Laccase production in bioreactor scale under saline condition by the marine-derived basidiomycete Peniophora sp. CBMAI 1063

Laccase production in saline conditions is still poorly studied. The aim of the present study was to investigate the production of laccase in two different types of bioreactors by the marine-derived basidiomycete Peniophora sp. CBMAI 1063. The highest laccase activity and productivity were obtained in the Stirred Tank (ST) bioreactor, while the highest biomass concentration in Air-lift (AL) bioreactor. The main laccase produced was purified by ion exchange and size exclusion chromatography and appeared to be monomeric with molecular weight of approximately 55 kDa. The optimum oxidation activity was obtained at pH 5.0. The thermal stability of the enzyme ranged from 30 to 50 °C (120 min). The Far-UV Circular Dichroism revealed the presence of high β-sheet and low α-helical conformation in the protein structure. Additional experiments carried out in flask scale showed that the marine-derived fungus was able to produce laccase only in the presence of artificial seawater and copper sulfate. Results from the present study confirmed the fungal adaptation to marine conditions and its potential for being used in saline environments and/or processes.     

Evaluating the role of morphological characters in the phylogeny of some trypanosomatid genera (Excavata,Kinetoplastea, Trypanosomatida)

Over the last three decades, it has been progressively assumed that morphology has become obsolete for trypanosomatid systematics. Traditional taxonomy, based on the occurrence of specific kinds of cell morphotypes during life cycles and the morphometry of such cells, is often rejected by molecular phylogenies inferred mostly from 18S rDNA alone or combined with GAPDH. In such context, we hypothesized the affinities of 35 representatives of seven trypanosomatid genera from separated and combined cladistics analyses of morphological and 18S matrices. Morphology is shown to be more consistent and to have stronger synapomorphy retention than the 18S data. The strict consensus of cladograms from separated analyses was mostly unresolved, while combined analysis produced a meaningful and robust phylogenetic pattern, as evidenced by partition congruence index, Bremer support and frequencies of groups present/contradicted. The results (1) corroborate the separation of Angomonas and Strigomonas from Crithidia, which is now shown to be monophyletic, (2) support the revalidation of the genus Wallaceina, and (3) place the genera Herpetomonas, Leptomonas and Phytomonas within a single clade. Overall, we demonstrate the belief that morphological characters are inferior to molecular ones for trypanosomatid systematics is a consequence of neglecting their inclusion in phylogenetic analyses.     

Spin-labeled extracellular loop from a seven-transmembrane helix receptor: studies in solution and interaction with model membranes

A spin-labeled pentadecapeptide was synthesized containing 2,2,6,6-tetramethylpiperidine-N-oxyl-4-amino-4-carboxylic acid (TOAC) as the N-terminal amino acid and residues 253-266 (EYWSTFGNLHHISL) of the mass oncogene receptor, a membrane-bound protein from the G-protein coupled receptors family. According to predictions, this protein folds into seven transmembrane helices connected by three extra- and three intracellular loops, and the peptide encompasses part of the third extracellular loop and part of the seventh helix. Electron paramagnetic resonance (EPR) spectra of the spin-labeled peptide (TOAC-14) were obtained in aqueous solution as a function of pH and temperature, in a secondary structure-inducing solvent [trifluoroethanol (TFE)], and in the presence of detergent micelles and phospholipid bilayers. The charged and uncharged amino groups of TOAC and TOAC-14 yielded spectra with different isotropic hyperfine splittings (aN). The slow exchange between protonated and unprotonated forms in the EPR time scale gave rise to composite spectra weighted by the Henderson-Hasselbalch equation. Plots of aN vs pH allowed the determination of the amino group pK values (8.4 and 4.5, for TOAC and TOAC-14, respectively). A small change in aN centered at pH 6.5 was ascribed to the titration of the histidines. Values of calculated rotational correlation times were indicative of a pH-induced conformational change. A conformational change was also observed in TFE. TOAC-14 bound to micelles irrespective of peptide and detergent head group charge. In contrast, the peptide bound to phospholipid bilayers only when both carried opposite charges. The slow exchange (in the EPR time scale) between membrane-bound and free TOAC-14 allowed the calculation of the peptide's partition coefficient. The spectral line shapes were affected by aggregate size and degree of packing of the constituent molecules. It is proposed that pH, polarity, and lipid environment can affect the conformation of water-exposed regions of membrane-bound receptors, thereby playing a role in the mechanism of signal transduction.     

Responses of a Specialized Insectivorous Mammal (Myrmecophaga tridactyla) to Variation in Ambient Temperature1

This study reports the effect of daily mean ambient temperature on daily activity and habitat use by the giant anteater (Myrmecophaga tridactyla) in the Pantanal wetlands of Brazil. We equipped 11 giant anteaters with a global position system collar programmed to acquire a location every 10 min for 15 d. A temperature data logger left in the meteorological station at the study site registered data on ambient temperature. Giant anteaters were mainly active in open fields except during the coldest days, when they sought protection inside forests. Giant anteaters also used open habitats for resting but during the hottest or chilliest days they sheltered in forests. In the Pantanal, where temperatures are usually high, giant anteaters were active all night long, but as daily average ambient temperature decreased anteaters began and finished activity progressively earlier and reduced total activity. As a consequence, time spent active at daylight increased progressively and time spent active during the night decreased progressively, probably to allow the anteaters to expose themselves to solar radiation and to avoid heat loss during the night.     

Stochastic simulation of the gas diffusion in the air phase of the human lung

The solution of the diffusion equation in the gas phase of the human lung is very difficult because of the structure of the bronchial tree. It is shown by means of physical arguments, how one can reduce the diffusion equation to a simple one-dimensional form. The solution is then obtained by a stochastic simulation, which is easily realized on a digital computer.     

Interacting effects of spatial gradients and fishing gears on characterization of fish assemblages in large reservoirs

Understanding the variation of fish assemblages in reservoirs is crucial for precise interpretations of ecological processes. However, representative samples in such environments are subjected both to spatial zonation (longitudinal and transversal gradients) and limitations of fishing gears. We used a large zoned reservoir (Itaipu, Brazil) to perform a quasi-experiment that assessed the relative role of spatial gradients and fishing methods on fish diversity and composition. Active (cast nets, electrofishing and seines) and passive (gillnets and longlines) fishing gears were employed in lotic, transitional and lentic habitats of the reservoir and three large tributaries. In total 6281 individuals of 101 species were captured. Sampling methods and spatial gradients had an influence on species composition, with significant interactions between these factors. In addition, results produced by a variation partitioning analysis suggested that 21 % of the species richness was explained by the sampling methods and 7 % by the longitudinal gradient; whereas for species composition, 12 and 3 % corresponded to sampling methods and transversal gradients, respectively. Overall, our results suggest that variations among fishing gears is greater than across strong spatial gradients, emphasizing the importance of using multiple and complementary fishing methods in reservoirs for characterization of fish assemblages. The interactions between spatial gradients and sampling methods were higher with transversal than with longitudinal gradients, suggesting that substantial differences in species richness and composition among sampling gears depend to some extent on habitat. The approach used here would be applicable to any large zoned inland waterscape.     

Distribution,habitat and trophic ecology of Antarctic squid Kondakovia longimana and Moroteuthis knipovitchi: inferences from predators and stable isotopes

Cephalopods have a key role in the marine environment though knowledge of their distribution and trophic ecology is limited by a lack of observations. This is particularly true for Antarctic species. Toothfish species are key predators of cephalopods and may be viewed as ideal biological samplers of these species. A total of 256 cephalopod lower beaks were identified from the stomachs of Patagonian toothfish (Dissostichus eleginoides) and Antarctic toothfish (Dissostichus mawsoni), captured in fisheries of South Georgia and the South Sandwich Islands in the South Atlantic between March and April 2009. Long-armed octopus squid (Kondakovia longimana) and smooth-hooked squid (Moroteuthis knipovitchi) were the main cephalopod prey and both were predated upon wherever toothfish were captured, though this cephalopod species appear to inhabit deeper waters at the South Sandwich Islands than at South Georgia. Measurements of δ¹³C from beak material indicated a clear segregation of habitat use comparing adult and sub-adult sized K. longimana. Variation in δ¹⁵N with size indicated an ontogenetic shift in the diet of cephalopods and also suggested some trophic plasticity among years. This study provides new insights into the private life of some elusive Antarctic cephalopods in an underexplored region of the South Atlantic.     

The molecular phylogeny of the sea star Echinaster (Asteroidea: Echinasteridae) provides insights for genus taxonomy

In this study, we used sequences of two mitochondrial genes, cytochrome c oxidase I (COI) and 16S rRNA, and one nuclear gene, 28S rRNA, to test the monophyly of the sea star genus Echinaster, and understand the phylogenetic relationships among species and subgenera within this genus. Phylogenetic analyses based on Bayesian inference and maximum likelihood methods revealed three clades with high values of genetic divergence among them (K2P distances for COI over 23%). One of the clades grouped all Echinaster (Othilia) species, and the other two clades included Echinaster (non‐Othilia) species and Henricia species, respectively. Although the relationships among Henricia, Othilia, and Echinaster could not be completely clarified, the Othilia clade was a well‐supported group with shared diagnostic morphological characters. Moreover, the approximately unbiased test applied to the phylogenetic reconstruction rejected the hypothesis of the genus Echinaster as a monophyletic group. According to these results, we suggest the revalidation of Othilia as a genus instead of a subgenus within Echinaster. Our study clarifies important points about the phylogenetic relationships among species of Echinaster. Other important systematic questions about the taxonomic classification of Echinaster and Henricia still remain open, but this molecular study provides bases for future research on the topic.