycle and size. The crab species Arenaeus cribrarius Lamarck, 1818, Callinectes danae Smith, 1869, Callinectes ornatus Ordway, 1863, Hepatus pudibundus Herbst, 1785, Libinia ferreirae Brito Capello, 1871, and Persephona punctata Linnaeus, 1758 were sampled and found to be infested by O. lowei. No juvenile crabs were infested. The prevalence of infestation by O. lowei was significantly different among C. danae, C. ornatus, and H. pudibundus males and females. All infested hosts were in the intermolt period. The mean size of infested crabs was larger than that observed for non-infested individuals. Internally, stalked barnacles were concentrated on the central gills or walls and floor of branchial chambers, suggesting that these gills provide more favorable conditions for the settlement and development of these epibionts. These results highlight the relationship between epibiont infestation and host biology, as well as the role of decapod crustaceans as a suitable substrate for the development of stalked barnacle O. lowei. © 2013 Sociedade Brasileira de Zoologia All rights reserved.