Population biology of Excirolana armata (Dana, 1853) (Isopoda, Cirolanidae) on an exposed sandy beach in Southeastern Brazil

The reproductive biology and population dynamics of the cirolanid isopod Excirolana armata (Dana, 1853) were analysed through monthly samples from December 2003 to November 2005 on Una beach, Sao Paulo state (24 degrees S), in Southeastern Brazil. Sampling was performed along three transects established from the base of foredunes to the waterline. On Una beach, E. armata showed continuous reproduction with higher abundances of ovigerous females in winter and spring (July-November) with a higher peak of juveniles in spring (November 2004). The fecundity ranged from 2 to 18 eggs/embryos per female, depending on the female length. The incubation period was estimated as 2 months. The life span of males and females was nearly 1 year. The short life span and the high energetic expenditure inherent to reproduction with maternal care, probably kept females from producing more than one brood in their lifetime. When comparing the population of E. armata on Una beach (24 degrees S) with populations in Southern Brazil (32 degrees S), Uruguay (34 degrees S) and Argentina (36 degrees S), it was verified that several biological population traits (length of the smallest juvenile, length of the largest individual, length of the smallest and largest ovigerous females, range of fecundity and life span) tended to increase at higher latitudes, whereas other traits (instantaneous rate of mortality and the curvature parameter of von Bertalanffy growth function) tended to decrease. However, comparing E. armata on Una beach (24 degrees S) with a population situated at a close latitude (25 degrees S), unexpected differences in relation to population structure and to growth demonstrated and reinforced the importance of density-dependent factors over life history traits of E. armata on dissipative beaches.

Prediction of apparent protein digestibility of ingredients and diets by in vitro pH-stat degree of protein hydrolysis with species-specific enzymes for juvenile Pacific white shrimp Litopenaeus vannamei

Aquafeed production faces global issues related to availability of feed ingredients. Feed manufacturers require greater flexibility in order to develop nutritional and cost-effective formulations that take into account nutrient content and availability of ingredients. The search for appropriate ingredients requires detailed screening of their potential nutritional value and variability at the industrial level. In vitro digestion of feedstuffs by enzymes extracted from the target species has been correlated with apparent protein digestibility (APD) in fish and shrimp species. The present study verified the relationship between APD and in vitro degree of protein hydrolysis (DH) with Litopenaeus vannamei hepatopancreas enzymes in several different ingredients (n = 26): blood meals, casein, corn gluten meal, crab meal, distiller`s dried grains with solubles, feather meal, fish meals, gelatin, krill meals, poultry by-product meal, soybean meals, squid meals and wheat gluten. The relationship between APD and DH was further verified in diets formulated with these ingredients at 30% inclusion into a reference diet. APD was determined in vivo (30.1 +/- 0.5 degrees C, 32.2 +/- 0.4%.) with juvenile L vannamei (9 to 12 g) after placement of test ingredients into a reference diet (35 g kg(-1) CP: 8.03 g kg(-1) lipid; 2.01 kcal g(-1)) with chromic oxide as the inert marker. In vitro DH was assessed in ingredients and diets with standardized hepatopancreas enzymes extracted from pond-reared shrimp. The DH of ingredients was determined under different assay conditions to check for the most suitable in vitro protocol for APD prediction: different batches of enzyme extracts (HPf5 or HPf6), temperatures (25 or 30 degrees C) and enzyme activity (azocasein): crude protein ratios (4 U: 80 mg CP or 4 U: 40 mg CP). DH was not affected by ingredient proximate composition. APD was significantly correlated to DH in regressions considering either ingredients or diets. The relationships between APD and DH of the ingredients could be suitably adjusted to a Rational Function (y = (a + bx)/(1 + cx + dx2), n = 26. Best in vitro APD predictions were obtained at 25 degrees C, 4 U: 80 mg CP both for ingredients (R(2) = 0.86: P = 0.001) and test diets (R(2) = 0.96; P = 0.007). The regression model including all 26 ingredients generated higher prediction residuals (i.e., predicted APD - determined APD) for corn gluten meal, feather meal. poultry by-product meal and krill flour. The remaining test ingredients presented mean prediction residuals of 3.5 points. A model including only ingredients with APD>80% showed higher prediction precision (R(2) = 0.98: P = 0.000004; n = 20) with average residual of 1.8 points. Predictive models including only ingredients from the same origin (e.g., marine-based, R(2) = 0.98; P = 0.033) also displayed low residuals. Since in vitro techniques have been usually validated through regressions against in vivo APD, the DH predictive capacity may depend on the consistency of the in vivo methodology. Regressions between APD and DH suggested a close relationship between peptide bond breakage by hepatopancreas digestive proteases and the apparent nitrogen assimilation in shrimp, and this may be a useful tool to provide rapid nutritional information. (C) 2009 Elsevier B.V. All rights reserved.

Are the preference and selection patterns of hermit crabs for gastropod shells species- or site-specific?

Experimental analyses of hermit crabs and their preferences for shells are essential to understand the intrinsic relationship of the crabs` dependence on shells, and may be useful to explain their shell use pattern in nature. The aim of this study was to evaluate the effect of crab species and site on the pattern of shell use, selection, and preference in the south-western Atlantic hermit crabs Pagurus brevidactylus and Pagurus criniticornis, comparing sympatric and allopatric populations. Differently from the traditional approach to evaluate shell preference by simply determining the shell selection pattern (i.e., the number of shells of each type selected), preference was defined (according to [Liszka, D., Underwood, AJ., 1990. An experimental design to determine preferences for gastropod shells by a hermit-crab. J. Exp. Mar. Biol. Ecol., 137(1), 47-62]) by the comparison of the number of crabs changing for a particular shell type when three options were given (Cerithium atratum, Morula nodulosa, and Tegula viridula) with the number of crabs changing for this same type when only this type was offered. The effect of crab species was tested at Cabelo Gordo Beach, where P. brevidacrylus was found occupying shells of C. atratum, M. nodulosa, and T viridula in similar frequencies, whereas P. criniticornis occupied predominantly shells of C atratum. In laboratory experiments the selection patterns of the two hermit-crab species for these three gastropods were different, with P criniticornis selecting mainly shells of C atratum, and R brevidactylus selecting more shells of M. nodulosa. The shell preference was also dependent on crab species, with P. criniticornis showing a clear preference for shells of C atratum, whereas P. brevidactylus did not show a preference for any of the tested shells. The effect of site was tested for the two species comparing data from Cabelo Gordo to Preta (P brevidactylus) and Araca beaches (P. criniticornis). The pattern of shell use, selection, and preference was demonstrated to be dependent on site only for P. brevidactylus. The results also showed that the shell use pattern of P criniticornis can be explained by its preference at both sites, whereas for P. brevidactylus it occurred only at Cabelo Gordo, where the absence of preference was correlated with the similar use of the three gastropod species studied. Finally, the results showed that the shell selection pattern cannot be considered as a measure of shell preference, since it overestimates crab selectivity. (C) 2009 Elsevier B.V. All rights reserved.

Development stages of the reproductive tract of male of Panulirus echinatus Smith (Decapoda : Palinuridae)

This paper characterizes the developmental stages of the testes and vasa deferentia of the Panulirus echinatus Smith, 1869 through comparisons between microscopic findings, macroscopic aspects, and gonadosomatic index (GSR). The lobsters were sampled monthly (November 1999 to October 2000) using seine nets and a total of 1716 males were obtained at Tamandare Bay. Each carapace was cut to allow evaluation of the reproductive organs; the testes and vasa deferentia were dissected, weighed, fixed in Bouin`s solution up to 12 hours and submitted for histological analysis to determine the presence and/or absence of spermatozoa. These measures, along with change in color, size, diameter, development of the spermatophores and the GSR allowed the caracterization of three development stages: immature, intermediate and ripe. In conclusion, the maturity of the testes precedes the maturity of the vasa deferentia. To evaluate if gonadosomatic relation was a good quantitative indicator of the maturity stage, t tests (alpha = 0,05) were used and verified significant difference in the averages of GSR. The statistics corroborated that GSR can be used as indicative of the developmental stages for P. echinatus.

Investigating South American biogeographic history using patterns of skull shape variation on Cerdocyon thous (Mammalia: Canidae)

Patterns of geographic variation of the canid Cerdocyon thous have historically been obscured by its remarkable intraspecific morphological variability. The observed distribution is highly associated with phytophysiognomy, a feature considered highly dynamic along geological time. In the present study, we tested whether vegetation distribution during the Holocene Glacial Maximum of South America (HGM) explains the patterns of morphological variation within Cerdocyon thous. The species was divided in groups according to paleohabitats that could support their presence during the HGM, and then tested for differences in skull morphometrics. The results obtained demonstrate that the climatic changes during the HGM influenced the population structure of this species, resulting in the establishment of geographical groups with different degrees of morphological cohesion. Higher morphological cohesion found in the Northern group might be explained by the marked discontinuity between its geographical range and the rest of the species`distribution. The Eastern and Southern morphological divergence is less striking and, although this could be related to past vegetation distribution, the disappearance of those barriers leads to a population structure that could be slowly breaking down. (C) 2009 The Linnean Society of London, Biological Journal of the Linnean Society, 2009, 98, 77-84.

Kinetic Analysis of Gill (Na+,K+)-ATPase Activity in Selected Ontogenetic Stages of the Amazon River Shrimp, (Decapoda, Palaemonidae): Interactions at ATP- and Cation-Binding Sites

We investigated modulation by ATP, Mg2+, Na+, K+ and NH4 (+) and inhibition by ouabain of (Na+,K+)-ATPase activity in microsomal homogenates of whole zoeae I and decapodid III (formerly zoea IX) and whole-body and gill homogenates of juvenile and adult Amazon River shrimps, . (Na+,K+)-ATPase-specific activity was increased twofold in decapodid III compared to zoea I, juveniles and adults, suggesting an important role in this ontogenetic stage. The apparent affinity for ATP ( (M) = 0.09 +/- A 0.01 mmol L-1) of the decapodid III (Na+,K+)-ATPase, about twofold greater than the other stages, further highlights this relevance. Modulation of (Na+,K+)-ATPase activity by K+ also revealed a threefold greater affinity for K+ ( (0.5) = 0.91 +/- A 0.04 mmol L-1) in decapodid III than in other stages; NH4 (+) had no modulatory effect. The affinity for Na+ ( (0.5) = 13.2 +/- A 0.6 mmol L-1) of zoea I (Na+,K+)-ATPase was fourfold less than other stages. Modulation by Na+, Mg2+ and NH4 (+) obeyed cooperative kinetics, while K+ modulation exhibited Michaelis-Menten behavior. Rates of maximal Mg2+ stimulation of ouabain-insensitive ATPase activity differed in each ontogenetic stage, suggesting that Mg2+-stimulated ATPases other than (Na+,K+)-ATPase are present. Ouabain inhibition suggests that, among the various ATPase activities present in the different stages, Na+-ATPase may be involved in the ontogeny of osmoregulation in larval The NH4 (+)-stimulated, ouabain-insensitive ATPase activity seen in zoea I and decapodid III may reflect a stage-specific means of ammonia excretion since functional gills are absent in the early larval stages.

PCBs, PBDEs and organochlorine pesticides in crabs Hepatus pudibundus and Callinectes danae from Santos Bay, State of Sao Paulo, Brazil

The occurrence of persistent organic pollutants (POPS) as polychlorinated biphenyls (PCBs), organochlorine pesticides (OCPs) and polybrominated diphenyl ethers (PBDE) in crabs Hepatus pudibundus and Callinectes danae was assessed from two different places inside of the Santos Bay and Moela Island near one of the most economically important metropolitan areas in Southern Brazil. Among POPs analyzed, Sigma PCBs (222-923 ng g(-1) lipid weight) and Sigma DDTs (154-410 ng g(-1) lw) exhibited the highest concentrations in the crabs. Sigma HCHs ranged from 10.3 to 30.9 ng g(-1), lw and were found in all individuals. Other OCPs found in lower concentration was Mirex (7.6-41.6 ng g(-1) lw) and HCB (5.83-16.9 ng g(-1) lw). Sigma PBDEs (24.1 ng g(-1) lw) were only found in one male individual from the species C. danae collected near to the submarine sewage of Santos. Male crabs showed higher POP concentrations than female crabs for those two species. (C) 2011 Elsevier Ltd. All rights reserved.

Illustrated key for the identification of the known zoeal stages of brachyuran crabs (Crustacea: Decapoda) from tropical and subtropical Brazil, southwestern Atlantic

Approximately 370 brachyuran species have so far been recorded from the Brazilian coast, 123 of which have had their larval stages fully or partially described. The pictorial guide allows the identification of the first zoea of 110 species. The remaining 13 species with known larval stages are treated to the genus level because of difficulties in the morphological differentiation of closely related species.

Peptide fingerprinting of the neurotoxic fractions isolated from the secretions of sea anemones Stichodactyla helianthus and Bunodosoma granulifera. New members of the APETx-like family identified by a 454 pyrosequencing approach

Sea anemones are known to contain a wide diversity of biologically active peptides, mostly unexplored according to recent peptidomic and transcriptomic studies. In the present work, the neurotoxic fractions from the exudates of Stichodactyla helianthus and Bunodosoma granulifera were analyzed by reversed-phase chromatography and mass spectrometry. The first peptide fingerprints of these sea anemones were assessed, revealing the largest number of peptide components (156) so far found in sea anemone species, as well as the richer peptide diversity of B. granulifera in relation to S. helianthus. The transcriptomic analysis of B. granulifera, performed by massive cDNA sequencing with 454 pyrosequencing approach allowed the discovery of five new APETx-like peptides (U-AITX-Bg1a-e - including the full sequences of their precursors for four of them), which together with type 1 sea anemone sodium channel toxins constitute a very distinguishable feature of studied sea anemone species belonging to genus Bunodosoma. The molecular modeling of these new APETx-like peptides showed a distribution of positively charged and aromatic residues in putative contact surfaces as observed in other animal toxins. On the other hand, they also showed variable electrostatic potentials, thus suggesting a docking onto their targeted channels in different spatial orientations. Moreover several crab paralyzing toxins (other than U-AITX-Bg1a-e), which induce a variety of symptoms in crabs, were isolated. Some of them presumably belong to new classes of crab-paralyzing peptide toxins, especially those with molecular masses below 2 kDa, which represent the smallest peptide toxins found in sea anemones. (C) 2011 Elsevier Inc. All rights reserved.

Tributyltin in Crustacean Tissues: Analytical Performance and Validation of Method

The hermit crab Clibanarius vittatus is a typical organism from intertidal regions being considered as a good bioindicator of tributyltin presence at these environments. Thus this study presents the analytical performance and validation method for TBT quantification in tissues of C. vittatus by gas chromatography with pulsed flame photometric detector (GC-PFPD) after extraction with an apolar solvent (toluene) and Grignard derivatization. The limits of detection of the method (LOD) were 2.0 and 2.8 ng g(-1) for TBT and DBT (dibutyltin), respectively, and its limits of quantification (LOQ) were 6.6 and 8.9 ng g(-1) for TBT and DBT, respectively. The method was applied to samples from Santos Estuary, Sao Paulo State, Brazil. TBT and DBT concentrations ranged from 26.7 to 175.0 ng g(-1) and from 46.2 to 156.0 ng g(-1), respectively. These concentrations are worrisome since toxic effects (such as endocrine disruption) have been reported for other organisms even under lower levels of registred at this study.

Effects of tributyltin exposure in hermit crabs: Clibanarius vittatus as a model

Tributyltin (TBT) contamination affects the reproductive system of many species of invertebrates worldwide. The present study was designed to evaluate the effects of exposure to TBT pollution on the reproduction of the hermit crab Clibanarius vittatus. An orthogonal experiment was designed with two treatments: contamination (with or without TBT in the food) and crab sex (males and females). The animals were reared in the laboratory for nine months, and macroscopic and histological analyses of reproductive organs were carried out after the end of the experiment. Tributyltin was recorded in exposed crabs, but no morphological alterations were detected in the gonads of males, regardless of whether they were exposed to TBT. In contrast, females exposed to TBT displayed disorganization and atrophy of their ovaries, thus directly affecting reproduction in this hermit crab species. This effect observed in female hermit crabs may harm populations located in harbor regions, where TBT concentration is high, even after the worldwide TBT ban.

Signaling Events During Cyclic Guanosine Monophosphate-Regulated Pigment Aggregation in Freshwater Shrimp Chromatophores

Crustacean color change results partly from granule aggregation induced by red pigment concentrating hormone (RPCH). In shrimp chromatophores, both the cyclic GMP (3', 5'-guanosine monophosphate) and Ca2+ cascades mediate pigment aggregation. However, the signaling elements upstream and downstream from cGMP synthesis by GC-S (cytosolic guanylyl cyclase) remain obscure. We investigate post-RPCH binding events in perfused red ovarian chromatophores to disclose the steps modulating cGMP concentration, which regulates granule translocation. The inhibition of calcium/calmodulin complex (Ca2+/CaM) by N-(6-aminohexyl)-5-chloro-1-naphthalenesulphonamide (W7) induces spontaneous aggregation but inhibits RPCH-triggered aggregation, suggesting a role in pigment aggregation and dispersion. Nitric oxide synthase inhibition by N omega-nitro-L-arginine methyl ester hydrochloride (L-NAME) strongly diminishes RPCH-induced aggregation; protein kinase G inhibition (by rp-cGMPs-triethylamine) reduces RPCH-triggered aggregation and provokes spontaneous dispersion, disclosing NO/PKG participation in aggregation signaling. Myosin light chain phosphatase inhibition (by cantharidin) accelerates RPCH-triggered aggregation, whereas Rho-associated protein kinase inhibition (by Y-27632, H-11522) reduces RPCH-induced aggregation and accelerates dispersion. MLCP (myosin light chain kinase) and ROCK (Rho-associated protein kinase) may antagonistically regulate myosin light chain (MLC) dephosphorylation/phosphorylation during pigment dispersion/aggregation. We propose the following general hypothesis for the cGMP/Ca2+ cascades that regulate pigment aggregation in crustacean chromatophores: RPCH binding increases Ca2+ (int), activating the Ca2+/CaM complex, releasing NOS-produced nitric oxide, and causing GC-S to synthesize cGMP that activates PKG, which phosphorylates an MLC activation site. Myosin motor activity is initiated by phosphorylation of an MLC regulatory site by ROCK activity and terminated by MLCP-mediated dephosphorylation. Qualitative comparison reveals that this signaling pathway is conserved in vertebrate and invertebrate chromatophores alike.

Alien shrimps in evidence: new records of the genus Athanas Leach, 1814 on the coast of So Paulo, southern Brazil (Caridea: Alpheidae)

The occurrence of two alien alpheid shrimps of the genus Athanas Leach, 1814 [in Leach 1813-1814], the Indo-West Pacific A. dimorphus Ortmann, 1894 and the Eastern Atlantic A. nitescens (Leach, 1813 [in Leach 1813-1814]), on the coast of the state of So Paulo, Brazil, is reported. The presence of A. dimorphus extends the range of this species in the western Atlantic farther to the south in Brazil, whereas A. nitescens is reported for the first time in the western Atlantic, representing the second alien alpheid species on this side of the Atlantic and the twenty-first decapod crustacean introduced in Brazil. We provide morphological accounts of the material examined and illustrate the most important diagnostic characters of both species. An overview of the possible mechanisms of their introduction on the coast of So Paulo is also provided.

Is shell partitioning between the hermit crabs Pagurus brevidactylus and Pagurus criniticornis explained by interference and/or exploitation competition?

The effect of crab behaviour on shell-use dynamics was analysed, comparing both interference and exploitation behaviours between the hermit crabs Pagurus criniticornis and Pagurus brevidactylus. Although these species exhibited microhabitat separation, with P. criniticornis dominating (100%) in sandy substrates and P. brevidactylus (80%) on rocky shores, they overlapped in the rocky shore/sand interface (P. criniticornis, 53%; P. brevidactylus, 43%). Pagurus criniticornis occupied shells of Cerithium atratum in higher frequencies (84%) than P. brevidactylus (37%), which was hypothesized to be a consequence of competitive interactions combined with their ability to acquire and/or retain this resource. The species P. criniticornis was attracted in larger numbers to simulated gastropod predation events than was P. brevidactylus, which, on the few occasions that it moved before P. criniticornis, tended to be attracted more rapidly. Interspecific shell exchanges between these species were few, suggesting the absence of dominance relationships. The shell-use pattern in this species pair is thus defined by exploitation competition, which is presumed to be intensified in areas of microsympatry. These results differ from other studies, which found that interference competition through interspecific exchanges shapes shell use, indicating that shell partitioning in hermit crabs is dependent on the behaviour of the species involved in the contests.

Hermit crabs as bioindicators of recent tributyltin (TBT) contamination

Evaluation of the assimilation pathway and depuration time of a given pollutant by aquatic species is important to understand the dynamics of this substance in the biota, and to search for potential ecological indicators. In the present study, the uptake pathway and depuration time and rate of the pollutant tributyltin (TBT) were investigated in the omnivorous hermit crab (Clibanarius vittatus). The assimilation and uptake pathway were investigated using hermit crabs collected in an area free of TBT. The crabs were held in the laboratory for 45 days, under one of four treatments: procedural control (PC) - water and food without TBT; T1 - water with and food without TBT; 12 - water without and food with Tiff; and 13 water and food with TBT. To determine the depuration time, the crabs were collected in a contaminated area, maintained in the laboratory with clean water, and removed every 15 days for 120 days. The concentrations of TBT and DBT (dibutyltin) were determined by chromatographic analysis. The TBT was taken up by the crabs mainly via food, and the presence of DBT in crab tissues was hypothesized to result from internal TBT degradation. TBT (as well as DBT) was depurated rapidly by C. vittatus. After approximately 30 days, the initial concentration of 111 +/- 36 ng Sn g(-1) w. w. decreased to 3 +/- 3 ng Sn g(-1) w. w., and after 75 days the TBT concentration was below the detection limit. The same pattern was recorded for DBT, which showed a higher deputation rate than TBT. The rapid TBT and DBT deputation is useful information, since C. vittatus and possibly other hermit crabs may be used as indicators of recent or recycled environmental contamination.