Culture conditions affect virulence and production of insect toxic proteins in the entomopathogenic fungus Metarhizium anisopliae

Conidial spores are often used as the infectious agent during insect biocontrol applications of entomopathogenic fungi. Here we show differential virulence of conidia derived from Metarhizium anisopliae strain EAMa 01/58-Su depending upon the solid substrata used for cultivation, where LC₅₀ values differed by up to ~10-fold (5.3×10⁶?4.5×10⁵ conidia/ml) and LT₅₀ values by ~40% (9.8?7.1 d). This fungal strain is also known to secrete proteins that are toxic towards adult Mediterranean fruit flies, Ceratitis capitata, and the Greater wax moth, Galleria mellonella, larvae. In vitro production and intrahemoceol injection using G. mellonella as the host was used to test fractions during purification of the protein toxins, demonstrating that they elicited defence-related responses including melanisation and tissue necrosis. Production of these proteins/peptides along with a number of potential cuticle degrading enzymes was confirmed both in vitro and during the infection process (in vivo). Two-dimensional gel electrophoresis, followed by gel elution and bioassay, was used to identify at least three proteins or peptides (molecular mass=11, 15 and 15 kDa) as mediating the observed insect toxicity. These data demonstrate that in vitro screening for insect toxins can mimic in vivo (i.e. during the infection process) secretion and applies the use of proteomics to invertebrate pathology.     

Fractionation of subcellular membranes of the secretory pathway from the peroxidase-producing white-rot fungus Phanerochaete chrysosporium

Abstract Mycelia from the basidiomycete Phanerochaete chrysosporium , producing lignin and manganese peroxidases, were homogenized and fractionated on a sucrose gradient. The main subcellular fungal membrane fractions were successfully separated. Lipid composition analyses of the isolated membranes as well as associated marker enzymes distribution gave evidence to similarities with membranes originating from plants. Lignin and manganese peroxidases were investigated by immunodetection in subcellular fractions. Our results show that lignin and manganese peroxidases are mainly associated with Golgi apparatus vesicles and, to a lesser extent, with endoplasmic reticulum and light density vesicles, but not with plasma membranes.     

Acremonoside,a phenolic glucoside from the sea fan-derived fungus Acremonium polychromum PSU-F125

A new phenolic glucoside, acremonoside (1), along with two known compounds, F-11334 A₂ and 2,2-dimethyl-2H-chromen-6-ol, were isolated from the sea fan-derived fungus Acremonium polychromum PSU-F125. The structure of 1 was elucidated by spectroscopic techniques, acid hydrolysis and X-ray crystallographic analysis. The isolated compounds were tested for antibacterial, antimalarial, antimycobacterial and cytotoxic activities.     

三株亚肉座菌乙酸乙酯提取物生物活性及GC-MS分析

采用乙酸乙酯提取3株亚肉座菌菌丝体,测试虫生真菌乙酸乙酯提取物（EAE）的抗肿瘤、抗菌和抗氧化活性,并借助GC-MS方法分析各提取物中的化学成分。结果发现2株亚肉座菌的菌丝体EAE对HepG2细胞的抑制活性较强,IC₅₀均小于9μmol/L;抗菌结果表明2株真菌的提取物具有抑制细菌生长的作用;1株供试菌的EAE表现出较强的DPPH自由基清除活性（清除率可达85%）。GC-MS分析表明从亚肉座菌JXJG201717、JXJG201720和ARSEF7697的EAE中分别鉴定出21、35和39种成分,主要成分为酯类、醇类和酸类;盘状亚肉座菌JXJG201720与ARSEF7697有相同化合物13个,与暹罗亚肉座菌JXJG201717存在7个相同化合物。本研究表明虫生亚肉座菌具有产生丰富活性成分的能力,彰显出多种利用价值。     

Post‐fire development of faunal habitat depends on plant regeneration traits

The concept that vegetation structure (and faunal habitat) develops predictably with time since fire has been central to understanding the relationship between fire and fauna. However, because plants regenerate after fire in different ways (e.g. resprouting from above‐ground stems vs. underground lignotubers), use of simple categories based on time since fire might not adequately represent post‐fire habitat development in all ecosystems. We tested the hypothesis that the post‐fire development of faunal habitat structure differs between ecosystems, depending on fire regeneration traits of the dominant canopy trees. We measured 12 habitat components at sites in foothill forests (n = 38), heathy woodlands (n = 38) and mallee woodlands (n = 98) in Victoria, Australia, and used generalised additive models to predict changes in each variable with time since fire. A greater percentage of faunal habitat variables responded significantly to time since fire in mallee woodlands, where fires typically are stand‐replacing, than in foothill forests and heathy woodlands, where canopy tree stems generally persist through fire. In the ecosystem with the highest proportion of epicormic resprouters (foothill forests), only ground cover and understorey vegetation responded significantly to time since fire, compared with all but one variable in the ecosystem dominated by basal resprouters (mallee woodlands). These differences between ecosystems in the post‐fire development of key habitat components suggest there may also be fundamental differences in the role of fire in shaping the distribution of fauna. If so, this challenges the way in which many fire‐prone ecosystems currently are categorised and managed, especially the level of dependence on time since fire and other temporal surrogates such as age‐classes and successional states. Where time since fire is a poor surrogate for habitat structural development, additional complexity (e.g. fire severity, topography and prior land‐use history) could better capture processes that determine faunal occurrence in fire‐prone ecosystems.     

Disease hotspots or hot species? Infection dynamics in multi‐host metacommunities controlled by species identity,not source location

Pathogen persistence in host communities is influenced by processes operating at the individual host to landscape‐level scale, but isolating the relative contributions of these processes is challenging. We developed theory to partition the influence of host species, habitat patches and landscape connectivity on pathogen persistence within metacommunities of hosts and pathogens. We used this framework to quantify the contributions of host species composition and habitat patch identity on the persistence of an amphibian pathogen across the landscape. By sampling over 11 000 hosts of six amphibian species, we found that a single host species could maintain the pathogen in 91% of observed metacommunities. Moreover, this dominant maintenance species contributed, on average, twice as much to landscape‐level pathogen persistence compared to the most influential source patch in a metacommunity. Our analysis demonstrates substantial inequality in how species and patches contribute to pathogen persistence, with important implications for targeted disease management.     

Micosis cutánea por Curvularia pallescens en un paciente trasplantado pulmonar: primer caso descrito en España

BackgroundCurvularia is a filamentous dematiaceous fungus increasingly recognized as a pathogen in immunocompromised patients. The most common clinical entities associated with this fungus are allergic sinusitis, cutaneous infection and keratitis. In this article, a report on the first clinical case of Curvularia pallescens cutaneous infection in Spain and its treatment is described.Case reportA 68 year-old man with a history of lung transplantation presented to Dermatology Unit due to a skin lesion in the knee that had been evolving for 6 months. A skin biopsy was performed for its study. In the histopathological study, an intense and non-specific inflammatory reaction in the dermis was observed, and with Grocott stain and periodic acid Schiff abundant septate hyphae and spores were found in the dermis. The culture of the sample revealed a filamentous fungus whose microscopic examination allowed to identify the genus as Curvularia. Using MALDI-TOF mass spectrometry and molecular identification, the fungus was finally identified as Curvularia pallescens. The patient underwent surgical resection of the lesion and was treated with posaconazole, evolving favorably.ConclusionsThe species of Curvularia should be considered causal agents of fungal skin infections in immunosuppressed patients. This clinical case, which showed good clinical response after surgical resection and treatment with posaconazole, is the first described in Spain due to this species.     

Integrating management techniques to restore subtropical forests invaded by Hedychium coronarium J. Köenig (Zingiberaceae) in a biodiversity hotspot

The restoration of areas invaded by non‐native plants is challenging as invasive plants may affect both biotic and abiotic components of ecosystems, leading to impacts that constrain recolonization by native species after invaders are eliminated. In such a scenario, restoration techniques as topsoil transposition might accelerate colonization by native species in forests. Hedychium coronarium J. Koenig (Zingiberaceae) is a Himalayan herbaceous rhizomatous plant recognized as invasive in several countries. This study aimed to experimentally evaluate the response of plant assemblages to topsoil transposition on a site invaded by H. coronarium after chemical control. Four treatments were applied: chemical control integrated with topsoil transposition, chemical control of H. coronarium alone, topsoil transposition alone, and no intervention (control). Plots were evaluated prior to the application of treatments and then monthly for 11 months after treatments. Parameters were measured for H. coronarium (number of ramets, ramet height, and cover) and other species (species richness, abundance, and cover). Plots treated with chemical control (regardless of topsoil transposition) were similar in terms of all parameters measured and species composition, with dominance of herbs and shrubs. Plots managed solely with topsoil transposition had lower species richness, abundance, and cover, but more diverse life‐forms, being equally rich in climbers, trees, and herbs. Chemical control was effective to control invasion by H. coronarium and increase species richness and abundance on the managed site. Topsoil transposition promoted colonization by species that might accelerate restoration.