Development of microsatellite markers for the Neotropical endemic Brazilian Guanabara frog, Euparkerella brasiliensis, through 454 shotgun pyrosequencing

The new-generation 454 GS-FLX Titanium pyrosequencing was used to isolate microsatellite markers for the Brazilian Guanabara frog, Euparkerella brasiliensis, an Atlantic forest endemic species. Three multiplex polymerase chain reaction sets were optimized for genotyping of 11 polymorphic (di- and tetranucleotide) microsatellite markers. Genetic diversity was assessed in 21 individuals from a population (Reserva Ecológica de Guapiaçu, REGUA) locatedin the central region of the Rio de Janeiro State, in Brazil. The mean number of alleles per locus ranged from 3 to 12. Observed and expected heterozygosities ranged from 0.095 to 0.905 and from 0.094 to 0.904, respectively. After using the Bonferroni correction for multiple tests, there was no evidence of linkage disequilibrium between pairs of loci but deviations for Hardy-Weinberg equilibrium were found in 4 loci. We found no evidence for allele dropouts or stuttering, but we detected the presence of null alleles at loci Eb10 and Eb36. These markers will be useful for analyses of fine-scale population structure and determination of relative effects of habitat loss and fragmentation on population genetic variability within species. © FUNPEC-RP.

Phenotypic changes and small mammal impoverishment on a Brazilian Atlantic Forest Island

Faunal impoverishment and distorted species compositions are common phenomena in oceanic islands; however, many land-bridge islands are poorly inventoried, especially in the Neotropics. We sampled a small mammal community on a land-bridge island (Anchieta Island) along the Brazilian coast. We found only one marsupial Didelphis aurita (Wied-Neuwied, 1826) and two rodent species Oligoryzomys nigripes (Olfers, 1818) and Trinomys iheringi (Thomas, 1911) during 12 months of live trapping and 9195 trap-nights. The diversity of rodents and marsupials was not explained by species-area relations, indicating possible past extinctions. The abundance of D. aurita and O. nigripes was approximately three times higher, while the abundance of T. iheringi was approximately four times lower than abundances reported from other Brazilian Atlantic Forest sites. The population of D. aurita exhibited many phenotypic changes; males were on average 8 % smaller and females produced 30 % less litters than those from the mainland and other land-bridge islands. The long history of forest disturbance, habitat loss, reduction in forest productivity, and the recent introduction of mesopredators may be the major drivers that explain the small mammal community composition on this island. © 2013 Walter de Gruyter GmbH, Berlin/Boston.

Water availability determines the richness and density of fig trees within Brazilian semideciduous forest landscapes

The success of fig trees in tropical ecosystems is evidenced by the great diversity (+750 species) and wide geographic distribution of the genus. We assessed the contribution of environmental variables on the species richness and density of fig trees in fragments of seasonal semideciduous forest (SSF) in Brazil. We assessed 20 forest fragments in three regions in Sao Paulo State, Brazil. Fig tree richness and density was estimated in rectangular plots, comprising 31.4 ha sampled. Both richness and fig tree density were linearly modeled as function of variables representing (1) fragment metrics, (2) forest structure, and (3) landscape metrics expressing water drainage in the fragments. Model selection was performed by comparing the AIC values (Akaike Information Criterion) and the relative weight of each model (wAIC). Both species richness and fig tree density were better explained by the water availability in the fragment (meter of streams/ha): wAICrichness = 0.45, wAICdensity = 0.96. The remaining variables related to anthropic perturbation and forest structure were of little weight in the models. The rainfall seasonality in SSF seems to select for both establishment strategies and morphological adaptations in the hemiepiphytic fig tree species. In the studied SSF, hemiepiphytes established at lower heights in their host trees than reported for fig trees in evergreen rainforests. Some hemiepiphytic fig species evolved superficial roots extending up to 100 m from their trunks, resulting in hectare-scale root zones that allow them to efficiently forage water and soil nutrients. The community of fig trees was robust to variation in forest structure and conservation level of SSF fragments, making this group of plants an important element for the functioning of seasonal tropical forests. © 2013 Elsevier Masson SAS. All rights reserved.

Development of an analytical method for the quantification of pfaffic acid in Brazilian ginseng (Hebanthe eriantha)

Hebanthe eriantha (Poir.) Pedersen (Amaranthaceae), which is known as Brazilian ginseng is widely used in folk medicine as an aphrodisiac and antidiabetic tonic. The anti-tumor activity, attributed to the pfaffic acid present in roots of H. eriantha, is responsible for the great interest in the commercialization of this species. In Brazil, the species H. eriantha is mainly used in commercial preparations, although other plants of the genus Pfaffia and Hebanthe have been marketed as Pfaffia paniculata or Brazilian ginseng. The pfaffic acid present in the roots is mainly conjugated with sugars (pfaffosides) and can be used as an active marker of H. eriantha, which helps to differentiate this species from others marketed as Brazilian ginseng. The main objective of this study was to develop and validate a liquid chromatographic method to quantify pfaffic acid in the roots of H. eriantha. The extraction and hydrolysis conditions were optimized using an univariate and experimental design, respectively, and the quantification of pfaffic acid by high performance liquid chromatography with diode-array detection (HPLC-DAD) was validated. This method was used to evaluate the pfaffic acid content in 30 different genotypes of the species from a germplasm collection. The content of pfaffic acid ranged from 0.97 to 4.29% (w/w) on a dry weight basis. © 2013 Elsevier B.V.

Distribution related to temperature and salinity of the shrimps Acetes americanus and Peisos petrunkevitchi (Crustacea: Sergestoidea) in the south-eastern Brazilian littoral zone

The abundance and ecological distribution of Acetes americanus and Peisos petrunkevitchi were investigated from July 2006 to June 2007, in Ubatuba, Brazil. Eight transects were identified and sampled monthly: six of these transects were located in Ubatuba bay, with depths reaching 21 m, and the other two transects were in estuarine environments. A total of 33,888 A. americanus shrimp were captured, with the majority coming from the shallower transects (up to 10 m). Conversely, 6,173 of the P. petrunkevitchi shrimps were captured in deeper areas (from 9 to 21 m). No individuals from either species were found in the estuary. The highest abundances obtained for both species were sampled during the summer. Canonical correlation analysis resulted in a coefficient value of 0.68 (P = 0.00). The abundance of both species was strongly correlated with depth. Variations in temperature and salinity values were also informative in predicting the seasonal presence of P. petrunkevitchi in deeper areas and A. americanus in the shallower areas of the bay. It is conceivable that the shrimp adjust their ecological distribution according to their intrinsic physiological limitations. © 2012 Marine Biological Association of the United Kingdom.

Population structure and canopy use by Coussapoa microcarpa, a strangler hemiepiphyte from the Brazilian Atlantic Forest

We studied density, size structure, and establishment sites of Coussapoa microcarpa in the Brazilian Atlantic rain forest. This species is a hemiepiphyte that begins its life in the tops of trees and survives the death of its host to become a free-standing tree. All individuals of C. microcarpa already rooted in the ground were recorded in a 3.43 ha (1.75 ha in lowland and 1.68 ha in submontane) sample of forest plots. Data on total height, root diameter at breast height, host height and diameter at breast height, as well as height, type and diameter of the establishment site were collected. Coussapoa microcarpa present a high density (36.5 ind. ha-1) and the population studied was composed mainly of young individuals. Young and adults differed in establishment sites. The diameter of establishment sites of young was narrower than the diameter of establishment sites of adults, which points out to a limiting factor (diameter of establishment site) regulating the establishment of C. microcarpa. © 2013 Cambridge University Press.

First cytogenetic information for Drymoreomys albimaculatus (Rodentia, Cricetidae), a recently described genus from Brazilian Atlantic forest

The recently described taxon Drymoreomys albimaculatus is endemic to the Brazilian Atlantic Forest and its biology and genetics are still poorly known. Herein, we present, for the first time, the karyotype of the species using classical and molecular cytogenetics, which showed 2n=62, FN=62, and interstitial telomeric signals at the sex chromosomes. Nuclear and mitochondrial DNA sequences from the two karyotyped individuals verify the taxonomic identity as the recently described D. albimaculatus and confirm the relationship of the species with other Oryzomyini. Additionally, external morphological information is provided. © Elkin Y. Suárez-Villota et al.

No evidence for association of the CD40, CD40L and BLYS polymorphisms, B-cell co-stimulatory molecules, with Brazilian endemic Plasmodium vivax malaria

Background: Plasmodium vivax is the most prevalent malaria species in Brazil. The parasite-host coevolutionary process can be viewed as an 'arms race', in which adaptive genetic changes in one are eventually matched by alterations in the other. Methods: Following the candidate gene approach we analyzed the CD40, CD40L and BLYS genes that participate in B-cell co-stimulation, for associations with P. vivax malaria. The study sample included 97 patients and 103 controls. We extracted DNA using the extraction and purification commercial kit and identified the following SNPs: 21C.T in the CD40 gene, 2726T.C in the CD40L gene and the 2871C.T in the BLyS gene using PCR-RFLP. We analyzed the genotype and allele frequencies by direct counting. We also compared the observed with the expected genotype frequencies using the Hardy-Weinberg equilibrium. Results: The allele and genotype frequencies for these SNPs did not differ statistically between patient and control groups. Gene-gene interactions were not observed between the CD40 and BLYS and between the CD40L and BLYS genes. Overall, the genes were in Hardy-Weinberg equilibrium. Significant differences were not observed among the frequencies of antibody responses against P. vivax sporozoite and erythrocytic antigens and the CD40 and BLYS genotypes. Conclusions: The results of this study show that, although the investigated CD40, CD40L and BLYS alleles differ functionally, this variation does not alter the functionality of the molecules in a way that would interfere in susceptibility to the disease. The variants of these genes may influence the clinical course rather than simply increase or decrease susceptibility. © Royal Society of Tropical Medicine and Hygiene 2013. All rights reserved.

Local and regional ecological drivers of fish assemblages in Brazilian estuaries

Spatial patterns in assemblage structures are generated by ecological processes that occur on multiple scales. Identifying these processes is important for the prediction of impact, for restoration and for conservation of biodiversity. This study used a hierarchical sampling design to quantify variations in assemblage structures of Brazilian estuarine fish across 2 spatial scales and to reveal the ecological processes underlying the patterns observed. Eight areas separated by 0.7 to 25 km (local scale) were sampled in 5 estuaries separated by 970 to 6000 km (regional scale) along the coast, encompassing both tropical and subtropical regions. The assemblage structure varied significantly in terms of relative biomass and presence/absence of species on both scales, but the regional variation was greater than the local variation for either dataset. However, the 5 estuaries sampled segregated into 2 major groups largely congruent with the Brazilian and Argentinian biogeographic provinces. Three environmental variables (mean temperature of the coldest month, mangrove area and mean annual precipitation) and distance between estuaries explained 44.8 and 16.3%, respectively, of the regional-scale variability in the species relative biomass. At the local scale, the importance of environmental predictors for the spatial structure of the assemblages differed between estuarine systems. Overall, these results support the idea that on a regional scale, the composition of fish assemblages is simultaneously determined by environmental filters and species dispersal capacity, while on a local scale, the effect of environmental factors should vary depending on estuary-specific physical and hydrological characteristics © 2013 Inter-Research.

Attitude of Brazilian veterinarians in the recognition and treatment of pain in horses and cattle

Objective: The objective of this study was to assess the use of analgesics, describe the attitudes of Brazilian veterinarians towards pain relief in horses and cattle and evaluate the differences due to gender, year of graduation and type of practice. Study design: Prospective survey. Methods: Questionnaires were sent to 1000 large animal veterinarians by mail, internet and delivered in person during national meetings. The survey investigated the attitudes of Brazilian veterinarians to the recognition and treatment of pain in large animals and consisted of sections asking about demographic data, use of analgesic drugs, attitudes to pain relief and to the assessment of pain. Descriptive statistics were used to analyze frequencies. Simple post hoc comparisons were performed using the chi-square test. Results: Eight hundred questionnaires were collected, but 87 were discarded because they were incomplete or blank. The opioid of choice for use in large animals was butorphanol (43.4%) followed by tramadol (39%). Flunixin (83.2%) and ketoprofen (67.6%) were the most frequently used NSAIDs by Brazilian veterinarians. Respondents indicated that horses received preoperative analgesics for laparotomy more frequently (72.9%) than cattle (58.5%). The most frequently administered preoperative drugs for laparotomy in horses were flunixin (38.4%) and xylazine (23.6%), whereas the preoperative drugs for the same surgical procedure in cattle were xylazine (31.8%) and the local administration of lidocaine (48%). Fracture repair was considered the most painful surgical procedure for both species. Most veterinarians (84.1%) believed that their knowledge in this area was not adequate. Conclusions and clinical relevance: Although these Brazilian veterinarians thought that their knowledge on recognition and treatment of pain was not adequate, the use of analgesic in large animals was similar in Brazil to that reported in other countries. © 2013 Association of Veterinary Anaesthetists and the American College of Veterinary Anesthesia and Analgesia.

Source-driven remobilizations of nutrients within stem wood in Eucalyptus grandis plantations

Nutrient remobilizations in tree ligneous components have been little studied in tropical forests. A complete randomized block design was installed in Brazilian eucalypt plantations to quantify the remobilizations of phosphorus (P), potassium (K), calcium (Ca), magnesium (Mg), and sodium (Na) within stem wood. Three treatments were studied: control with neither K nor Na addition (C), 3 kmol ha-1 K applied (+K), and 3 kmol ha-1 Na applied (+Na). Biomass and nutrient contents were measured in the stem wood of eight trees destructively sampled at 1, 2, 3 and 4 years after planting in each treatment and annual rings were localized on discs of wood sampled every 3 m in half of the trees. Chemical analyses and wood density measurements were performed individually for each ring per level and per tree sampled. Nutrient remobilizations in annual rings were calculated through mass balance between two successive ages. Our results show that nutrient remobilizations within stem wood were mainly source-driven. Potassium and Na additions largely increased their concentration in the outer rings as well as the amounts remobilized in the first 2 years after the wood formation. The amount of Na remobilized in annual rings was 15 % higher in +Na than in +K the fourth year after planting despite a 34 % higher production of stem wood in +K leading to a much higher nutrient sink. A partial substitution of K by Na in the remobilizations within stem wood might contribute to enhancing Eucalyptus grandis growth in K-depleted soils. © 2013 Springer-Verlag Berlin Heidelberg.

Study of Brazilian commercial Oriented Strand Board panels using stress wave

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Instantaneously measured traits may detect non-plastic ecophysiological performances in response to drought, explaining distributions of Styrax species in the Cerrado

We analyzed the differences between irrigated and non-irrigated plants of three congeneric Styrax species that present distinct distribution patterns in the physiognomies of the Cerrado vegetation in Brazil. Styrax ferrugineus showed a stomatal conductance (gs) unresponsive to soil water deficit in potted plants. This may explain the high gas exchange and photochemical efficiency found in this species, which is well adapted to the Cerrado sensu stricto (s. str.), a savanna-type vegetation. S. camporum, which is widely distributed in the Cerrado sensu lato (s. l.) areas, was the only species that exhibited increased intrinsic water use efficiency on the days of maximum water deficit. This result distinguishes S. camporum from S. pohlii, which is a forest species, since the gs of both species decreased during the days of maximum water stress. In contrast to other studies, we propose that instantaneously measured traits, such as leaf gas exchange rates and chlorophyll fluorescence, may be used to detect non-plastic performances in response to environmental stress, helping explain distinct geographical distributions of congeneric species in the Cerrado vegetation. © 2013 Springer-Verlag Berlin Heidelberg.

Yeast diversity isolated from grape musts during spontaneous fermentation from a brazilian winery

Saccharomyces and non-Saccharomyces yeast species from a winery located in Brazil were identified by ribosomal gene-sequencing analysis. A total of 130 yeast strains were isolated from grape surfaces and musts during alcoholic fermentation from Isabel, Bordeaux, and Cabernet Sauvignon varieties. Samples were submitted to PCR-RFLP analysis and genomic sequencing. Thirteen species were identified: Candida quercitrusa, Candida stellata, Cryptococcus flavescens, Cryptococcus laurentii, Hanseniaspora uvarum, Issatchenkia occidentalis, Issatchenkia orientalis, Issatchenkia terricola, Pichia kluyveri, Pichia guilliermondii, Pichia sp., Saccharomyces cerevisiae, and Sporidiobolus pararoseus. A sequential substitution of species during the different stages of fermentation, with a dominance of non-Saccharomyces yeasts at the beginning, and a successive replacement of species by S. cerevisiae strains at the final steps were observed. This is the first report about the yeast distribution present throughout the alcoholic fermentation in a Brazilian winery, providing supportive information for future studies on their contribution to wine quality. © 2013 Springer Science+Business Media New York.

Relative growth, sexual maturity, and breeding season of three species of the genus Persephona (Decapoda: Brachyura: Leucosiidae): A comparative study

The relative growths of Persephona lichtensteinii, P. mediterranea, and P. punctata were investigated on the south-eastern Brazilian coast, focusing on differences in the growth rates between immature and mature phases, the onset of morphological sexual maturity, and the breeding seasons of these species. Crabs were collected every two months from January 1991 through to November 1992, from a shrimp fishing boat equipped with two otter-trawl nets. Significant differences in the patterns of body growth were observed between immature and mature phases of all three species. Changes in the growth rates of the chelipeds (males) and abdomen (females) observed for P. lichtensteinii, P. mediterranea, and P. punctata, seem to be related to the puberty moult for both sexes. Males of P. mediterranea and P. punctata reached larger mean sizes of carapace width than females, whereas no difference was recorded for P. lichtensteinii. The body size at which 50% of males attained sexual maturity was also larger in P. mediterranea and P. punctata, and smaller in P. lichtensteinii. The absence of a pronounced sexual dimorphism and the size at the onset of sexual maturity observed only for P. lichtensteinii might be explained by distinct reproductive strategies of males. The presence of ovigerous females during the entire sampling period suggests that all three species have a continuous reproduction pattern at the Ubatuba region. Future studies on the population structure, functional maturity, and mating system should improve the understanding of factors driving the biology and ecology of these species at a subtropical region. © Marine Biological Association of the United Kingdom 2013.