Comparative Live-Cell Imaging Analyses of SPA-2, BUD-6 and BNI-1 in Neurospora crassa Reveal Novel Features of the Filamentous Fungal Polarisome

A key multiprotein complex involved in regulating the actin cytoskeleton and secretory machinery required for polarized growth in fungi, is the polarisome. Recognized core constituents in budding yeast are the proteins Spa2, Pea2, Aip3/Bud6, and the key effector Bni1. Multicellular fungi display a more complex polarized morphogenesis than yeasts, suggesting that the filamentous fungal polarisome might fulfill additional functions. In this study, we compared the subcellular organization and dynamics of the putative polarisome components BUD-6 and BNI-1 with those of the bona fide polarisome marker SPA-2 at various developmental stages of Neurospora crassa. All three proteins exhibited a yeast-like polarisome configuration during polarized germ tube growth, cell fusion, septal pore plugging and tip repolarization. However, the localization patterns of all three proteins showed spatiotemporally distinct characteristics during the establishment of new polar axes, septum formation and cytokinesis, and maintained hyphal tip growth. Most notably, in vegetative hyphal tips BUD-6 accumulated as a subapical cloud excluded from the Spitzenkörper (Spk), whereas BNI-1 and SPA-2 partially colocalized with the Spk and the tip apex. Novel roles during septal plugging and cytokinesis, connected to the reinitiation of tip growth upon physical injury and conidial maturation, were identified for BUD-6 and BNI-1, respectively. Phenotypic analyses of gene deletion mutants revealed additional functions for BUD-6 and BNI-1 in cell fusion regulation, and the maintenance of Spk integrity. Considered together, our findings reveal novel polarisome-independent functions of BUD-6 and BNI-1 in Neurospora, but also suggest that all three proteins cooperate at plugged septal pores, and their complex arrangement within the apical dome of mature hypha might represent a novel aspect of filamentous fungal polarisome architecture

Importancia de la genómica en aves para entender su interacción con patógenos

Anthropogenic pressure and global change are favouring the increase of infectious diseases not only in humans but also in wildlife. The negative effects of pathogens can be devastating, affecting natural populations and even causing the extinction of species. Therefore, it is fundamental to understand the mechanisms and strategies that these agents develop during their life cycle, as well as the immunological and biochemical responses that natural hosts show during host-pathogen interaction. Development of new molecular tools such as genomics or the analysis of transcriptomes have provided the scientific community significant information about host-pathogen interactions, including the biochemical cascades and immunological responses that occur. This article focuses on showing the advances in this field in birds infected with avian Plasmodium and West Nile Virus. In addition, we also review the problems that scientists must face in order to better understand the host-pathogen dynamics

Host escape behavior and blood parasite infections in birds

Active and risk-taking behavior may bring animals into contact with predators but can also result in frequent encounters with parasites and vectors via the exploration of risky or diverse habitats. Therefore, we predicted that antipredator behavior, here measured as escape behavior when captured by a human, would correlate with risk of parasite infection at the interspecific level with bolder species having more parasites than risk-averse species. Here we tested whether species with more active escape behavior also tended to have high prevalence of blood parasites, specifically hemosporidian parasites. Focusing on effect sizes we found that escape behavior was intermediately and positively related to prevalence of infection with Haemoproteus and Leucocytozoon, whereas that was not the case for the more virulent Plasmodium. Species that were habitat generalists and hence encountered a greater diversity of habitats had higher prevalence of blood parasites than specialists. In addition, some components of escape behavior were correlated at an intermediate magnitude with habitat exploration, as reflected by the relative frequency of feeding innovations, and coloniality. We failed to find considerable patterns of correlations between most of the behavioral variables and flight initiation distance, another commonly used antipredator behavior. Therefore, behavioral responses to an approaching predator and to being caught by a human likely represent 2 independent axes of antipredator behavior that do not evolve in concert. These findings are consistent with the hypothesis that escape behavior is related to risk of infection with blood parasites partially mediated by the effect of habitat generalism. © 2014 The Author.Peer Reviewe

El volumen de la glándula uropigial y la infección por malaria varía entre hábitats urbano-rural en el gorrión común

All the organisms have developed defensive against different (parasites, harsh environmental conditions or contaminants) to which they should face in their environments. To deal with these selective factors, birds have developed defensive mechanisms such as the uropygial gland, which produces a secretion acting as a physico-chemical defensive barrier against a high diversity of ectoparasites and prevent the potential transmission of blood parasites. Therefore, this secretion plays a fundamental role on the feather resistance to abrasion. However, there is still little knowledge about the functional adaptations of this gland to different environments. Here, we analysed the variation in uropygial gland volume in relation to different habitats (rural vs. urban) and to malaria infection in house sparrows (Passer domesticus). Our results showed that rural sparrows infected with malaria had the lowest uropygial volume. Furthermore, we found a positive relationship between uropygial gland volume and body condition, regardless of habitat, sex or prevalence of infection (infected or uninfected). These findings highlight the important role of this gland in the avian body condition and on its adaptation to different environments

Immune gene expression in the mosquito vector Culex quinquefasciatus during an avian malaria infection

Plasmodium relictum is the most widespread avian malaria parasite in the world. It is listed as one of the 100 most dangerous invasive species, having been responsible for the extinction of several endemic bird species, and the near-demise of several others. Here we present the first transcriptomic study focused on the effect of P. relictum on the immune system of its vector (the mosquito Culex quinquefasciatus) at different times post-infection. We show that over 50% of immune genes identified as being part of the Toll pathway and 30-40% of the immune genes identified within the Imd pathway are overexpressed during the critical period spanning the parasite's oocyst and sporozoite formation (8-12 days), revealing the crucial role played by both these pathways in this natural mosquito-Plasmodium combination. Comparison of infected mosquitoes with their uninfected counterparts also revealed some unexpected immune RNA expression patterns earlier and later in the infection: Significant differences in expression of several immune effectors were observed as early as 30 minutes after the ingestion of the infected blood meal. In addition, in the later stages of the infection (towards the end of the mosquito lifespan), we observed an unexpected increase in immune investment in uninfected, but not in infected, mosquitoes. In conclusion, our work extends the comparative transcriptomic analyses of malaria-infected mosquitoes beyond human and rodent parasites and provides insights into the degree of conservation of immune pathways and into the selective pressures exerted by Plasmodium parasites on their vectors

Application of Neutron Activation Analysis for Determination of As, Cr, Hg, and Se in Mosses in the Metropolitan Area of the Valley of Toluca, Mexico

This research presents a study of environmental monitoring at different sampling sites from the Metropolitan Area of the Valley of Toluca (MAVT), Mexico, using mosses (Leskea angustata (Tayl.) and Fabronia ciliaris (Brid.)) and soil samples. The epiphytic mosses and soils were sampled in two campaigns within two periods of the year, a rainy and dry-cold season. The selected sampling sites included urban regions (UR), transitional regions (TR), and protected natural areas (PA). The samples were analyzed by the Instrumental Neutron Activation Analysis (INAA) to determine As, Cr, Hg, and Se principally. However, due to the versatility of the analytic technique used, other elements including Cs, Co, Sc, Sb, Rb, Ce, La, Eu, and Yb were also detected. Statistical analysis (As, Cr, Hg, and Se) was carried out with principal components and cluster analysis methods; this revealed that a good correlation exists between metal content in mosses and the degree of pollution in the areas sampled. The obtained results in mosses showed that the concentrations of As, Cr, Co, Cs, Rb, Ce, La, and Yb increased with respect to the concentrations obtained during the first sampling, whereas Se, Sc, Sb and Eu, concentrations were decreased. For As and Hg, the concentrations were similar in both sampling periods. The soil samples present the most significant concentration

Preparation and Characterization of High Purity Anhydrous β-Lactose from α-Lactose Monohydrate at Mild Temperature

Lactose is a disaccharide of importance in humans dietary, food products, and the pharmaceutical industry. From the existing isomeric forms, β-lactose is rarely found in nature. Thus, in this work, a simple methodology to obtain anhydrous β-lactose (βL) from α-lactose monohydrate (αL·H2O) is presented. The αL·H2O powder was dispersed into a basic alcoholic solution (72 hours), at controlled conditions of temperature (27, 29, 31, and 32°C), without stirring. The slurry was dried at room temperature and characterized. Fourier transform infrared spectroscopy showed the formation of βL for the samples prepared at 29 and 32°C. Raman spectroscopy confirmed this result and suggested the occurrence of crystalline βL. Rietveld refinement of the X-ray diffraction patterns was employed to identify and quantify the composition of the isomers. The samples prepared at 29 and 31°C showed the formation of pure βL, while those at 27 and 32°C showed the presence of αL·H2O and a mixture of the two isomers, respectively. The morphology of the powders was studied by scanning electron microscopy, observing the formation of irregular shape αL·H2O particles and axe-like βL particles. Clearly, with this methodology, it was possible to obtain pure, crystalline, and anhydrous βL at mild temperature

First Report of Powdery Mildew (Pseudoidium anacardii) of Mango Trees in Sinaloa, Mexico

Powdery mildew of mango is an important disease in Mexico's northern Sinaloa state. Identification of the causal fungal agent has been hindered by the absence of information regarding its teleomorph, as well as a detailed morphometric analysis of the anamorph and molecular characterization. The first symptoms of the disease appear in mango inflorescences of early February, and it subsequently affects young fruits. The disease progresses during March and early April, causing significant fruit abortion and a scabby appearance in a high percentage of fruits that remain attached to the trees. We observed the disease on inflorescences but not in leaves during our sampling period. Powdery mildew specimens were collected during 2011 and 2012 and included Kent and Keith varieties from commercial orchards, and creole materials from backyards of private residences in the Ahome and Fuerte Counties of northern Sinaloa, Mexico. Symptomatic inflorescences were analyzed morphologically. Conidiophores and conidia were prepared by touching the whitish lesions with clear adhesive tape, which was then placed over microscope slides with a drop of distilled water and observed under a compound microscope. The anamorph structures of the pathogen were measured. The mycelium was septate and ramified on the surface of the host, forming a dense coat of branching hyphae. The mycelium had a diameter of 2.5 to 8.7 μm; conidiophores (Pseudoidium type) emerged from the superficial mycelium, were unbranched, and consisted of 1 to 3 cells with conidia forming singly from the apex. The length of the conidiophores varied from 30.0 to 77.5 μm; the foot cell of the conidiophores was straight, 10.0 to 47.5 μm long and with a diameter of 5.0 to 15.5 μm across its midpoint. Conidia without fibrosin bodies were borne singly, and were ellipsoid/ovoid, 22.5 to 46.2 μm long and 15.0 to 27.5 μm wide. Eighty percent of the germ tubes were forked (lobed); the rest were simple, emerged from the end, and were occasionally on the side of the conidia. Germ tubes ranged from 2.0 to 7.2 μm at the midpoint. The surface of the conidia appeared smooth under the scanning electron microscope, and elliptical conidia appeared constricted at their ends; this, however, was not observed in the ovoid conidia. In both cases, the terminal end of the conidia was smooth. The teleomorph was not found. Molecular and phylogenetic analysis of the ITS rDNA (2) region showed that samples are closely related to specimens of Pseudoidium anacardii (1) (teleomorph: Erysiphe quercicola [4]) collected from mango trees in diverse countries. Measurements of somatic and asexual structures are in agreement with descriptions of P. anachardii (formerly known as Oidium mangiferae) from India (3). The nucleotide sequences derived from this research were deposited in GenBank (Accession Nos. JX893951 to JX893957). To our knowledge, this is the first report of P. anacardii associated to mango inflorescences in Sinaloa, Mexico. Due to the economic importance of powdery mildew of mango trees in Sinaloa, future research directions should focus on finding the teleomorph of the fungus to support its identity