Temporal, diel and spatial variability of decapod larvae from St Paul`s Rocks, an equatorial oceanic island of Brazil

Temporal, spatial and diel variation in the distribution and abundance of organisms is an inherent property of ecological systems. The present study describes these variations and the composition of decapod larvae from the surface waters of St Paul`s Rocks. The expeditions to the archipelago were carried out in April, August and November 2003, March 2004 and May 2005. Surface plankton samples were collected during the morning and dusk periods, inside the inlet and in increasing distances around the archipelago (similar to 150, 700 and 1500 m). The identification resulted in 51 taxa. Seven species, six genera and larvae of the families Pandalidae and Portunidae were identified for the first time in the area. The mean larval density varied from zero to 150.2 +/- 69.6 individuals 100 m(-3) in the waters surrounding the archipelago and from 1.7 +/- 3.0 to 12,827 +/- 15,073 individuals 100 m(-3) inside the inlet. Significant differences on larval density were verified between months and period of the day, but not among the three sites around the archipelago. Cluster and non-metric multidimensional scaling analysis indicated that the decapod larvae community was divided into benthic and pelagic assemblages. Indicator species analysis (ISA) showed that six Brachyura taxa were good indicators for the inlet, while three sergestids were the main species from the waters around the archipelago. These results suggest that St Paul`s Rocks can be divided into two habitats, based on larval composition, density and diversity values: the inlet and the waters surrounding the archipelago.

Vertical distribution of benthic invertebrate larvae during an upwelling event along a transect off the tropical Brazilian continental margin

Abundance and composition of marine benthic communities have been relatively well studied in the SE Brazilian coast, but little is known on patterns controlling the distribution of their planktonic larval stages. A survey of larval abundance in the continental margin, using a Multi-Plankton Sampler, was conducted in a cross-shelf transect off Cabo Frio (23 degrees S and 42 degrees W) during a costal upwelling event. Hydrographic conditions were monitored through discrete CDT casts. Chlorophyll-a in the top 100 m of the water column was determined and changes in surface chlorophyll-a was estimated using SeaWiFS images. Based on the larval abundances and the meso-scale hydrodynamics scenario, our results suggest two different processes affecting larval distributions. High larval densities were found nearshore due to the upwelling event associated with high chlorophyll a and strong along shore current. On the continental slope, high larval abundance was associated with a clockwise rotating meander, which may have entrapped larvae from a region located further north (Cabo de Sao Tome, 22 degrees S and 41 degrees W). In mid-shelf areas, our data suggests that vertical migration may likely occur as a response to avoid offshore transport by upwelling plumes and/or cyclonic meanders. The hydrodynamic scenario observed in the study area has two distinct yet extremely important consequences: larval retention on food-rich upwelling areas and the broadening of the tropical domain to southernmost subtropical areas. (C) 2009 Elsevier B.V. All rights reserved.

Podochela meloi Sankarankutty, Ferreira & Cunha, 2001, a junior synonym of the spider crab Inachoides forceps A. Milne-Edwards, 1879 (Crustacea: Brachyura: Inachoididae)

Podochela meloi Sankarankutty, Ferreira & Cunha, 2001, originally described in the Inachidae MacLeay, 1838, was recently transferred to the Inachoididae genus Inachoides H. Milne Edwards & Lucas, 1842, based upon overall similarities. Placement of P. meloi in both Inachoididae and Inachoides is found to be supported by a number of synapomorphies as shown herein. Podochela meloi is shown to be a junior synonym of Inachoides forceps A. Milne-Edwards, 1879.

Reproductive migration and population dynamics of the blue crab Callinectes danae in an estuary in southeastern Brazil

Portunid crabs are an important resource in estuaries, and require appropriate management to guarantee their long-term availability. We investigated the population dynamics and reproduction of Callinectes danae in the Estuarine-Bay Complex of Sao Vicente, Sao Paulo, Brazil, to provide basic biological information for public policies for the management of this fishery. Monthly samples were obtained from March 2007 to February 2008 on eight transects, four in the estuary and four in the bay. A total of 2261 specimens (403 males, 1288 females, of which 570 were ovigerous) were collected. Males were significantly larger than females, and the size-frequency distribution was unimodal for males, females and ovigerous females. The sex ratio was nearly always skewed toward females (M:F - 1:4.6). C. danae showed seasonal-continuous reproduction, with high reproductive activity in the warmer season. C. danae breeds in the estuarine-bay complex, with males and juvenile females concentrated in the estuary. After copulation, fertilized females migrate to the estuary entrance and the bay, where ovigerous females are commonly found spawning in high-salinity areas. Therefore, to manage this important economic resource, both the estuary and the bay should be considered for protection, but special attention should be given to the estuary entrance during the summer months, when ovigerous females concentrate.

Molecular barcode and morphological analyses reveal the taxonomic and biogeographical status of the striped-legged hermit crab species Clibanarius sclopetarius (Herbst, 1796) and Clibanarius vittatus (Bosc, 1802) (Decapoda: Diogenidae)

The taxonomic status of the species Clibanarius sclopetarius (Herbst, 1796) and Clibanarius vittatus (Bosc, 1802), which have sympatric biogeographical distributions restricted to the western Atlantic Ocean, is based only on differences in the colour pattern of the walking legs of adults. Their morphological similarity led to the suggestion that they be synonymised. In order to investigate this hypothesis, we included species of Clibanarius Dana, 1892 in a molecular phylogenetic analysis of partial sequences of the mitochondrial 16S rDNA gene and the COI barcode region. In addition, we combined the molecular results with morphological observations obtained from several samples of these two species. The genetic divergences of the 16S rDNA and COI sequences between C. sclopetarius and C. vittatus ranged from 4.5 to 5.9% and 9.4 to 11.9%, which did not justify their synonymisation. Differences in the telson morphology, chela ornamentation, and coloration of the eyestalks and antennal peduncle provided support for the separation of the two species. Another interesting result was a considerable genetic difference found between populations of C. vittatus from Brazil and the Gulf of Mexico, which may indicate the existence of two homonymous species.

Analytical method for determination of nitric oxide in zebrafish larvae: Toxicological and pharmacological applications

Zebrafish are currently used at various stages of the drug discovery process and can be a useful and cost-effective alternative to some mammalian models. Nitric oxide (NO) plays an important role in physiology of zebrafish. The availability of appropriate analytical techniques to quantify the NO is crucial for studying its role in physiological and pathological conditions. This work aimed at establishing a high-performance liquid chromatography method for determination of NO levels in zebrafish larvae. Attempts were also made to assess the normal levels of NO at the first days postfertilization and the possible changes under pathological conditions. The method validation was quantitatively evaluated in terms of sensitivity, specificity, precision, accuracy, linearity, and recovery. NO levels from zebrafish larvae at the first days postfertilization and larvae challenged to N(G)-nitro-L-arginine methyl ester, sodium nitroprusside, Escherichia coil lipopolysaccharide, and copper sulfate were analyzed. The samples were derivatized with 2,3-diaminonaphthalene, and fluorescence detection was used for the indirect determination of NO. The method showed a good performance for all validation parameters evaluated and was efficient to monitor changes in NO concentration under physiological and pathophysiological conditions. This method might represent a powerful tool to be applied in NO studies with zebrafish larvae. (C) 2011 Elsevier Inc. All rights reserved.

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 Cubatao, state of Sao 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. (C) 2012 Elsevier Inc. All rights reserved.

Relative abundance and population biology of the non-indigenous crab Charybdis hellerii (Crustacea: Brachyura: Portunidae) in a southwestern Atlantic estuary-bay complex

Invasive species are potential threats to biodiversity, especially if they become established and outnumber native species. In this study, a population of the non-indigenous crab Charybdis hellerii was analyzed in an estuary-bay complex on the southeastern Brazilian coast, with respect to its abundance relative to sympatric native brachyuran species, as well as the size structure, sexual maturity, sex ratio, frequency of mutilation, reproductive period, and development of the reproductive system. Crabs were sampled monthly both in the intertidal zone of rocky shores and on sublittoral soft-bottom. Nine species were recorded on the rocky shores, where C. hellerii was the second most abundant species; only three individuals of C. hellerii were collected in the sublittoral samples. This population of C. hellerii showed a unimodal size structure composed mainly of mature individuals; males were larger than females, and the sex ratio was skewed toward males (3.1:1). About 46.9% of the individuals (75 of 160 crabs) had mutilated or regenerating appendages, more frequent in males (56.8%) than in females (28.2%), which may reflect both inter- and intraspecific agonistic interactions. A continuous reproductive pattern is suggested for this population, although ovigerous females occurred unevenly during the year, with 58.82% of them being collected in winter. There was evidence of multiple spawning, since the ovigerous females with an initial egg mass showed mature ovaries as well as seminal receptacles filled with sperm. C. hellerii is well established in the estuary-bay complex, but is concentrated in intertidal and shallow subtidal rocky shores, where it may compete with and replace other species such as the portunid Cronius ruber. This study also highlights the importance of systematic monitoring studies to evaluate the effects of the introduction of non-indigenous species on ecologically similar natives.

Hemolymph ion regulation and kinetic characteristics of the gill (Na+, K+)-ATPase in the hermit crab Clibanarius vittatus (Decapoda, Anomura) acclimated to high salinity

We examine hemolymph ion regulation and the kinetic properties of a gill microsomal (Na+, K+)-ATPase from the intertidal hermit crab, Clibanarius vittatus, acclimated to 45 parts per thousand salinity for 10 days. Hemolymph osmolality is hypo-regulated (1102.5 +/- 22.1 mOsm kg(-1) H2O) at 45 parts per thousand but elevated compared to fresh-caught crabs (801.0 +/- 40.1 mOsm kg(-1) H2O). Hemolymph [Na+ (323.0 +/- 2.5 mmol L-1) and [Me2+) (34.6 +/- 1.0 mmol L-1) are hypo-regulated while [Ca2+] (22.5 +/- 0.7 mmol L-1) is hyper-regulated; [K+] is hyper-regulated in fresh-caught crabs (17.4 +/- 0.5 mmol L-1) but hypo-regulated (6.2 +/- 0.7 mmol L-1) at 45 parts per thousand. Protein expression patterns are altered in the 45 parts per thousand-acclimated crabs, although Western blot analyses reveal just a single immunoreactive band, suggesting a single (Na+, K+)-ATPase alpha-subunit isoform, distributed in different density membrane fractions. A high-affinity (Vm = 46.5 +/- 3.5 U mg(-1); K-0.5 = 7.07 +/- 0.01 mu mol L-1) and a low-affinity ATP binding site (Vm = 108.1 +/- 2.5 U mg(-1); K-0.5 = 0.11 +/- 0.3 mmol L-1), both obeying cooperative kinetics, were disclosed. Modulation of (Na+, K+)-ATPase activity by Mg2+, K+ and NH4+ also exhibits site-site interactions, but modulation by Na+ shows Michaelis-Menten kinetics. (Na+, K+)-ATPase activity is synergistically stimulated up to 45% by NH4+ plus K+. Enzyme catalytic efficiency for variable [K+] and fixed [NH4+] is 10-fold greater than for variable [NH4+] and fixed [K+]. Ouabain inhibited approximate to 80% of total ATPase activity (K-I=464.7 +/- 23.2 mu mol L-1), suggesting that ATPases other than (Na+, K+)-ATPase are present. While (Na+, K+)-ATPase activities are similar in fresh-caught (around 142 nmol Pi min(-1) mg(-1)) and 45 parts per thousand-acclimated crabs (around 154 nmol Pi min(-1) mg(-1)), ATP affinity decreases 110-fold and Na+ and K+ affinities increase 2-3-fold in 45 parts per thousand-acclimated crabs. (C) 2012 Elsevier Inc. All rights reserved.

Comparative analysis of shell occupation by two southern populations of the hermit crab Loxopagurus loxochelis (Decapoda, Diogenidae)

The present study aimed to comparatively verify the relation between the hermit crabs and the shells they use in two populations of Loxopagurus loxochelis. Samples were collected monthly from July 2002 to June 2003, at Caraguatatuba and Ubatuba Bay, Sao Paulo, Brazil. The animals sampled had their sex identified, were weighed and measured; their shells were identified, measured and weighed, and their internal volume determined. To relate the hermit crab's characteristics and the shells' variables, principal component analysis (PCA) and a regression tree were used. According to the PCA analysis, the three gastropod shells most frequently used by L. loxochelis varied in size. The regression tree successfully explained the relationship between the hermit crab's characteristics and the internal volume of the inhabited shell. It can be inferred that the relationship between the morphometry of an individual hermit crab and its shell is not straightforward and it is impossible to explain only on the basis of direct correlations between the body's and the shell's attributes. Several factors (such as the morphometry and the availability of the shell, environmental conditions and inter- and intraspecific competition) interact and seem to be taken into consideration by the hermit crabs when they choose a shell, resulting in the diversified pattern of shell occupancy shown here and elsewhere.

The Laboratory Efficacy of Cypermethrin Dust Against Lesser Mealworm Larvae and Adults, Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae)

Cypermethrin dust was evaluated as a tool for the integrated management of lesser mealwonns (also called the darkling beetle), Alphitobius diaperinus (Panzer). This experiment examined the efficacy of the cypermethrin against adult and late instar lesser mealwonns under laboratory conditions. Two bioassay methods were evaluated, using either a petri plate or a covered plastic container simulating poultry house conditions. In the simulated conditions, two different samples were used and cypermethrin was either dusted onto the surface of the container or was directly dusted onto the bottom. The LC50 for adults was 636.6 ppm, however, 929.7 ppm of cypermethrin dust was needed to achieve a 50% mortality rate in late instar larvae 24 h after the administration of the insecticide. A similar trend was observed in the simulated poultry houses when the adult mortality was > 90% while effectiveness in late in. star larvae was decreased, i.e., between 50 and 85%. Significant differences in the toxicity profiles were observed in larvae mortality when cypermethrin it was dusted directly onto the litter surface, compared to the bottom of the container. We have verified that cypermethrin dust is available for use in poultry houses however, toxicity profiles of lesser mealworm may depend on the beetle's stage of development and method of application

Reproductive apparatus of the male giant hermit crab Petrochirus diogenes (Anomura, Diogenidae): morphology and phylogenetic implications

We describe the male reproductive apparatus of the giant hermit crab Petrochirus diogenes, with morphological and biometric analyses of the spermatophore, the gonopore and the ultrastructure of the spermatozoa. Specimens were collected from the southern coast of Sao Paulo, Brazil. Morphological analyses were done using stereoscopic, light, transmission and scanning electron microscopy. The reproductive system of this hermit crab is composed of elongate and lobular testes followed by vasa deferentia that connect to the exterior via gonopores. The gonopores are ovoid and surrounded by setae, and each gonopore is composed of a membranous operculum that forms a depression constituting the gonopore opening. The gonopore constitutes a unique structure among the Diogenidae due to its number of setae. The spermatophores are tripartite, composed of a sperm-containing ampulla, a peduncle and a proximal foot. The spermatozoon has 3 main regions (acrosomal vesicle, nucleus and cytoplasm). The structure of the spermatophore indicates that this species can be considered an exception within Diogenidae with regard to spermatophore morphology and can therefore be used for phylogenetic inferences.

Dexamethasone reduces bronchial wall remodeling during pulmonary migration of Strongyloides venezuelensis larvae in rats

Strongyloidiasis is an intestinal parasitosis with an obligatory pulmonary cycle. A Th2-type immune response is induced and amplifies the cellular response through the secretion of inflammatory mediators. Although this response has been described as being similar to asthma, airway remodeling during pulmonary migration of larvae has not yet been established. The aim of this study was to identify the occurrence of airway remodeling during Strongyloides venezuelensis (S. v.) infection and to determine the ability of dexamethasone treatment to interfere with the mechanisms involved in this process. Rats were inoculated with 9,000 S. v. larvae, treated with dexamethasone (2 mg/kg) and killed at 1, 3, 5, 7, 14 and 21 days. Morphological and morphometric analyzes with routine stains and immunohistochemistry were conducted, and some inflammatory mediators were evaluated using ELISA. Goblet cell hyperplasia and increased bronchiolar thickness, characterized by edema, neovascularization, inflammatory infiltrate, collagen deposition and enlargement of the smooth muscle cell layer were observed. VEGF, IL1-beta and IL-4 levels were elevated throughout the course of the infection. The morphological findings and the immunomodulatory response to the infection were drastically reduced in dexamethasone-treated rats. The pulmonary migration of S. venezuelensis larvae produced a transitory, but significant amount of airway remodeling with a slight residual bronchiolar fibrosis. The exact mechanisms involved in this process require further study. (C) 2012 Elsevier Ireland Ltd. All rights reserved.

NEW RECORD OF CALYPTRAEID HOSTS FOR THE PEA CRAB CALYPTRAEOTHERES GARTHI (FENUCCI, 1975) (DECAPODA, PINNOTHERIDAE) IN ARGENTINA

Calyptraeotheres garthi (Fenucci, 1975) is one of nine species of Pinnotheridae occurring in Argentina and the only species of that genus located in this region. This species can be found from the San Maths Gulf, Argentina to Rio Grande do Sul, Brazil where it lives in symbiotic association with limpets of the family Calyptraeidae. Currently, two species of limpets have been reported as hosts of this species: Crepidula protea (d'Orbigny, 1841) and Crepidula argentina Simone, Pastorino & Penchaszadeh, 2000. We report here four more species of limpets serving as hosts of C. garthi, namely Bostrycapulus odites Collin, 2005, Crepidula cachimilla Cledon, Simone & Penchaszadeh, 2004, Crepidula plana Say, 1822 and Trochita pileus (Lamarck, 1822). Thus, this pinnotherid appears to be, as others members of the family, a generalist symbiotic species since it dwells on almost all the potential limpet species reported for Argentina. However, there are notable differences in prevalence of C. garthi on each host, which suggest that the symbiont might prefer species of Crepidula over the others genera (Trochita and Bostrycapulus). Additionally, there exists an apparent relationship between host size and prevalence (e.g., Crepidula cachimilla, the limpet with the highest prevalence, is clearly larger than the others hosts). Perhaps it is premature to conclude that the size of the host might be of benefit for the symbiont, but it is a hypothesis worthy of being explored.

Fatty acids as a tool to compare cachara (Pseudoplatystoma reticulatum) (Siluriformes: Pimelodidae) and hybrid (Pseudoplatystoma corruscans x Pseudoplatystoma reticulatum) larvae during early development

In this study, we investigated the physiological alterations during ontogeny for cachara (Pseudoplatystoma reticulatum) and their hybrid larvae (Pseudoplatystoma corruscans x P. reticulatum) using lipids and fatty acids as physiological tools to elucidate the basis for differences in these groups' productivity in an industrial setting. Eggs and larvae samples were collected during January and February of 2008 in the city of Bandeirantes, MS, and were divided into three primary phases: phase I (0-16 h after fertilization); phase II (24 h after fertilization to 6 days after fertilization); and phase III (7-25 days after fertilization). The larvae of both groups showed a high degree of similarity, suggesting that the hybrid larvae showed a high level of heritability from the cachara broodstock. Analysis of the total lipid content provided evidence that there is no alteration in lipid concentration during ontogeny for both groups (i.e., the cachara and hybrids). However, the fatty acid profile showed that during the endogenous feeding period (phase II), when the larvae must use the energy reserves from the mother, the cachara larvae used mainly monounsaturated fatty acids for development. This is typical for most fish species, though notably, the hybrids preferentially used saturated fatty acids. Furthermore, certain specific changes demonstrate unique patterns of energy utilization and structural substrates, which may aid in elucidating the empirical differences reported by fish farmers (i.e., that the hybrids perform better than cacharas in captivity).