Capybara (Hydrochoerus hydrochaeris) distribution in agroecosystems: a cross-scale habitat analysis

Aim The aim of this study was to understand the spatial distribution of capybara (Hydrochoerus hydrochaeris) according to habitat attributes, using a multiscale approach based on fine‐ and broad‐scale variables in agroecosystems. Location Piracicaba river basin, south‐eastern Brazil (22°00′–23°30′ S; 45°45′–48°30′ W). Methods Potential habitats for capybara were selected in order to evaluate species presence/absence from October 2001 to December 2002. In each site, habitat attributes were sampled in the field (fine scale) and from GIS maps (broad scale) in terms of their presence or absence close to water. The variability of land cover between study sites was described by principal components analysis. Chi‐square tests were calculated for capybara presence/absence and the presence of each habitat attribute. A linear discriminant function analysis was used to describe to what extent the species’ presence could be explained by habitat attributes. Results The species presence was predominantly related to flat open areas (slope ranging from 0% to 6%) (χ² = 37.054, d.f. = 4, P < 0.001), covered by sugar cane or cultivated pasture (χ² = 84.814, d.f. = 9, P < 0.001). Terrain curvature, water meadows, aquatic vegetation, forest cover and open areas resulted in the best combination of variables, explaining 69.7% of capybara occurrence in the study sites in this river basin. Main conclusions Capybaras are widespread in the Piracicaba river basin, except in elevated areas. The spatial distribution of capybara was associated with the main types of land cover in the river basin – sugar cane plantations or pasture – both key food sources for capybara. This probably explains the species’ recent abundance in the region, since an intensive process of landscape alteration has taken place in this region owing to the expansion of agriculture in recent decades. These results may be useful in understanding the relationship between recent landscape modifications and the species’ population expansion in agroecosystems.     

Evaluation of the Stability of Concentrated Emulsions for Lemon Beverages Using Sequential Experimental Designs

The study of the stability of concentrated oil-in-water emulsions is imperative to provide a scientific approach for an important problem in the beverage industry, contributing to abolish the empiricism still present nowadays. The use of these emulsions would directly imply a reduction of transportation costs between production and the sales points, where dilution takes place. The goal of this research was to evaluate the influence of the main components of a lemon emulsion on its stability, aiming to maximize the concentration of oil in the beverage and to correlate its physicochemical characteristics to product stability, allowing an increase of shelf life of the final product. For this purpose, analyses of surface and interface tension, electrokinetic potential, particle size and rheological properties of the emulsions were conducted. A 2^4-1 fractional factorial design was performed with the following variables: lemon oil/water ratio (30% to 50%), starch and Arabic gum concentrations (0% to 30%) and dioctyl sodium sulfosuccinate (0 mg/L to 100 mg/L), including an evaluation of the responses at the central conditions of each variable. Sequentially, a full design was prepared to evaluate the two most influential variables obtained in the first plan, in which concentration of starch and gum ranged from 0% to 20%, while concentration of lemon oil/water ratio was fixed at 50%, without dioctyl sodium sulfosuccinate. Concentrated emulsions with stability superior to 15 days were obtained with either starch or Arabic gum and 50% lemon oil. The most stable formulations presented viscosity over 100 cP and ratio between the surface tension of the emulsion and the mucilage of over 1. These two answers were selected, since they better represent the behavior of emulsions in terms of stability and could be used as tools for an initial selection of the most promising formulations.     

Ponseti Method After Walking Age – A Multi-Centric Study of 429 Feet: Results,Possible Treatment Modifications and Outcomes According to Age Groups

ObjectivePonseti method is suitable to treat neglected clubfoot after the walking age. However, limited evidence exists on its effectiveness, outcomes and rate of relapse. Methods: 429 clubfeet in 303 patients with no previous treatment and older than one-year were treated with the Ponseti method in 15 centers from seven countries. The median age at treatment onset was three years, and the median follow-up of 1.3 years. Standard Ponseti Method was applied. Bilateral abduction brace was recommended after casting. Patients were classified according to group ages (<2 years, 2-4 years, >4-8years, >8 years). Feet were evaluated by Pirani score and a clinical outcome classification. Relapses were described in a subset of 103 clubfeet with minimal follow-up of two years.ResultsPonseti method was able to correct the deformity in 87% (373 of 429) of neglected clubfeet, after a mean of 6.8 casts. Residual equinus was treated with percutaneous sectioning of the Achilles tendon in 356 (83%) of 429 clubfeet. A bilateral foot abduction brace was prescribed and used in 70% of children. Relapses occurred in 31% (32 of 103) of clubfeet and were associated with age less than 4 years at treatment onset, and bracing noncompliance.ConclusionThe Ponseti method is effective to correct neglected clubfeet. Relapses occurred in one-third of clubfeet, mainly in children younger than four years and in noncompliance with the brace. Our study reinforces the recommendation for the Ponseti method with no major modification to treat neglected clubfoot in patients after walking age.Level of Evidence: IV     

First Microsatellite Markers Developed from Cupuassu ESTs: Application in Diversity Analysis and Cross-Species Transferability to Cacao

The cupuassu tree (Theobroma grandiflorum) (Willd. ex Spreng.) Schum. is a fruitful species from the Amazon with great economical potential, due to the multiple uses of its fruit´s pulp and seeds in the food and cosmetic industries, including the production of cupulate, an alternative to chocolate. In order to support the cupuassu breeding program and to select plants presenting both pulp/seed quality and fungal disease resistance, SSRs from Next Generation Sequencing ESTs were obtained and used in diversity analysis. From 8,330 ESTs, 1,517 contained one or more SSRs (1,899 SSRs identified). The most abundant motifs identified in the EST-SSRs were hepta- and trinucleotides, and they were found with a minimum and maximum of 2 and 19 repeats, respectively. From the 1,517 ESTs containing SSRs, 70 ESTs were selected based on their functional annotation, focusing on pulp and seed quality, as well as resistance to pathogens. The 70 ESTs selected contained 77 SSRs, and among which, 11 were polymorphic in cupuassu genotypes. These EST-SSRs were able to discriminate the cupuassu genotype in relation to resistance/susceptibility to witches’ broom disease, as well as to pulp quality (SST/ATT values). Finally, we showed that these markers were transferable to cacao genotypes, and that genome availability might be used as a predictive tool for polymorphism detection and primer design useful for both Theobroma species. To our knowledge, this is the first report involving EST-SSRs from cupuassu and is also a pioneer in the analysis of marker transferability from cupuassu to cacao. Moreover, these markers might contribute to develop or saturate the cupuassu and cacao genetic maps, respectively.     

The effect of agriculture on the seasonal dynamics and functional diversity of benthic biofilm in tropical headwater streams

Tropical streams are one of the most endangered ecosystems in the world due to the constant pressures from human activities. Among these activities, agriculture represents a land use that is crucial for human development but also a key driver of stream degradation and biodiversity decline in the tropics. Against this background, we investigated indirect effects of agriculture (alterations in stream flow and nutrient availability) and climate characteristics (water temperature) on benthic biofilm communities in tropical streams (São Paulo State, Brazil). Three first‐order streams draining catchments dominated by agricultural land use (sugarcane for bioenergy production, pasture) with some remaining riparian forest were studied for 1 year. We focused on the relationships of benthic biofilm biomass, algal biomass, diatom community, and functional structure with streamflow dynamics, nitrate concentrations, and water temperature. Our results indicate that these biological responses were mainly mediated by flow and water temperature and not by resource availability in the studied headwater streams. This result could be explained by the heavy rains and elevated runoff generation in these tropical catchments under agricultural influence, which might override the known effects of nutrient enrichment on benthic biofilm communities. Considering forecast climate and land‐use changes in tropical streams, our findings may suggest potential shifts in benthic biofilm communities, with functional consequences for aquatic food webs in these environments. Abstract in Brazilian Portuguese is available with online material.     

Disruption of TNF-α signaling improves osteoblastic differentiation of adipose-derived mesenchymal stem cells and bone repair

Adipose tissue is an attractive source of mesenchymal stem cells (at-MSCs), but their low osteogenic potential limits their use in bone regeneration. Adipose tissue plays a role in pro-inflammatory diseases by releasing cytokines with a catabolic effect on bone, such as tumor necrosis factor-alpha (TNF-α). Thus, we hypothesized that endogenous TNF-α could have a negative effect on at-MSC differentiation into osteoblasts. Short interfering RNAs (siRNAs) targeting TNF-α receptors (siR1, siR2, and si1R/R2) were transfected into at-MSCs, and cell differentiation was assessed by measuring the expression of bone markers, ALP activity, and mineralized matrix. Scrambled was used as Control. Knockout at-MSCs (KOR1/R2) was injected in mice calvaria defects, and bone formation was evaluated by microtomography and histological analysis. Data were compared by Kruskal–Wallis or analysis of variance (5%). The expression of bone markers confirmed that at-MSCs differentiate less than bone marrow MSCs. In silenced cells, the expression of Alp, Runx2, and Opn was generally higher compared to Control. ALP, RUNX2, and OPN were expressed at elevated levels in silenced groups, most notably at-MSCs-siR1/R2. ALP was detected at high levels in at-MSCs-siR1/R2 and in-MSCs-siR1, followed by an increase in mineralized nodules in at-MSCs-siR1/R2. As the morphometric parameters increased, the groups treated with KOR1/R2 exhibited slight bone formation near the edges of the defects. Endogenous TNF-α inhibits osteoblast differentiation and activity in at-MSCs, and its disruption increases bone formation. While opening a path of investigation, that may lead to the development of new treatments for bone regeneration using at-MSC-based therapies.     

Detection of Wuchereria bancrofti DNA in paired serum and urine samples using polymerase chain reaction-based systems

The Global Program for the Elimination of Lymphatic Filariasis (GPELF) aims toeliminate this disease by the year 2020. However, the development of more specificand sensitive tests is important for the success of the GPELF. The present studyaimed to standardise polymerase chain reaction (PCR)-based systems for the diagnosisof filariasis in serum and urine. Twenty paired biological urine and serum samplesfrom individuals already known to be positive for Wuchereria bancroftiwere collected during the day. Conventional PCR and semi-nested PCR assayswere optimised. The detection limit of the technique for purified W.bancrofti DNA extracted from adult worms was 10 fg for the internalsystems (WbF/Wb2) and 0.1 fg by using semi-nested PCR. The specificity of the primerswas confirmed experimentally by amplification of 1 ng of purified genomic DNA fromother species of parasites. Evaluation of the paired urine and serum samples by thesemi-nested PCR technique indicated only two of the 20 tested individuals werepositive, whereas the simple internal PCR system (WbF/Wb2), which has highlypromising performance, revealed that all the patients were positive using bothsamples. This study successfully demonstrated the possibility of using the PCRtechnique on urine for the diagnosis of W. bancrofti infection.     

Changes in γ-aminobutyric acid concentration,gas exchange,and leaf anatomy in <Emphasis Type="Italic">Eucalyptus</Emphasis> clones under drought stress and rewatering

Drought stress promotes biochemical and physiological alterations in plant metabolism that limit growth and yield. This study investigated the accumulation of γ-aminobutyric acid (GABA) in plant tissue, the stomatal conductance (gs) and changes in leaf anatomy in Eucalyptus following drought stress situation. In this study, eight Eucalyptus clones were evaluated under normal water supply (control) and drought stress conditions (stress). For the control treatment, plants were irrigated every day with an automated system until the soil was saturated, and for the stress treatment, drought stress was imposed by non-irrigation of plants, and pots were covered using plastic sheeting to avoid rainfall and humidity. This study has shown that: (1) all clones decreased gs with increasing vapor pressure deficit (D) in both treatments. All plastics and drought-tolerant clones (except GG) presented lower stomatal sensitivity to D under stress conditions than drought-sensitive clones; (2) GABA concentrations increased fast after drought stress, but we could not find correlation with these changes and resistance to water stress; and (3) all clones increased the number of stomata and reduced leaf thickness after water stress. The finding is that GABA is a fast stress-signaling molecule in Eucalyptus, but the response of gs to D is a best physiological variable to differentiate drought-tolerant and drought-sensitive Eucalyptus clones.     

Consequences of excessive glucosylsphingosine in glucocerebrosidase-deficient zebrafish.

In Gaucher disease (GD), the deficiency of glucocerebrosidase causes lysosomal accumulation of glucosylceramide (GlcCer), which is partly converted by acid ceramidase to glucosylsphingosine (GlcSph) in the lysosome. Chronically elevated blood and tissue GlcSph is thought to contribute to symptoms in GD patients as well as to increased risk for Parkinson’s disease. On the other hand, formation of GlcSph may be beneficial since the water soluble sphingoid base is excreted via urine and bile. To study the role of excessive GlcSph formation during glucocerebrosidase deficiency, we studied zebrafish that have two orthologs of acid ceramidase, Asah1a and Asah1b. Only the latter is involved in the formation of GlcSph in glucocerebrosidase-deficient zebrafish as revealed by knockouts of Asah1a or Asah1b with glucocerebrosidase deficiency (either pharmacologically induced or genetic). Comparison of zebrafish with excessive GlcSph (gba1^-/- fish) and without GlcSph (gba1^-/-:asah1b^-/- fish) allowed us to study the consequences of chronic high levels of GlcSph. Prevention of excessive GlcSph in gba1^-/-:asah1b^-/- fish did not restrict storage cells, GlcCer accumulation, or neuroinflammation. However, GD fish lacking excessive GlcSph show an ameliorated course of disease reflected by significantly increased lifespan, delayed locomotor abnormality, and delayed development of an abnormal curved back posture. The loss of tyrosine hydroxylase 1 (th1) mRNA, a marker of dopaminergic neurons, is slowed down in brain of GD fish lacking excessive GlcSph. In conclusion, in the zebrafish GD model, excess GlcSph has little impact on (neuro)inflammation or the presence of GlcCer-laden macrophages but rather seems harmful to th1-positive dopaminergic neurons.     

Mimetic Membrane System to Carry Multiple Antigenic Proteins from Leishmania amazonensis

Liposomes have long been used as models for lipid membranes and for the reconstitution of a single or multiple proteins. Also, liposomes have adjuvant activity in vaccines against several protozoan or bacterial organisms. Thus, the main objective of the present study was to obtain a crude extract of detergent-solubilized proteins of Leishmania amazonensis amastigotes and reconstitute them into liposomes. Neutral and zwiterionic detergents were less efficient than an ionic detergent. In order to obtain efficient solubilization using only sodium dodecyl sulfate (SDS), the effects of detergent and protein concentration and incubation time were studied. The maximum of solubilized proteins was obtained instantaneously using a ratio of 0.5 mg/ml of protein to 0.1% (w/v) detergent at 4°C. Dipalmitoylphosphatidylcholine (DPPC), dipalmitoylphosphatidylserine (DPPS) and cholesterol in a weight ratio of 5:1:4 were used for protein reconstitution into liposomes using the cosolubilization method, yielding 60% of incorporation. The incorporation of multiple parasite proteins results in a vesicular diameter of proteoliposomes of about 140 nm, presenting a final lipid weight ratio for DPPC, DPPS and cholesterol of 1:1:5, with high stability. The detergent-solubilized proteins of L. amazonensis amastigotes present in the proteoliposome, when analyzed by SDS-polyacrylamide gel electrophoresis, include a wide range of parasite-incorporated proteins. BALB/c mice inoculated with these proteoliposomes were able to produce antibodies against the proteins reconstituted in DPPC:DPPS:cholesterol liposomes and were partially resistant to infection with L. amazonensis promastigotes. These results indicate that this system can be used as a possible vaccine against L. amazonensis.