Antiproliferative Activity and Ultrastructural Changes in Promastigote and Amastigote forms of Leishmania amazonensis Caused by Limonene-Acylthiosemicarbazide Hybrids

Leishmaniasis is a tropical zoonotic disease. It is found in 98 countries, with an estimated 1.3 million people being affected annually. During the life cycle, the Leishmania parasite alternates between promastigote and amastigote forms. The first line treatment for leishmaniasis are the pentavalent antimonials, such as N-methylglucamine antimoniate (Glucantime®) and sodium stibogluconate (Pentostam®). These drugs are commonly related to be associated with dangerous side effects such as cardiotoxicity, nephrotoxicity, hepatotoxicity, and pancreatitis. Considering these aspects, this work aimed to obtain a new series of limonene-acylthiosemicarbazides hybrids as an alternative for the treatment of leishmaniasis. For this, promastigotes, axenic amastigotes, and intracellular amastigotes of Leishmania amazonensis were used in the antiproliferative assay; J774-A1 macrophages for the cytotoxicity assay; and electron microscopy techniques were performed to analyze the morphology and ultrastructure of parasites. ATZ−S-04 compound showed the best result in both tests. Its IC₅₀, in promastigotes, axenic amastigotes and intracellular amastigotes was 0.35±0.08 μM, 0.49±0.06 μM, and 15.90±2.88 μM, respectively. Cytotoxicity assay determined a CC₅₀ of 16.10±1.76 μM for the same compound. By electron microscopy, it was observed that ATZ−S-04 affected mainly the Golgi complex, in addition to morphological changes in promastigote forms of L. amazonensis.     

Application of nonspecific commercial AMF inocula results in poor mycorrhization in Camellia japonica L.

Camellia japonica L. (Theaceae, Theales) is an acidophilic evergreen flowering shrub and is traded worldwide. Symbiotic associations between the roots of this plant species and arbuscular mycorrhizal fungi (AMF), commonly recognized as natural biofertilizers and biocontrol agents, have been poorly studied so far. The aim of our study was to verify whether the application of nonspecific commercial AMF-based inocula could succeed and improve the growth of C. japonica. An experiment was conducted concerning the application of commercial inocula constituted by a specific AMF isolate (Funneliformis mosseae) or a consortium of different fungi and bacteria as alternatives to fertilization in pot cultivated C. japonica ‘Dr. Burnside’. Several growth parameters, plant nutrition and mycorrhization levels were monitored at the end of plant cultivation. Generally, increases in some macroelements (Ca, Mg, K) and microelements (Cu, Mn, Fe, Zn) were detected in the root system of inoculated camellias, while only Cu and Mn accumulation was increased in the leaves. Plants inoculated with the consortium inoculum exhibited increased chlorophyll content and foliage diameter while plants inoculated with F. mosseae exhibited increased number of flowers. Based on the polymorphism of an 18S rDNA region, we assessed the inoculated AMF that colonized C. japonica roots. The experiments showed that the applied AMF poorly colonized the root system of C. japonica. We suggest that commercial AMF formulations should be more targeted and host-specific in order to successfully colonize the host root and, potentially, fully express their benefits.     

Livestock Management Within a Traditional Agrosilvopastoral System in Northern Minas Gerais,Brazil: A Model for Reconciling Livelihoods and Conservation at a Time of Environmental Change

Human Ecology - We examine the ongoing transformation of the livestock raising component of a complex agrosilvopastoral management system associated with the Geraizeiro of northern Minas Gerais...     

The fungicide Mancozeb induces metacaspase-dependent apoptotic cell death in <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> BY4741

Mancozeb (MZ), a mixture of ethylene-bis-dithiocarbamate manganese and zinc salts, is one of the most widely used fungicides in agriculture. Toxicologic studies in mammals and mammalian cells indicate that this fungicide can cause neurological and cytological disorders, putatively associated with pro-oxidant and apoptotic effects. Yeast adaptation to sub-inhibitory concentrations of MZ has been correlated with oxidative response, proteins degradation, and energy metabolism, and its main effect on yeast has been attributed to its high reactivity with thiol groups in proteins. Herein, we show that acute MZ treatments on aerobic exponentially growing yeast of wild type (BY4741) and deletion mutant strains, coupled with multiplex flow cytometry analysis, conclusively demonstrated that MZ displays the typical features of pro-oxidant activity on Saccharomyces, elevating mitochondrial ROS, and causing hyper-polarization of mitochondrial membranes leading to apoptosis. A drastic reduction of cellular viability associated with the maintenance of cell membrane integrity, as well as phosphatidyl serine externalization on yeast cells exposed to MZ, also supports an apoptotic mode of action. Moreover, abrogation of the apoptotic response in yca1 deficient mutants indicates that metacaspase-1 is involved in the programmed cell death mechanism induced by MZ in yeast.     

Direct seeding of dry forest tree species in abandoned pastures: effects of grass canopy and seed burial on germination

Natural tree regeneration in abandoned pastures can be hampered by various biotic and abiotic filters, including seed removal and predation. We tested the effects of maintenance and removal of grass and seed deposition (buried and unburied) on seed germination of 12 tree species in dry forest pastures. We obtained evidence supporting the hypothesis that seeds attain higher germination under a grass canopy than on bare ground. For most species, grass cover provides safety from seed predators and facilitates germination by providing a suitable microclimate with soil humidity similar to the forest. The hypothesis that buried seeds attain higher germination was not supported by our data. Predation and removal of unburied seeds ranged from 0 to 77 % and, alone or together, were the major causes of non-germination. Direct seeding is a promising technique for revegetation of recently abandoned pastures in areas originally covered by tropical dry forests. The high germination rate of seeds deposited on the ground and under grass reduces costs during initial restoration stages, potentially facilitating the spread and use of this technique.     

Flowering phenology of a palm community in a central Amazon forest

The flowering phenology of 27 taxa of palms in a lowland moist forest in the central Amazon was recorded over a 40 month period. Phenological data were transformed into measures, of synchrony, regularity, and duration. Flowering was observed at all times of year and there was no correlation with rainfall either for the current month or for any monthly lag interval. The 27 taxa were divided into three pollination groups—weevil, bee/fly, or beetle—based on their behavior at anthesis. Phenology was significantly correlated with pollination groups. Weevilpollinated palms had higher synchrony and shorter duration of flowering than other groups. Bee/fly-pollinated palms had lower synchrony and longer duration of flowering. The beetle-pollinated palms were intermediate between the two other groups. Seasonal regularity of flowering was similar in all three groups. We suggest that, at least inBactris, taxa exhibit staggered flowering.     

Does Restoration Enhance Regeneration of Seasonal Deciduous Forests in Pastures in Central Brazil?

The goal of restoration is to accelerate ecosystem recovery, but in ecosystems that naturally regenerate rapidly restoration techniques need to be selected carefully to facilitate rather than impede natural recovery. We compared the effects of five restoration techniques, such as plowing the soil, removing grasses, adding forest litter, seeding, and planting nursery‐growing seedlings, on the regeneration of seasonal deciduous forest trees in four abandoned pastures in central Brazil. We monitored all woody stems immediately prior to treatments and again 14 months after the treatments. We recorded an average of 16,663 tree stems per hectare and a total of 83 species before implementing treatments. Planting strongly increased species richness; adding litter and seeding had weaker positive effects on richness; and plowing and grass removal had no effect. Plowing substantially reduced the density of naturally established stems. Despite the high survival of planted seedlings, stem density in planting treatments did not change because the tractor and digging holes to plant seedlings caused mortality of naturally regenerating seedlings. Tree stems grew more in the grass release plots than in the control plots. Our results suggest that early succession of seasonal deciduous forest in pastures in the region studied does not need to be stimulated once the perturbation is stopped and that intensive restoration efforts may actually slow recovery. We recommend only enrichment planting of seedlings that are not able to resprout.     

Development and evaluation of microsatellite markers in Phoenix dactylifera L. and their transferability to other Phoenix species

Forty one simple sequence repeats were isolated from two microsatellite enriched libraries of date palm (Phoenix dactylifera L.). After screening, 17 selected microsatellite loci were characterized and evaluated on a set of 31 cultivars and clones from Algerian and Californian germplasm. All primer pairs produced an amplification product of the expected size and detected high polymorphism among the analysed samples. These nuclear simple sequence repeat (SSR) markers are expected to be a very effective tool for evaluating genetic diversity in date palm germplasm. Acrosstaxa amplification showed the usefulness of most SSR markers in 14 other species across the genus Phoenix.     

Effects of Habitat, Cattle Grazing and Selective Logging on Seedling Survival and Growth in Dry Forests of Central Brazil

We studied the effect of forest gaps and cattle grazing on survival and growth of seedlings of seven native tree species, planted in dry forest fragments with different intensities of past logging in Central Brazil. Seedling survival after 12 mo ranged from 35 to 77 percent among species. Survival of most species was similar in gaps and understory in minimally disturbed forest remnants, but was lower in the gaps of more heavily logged forests. In contrast, growth was much higher in gaps than in understory in minimally disturbed forests, but for most species was similar in gaps and understory in more heavily logged forests. We did not detect an effect of cattle on seedling survival. Seedlings of the most commonly logged species show a high potential to survive and grow when planted in logged forests.