Where to Live? How Morphology and Evolutionary History Predict Microhabitat Choice by Tropical Tadpoles

Tadpoles have diverse morphologies and occupy diverse habitats. The morphological differences between tadpoles can be represented by linear and geometric measurements and used to explain the organization of tadpole assemblages. However, the effects of evolutionary history must be isolated from the morphological differences before we can determine which patterns result from the use and sharing of common ecological resources. Here, we aimed to determine how morphological similarities and phylogenetic distances affect microhabitat choice by tadpoles. We analyzed the tadpoles of 101 anuran species and classified them according to ecomorphological guild, habitat use, position in the water column, and floor substrate. We used geometric and traditional morphometric approaches to describe the morphological variation among tadpoles and calculated the patristic distance for each species. Afterwards, we used morphometric and phylogenetic matrices as predictors of the variance in the ecological matrix, using a partial redundancy analysis. When we used traditional morphometric data, phylogeny explained a large amount of the ecological variation. By contrast, when we used geometric morphometric data, morphology and phylogeny explained similar amounts of the ecological variation, showing that the technique used to extract morphological variation affects the results. We provide evidence that both morphology, as a surrogate for contemporary factors, and evolutionary inertia are important in determining the behavior of tadpoles. Thus, niche conservatism can be important in modeling the behavior of tadpoles, but does not explain all the preferences of tadpoles.     

Phenylhydrazides as inhibitors of Leishmania amazonensis arginase and antileishmanial activity

Searching for new substances with antileishmanial activity, we synthesized and evaluated a series of α,α-difluorohydrazide and α,α-difluoramides against Leishmania amazonensis arginase (LaArg). Four α,α-difluorohydrazide derivatives showed activity against LaArg with K_i in the range of 1.3–26 μM. The study of the kinetics of LaArg inhibition showed that these substances might act via different inhibitory mechanisms or even by a combination of these. The compounds were tested against L. amazonensis promastigotes and the best result was obtained to the compound 4 (EC₅₀ of 12.7 ± 0.3 μM). In addition, in order to obtain further insight into the binding mode of such compounds, molecular docking studies were performed to obtain additional validation of experimental results. Considering these results, it is possible to conclude that α,α-difluorohydrazide derivatives are a promising scaffold in the development of new substances against the etiological agent of leishmaniasis by targeting LaArg.     

The parasitoid Gonatopus bartletti reduces presence of plant-pathogenic Spiroplasma kunkelii within the leafhopper vector Dalbulus maidis

Homopteran vectors (e.g., leafhoppers) of plant pathogens are vessels for reproduction of cell wall‐free bacteria. These vectors also serve as hosts for larval parasitoid dipterans, hymenopterans, and strepsipterans. However, no study has explored the relationship among these wall‐free bacteria and parasitoid larvae within the insect host. We studied the corn stunt spiroplasma (CSS), Spiroplasma kunkelii Whitcomb (Mycoplasmatales: Spiroplasmataceae), a bacterium that originated from secondary symbionts that cause corn stunt disease in maize, Zea mays L., and its reproduction in the haemolymph of the corn leafhopper, Dalbulus maidis (Delong and Wolcott) (Homoptera: Cicadellidae). We also studied the dryinid parasitoid Gonatopus bartletti Olmi (Hymenoptera: Dryinidae), the larva of which feeds in the corn leafhopper haemolymph. Our results showed that when CSS and the wasp coexisted in D. maidis, the development of the parasitoid was not affected by S. kunkelii. Parasitoid development was successfully completed when leafhoppers acquired S. kunkelii before or after parasitism and when CSS had median (10 days) and long (20 days) incubation periods in the leafhopper before parasitization. The presence of S. kunkelii did not affect parasitoid development to the adult stage. However, polymerase chain reaction showed that the presence (survival) of S. kunkelii in the leafhopper was negatively affected by the parasitoid larva. Fewer leafhoppers had CSS before and after parasitization compared with leafhoppers that only acquired the CSS. This negative effect helps to explain the high parasitism rate by G. bartletti in D. maidis and the low presence of S. kunkelii in the corn leafhopper when CSS and the wasp parasitoid overlap throughout their geographic distribution. The parasitoid larva may negatively affect S. kunkelii by (1) producing antibacterial peptides that are toxic to CSS; (2) producing teratocytes that take nutrients from the host for larval development, but these nutrients are required by CSS; (3) affecting, indirectly, CSS through other symbiotic microorganisms; and (4) producing proteins with antibacterial activity that are present in the venom of the wasp parasitoid.     

Mapping the anatomy of a Plasmodium falciparum MSP-1 epitope using pseudopeptide-induced mono- and polyclonal antibodies and CD and NMR conformation analysis

Antigen structure modulation represents an approach towards designing subunit malaria vaccines. A specific epitope's alpha carbon stereochemistry, as well as its backbone topochemistry, was assessed for obtaining novel malarial immunogens. A variety of MSP-1(38-61) Plasmodium falciparum epitope pseudopeptides derived were synthesised, based on solid-phase pseudopeptide chemistry strategies; these included all-L, all-D, partially-D substituted, all-Psi-[NH-CO]-Retro, all-Psi-[NH-CO]-Retro-inverso, and Psi-[CH2NH] reduced amide surrogates. We demonstrate that specific recombinant MSP-1(34-469) fragment binding to red blood cells (RBCs) is specifically inhibited by non-modified MSP-1(42-61), as well as by its V52-L53, M51-V52 reduced amide surrogates and partial-D substitutions in K48 and E49. In vivo tests revealed that reduced amide pseudopeptide-immunised Aotus monkeys induced neutralising antibodies specifically recognising the MSP-1 N-terminus region. These findings support the role of molecular conformation in malaria vaccine development.     

Worldwide genomic diversity of the high-risk human papillomavirus types 31, 35, 52, and 58, four close relatives of human papillomavirus type 16

Among the more than one hundred formally described human papillomavirus (HPV) types, 18 are referred to as high-risk HPV types due to their association with anogenital cancer. Despite pathogenic similarities, these types form three remotely related taxonomic groups. One of these groups is called HPV species 9 and is formed by HPV-16, the most common and best-studied type, together with HPV-31, -33, -35, -52, -58, and -67. Previous worldwide comparisons of HPV-16 samples showed about 2% nucleotide diversity between isolates, which were subsequently termed variants. The distribution of divergent variants has been found to correlate frequently with the geographic origin and the ethnicity of the infected patients and led to the concept of unique African, European, Asian, and Native American HPV-16 variants. In the current study, we address the question of whether geography and ethnicity also correlate with sequence variations found for HPV-31, -35, -52, and -58. This was done by sequencing the long control region in samples derived from Europe, Asia, and Africa, and from immigrant populations in North and South America. We observed maximal divergence between any two variants within each of these four HPV types ranging from 1.8 to 3.6% based on nucleotide exchanges and, occasionally, on insertions and deletions. Similar to the case with HPV-16, these mutations are not random but indicate a relationship between the variants in form of phylogenetic trees. An interesting example is presented by a 16-bp insert in select variants of HPV-35, which appears to have given rise to additional variants by nucleotide exchanges within the insert. All trees showed distinct phylogenetic topologies, ranging from dichotomic branching in the case of HPV-31 to star phylogenies of the other three types. No clear similarities between these types or between these types and HPV-16 exist. While variant branches in some types were specific for Europe, Africa, or East Asia, none of the four trees reflected human evolution and spread to the extent illustrated by HPV-16. One possible explanation is that the rare HPV types that we studied spread and thereby diversified more slowly than the more abundant HPV-16 and may have established much of today's variant diversity already before the worldwide spread of humans 100,000 years ago. Most variants had prototypic amino acid sequences within the E6 oncoprotein and a segment of the L1 capsid protein. Some had one, two, or three amino acid substitutions in these regions, which might indicate biological and pathogenic diversity between the variants of each HPV type.     

The Role of the Anaphase-Promoting Complex/Cyclosome in Plant Growth

The anaphase-promoting complex/cyclosome (APC/C) is a multi-subunit E3 ubiquitin ligase that plays a major role in the progression of the eukaryotic cell cycle. This unusual protein complex targets key cell cycle regulators, such as mitotic cyclins and securins, for degradation via the 26S proteasome by ubiquitination, triggering the metaphase-to-anaphase transition and exit from mitosis. Because of its essential role in cell cycle regulation, the APC/C has been extensively studied in mammals and yeasts, but relatively less in plants. Evidence shows that, besides its well-known role in cell cycle regulation, the APC/C also has functions beyond the cell cycle. In metazoans, the APC/C has been implicated in cell differentiation, disease control, basic metabolism and neuronal survival. Recent studies also have shed light on specific functions of the APC/C during plant development. Plant APC/C subunits and activators have been reported to play a role in cellular differentiation, vascular development, shoot branching, female and male gametophyte development and embryogenesis. Here, we discuss our current understanding of the APC/C controlling plant growth.     

Two nucleotide positions in the Citrus exocortis viroid RNA associated with symptom expression in Etrog citron but not in experimental herbaceous hosts

Citrus exocortis viroid (CEVd) is the causal agent of exocortis disease of citrus. CEVd has a wide host range that includes woody and herbaceous species. A new CEVd strain (CEVd(COL)), phylogenetically clustering with CEVd variants of Class A inducing severe symptoms in tomato, was identified in Colombia and shown to induce only extremely mild symptoms in Etrog citron indicator plants. Using site-directed mutagenesis, two nucleotide substitutions (314A → G and 315U → A) in the lower strand of the P domain of the predicted CEVd(COL) secondary structure resulted in a severe artificial CEVd(MCOL) variant. Conversely, two nucleotide exchanges (314G → A and 315A → U) in the same region of the severe variant CEVd(E-117) resulted in a symptomless artificial CEVd(ME-117) variant. Infectivity assays conducted with the natural and mutated variants showed that all induced severe symptoms in Gynura aurantiaca, tomato and chrysanthemum. This is the first report of the identification of pathogenic determinants of CEVd in citrus, and shows that these pathogenicity determinants are host dependent.     

Lowland tapirs facilitate seed dispersal in degraded Amazonian forests

The forests of southeastern Amazonia are highly threatened by disturbances such as fragmentation, understory fires, and extreme climatic events. Large‐bodied frugivores such as the lowland tapir (Tapirus terrestris) have the potential to offset this process, supporting natural forest regeneration by dispersing a variety of seeds over long distances to disturbed forests. However, we know little about their effectiveness as seed dispersers in degraded forest landscapes. Here, we investigate the seed dispersal function of lowland tapirs in Amazonian forests subject to a range of human (fire and fragmentation) and natural (extreme droughts and windstorms) disturbances, using a combination of field observations, camera traps, and light detection and ranging (LiDAR) data. Tapirs travel and defecate more often in degraded forests, dispersing much more seeds in these areas [9,822 seeds per ha/year (CI_95% = 9,106; 11,838)] than in undisturbed forests [2,950 seeds per ha/year (CI_95% = 2,961; 3,771)]. By effectively dispersing seeds across disturbed forests, tapirs may contribute to natural forest regeneration—the cheapest and usually the most feasible way to achieve large‐scale restoration of tropical forests. Through the dispersal of large‐seeded species that eventually become large trees, such frugivores also contribute indirectly to maintaining forest carbon stocks. These functions may be critical in helping tropical countries to achieve their goals to maintain and restore biodiversity and its ecosystem services. Ultimately, preserving these animals along with their habitats may help in the process of natural recovery of degraded forests throughout the tropics. Abstract in Portuguese is available with online material.