3-Hydroxypropionic Acid as an Antibacterial Agent from Endophytic Fungi Diaporthe phaseolorum

Endophytic fungi are considered a rich source of active compounds resulting from their secondary metabolism. Fungi from marine environment grow in a habitat with unique conditions that can contribute to the activation of metabolic pathways of synthesis of different unknown molecules. The production of these compounds may support the adaptation and survival of the fungi in the marine ecosystem. Mangroves are ecosystems situated between land and sea. They are frequently found in tropical and subtropical areas and enclose approximately 18.1 million hectares of the planet. The great biodiversity found in these ecosystems shows the importance of researching them, including studies regarding new compounds derived from the endophytic fungi that inhabit these ecosystems. 3-hydroxypropionic acid (3-HPA) has been isolated from the mangrove endophytic fungus Diaporthe phaseolorum, which was obtained from branches of Laguncularia racemosa. The structure of this compound was elucidated by spectroscopic methods, mainly 1D and 2D NMR. In bioassays, 3-HPA showed antimicrobial activities against both Staphylococcus aureus and Salmonella typhi. The structure of this antibiotic was modified by the chemical reaction of Fischer-Speier esterification to evaluate the biologic activity of its chemical analog. The esterified product, 3-hydroxypropanoic ethyl ester, did not exhibit antibiotic activity, suggesting that the free carboxylic acid group is important to the pharmacological activity. The antibiotic-producing strain was identified with internal transcribed spacer sequence data. To the best of our knowledge, this is the first report of antibacterial activity by 3-HPA against the growth of medically important pathogens.

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.

The Dynamics of a frictionally-dominated Amazonian estuary

The hydrodynamics, morphology and sedimentology of the Taperacu estuary were investigated. This is one of several estuaries located within the largest mangrove fringe in the world, bordering the Amazon region, subject to a macrotidal regime and regionally atypical negligible fresh water supply. The results reveal widespread sand banks that occupy the central portion of the estuarine cross-section. Well-sorted very fine sandy sediments of marine origin prevail. Shorter flood phases, with substantially higher current velocities, were observed in the upper sector of Taperacu, as expected for a shallow, friction-dominated estuary. However, ebb domination can be expected for estuaries with large associated mangrove areas and substantial estuarine infilling, both of which situations occur on the Taperacu. The tidal asymmetry favoring flood currents could be the result of the absence of an effective fluvial discharge. Furthermore, it was observed that the Taperacu is connected by tidal creeks to the neighboring Caete estuary, allowing a stronger flux during the flood and intensifying the higher flood currents. As a whole, the results have shown a complex interaction of morphological aspects (friction, fluvial drainage, connections with neighbor estuaries, infilling and large storage area) in determining hydrodynamic patterns, thus improving the understanding of Amazon estuaries.

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.

Reproductive biology of the ghost shrimp Lepidophthalmus bocourti (A. Milne-Edwards, 1870) (Decapoda: Axiidea: Callianassidae): A tropical species with a seasonal reproduction

Based on the hypothesis that reproduction is a continuous process in tropical habitats, we analysed reproductive periodicity and egg production in the callianassid ghost shrimp Lepidophthalmus bocourti, one of the most common species in mangrove systems along the Pacific coast of Central America. During one year (May 2008 to April 2009), individuals of L. bocourti (N = 499) were collected nearshore Gulf of Nicoya, Pacific coast of Costa Rica. Observations were made on presence or absence of incubated embryos, and gonad activity of females was analysed as gonadosomatic index (GSI). Our results revealed that L. bocourti has a marked seasonal breeding period, which contradicts previous reports regarding coastal marine decapods from the tropics. Ovigerous females were found only from June to August, while high GSI values were obtained from March to July. The increase of GSI and appearance of ovigerous females were associated with a concomitant decrease of salinity, but not with temperature. We assume that reproduction of L. bocourti is adapted to local changes of environmental conditions, and that a decrease in salinity during rainy season may serve as a triggering factor for ovarian development. Compared to other ghost shrimps, L. bocourti produced on average more (2002 +/- 1365) and smaller (0.87 +/- 0.109 mm) eggs, which seems to suggest that this species does not have an abbreviated larval development as reported for other species of genus. The deviation from the generalization of constant reproduction in the tropics for shallow water marine invertebrates and its probable cause are adequately discussed.

Brazilian research on extremophiles in the context of astrobiology

Extremophiles are organisms adapted to grow at extreme ranges of environmental variables, such as high or low temperatures, acid or alkaline medium, high salt concentration, high pressures and so forth. Most extremophiles are micro-organisms that belong to the Archaea and Bacteria domains, and are widely spread across the world, which include the polar regions, volcanoes, deserts, deep oceanic sediments, hydrothermal vents, hypersaline lakes, acid and alkaline water bodies, and other extreme environments considered hostile to human life. Despite the tropical climate, Brazil has a wide range of ecosystems which include some permanent or seasonally extreme environments. For example, the Cerrado is a biome with very low soil pH with high Al+3 concentration, the mangroves in the Brazilian coast are anaerobic and saline, Pantanal has thousands of alkaline-saline lakes, the Caatinga arid and hot soils and the deep sea sediments in the Brazilian ocean shelf. These environments harbour extremophilic organisms that, coupled with the high natural biodiversity in Brazil, could be explored for different purposes. However, only a few projects in Brazil intended to study the extremophiles. In the frame of astrobiology, for example, these organisms could provide important models for defining the limits of life and hypothesize about life outside Earth. Brazilian microbiologists have, however, studied the extremophilic micro-organisms inhabiting non-Brazilian environments, such as the Antarctic continent. The experience and previous results obtained from the Brazilian Antarctic Program (PROANTAR) provide important results that are directly related to astrobiology. This article is a brief synopsis of the Brazilian experience in researching extremophiles, indicating the most important results related to astrobiology and some future perspectives in this area. Received 29 February 2012, accepted 25 May 2012, first published online 11 July 2012

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.