Temporal Diversity Patterns and Phenology in Fruit-feeding Butterflies in the Atlantic Forest

The Atlantic Forest deserves special attention due to its high level of species endemism and degree of threat. As in other tropical biomes, there is little information about the ecology of the organisms that occur there. The objectives of this study were to verify how fruit-feeding butterflies are distributed through time, and the relation with meteorological conditions. Species richness and Shannon index were partitioned additively at the monthly level, and beta diversity, used as a hierarchical measure of temporal species turnover, was calculated among months, trimesters, and semesters. Circular analysis was used to verify how butterflies are distributed along seasons and its relation with meteorological conditions. We sampled 6488 individuals of 73 species. Temporal diversity of butterflies was more grouped than expected by chance among the months of each trimester. Circular analyses revealed that diversity is concentrated in hot months (September-March), with the subfamily Brassolinae strongly concentrated in February-March. Average temperature was correlated with total abundance of butterflies, abundance of Biblidinae, Brassolinae and Morphinae, and richness of Satyrinae. The present results show that 3mo of sampling between September and March is enough to produce a nonbiased sample of the local assemblage of butterflies, containing at least 70 percent of the richness and 25 percent of abundance. The influence of temperature on sampling is probably related to butterfly physiology. Moreover, temperature affects resource availability for larvae and adults, which is higher in hot months. The difference in seasonality patterns among subfamilies is probably a consequence of different evolutionary pressures through time.

Proteomics of the neurotoxic fraction from the sea anemone Bunodosoma cangicum venom: Novel peptides belonging to new classes of toxins

In contrast to the many studies on the venoms of scorpions, spiders, snakes and cone snails, tip to now there has been no report of the proteomic analysis of sea anemones venoms. In this work we report for the first time the peptide mass fingerprint and some novel peptides in the neurotoxic fraction (Fr III) of the sea anemone Bunodosoma cangicum venom. Fr III is neurotoxic to crabs and was purified by rp-HPLC in a C-18 column, yielding 41 fractions. By checking their molecular masses by ESI-Q-Tof and MALDI-Tof MS we found 81 components ranging from near 250 amu to approximately 6000 amu. Some of the peptidic molecules were partially sequenced through the automated Edman technique. Three of them are peptides with near 4500 amu belonging to the class of the BcIV, BDS-I, BDS-II, APETx1, APETx2 and Am-II toxins. Another three peptides represent a novel group of toxins (similar to 3200 amu). A further three molecules (similar to similar to 4900 amu) belong to the group of type 1 sodium channel neurotoxins. When assayed over the crab leg nerve compound action potentials, one of the BcIV- and APETx-like peptides exhibits an action similar to the type 1 sodium channel toxins in this preparation, suggesting the same target in this assay. On the other hand one of the novel peptides, with 3176 amu, displayed an action similar to potassium channel blockage in this experiment. In summary, the proteomic analysis and mass fingerprint of fractions from sea anemone venoms through MS are valuable tools, allowing us to rapidly predict the occurrence of different groups of toxins and facilitating the search and characterization of novel molecules without the need of full characterization of individual components by broader assays and bioassay-guided purifications. It also shows that sea anemones employ dozens of components for prey capture and defense. (C) 2008 Elsevier Inc. All rights reserved.

Revisiting cangitoxin, a sea anemone peptide: Purification and characterization of cangitoxins II and III from the venom of Bunodosoma cangicum

Sodium channel toxins from sea anemones are employed as tools for dissecting the biophysical properties of inactivation in voltage-gated sodium channels. Cangitoxin (CGTX) is a peptide containing 48 amino acid residues and was formerly purified from Bunodosoma cangicum. Nevertheless, previous works reporting, the isolation procedures for such peptide from B. cangicum secretions are controversial and may lead to incorrect information. In this paper, we report a simple and rapid procedure, consisting of two chromatographic steps, in order to obtain a CGTX analog directly from sea anemone venom. We also report a substitution of N16D in this peptide sample and the co-elution of an inseparable minor isoform presenting the R14H substitution. Peptides are named as CGTX-II and CGTX-III, and their effects over Nav1.1 channels in patch clamp experiments are demonstrated. (c) 2008 Elsevier Ltd. All rights reserved.

Bunodosine 391: An Analgesic Acylamino Acid from the Venom of the Sea Anemone Bunodosoma cangicum

A new acylamino acid, bunodosine 391 (BDS 391), was isolated from the venom of the sea anemone Bunodosoma cangicum. The structure was elucidated by spectroscopic analyses (2D NMR, ESIMS/MS) and verified by its synthesis. Intraplantar injection of BDS 391 into the hind paw of a rat induced a potent analgesic effect. This effect was not altered by naloxone (an opioid receptor antagonist), but was completely reversed by methysergide (a serotonin receptor antagonist), indicating that the effect is mediated by activation of serotonin receptors:

Sequence and function of lysosomal and digestive cathepsin D-like proteinases of Musca domestica midgut

Musca domestica larvae display in anterior and middle midgut contents, a proteolytic activity with pH optimum of 3.0-3.5 and kinetic properties like cathepsin D. Three cDNAs coding for preprocathepsin D-like proteinases (ppCAD 1, ppCAD 2, ppCAD 3) were cloned from a M. domestica midgut cDNA library. The coded protein sequences included the signal peptide, propeptide and mature enzyme that has all conserved catalytic and substrate binding residues found in bovine lysosomal cathepsin D. Nevertheless, ppCAD 2 and ppCAD 3 lack the characteristic proline loop and glycosylation sites. A comparison among the sequences of cathepsin D-like enzymes from some vertebrates and those found in M. domestica and in the genomes of Aedes aegypti, Drosophila melanogaster, Tribolium castaneum, and Bombyx mori showed that only flies have enzymes lacking the proline loop (as defined by the motif: DxPxPx(G/A)P), thus resembling vertebrate pepsin. ppCAD 3 should correspond to the digestive cathepsin D-like proteinase (CAD) found in enzyme assays because: (1) it seems to be the most expressed CAD, based on the frequency of ESTs found. (2) The mRNA for CAD 3 is expressed only in the anterior and proximal middle midgut. (3) Recombinant procathepsin D-like proteinase (pCAD 3), after auto-activation has a pH optimum of 2.5-3.0 that is close to the luminal pH of M. domestica midgut. (4) Immunoblots of proteins from different tissues revealed with anti-pCAD 3 serum were positive only in samples of anterior and middle midgut tissue and contents. (5) CAD 3 is localized with immunogold inside secretory vesicles and around microvilli in anterior and middle midguit cells. The data support the view that on adapting to deal with a bacteria-rich food in an acid midgut region, M. domestica digestive CAD resulted from the same archetypical gene as the intracellular cathepsin D, paralleling what happened with vertebrates. The lack of the proline loop may be somehow associated with the extracellular role of both pepsin and digestive CAD 3. (C) 2009 Elsevier Ltd. All rights reserved.

Wolbachia in Anastrepha Fruit Flies (Diptera: Tephritidae)

Endosymbiotic bacteria of the genus Wolbachia are widespread among arthropods and cause a variety of reproductive abnormalities, such as cytoplasmic incompatibility, thelytokous parthenogenesis, male-killing, and host feminization. In this study, we used three sets of Wolbachia-specific primers (16S rDNA, ftsZ, and wsp) in conjunction with the polymerase chain reaction (PCR), cloning and sequencing to study the infection of fruit flies (Anastrepha spp. and Ceratitis capitata) by Wolbachia. The flies were collected at several localities in Brazil and at Guayaquil, Ecuador. All of the fruit flies studied were infected with Wolbachia supergroup A, in agreement with the high prevalence of this group in South America. Phylogenetic analysis showed that the wsp gene was the most sensitive gene for studying the relationships among Wolbachia strains. The Wolbachia sequences detected in these fruit flies were similar to those such as wMel reported for other fruit flies. These results show that the infection of Anastrepha fruit flies by Wolbachia is much more widespread than previously thought.

Characterizations of juvenile stages of some semaeostome Scyphozoa (Cnidaria), with recognition of a new family (Phacellophoridae)

Phacellophora camtschatica has long been assigned to the semaeostome scyphozoan family Ulmaridae. Early stages (scyphistomae, strobilae, ephyrae, postephyrae, and young medusae) of the species were compared with those of several other semaeostomes currently assigned to Ulmaridae, Pelagiidae, and Cyaneidae. Juveniles of P. camtschatica did not strictly conform with characters of those of any of these families, and appeared intermediate between Cyaneidae and Ulmaridae. A new family, Phacellophoridae, is proposed to accommodate P. camtschatica.

New terebellids (Polychaeta: Terebellidae) from northeastern Brazil

Four new species of terebellid polychaetes were collected at the Reserva de Desenvolvimento Sustentavel da Ponta de Tubarao, State of Rio Grande do Norte, northeastern Brazil. These species are Streblosoma patriciae sp. nov., Terebella leslieae sp. nov., Nicolea ceciliae sp. nov., and Pista alonsae sp. nov. All these species are herein described and compared with the morphologically most similar congeners.

Is there any risk in a symbiotic species associating with an endangered one? A case of a phoronid worm growing on a Ceriantheomorphe tube

The conservation of emblematic threatened species is in highlight nowadays. Interestingly, few invertebrate groups attract scientific attention on this issue while they constitute the vast majority of animal biodiversity. Nevertheless, many invertebrate species are nowadays at risk of extinction. This means that plenty of species are currently disappearing out of sight. During a survey in the southwestern Atlantic Ocean tubes of an endangered species of cerianthid were sampled. This study reports for the very first time the occurrence of the species Phoronis australis in southwestern Atlantic waters and the association of phoronids with the genus Ceriantheomorphe. This raises questions on mutual extinction risks for symbiotic species and also on the criteria for their inclusion on Red Lists.

The taxonomic position of the pelagic `staurozoan` Tessera gemmaria as a ceriantharian larva

Based on 16 specimens from the Southwestern Atlantic coast (Argentina and Brazil) we reinterpret the taxonomic position of Tessera gemmaria Goy, 1979, a stauromedusa considered as incertae sedis for a long time. Using external morphology, histological preparations and molecular data (16S and COI) we conclude that T. gemmaria is an early stage of a cerinula, the long-lived planktonic larval stage of the Ceriantharia (Anthozoa).

Food resources influencing the asexual reproductive cycle of coronate Scyphozoa

The study of the strobilation process, a feature unique in the class Scyphozoa, is an issue that helps understanding the patterns of asexual reproduction in sessile invertebrates. Many inducers of asexual reproduction are known for scyphozoans. However, the influence of food resources on the strobilation of Coronate Scyphozoa has never been tested. WO observed strobilation of a large number of polyps of Nausithoe aurea, from a wide sampling area along the South Atlantic coast of Brazil, through the administration of controlled number of hatched nauplii of Artemia franciscana under a previous tested starvation and feeding protocol. The number of strobilations between and within groups varied and the fate and shape of strobilation deviated from the biology reported in the original description. Artificial seawater was used to reduce the influence of dissolved organic matter as likely important alternative nourishment.

The spider genus Chrysometa (Araneae, Tetragnathidae) from the Pico da Neblina and Serra do Tapirapeco mountains (Amazonas, Brazil): new species, new records, diversity and distribution along two altitudinal gradients

Eight new species of the spider genus Chrysometa Simon, 1894 (Araneae, Tetragnathidae) are described and illustrated. Chrysometa nubigena n. sp., C. waikoxi n. sp., C. petrasierwaldae n. sp., C. santosi n. sp., C. yanomami n. sp., C. candianii n. sp., C. lomanhungae n. sp., and C. saci n. sp. Those species were collected in a study on the diversity of spider communities along altitudinal gradients in Brazilian Amazonia. C. saci was captured at the Serra do Tapirapeco (Barcelos), while all the other species are from the Pico da Neblina (Sao Gabriel da Cachoeira), the highest mountain in Brazil. We provide new records for C. boraceia, C. flava, C. guttata, C. minuta and C. opulenta, and we describe the male of C. minuta for the first time. We also present the first results on the diversity and altitudinal distribution of the species of Chrysometa at the Pico da Neblina and Serra do Tapirapeco. We sampled the first locality at six different elevations, and obtained 336 specimens distributed in 12 species. Richness and abundance, as well as relative importance peaked at the highest sites sampled (2,000 and 2,400 m). The three most abundant species showed a segregated distribution, being dominant or exclusively distributed in different altitudes. At the Serra do Tapirapeco, sampling at four different elevations up to 1200 m, we only obtained 40 individuals divided in four species, and there was no clear relation to altitude. Most of the new species were found at mid and high altitude sites, while species from lower altitude sites represented widespread species. The comparison with other neotropical spiders inventories highlights the high diversity recorded at Pico da Neblina, which could be assigned to the large environmental variation covered in this work and to the sampling of high-altitude environments. Inventories in the Andean region and other information in the literature also seem to support the association of Chrysometa with high altitude environments.

Natural History of the Lutz`s Frog Cycloramphus lutzorum Heyer, 1983 (Anura: Cycloramphidae) in the Brazilian Atlantic Forest: Description of the Advertisement Call, Tadpole, and Karyotype

We describe the advertisement call, tadpole, karyotype, and additional information on the natural history of Cycloramphus lutzorum from southern Brazil. Sonograms were generated from digitally recorded calls. Tadpoles were collected in the field for description in the lab, and an adult was collected for karyotyping. Data on seasonal activity were gathered monthly from November 2005 to November 2007. All tadpoles (N = 21), juveniles (N = 18), and adults (N = 52) were found exclusively in streams. Reproduction, as identified by calling frogs, occurred from July through November. Frogs call all day long, but mostly at dusk, from rock crevices inside the stream edges near the splash zone. The call is short and loud, with 11 pulsed notes, of 491-641 ms, with a dominant frequency of 0.98-1.39 kHz. We describe the exotrophic and semiterrestrial tadpoles, always found in constantly humid vertical rock walls in the stream. Tadpoles of C. lutzorum are recognized by differences in labial tooth row formula, eye diameter, body shape, position of nares, and development of tail. Like congeneric species, the karyotype of C. lutzorum comprises 26 metacentric and submetacentric chromosomes. Cycloramphus lutzorum is restricted to and adapted for living in fast flowing streams, many of which are threatened by deforestation, pollution, and habitat loss. Therefore, we recommend the status of C. lutzorum be changed from its current ""Data Deficient"" to ""Near Threatened (NT)"" in the IUCN species red list.

Craniometric Similarities Within and Between Human Populations in Comparison with Neutral Genetic Data

The statement that pairs of individuals from different populations are often more genetically similar than pairs from the same population is a widespread idea inside and outside the scientific community. Witherspoon et al. [""Genetic similarities within and between human populations,"" Genetics 176:351-359 (2007)] proposed an index called the dissimilarity fraction (omega) to access in a quantitative way the validity of this statement for genetic systems. Witherspoon demonstrated that, as the number of loci increases, omega decreases to a point where, when enough sampling is available, the statement is false. In this study, we applied the dissimilarity fraction to Howells`s craniometric database to establish whether or not similar results are obtained for cranial morphological traits. Although in genetic studies thousands of loci are available, Howells`s database provides no more than 55 metric traits, making the contribution of each variable important. To cope with this limitation, we developed a routine that takes this effect into consideration when calculating. omega Contrary to what was observed for the genetic data, our results show that cranial morphology asymptotically approaches a mean omega of 0.3 and therefore supports the initial statement-that is, that individuals from the same geographic region do not form clear and discrete clusters-further questioning the idea of the existence of discrete biological clusters in the human species. Finally, by assuming that cranial morphology is under an additive polygenetic model, we can say that the population history signal of human craniometric traits presents the same resolution as a neutral genetic system dependent on no more than 20 loci.

mRNA expression of corticotropin-releasing factor and urocortin 1 after restraint and foot shock together with alprazolam administration

Corticotropin-releasing factor (CRF) is expressed in the paraventricular nucleus of the hypothalamus (PVN), and act centrally to provoke stress-like autonomic and behavioral responses. Urocortins 1-3 are additional ligands to the CRF receptors 1 and 2. Ucn 1 neurons are primarily concentrated in the Edinger-Westphal (EW) nucleus and also have been associated with stress responses. It is also known that UCN 1 respond in different ways depending on the stressor presented. Benzodiazepines can act via the CRF peptidergic system and chronic administration of alprazolam does not interfere with CRF mRNA expression in the PVN, but significantly increase Ucn 1 mRNA expression in the EW. The aim of our study was to investigate the relationship between different stressor stimuli, foot shock (FS) and restraint (R), and the mRNA expression of CRF and Ucn 1 in the PVN and EW using alprazolam (A). We employed fos activation and in situ hybridization. Restraint group presented increased fos-ir and CRF mRNA expression in the PVN compared to FS group. The stress responses of R group were prevented by A. In the EW,fos-ir was higher in the FS group than in the R group, whereas Ucn 1 mRNA expression was higher in the R group than in the FS group. Alprazolam significantly increased fos-ir and Ucn 1 mRNA expression in both groups. Our results show that PVN and EW respond in different ways to the same stressors. Furthermore, EW of stressed animals replies in a complementary way comparing to PVN with the use of Alprazolam. (C) 2010 Elsevier Inc. All rights reserved.