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Sample records for deuteromycota hyphomycetes conidia

  1. Impact of two treatments of a formulation of Beauveria bassiana (Deuteromycota: Hyphomycetes) conidia on Varroa mites (Acari: Varroidae) and on honeybee (Hymenoptera: Apidae) colony health.

    PubMed

    Meikle, William G; Mercadier, Guy; Holst, Niels; Girod, Vincent

    2008-12-01

    Bee colonies in southern France were treated with conidia (asexual spores) from two strains of Beauveria bassiana, an entomopathogenic fungus. One strain was commercial (GHA) and the other had been isolated from Varroa mites in the region (Bb05002). Objectives were to evaluate treatment effect on colony weight, adult bee mass, capped brood, and on Varroa fall onto sticky boards. Treatments included conidia formulated with either carnauba or candelilla wax powder, candelilla wax powder alone, or control; in two treatment groups formulation was applied a second time after one week. Treatment did not affect colony health. Colonies treated twice with Bb05002 conidia and carnauba wax powder had significantly higher mite fall compared to colonies treated with blank candelilla wax powder. The proportion of fallen mites that were infected in both conidia treatments was higher than controls for 18 days after the second treatment. The number of fungal propagules on the bees themselves remained elevated for about 14 days after the second treatment. These results were compared to published results from previous experiments with regard to infection duration.

  2. Impact of two treatments of a formulation of Beauveria bassiana (Deuteromycota: Hyphomycetes) conidia on Varroa mites (Acari: Varroidae) and on honeybee (Hymenoptera: Apidae) colony health

    USDA-ARS?s Scientific Manuscript database

    Bee colonies in southern France were treated with conidia from 2 strains of Beauveria bassiana – one commercial (GHA) and the other isolated from Varroa mites in the region (Bb05002). Objectives were to evaluate treatment effect on colony weight, adult bee mass, capped brood, and on Varroa fall onto...

  3. Duration and spread of an entomopathogenic fungus, Beauveria bassiana (Deuteromycota: Hyphomycetes), used to treat varroa mites (Acari: Varroidae) in honey bee (Hymenoptera: Apidae) hives.

    PubMed

    Meikle, W G; Mercadier, G; Holst, N; Nansen, C; Girod, V

    2007-02-01

    A strain of the fungus Beauveria bassiana (Balsamo) Vuillemin (Deuteromycota: Hyphomycetes) isolated from varroa mites, Varroa destructor Anderson & Trueman (Acari: Varroidae), was used to treat honey bees, Apis mellifera L. (Hymenoptera: Apidae), against varroa mites in southern France. Fungal treatment caused a significant increase in the percentage of infected varroa mites compared with control treatments in two field experiments. In the first experiment, hives were treated with a formulation containing 0.37 g of B. bassiana conidia per hive and in the second experiment with a dose of 1.0 g of conidia per hive. The percentage of infected varroa mites also increased in the nontreated (control) hives, suggesting a movement of conidia, probably via bee drift, among the hives. Mite fall was significantly higher among treated hives compared with control hives on the sixth and eighth days after treatment in the first experiment. These days correspond to previously published data on the median survivorship of mites exposed to that fungal solate. The interaction of treatment and date was significant in the second experiment with respect to mite fall. Increases in colony-forming unit (cfu) density per bee were observed in all treatments but were significantly higher among bees from treated hives than control hives for at least a week after treatment. The relationship between cfu density per bee and proportion infected was modeled using a sigmoid curve. High levels of infection (>80%) were observed for cfu density per bee as low as 5 x 102 per bee, but the cfu density in hives treated with 0.37 g generally dropped below this level less than a week after treatment.

  4. Digitomyces, a new genus of hyphomycetes with cheiroid conidia.

    PubMed

    Mercado Sierra, Angel; Calduch, Misericordia; Gené, Josepa; Guarro, Josep; Delgado, Gregorio

    2003-01-01

    The new anamorphic genus Digitomyces, based on Dictyosporium verrucosum, is proposed. It is characterized by micronematous or semimacronematous conidiophores, integrated or discrete, holoblastic, determinate conidiogenous cells, and olivaceous to pale brown, verrucose, cheiroid conidia, with a truncate basal cell and (2-)3 arms. The new taxon is compared with morphologically similar anamorph genera.

  5. Effects of whole-stream nutrient enrichment on the concentration and abundance of aquatic hyphomycete conidia in transport

    Treesearch

    Vol. 96(1): 57-65 Mycologia

    2004-01-01

    The concentrations and relative abundances of aquatic hyphomycete conidia in water were followed during a three-year study in two headwater streams at Coweeta Hydrologic Laboratory, North Carolina, using the membrane-filtration technique. After a one-ye& pretreatment period, one of the streams was enriched continuously with inorganic nutrients (N+P) for two years...

  6. Effects of whole-stream nutrient enrichment on the concentration and abundance of aquatic hyphomycete conidia in transport.

    PubMed

    Gulis, Vladislav; Suberkropp, Keller

    2004-01-01

    The concentrations and relative abundances of aquatic hyphomycete conidia in water were followed during a three-year study in two headwater streams at Coweeta Hydrologic Laboratory, North Carolina, using the membrane-filtration technique. After a one-year pretreatment period, one of the streams was enriched continuously with inorganic nutrients (N+P) for two years while the other stream served as the reference. This ecosystem-level nutrient manipulation resulted in concentrations of aquatic hyphomycete conidia in the water of the treated stream that were 4.5-6.9 times higher than the concentrations observed during the pretreatment period and in the reference stream. Nutrient enrichment led to an increase in the number of fungal species detected on each sampling date. Changes in dominance patterns and relative abundances of individual species also were detected after treatment. Nutrient addition stimulates the reproductive activity of aquatic hyphomycetes, their colonization success and fungal-mediated leaf-litter decomposition. Such changes in the activity of the fungal community might affect higher trophic levels in lotic ecosystems.

  7. Effection of Ocneridia volxemi Bolivar (Pamphaginae, Orthoptera) hoppers and adults by Beauveria bassiana (Deuteromycotina, hyphomycetes) conidia in an oil formulation.

    PubMed

    Bounechada, M; Doumandji, S E

    2004-01-01

    Ocneridia volxemi Bolivar (Pamphaginae, Orthoptera) is one of the important insects pest in the heigh plains of north east of Algeria. Larval and adult stages caused severe damage to the leaves of cereals and an other various crops. Laboratory studies were conducted to the determine the potential of the fungus Beauveria bassiana for the control of the pest of larval and adults. Positive results were obtaved when larvae and adults were sprayed with different concentration of fungus conidia (D1: 2 x 10(6) conidia/ml, D2: 5 x 10(5) conidia/ml, D3: 8 x 10(4) conidia/ml, D4: 10(2) conidia/ml). Concentration at D1 and D2 in an oil formulation showed respectively 100% and 90 % larval mortality at the first day of treatment and 100%-9% adults mortality at the five day of treatement. The results revealed that the rates of infection and mortality were significantly related to the concentration of conidia in the corresponding samples. Beauveria bassiana can be considerd as a promising biocontrol agents of Ocneridia volxemi may be an other locuts, but field experimentation is necessary to subtantiate these finding.

  8. The effect of selected allelochemicals on germination of conidia and blastospores and mycelial growth of the entomopathogenic fungus, Paecilomyces fumosoroseus (Deuteromycotina: Hyphomycetes).

    PubMed

    Lacey, L A

    1998-01-01

    Selected allelochemicals that protect plants from invasion by plant pathogenic fungi were investigated for their activity against the entomopathogenic fungus, Paecilomyces fumosoroseus. The alkaloids tomatine, solanine, and camptothecin; the furanocoumarin, xanthotoxin; and the phenolic, tannic acid were tested for their effects on germination of conidia and blastospores and growth of mycelia. The LC50 values (corresponding to 50% inhibition of germination) for tomatine, solanine, camptothecin, xanthotoxin and tannic acid were 51.6, 95.9, 55.9, 83.0 and 72.8 mg/l respectively. When blastospores were placed on media containing a concentration of the individual allelochemicals that inhibit germination in approximately 50% of conidia, all but blastospores on tomatine had significantly less germination than did aerial conidia. Growth rates of mycelia were slowest in the camptothecin medium, followed by those of tomatine and xanthotoxin and were not significantly different from controls in the media containing solanine and tannic acid. A multitude of biotic and abiotic factors are responsible for specificity and degree of pathogenicity of entomopathogens. The effect of crop plant chemistry on the efficacy of entomopathogens should be quantified further in order to maximize their potential when used concomitantly with resistant plant varieties.

  9. The genera of Hyphomycetes - 2011 update.

    PubMed

    Seifert, K A; Gams, W

    2011-12-01

    This supplement to the taxonomic monograph The Genera of Hyphomycetes summarises information on 23 accepted new genera and c. 160 species described in 2011. These include three dematiaceous genera (Funbolia, Noosia, Pyrigemmula, all related to Dothideomycetes), a bulbil-producing genus, Spiroplana (Pleosporales), and two endophytic genera, the sterile Periglandula (Clavicipitaceae), and the hyaline, sympodial Micronematobotrys (Pyronemataceae). Slow-growing, morphologically-reduced, darkly pigmented fungi continue to be the source of new taxa, including the new genus Atramixtia (Dothioraceae). Eight new genera of darkly pigmented chlamydospore-like anamorphs were described from marine or subtidal environments (Glomerulispora, Halozoön, Hiogispora, Matsusporium, Moheitospora, Moleospora, Moromyces), mostly associated with subclades of the Lulworthiales. Several genera that are morphologically similar to but phylogenetically distinct from genera of the Capnodiales (Pseudopassalora, Scleroramularia) were introduced, as well as segregates from the classical concepts of Alternaria (Sinomyces), Chalara and Phialophora (Brachyalara, Infundichalara, Lasiadelphia), and Paecilomyces (Purpureocillium for the former Paecilomyces lilacinus complex). In addition, in anticipation of the new nomenclatural rules, newly configured formerly-teleomorph genera were proposed as segregates from classical hyphomycete genera in the Hypocreales, namely Acremonium (Cosmospora), Fusarium (Cyanonectria, Dialonectria, Geejayessia, Macroconia, Stylonectria), and Volutella (Pseudonectria) and the Trichocomaceae, Eurotiales, Penicillium (Talaromyces for the former Penicillium subg. Biverticillium). Standardized generic mini-diagnoses are provided for the accepted new genera, along with details of distribution, substrates, numbers of new species and phylogenetic affinities within the Dikarya. GenBank accession numbers for ITS DNA-barcodes are provided where available. New information on generic

  10. Preliminary Insights into the Phylogeography of Six Aquatic Hyphomycete Species

    PubMed Central

    Duarte, Sofia; Seena, Sahadevan; Bärlocher, Felix; Cássio, Fernanda; Pascoal, Cláudia

    2012-01-01

    Aquatic hyphomycetes occur worldwide on a wide range of plant substrates decomposing in freshwaters, and are known to play a key role in organic matter turnover. The presumed worldwide distribution of many aquatic hyphomycete species has been based on morphology-based taxonomy and identification, which may overlook cryptic species, and mask global-scale biogeographical patterns. This might be circumvented by using DNA sequence data. The internal transcribed spacer (ITS) region from rDNA was recently designated as the most suitable barcode for fungal identification. In this study, we generated ITS barcodes of 130 isolates belonging to 6 aquatic hyphomycete species (Anguillospora filiformis, Flagellospora penicillioides, Geniculospora grandis, Lunulospora curvula, Tetrachaetum elegans and Tricladium chaetocladium), and collected from streams of Southwest Europe (86 isolates) and East Australia (44 isolates). European and Australian populations of 4 species (A. filiformis, F. penicillioides, G. grandis and T. elegans) grouped into different clades, and molecular diversity indices supported significant differentiation. Continents did not share haplotypes, except for T. chaetocladium. Overall results show substantial population diversity for all tested species and suggests that the biogeography of aquatic hyphomycetes may be species-specific. PMID:23028909

  11. Nuclear staining of Paracoccidioides brasiliensis conidia.

    PubMed

    McEwen, J G; Restrepo, B I; Salazar, M E; Restrepo, A

    1987-10-01

    More than 80% of the conidia produced by two different isolates of Paracoccidioides brasiliensis, were found to be uninucleate; however, when they were incubated at 37 degrees C and began to transform into yeast cells, they became bi- or multi-nucleated, so that when most of the conidia had already transformed into yeast cells (72-96 h), there were at least four or five nuclei per cell in approximately 80% of the conidia examined.

  12. Failure of ticks to transmit Scopulariopsis brevicaulis (Deuteromycota), a common filamentous fungal commensal of ticks.

    PubMed

    Yoder, Jay A; Benoit, Joshua B; Rellinger, Eric J; Telford, Sam R

    2005-05-01

    The capacity of ticks to transmit a fungus was examined by analyzing tick saliva, host tissue from feeding sites, and host blood for presence of Scopulariopsis brevicaulis (Sacc.) Bainier, an internal mycosymbiont of the American dog tick, Dermacentor variabilis (Say), and lone star tick, Amnblyomma americanum (L.). Although >85% of ticks were infected with S. brevicaulis, conidia presence was low (0-5% of samples) in microscopic observations and mycological culturing of saliva expressed from larvae, nymphs, and adults. Additionally, the recovery of S. brevicaulis from blood and tissue feeding sites from a rabbit where S. brevicaulis-positive adult ticks had attached and fed was not increased compared with control tissue where no feeding occurred, indicating that transmission does not occur by the blood-feeding route. Tick mouthparts were found to contain S. brevicaulis in addition to Penicillium glabrum (Wehmer) Westling, but these agents were sparse in isolations from the feeding sites, which makes it unlikely that ticks act frequently as a mechanical fungal vector.

  13. Hyphomycetes on the Vochysiaceae from the Brazilian cerrado.

    PubMed

    Dornelo-Silva, Denise; Dianese, José C

    2003-01-01

    New hyphomycetes are described in association with leaves of native plants of the family Vochysiaceae, as part of studies of cerrado fungi. Six new species are described belonging to genera Alternaria (A. qualeae sp. nov.), Janetia (J. salvertiae sp. nov.), Passalora (P. qualeae sp. nov.) and Periconiella (P. longispora sp. nov., P. qualeae-grandiflorae sp. nov. and P. campo-grandensis sp. nov.). A key to the species of Periconiella on Qualea is provided.

  14. Aquatic hyphomycete communities as potential bioindicators for assessing anthropogenic stress.

    PubMed

    Solé, M; Fetzer, I; Wennrich, R; Sridhar, K R; Harms, H; Krauss, G

    2008-01-25

    With a profound knowledge of how physico-chemical parameters affect these communities, microbial communities could be used as indicators for environmental changes and for risk assessment studies. We studied aquatic hyphomycete communities in rivers and aquifers from sites shaped by intense mining activities (namely the "Mansfeld region") and chemical industry (cities of Halle and Bitterfeld) in Central Germany. Environmental stress factors such as high concentrations of heavy metals, sulphate, and nitrate as well as low concentrations of oxygen significantly reduced the diversity and biomass of hyphomycetes in the investigated samples. Redundancy analysis (RDA) indicates that variations in water chemistry cause a significant proportion of the change in fungal community structure (86.2%). Fungi were negatively correlated with high metal and nutrient concentrations. RDA also showed a strong influence of organic matter on individual species, with Anguillospora longissima (Sacc. et Syd.), Clavatospora longibrachiata (Ingold), Clavariopsis aquatica (De Wild), Flagellospora curvula (Ingold), Heliscus lugdunensis (Sacc. et Thérry), Tumularia aquatica (Ingold) and Lemonniera aquatica (De Wild) being most sensitive. We propose that aquatic hyphomycete communities can be used as sensitive and integrative indicators for freshwater quality.

  15. Proteomic profile of dormant Trichophyton Rubrum conidia

    PubMed Central

    Leng, Wenchuan; Liu, Tao; Li, Rui; Yang, Jian; Wei, Candong; Zhang, Wenliang; Jin, Qi

    2008-01-01

    Background Trichophyton rubrum is the most common dermatophyte causing fungal skin infections in humans. Asexual sporulation is an important means of propagation for T. rubrum, and conidia produced by this way are thought to be the primary cause of human infections. Despite their importance in pathogenesis, the conidia of T. rubrum remain understudied. We intend to intensively investigate the proteome of dormant T. rubrum conidia to characterize its molecular and cellular features and to enhance the development of novel therapeutic strategies. Results The proteome of T. rubrum conidia was analyzed by combining shotgun proteomics with sample prefractionation and multiple enzyme digestion. In total, 1026 proteins were identified. All identified proteins were compared to those in the NCBI non-redundant protein database, the eukaryotic orthologous groups database, and the gene ontology database to obtain functional annotation information. Functional classification revealed that the identified proteins covered nearly all major biological processes. Some proteins were spore specific and related to the survival and dispersal of T. rubrum conidia, and many proteins were important to conidial germination and response to environmental conditions. Conclusion Our results suggest that the proteome of T. rubrum conidia is considerably complex, and that the maintenance of conidial dormancy is an intricate and elaborate process. This data set provides the first global framework for the dormant T. rubrum conidia proteome and is a stepping stone on the way to further study of the molecular mechanisms of T. rubrum conidial germination and the maintenance of conidial dormancy. PMID:18578874

  16. Ergosterol-to-Biomass Conversion Factors for Aquatic Hyphomycetes

    PubMed Central

    Gessner, Mark O.; Chauvet, Eric

    1993-01-01

    Fourteen strains of aquatic hyphomycete species that are common on decaying leaves in running waters were grown in liquid culture and analyzed for total ergosterol contents. Media included an aqueous extract from senescent alder leaves, a malt extract broth, and a glucose-mineral salt solution. Concentrations of ergosterol in fungal mycelium ranged from 2.3 to 11.5 mg/g of dry mass. The overall average was 5.5 mg/g. Differences among both species and growth media were highly significant but followed no systematic pattern. Stationary-phase mycelium had ergosterol contents 10 to 12% lower or higher than mycelium harvested during the growth phase, but these differences were only significant for one of four species examined. Availability of plant sterols in the growth medium had no clear effect on ergosterol concentrations in two species tested. To convert ergosterol contents determined in field samples to biomass values of aquatic hyphomycetes, a general multiplicative factor of 182 is proposed. More accurate estimates would be obtained with species-specific factors. Using these in combination with estimates of the proportion of the dominant species in a naturally established community on leaves resulted in biomass estimates that were typically 20% lower than those obtained with the general conversion factor. Improvements of estimates with species-specific factors may be limited, however, by intraspecific variability in fungal ergosterol content. PMID:16348874

  17. Penzigomyces catalonicus, a new species of hyphomycetes from Spain.

    PubMed

    Mercado-Sierra, Angel; Gené, Josepa; Calduch, Misericordia; Guarro, Josep

    2004-01-01

    Penzigomyces catalonicus sp. nov., collected on dead branches of an unknown tree species in Spain, is described and illustrated. This fungus is characterized by short, dark brown, percurrent conidiophores, usually doliiform conidiogenous cells and ellipsoidal or slightly obclavate, (2-)3-4(-6) euseptate, thin-walled conidia. The new taxon is compared with species from other morphologically similar genera, and a key to the known species of Penzigomyces is provided.

  18. Elevated aluminium concentration in acidified headwater streams lowers aquatic hyphomycete diversity and impairs leaf-litter breakdown.

    PubMed

    Baudoin, J M; Guérold, F; Felten, V; Chauvet, E; Wagner, P; Rousselle, P

    2008-08-01

    Aquatic hyphomycetes play an essential role in the decomposition of allochthonous organic matter which is a fundamental process driving the functioning of forested headwater streams. We studied the effect of anthropogenic acidification on aquatic hyphomycetes associated with decaying leaves of Fagus sylvatica in six forested headwater streams (pH range, 4.3-7.1). Non-metric multidimensional scaling revealed marked differences in aquatic hyphomycete assemblages between acidified and reference streams. We found strong relationships between aquatic hyphomycete richness and mean Al concentration (r = -0.998, p < 0.0001) and mean pH (r = 0.962, p < 0.002), meaning that fungal diversity was severely depleted in acidified streams. By contrast, mean fungal biomass was not related to acidity. Leaf breakdown rate was drastically reduced under acidic conditions raising the issue of whether the functioning of headwater ecosystems could be impaired by a loss of aquatic hyphomycete species.

  19. Maturation of conidia on conidiophores of Aspergillus niger.

    PubMed

    Teertstra, Wieke R; Tegelaar, Martin; Dijksterhuis, Jan; Golovina, Elena A; Ohm, Robin A; Wösten, Han A B

    2017-01-01

    Conidia of Aspergillus niger are produced on conidiophores. Here, maturation of conidia on these asexual reproductive structures was studied. Pigmented conidia that had developed on conidiophores for 2, 5, and 8days were similarly resistant to heat and were metabolically active as shown by CO2 release and conversion of the metabolic probe Tempone. A total number of 645-2421 genes showed a ⩾2-fold change in expression when 2-day-old conidia were compared to 5- and 8-day-old spores. Melanin was extracted more easily from the cell wall of 2-day-old conidia when compared to the older spores. In addition, mannitol content and germination rate of the 2-day-old conidia were higher. Dispersal efficiency by water was lower in the case of the 8-day-old conidia but no differences were observed in dispersal by wind and a hydrophobic moving object. These data and the fact that only a minor fraction of the conidia on a conidiophore were dispersed in the assays imply that a single colony of A. niger releases a heterogeneous population of conidia. This heterogeneity would provide a selective advantage in environments with rapidly changing conditions such as availability of water. Copyright © 2016 The Authors. Published by Elsevier Inc. All rights reserved.

  20. Splash dispersal of Phyllosticta citricarpa conidia from infected citrus fruit

    PubMed Central

    Perryman, S. A. M.; Clark, S. J.; West, J. S.

    2014-01-01

    Rain-splash dispersal of Phyllosticta citricarpa (syn. Guignardia citricarpa) conidia (pycnidiospores) from infected oranges was studied in still air and combined with wind. High power microscopy demonstrated the presence of conidia in splash droplets from diseased oranges, which exuded conidia for over one hour during repeated wetting. The largest (5 mm) incident drops produced the highest splashes (up to 41.0 cm). A linear-by-quadratic surface model predicted highest splashes to be 41.91 cm at a horizontal distance of 25.97 cm from the target orange. Large splash droplets contained most conidia (4–5.5 mm splashes averaged 308 conidia), but were splashed <30 cm horizontal distance. Most (80–90%) splashes were <1 mm diameter but carried only 0–4 conidia per droplet. In multiple splash experiments, splashes combined to reach higher maxima (up to 61.7 cm; linear-by-quadratic surface model prediction, 62.1 cm) than in the single splash experiments. In combination with wind, higher wind speeds carried an increasing proportion of splashes downwind travelling horizontally at least 8 m at the highest wind speed tested (7 m/s), due to a small proportion of droplets (<1 mm) being aerosolised. These experiments suggest that P. citricarpa conidia can be dispersed from infected oranges by splashes of water in rainfall events. PMID:25298272

  1. Aquatic hyphomycete species are screened by the hyporheic zone of woodland streams.

    PubMed

    Cornut, Julien; Chauvet, Eric; Mermillod-Blondin, Florian; Assemat, Fiona; Elger, Arnaud

    2014-03-01

    Aquatic hyphomycetes strongly contribute to organic matter dynamics in streams, but their abilities to colonize leaf litter buried in streambed sediments remain unexplored. Here, we conducted field and laboratory experiments (slow-filtration columns and stream-simulating microcosms) to test the following hypotheses: (i) that the hyporheic habitat acting as a physical sieve for spores filters out unsuccessful strategists from a potential species pool, (ii) that decreased pore size in sediments reduces species dispersal efficiency in the interstitial water, and (iii) that the physicochemical conditions prevailing in the hyporheic habitat will influence fungal community structure. Our field study showed that spore abundance and species diversity were consistently reduced in the interstitial water compared with surface water within three differing streams. Significant differences occurred among aquatic hyphomycetes, with dispersal efficiency of filiform-spore species being much higher than those with compact or branched/tetraradiate spores. This pattern was remarkably consistent with those found in laboratory experiments that tested the influence of sediment pore size on spore dispersal in microcosms. Furthermore, leaves inoculated in a stream and incubated in slow-filtration columns exhibited a fungal assemblage dominated by only two species, while five species were codominant on leaves from the stream-simulating microcosms. Results of this study highlight that the hyporheic zone exerts two types of selection pressure on the aquatic hyphomycete community, a physiological stress and a physical screening of the benthic spore pool, both leading to drastic changes in the structure of fungal community.

  2. Aquatic Hyphomycete Species Are Screened by the Hyporheic Zone of Woodland Streams

    PubMed Central

    Chauvet, Eric; Mermillod-Blondin, Florian; Assemat, Fiona; Elger, Arnaud

    2014-01-01

    Aquatic hyphomycetes strongly contribute to organic matter dynamics in streams, but their abilities to colonize leaf litter buried in streambed sediments remain unexplored. Here, we conducted field and laboratory experiments (slow-filtration columns and stream-simulating microcosms) to test the following hypotheses: (i) that the hyporheic habitat acting as a physical sieve for spores filters out unsuccessful strategists from a potential species pool, (ii) that decreased pore size in sediments reduces species dispersal efficiency in the interstitial water, and (iii) that the physicochemical conditions prevailing in the hyporheic habitat will influence fungal community structure. Our field study showed that spore abundance and species diversity were consistently reduced in the interstitial water compared with surface water within three differing streams. Significant differences occurred among aquatic hyphomycetes, with dispersal efficiency of filiform-spore species being much higher than those with compact or branched/tetraradiate spores. This pattern was remarkably consistent with those found in laboratory experiments that tested the influence of sediment pore size on spore dispersal in microcosms. Furthermore, leaves inoculated in a stream and incubated in slow-filtration columns exhibited a fungal assemblage dominated by only two species, while five species were codominant on leaves from the stream-simulating microcosms. Results of this study highlight that the hyporheic zone exerts two types of selection pressure on the aquatic hyphomycete community, a physiological stress and a physical screening of the benthic spore pool, both leading to drastic changes in the structure of fungal community. PMID:24441154

  3. Development of Carrot Medium Suitable for Conidia Production of Venturia nashicola

    PubMed Central

    Choi, Eu Ddeum; Kim, Gyoung Hee; Lee, Young Sun; Jung, Jae Sung; Song, Jang Hoon; Koh, Young Jin

    2017-01-01

    The causal fungus of pear scab, Venturia nashicola, grows slowly and rarely produces conidia on artificial media in the laboratory, but it produced conidia on the Cheongah medium containing Cheongah powder. V. nashicola grew too slow to produce conidia until 15 days after cultivation but produced conidia with 4 × 104 conidia/plate 30 days after cultivation on the Cheongah medium containing 1% Cheongah powder. V. nashicola showed a peak production of conidia with 4.5 × 105 conidia/plate 60 days after cultivation on the carrot medium containing 2% carrot powder, one of the constituents of Cheongah powder. The carrot medium is considered to be the best medium to obtain conidia of V. nashicola in the laboratory until now. This is the first report on the development of a suitable medium for conidia production of V. nashicola, as far as we know. PMID:28167890

  4. Acquisition of lethal doses of Beauveria bassiana conidia by western flower thrips, Frankliniella occidentalis, exposed to foliar spray residues of formulated and unformulated conidia.

    PubMed

    Ugine, Todd A; Wraight, Stephen P; Sanderson, John P

    2005-09-01

    Secondary acquisition of Beauveria bassiana conidia was recorded on the whole bodies and selected body parts of second-instar nymphs and adult female western flower thrips exposed to foliar spray residues of three differently formulated conidial preparations, for 24 h. Conidia were formulated in emulsifiable oil or with clay (wettable powder), or were essentially unformulated conidia (technical grade powder suspended in water with a surfactant). Formulation had no significant effect on dose acquisition and no effect on virulence of acquired conidia. The mean nymphal LC50/LD50 was 116 conidia/mm2 and 52conidia/insect, respectively; the values for adults were 19 conidia/mm2 and 5conidia/insect. Greatest numbers of conidia were recorded on the legs and abdomens of nymphs and on the legs, wings, and thoraces of adults. As would be expected, numbers of conidia acquired increased with residue concentration (application rate). However, an inverse relationship was noted between acquisition rate (conidia acquired/total conidia applied) and residue concentration. The mechanism underlying this response was not determined. However, there was no indication that any body parts (e.g., tarsi) became saturated with spores, which suggests that either the thrips were repelled by the conidial residues or that as the concentrations of conidia on the substrate increased, conidia somehow became more difficult to acquire. Slopes of the LC probit regressions were lower than those of the LD regressions (mean 1.14 vs 1.78), suggesting that the low slopes often obtained in fungal pathogen assays could be partly an artifact of unequal rates of dose acquisition at low vs high application rates.

  5. [The carnivorous fungi hyphomycetes are natural regulators of the size of animal parasitic nematodes].

    PubMed

    Tepliakova, T V; Efremova, E A; Riabchikova, E I

    2005-01-01

    The carnivorous fungi hyphomycetes are natural enemies of soil nematodes. Laboratory tests examining the effect of the effective strain Duddingtonia flagrans T-89 on equine strongyle larvae have indicated that their size can be reduced 5-48-fold under the action of the fungus. Using helminth-infected mice as an example has ascertained that when the animals are fed a biopreparation, the chlamydial spores of the carnivorous fungus D. flagrans remain viable and continue their development in the excrements. The dead nematodes show cell structural impairments in all tissues and organs, which may be associated with the action of the substances contained in the cell envelope of the fungus.

  6. An overlooked source of fungal diversity: novel hyphomycete genera on trichomes of cerrado plants.

    PubMed

    Pereira-Carvalho, Rita C; Sepúlveda-Chavera, German; Armando, Eliane A S; Inácio, Carlos A; Dianese, José C

    2009-02-01

    Eight monotypic hyphomycete genera new to science are described from the trichomes of native plants growing in the cerrado of Brazil: Trichomatoclava cerradensis, Echinoconidiophorum cerradense, Globoconidiopsis cerradensis, Globoconidium cerradense, Helminthosporiomyces cerradensis, Trichomatosphaera [corrected] cerradensis , Phragmoconidium cerradense, and Vesiculohyphomyces cerradensis gens. spp. nov. Two of the new genera were found on hosts belonging in Myrtaceae, and one of each of the following families: Icacinaceae, Malphigiaceae, Fabaceae, Dilleniaceae, Chrysobalanaceae, and Caryocaraceae. These discoveries suggest that the trichomes of neotropical plants are an unexplored source of novel fungal diversity, and merit more attention in biodiversity surveys.

  7. Capillary electrophoresis of conidia from cultivated microscopic filamentous fungi.

    PubMed

    Horká, Marie; Růzicka, Filip; Kubesová, Anna; Holá, Veronika; Slais, Karel

    2009-05-15

    In immunocompromised people fungal agents are able to cause serious infections with high mortality rate. An early diagnosis can increase the chances of survival of the affected patients. Simultaneously, the fungi produce toxins and they are frequent cause of allergy. Currently, various methods are used for detection and identification of these pathogens. They use microscopic examination and growth characteristic of the fungi. New methods are based on the analysis of structural elements of the target microorganisms such as proteins, polysaccharides, glycoproteins, nucleic acids, etc. for the construction of antibodies, probes, and primers for detection. The above-mentioned methods are time-consuming and elaborate. Here hydrophobic conidia from the cultures of different strains of the filamentous fungi were focused and separated by capillary zone electrophoresis and capillary isoelectric focusing. The detection was optimized by dynamic modifying of conidia by the nonionogenic tenside on the basis of pyrenebutanoate. Down to 10 labeled conidia of the fungal strains were fluorometrically detected, and isoelectric points of conidia were determined. The observed isoelectric points were compared with those obtained from the separation of the cultured clinical samples, and they were found to be not host-specific.

  8. Modification of a pollen trap design to capture airborne conidia of Entomophaga maimaiga and detection of conidia by quantitative PCR

    Treesearch

    Tonya D. Bittner; Ann E. Hajek; Andrew M. Liebhold; Harold Thistle; Dan Cullen

    2017-01-01

    The goal of this study was to develop effective and practical field sampling methods for quantification of aerial deposition of airborne conidia of Entomophaga maimaiga over space and time. This important fungal pathogen is a major cause of larval death in invasive gypsy moth (Lymantria dispar) populations in the United States...

  9. Aquatic hyphomycete diversity and identity affect leaf litter decomposition in microcosms.

    PubMed

    Duarte, Sofia; Pascoal, Cláudia; Cássio, Fernanda; Bärlocher, Felix

    2006-04-01

    We conducted a microcosm experiment with monocultures and all possible combinations of four aquatic hyphomycete species, Articulospora tetracladia, Flagellospora curta, Geniculospora grandis and Heliscus submersus, to examine the potential effects of species richness on three functional aspects: leaf litter decomposition (leaf mass loss), fungal production (ergosterol buildup) and reproductive effort (released spores). Both species richness and identity significantly affected fungal biomass and conidial production (number and biomass of released spores), whereas only species identity had a significant effect on leaf mass loss. In mixed cultures, all measures of fungal functions were greater than expected from the weighted performances of participating species in monoculture. Mixed cultures outperformed the most active monoculture for biomass accumulation but not for leaf mass loss and conidial production. The three examined aspects of aquatic hyphomycete activity tended to increase with species richness, and a complementary effect was unequivocally demonstrated for fungal biomass. Our results also suggest that specific traits of certain species may have a greater influence on ecosystem functioning than species number.

  10. Optimization of Solid-State Fermentation for Improved Conidia Production of Beauveria bassiana as a Mycoinsecticide

    PubMed Central

    Pham, Tuan Anh; Kim, Jeong Jun

    2010-01-01

    The production of conidia of entomopathogenic Beauveria bassiana by solid-state fermentation was studied for the development of a biocontrol agent against aphid Myzus persicae. The optimal conditions for conidia production on polished white rice were 40% moisture content, 25℃ culture temperature, 2-day-old seeding culture grown in 3% corn meal, 2% rice bran, 2% corn steep powder medium, initial conidia concentration of 107 conidia/g in the wet rice, 10% inoculum size, and use of a polyethylene bag as a container. The polyethylene bag containing inoculated rice was hand-shaken every 12 hr during fermentation. Using optimal conditions, the maximum conidia production obtained was 4.05 g conidia/100 g dry rice after 14 days of cultivation, a rate 2.83 times higher than conidia yield of pre-optimization. PMID:23956642

  11. Kinetic studies on the aggregation of Aspergillus niger conidia.

    PubMed

    Grimm, L H; Kelly, S; Hengstler, J; Göbel, A; Krull, R; Hempel, D C

    2004-07-20

    Morphology has a crucial effect on productivity and the supply of substrate for cultures of filamentous fungi. However, cultivation parameters leading to the desired morphology are often chosen empirically as the mechanisms governing the processes involved are usually unknown. For coagulating microorganisms like Aspergillus niger the morphological development is considered to start with the aggregation of conidia right after inoculation. To elucidate the mechanism of this process, kinetic studies were carried out using an in-line particle size analyzer. Based on the data obtained from these experiments a model for conidial aggregation is proposed in this article. It consists of two separate aggregation steps. The first one takes place immediately after inoculation, but only leads to a small decrease of total particle concentration. Most suspended conidia aggregate after a second aggregation step triggered by germination and hyphal growth. Aggregation velocity of this second phase is linearly dependent on the particle growth rate.

  12. Heterogeneity in liquid shaken cultures of Aspergillus niger inoculated with melanised conidia or conidia of pigmentation mutants.

    PubMed

    van Veluw, G J; Teertstra, W R; de Bekker, C; Vinck, A; van Beek, N; Muller, W H; Arentshorst, M; van der Mei, H C; Ram, A F J; Dijksterhuis, J; Wösten, H A B

    2013-03-15

    Black pigmented conidia of Aspergillus niger give rise to micro-colonies when incubated in liquid shaken medium. These micro-colonies are heterogeneous with respect to gene expression and size. We here studied the biophysical properties of the conidia of a control strain and of strains in which the fwnA, olvA or brnA gene is inactivated. These strains form fawn-, olive-, and brown-coloured conidia, respectively. The ΔolvA strain produced larger conidia (3.8 μm) when compared to the other strains (3.2-3.3 μm). Moreover, the conidia of the ΔolvA strain were highly hydrophilic, whereas those of the other strains were hydrophobic. The zeta potential of the ΔolvA conidia in medium was also more negative when compared to the control strain. This was accompanied by the near absence of a rodlet layer of hydrophobins. Using the Complex Object Parametric Analyzer and Sorter it was shown that the ratio of individual hyphae and micro-colonies in liquid shaken cultures of the deletion strains was lower when compared to the control strain. The average size of the micro-colonies of the control strain was also smaller (628 μm) than that of the deletion strains (790-858 μm). The size distribution of the micro-colonies of the ΔfwnA strain was normally distributed, while that of the other strains could be explained by assuming a population of small and a population of large micro-colonies. In the last set of experiments it was shown that relative expression levels of gpdA, and AmyR and XlnR regulated genes correlate in individual hyphae at the periphery of micro-colonies. This indicates the existence of transcriptionally and translationally highly active and lowly active hyphae as was previously shown in macro-colonies. However, the existence of distinct populations of hyphae with high and low transcriptional and translational activity seems to be less robust when compared to macro-colonies grown on solid medium.

  13. Melanin dependent survival of Apergillus fumigatus conidia in lung epithelial cells.

    PubMed

    Amin, Shayista; Thywissen, Andreas; Heinekamp, Thorsten; Saluz, Hans Peter; Brakhage, Axel A

    2014-07-01

    Aspergillus fumigatus is the most important air-borne pathogenic fungus of humans. Upon inhalation of conidia, the fungus makes close contact with lung epithelial cells, which only possess low phagocytic activity. These cells are in particular interesting to address the question whether there is some form of persistence of conidia of A. fumigatus in the human host. Therefore, by also using uracil-auxotrophic mutant strains, we were able to investigate the interaction of A549 lung epithelial cells and A. fumigatus conidia in detail for long periods. Interestingly, unlike professional phagocytes, our study showed that the presence of conidial dihydroxynaphthalene (DHN) melanin enhanced the uptake of A. fumigatus conidia by epithelial cells when compared with non-pigmented pksP mutant conidia. Furthermore, conidia of A. fumigatus were able to survive within epithelial cells. This was due to the presence of DHN melanin in the cell wall of conidia, because melanised wild-type conidia showed a higher survival rate inside epithelial cells and led to inhibition of acidification of phagolysosomes. Both effects were not observed for white (non-melanised) conidia of the pksP mutant strain. Moreover, in contrast to pksP mutant conidia, melanised wild-type conidia were able to inhibit the extrinsic apoptotic pathway in A549 lung epithelial cells even for longer periods. The anti-apoptotic effect was not restricted to conidia, because both conidia-derived melanin ghosts (cell-free DHN melanin) and a different type of melanin, dihydroxyphenylalanine (DOPA) melanin, acted anti-apoptotically. Taken together, these data indicate the possibility of melanin-dependent persistence of conidia in lung epithelial cells.

  14. Photodynamic inactivation of Penicillium chrysogenum conidia by cationic porphyrins.

    PubMed

    Gomes, Maria C; Woranovicz-Barreira, Sandra M; Faustino, Maria A F; Fernandes, Rosa; Neves, Maria G P M S; Tomé, Augusto C; Gomes, Newton C M; Almeida, Adelaide; Cavaleiro, José A S; Cunha, Angela; Tomé, João P C

    2011-11-01

    This work reports the photophysical and biological evaluation of five cationic porphyrins as photosensitizers (PS) for the photodynamic inactivation (PDI) of Penicillium chrysogenum conidia. Two different cationic porphyrin groups were synthesized from 5,10,15,20-tetrakis(4-pyridyl)porphyrin and 5,10,15,20-tetrakis(pentafluorophenyl)porphyrin. The photostability and singlet oxygen generation studies showed that these molecules are photostable and efficient singlet oxygen generators. PDI experiments of P. chrysogenum conidia conducted with 50 μmol L(-1) of photosensitiser under white light at a fluence rate of 200 mW cm(-2) over 20 min showed that the most effective PS caused a 4.1 log reduction in the concentration of viable conidia. The present results show that porphyrins 1a and 1b are more efficient PSs than porphyrin 2a while porphyrins 1c and 2b show no inactivation of P. chrysogenum. It is also clear that the effectiveness of the molecule as PS for antifungal PDI is strongly related with the porphyrin substituent groups, and consequently their solubility in physiological media. The average amount of PS adsorbed per viable conidium was a determining factor in the photoinactivation efficiency and varied between the different studied PSs. Cationic PSs 1a and 1b might be promising anti-fungal PDI agents with potential applications in phytosanitation, biofilm control, bioremediation, and wastewater treatment.

  15. Preparation of fungal conidia impacts their susceptibility to inactivation by ethanol vapours.

    PubMed

    Dao, Thien; Dantigny, Philippe

    2009-11-15

    A common protocol employed for the preparation of conidia employs flooding a fungal colony grown on semi-solid media under optimum conditions with an aqueous solution. In contrast, conidia produced in a natural environment are usually not hydrated when disseminated in air and can be produced under water stress. In order to simulate the latter conditions, cultures were grown at different water activities and conidia were dry-harvested on the lid by turning the dishes upside-down then gently tapping the bottom of the box. This study aimed at assessing the effect of the preparation of fungal conidia on their inactivation by ethanol vapours. Firstly ethanol vapours (either 0.30 or 0.45 kPa) were applied to conidia obtained from the standardised protocol and to dry-harvested conidia for some species of Penicillium. While all dry-harvested conidia remained viable after 24 h of treatment, about 1.0, 3.5 and 2.5 log(10) reductions were observed for hydrated conidia of Penicillium chrysogenum, Penicillium digitatum and Penicillium italicum respectively. Secondly ethanol vapours (0.67 kPa) were applied to dry-harvested conidia obtained from cultures grown at 0.99 a(w) and at reduced water activities. For all species, the susceptibility to ethanol vapours of conidia obtained at 0.99 a(w) was significantly greater than that of conidia obtained at reduced water activities. Conidia produced in a natural environment under non-optimal conditions would be much more resistant to ethanol vapours than those produced in the laboratory. This phenomenon may be due to a reduced intracellular water activity of dry-harvested conidia.

  16. Trichothecene mycotoxins in aerosolized conidia of Stachybotrys atra.

    PubMed Central

    Sorenson, W G; Frazer, D G; Jarvis, B B; Simpson, J; Robinson, V A

    1987-01-01

    Stachybotrys atra is the etiologic agent of stachybotryotoxicosis, and this fungus and its trichothecene mycotoxins were recently implicated in an outbreak of unexplained illness in homes. S. atra was grown on sterile rice, autoclaved, dried, and then aerosolized by acoustic vibration. The distribution of particles (mass and number) was monitored on an aerodynamic particle sizer interfaced with a computer. Dust was collected on preweighed glass-fiber filters and extracted with 90% aqueous methanol. Extracts were tested for the ability to inhibit protein synthesis in rat alveolar macrophages, the ability to inhibit the proliferation of mouse thymocytes, and the presence of specific trichothecene mycotoxins. Virtually all of the particles were less than 15 micron in aerodynamic diameter, and the mass median diameter was 5 micron. Thus, most of the particles were respirable. Microscopic analysis of the generated dust revealed that ca. 85% of the dust particles were conidia of S. atra, another 6% were hyphal fragments, and the remainder of the particles were unidentifiable. Thus, greater than 90% of the particles were of fungal origin. The extracts strongly inhibited protein synthesis and thymocyte proliferation. Purified satratoxin H was also highly toxic in the same systems. Each of the individual filters contained satratoxin H (average, 9.5 ng/mg of dust). Satratoxin G and trichoverrols A and B were found in lesser amounts in some, but not all, of the filters. The limit of analysis is ca. 50 ng. These results establish that the conidia of S. atra contain trichothecene mycotoxins. In view of the potent toxicity of the trichothecenes, the inhalation of aerosols containing high concentrations of these conidia could be a potential hazard to health. PMID:3496850

  17. Paleomycology of the Princeton Chert I. Fossil hyphomycetes associated with the early Eocene aquatic angiosperm, Eorhiza arnoldii.

    PubMed

    Klymiuk, Ashley A; Taylor, Thomas N; Taylor, Edith L; Krings, Michael

    2013-01-01

    The Eocene (~ 48.7 Ma, Ypresian-Lutetian) Princeton Chert of British Columbia, Canada, has long been recognized as a significant paleobotanical locality, and a diverse assemblage of anatomically preserved fossil plants has been extensively documented. Co-occurring fossil fungi also have been observed, but the full scope of their diversity has yet to be comprehensively assessed. Here, we present the first of a series of investigations of fossilized fungi associated with the silicified plants of the Princeton Chert. This report focuses on saprotrophic, facultative-aquatic hyphomycetes observed in cortical aerenchyma tissue of an enigmatic angiosperm, Eorhiza arnoldii. Our use of paleontological thin sections provides the opportunity to observe and infer developmental features, making it possible to more accurately attribute two hyphomycetes that were observed in previous studies. These comprise multiseptate, holothallic, chlamydospore-like phragmoconidia most similar to extant Xylomyces giganteus and basipetal phragmospore-like chains of amerospores like those of extant Thielaviopsis basicola. We also describe a third hyphomycete that previously has not been recognized from this locality; biseptate, chlamydosporic phragmoconidia are distinguished by darkly melanized, inflated apical cells and are morphologically similar to Brachysporiella rhizoidea or Culcitalna achraspora.

  18. Transient transformation of Podosphaera xanthii by electroporation of conidia.

    PubMed

    Vela-Corcía, David; Romero, Diego; Torés, Juan Antonio; De Vicente, Antonio; Pérez-García, Alejandro

    2015-02-06

    Powdery mildew diseases are a major phytosanitary issue causing important yield and economic losses in agronomic, horticultural and ornamental crops. Powdery mildew fungi are obligate biotrophic parasites unable to grow on culture media, a fact that has significantly limited their genetic manipulation. In this work, we report a protocol based on the electroporation of fungal conidia, for the transient transformation of Podosphaera fusca (synonym Podosphaera xanthii), the main causal agent of cucurbit powdery mildew. To introduce DNA into P. xanthii conidia, we applied two square-wave pulses of 1.7 kV for 1 ms with an interval of 5 s. We tested these conditions with several plasmids bearing as selective markers hygromycin B resistance (hph), carbendazim resistance (TUB2) or GFP (gfp) under control of endogenous regulatory elements from Aspergillus nidulans, Neurospora crassa or P. xanthii to drive their expression. An in planta selection procedure using the MBC fungicide carbendazim permitted the propagation of transformants onto zucchini cotyledons and avoided the phytotoxicity associated with hygromycin B. This is the first report on the transformation of P. xanthii and the transformation of powdery mildew fungi using electroporation. Although the transformants are transient, this represents a feasible method for the genetic manipulation of this important group of plant pathogens.

  19. Evaluation of soyscreen in an oil-based formulation for UV protection of Beauveria bassiana conidia.

    PubMed

    Behle, Robert W; Compton, David L; Laszlo, Joseph A; Shapiro-Ilan, David I

    2009-10-01

    Soyscreen oil was studied as a formulation ingredient to protect Beauveria bassiana (Balsamo) Vuillemin conidia from UV degradation. Feruloylated soy glycerides, referred to as Soyscreen oil, are biobased UV-absorbing molecules made by combining molecules of soybean oil with ferulic acid. Conidia stored in Soyscreen oil for 28 wk at 25, 30, and 35 degrees C retained viability as well as conidia stored in sunflower oil, demonstrating that Soyscreen did not adversely affect viability with prolonged storage. For samples applied to glass and exposed to simulated sunlight (xenon light), conidia in sunflower oil with or without sunscreens (Soyscreen or oxyl methoxycinnimate) had similar conidia viability after exposure. These oil formulations retained conidia viability better than conidia applied as an aqueous treatment. However, the 10% Soyscreen oil formulation applied to field grown cabbage (Brassica oleracea L.) and bean (Phaseolus vulgaris L.) plants, did not improve residual insecticidal activity compared with aqueous applications of unformulated conidia or two commercial formulations when assayed against Trichoplusia ni (Hübner) larvae. Our results suggest that the oil applications lose UV protection because the oil was absorbed by the leaf. This conclusion was supported in subsequent laboratory exposures of conidia in oil-based formulations with UV screens applied to cabbage leaves or balsa wood, which lost protection as measured by decreased viability of conidia when exposed to simulated sunlight. As a result, additional formulation techniques such as encapsulation to prevent separation of the protective oil from the conidia may be required to extend protection when oil formulations are applied in the field.

  20. Microencapsuling aerial conidia of Trichoderma harzianum through spray drying at elevated temperatures

    USDA-ARS?s Scientific Manuscript database

    Trichoderma conidia are mostly produced by solid fermentation systems. Inoculum is produced by liquid culturing, and then transferred to solid substrate for aerial conidial production. Aerial conidia of T. harzianum are hydrophilic in nature, and it is difficult to separate them from the solid subst...

  1. Novel technique for quantifying adhesion of Metarhizium anisopliae conidia to the tick cuticle.

    PubMed

    Ment, Dana; Gindin, Galina; Rot, Asael; Soroker, Victoria; Glazer, Itamar; Barel, Shimon; Samish, Michael

    2010-06-01

    The present study describes an accurate quantitative method for quantifying the adherence of conidia to the arthropod cuticle and the dynamics of conidial germination on the host. The method was developed using conidia of Metarhizium anisopliae var. anisopliae (Metschn.) Sorokin (Hypocreales: Clavicipitaceae) and engorged Rhipicephalus annulatus (Say) (Arachnida: Ixodidae) females and was also verified for M. anisopliae var. acridum Driver et Milner (Hypocreales: Clavicipitaceae) and Alphitobius diaperinus (Panzer) (Coleoptera: Tenebrionidae) larvae. This novel method is based on using an organic solvent (dichloromethane [DCM]) to remove the adhered conidia from the tick cuticle, suspending the conidia in a detergent solution, and then counting them using a hemocytometer. To confirm the efficacy of the method, scanning electron microscopy (SEM) was used to observe the conidial adherence to and removal from the tick cuticle. As the concentration of conidia in the suspension increased, there were correlating increases in both the number of conidia adhering to engorged female R. annulatus and tick mortality. However, no correlation was observed between a tick's susceptibility to fungal infection and the amount of adhered conidia. These findings support the commonly accepted understanding of the nature of the adhesion process. The mechanism enabling the removal of the adhered conidia from the host cuticle is discussed.

  2. Fate of conidia of Paracoccidioides brasiliensis after ingestion by resident macrophages or cytokine-treated macrophages.

    PubMed Central

    Cano, L E; Brummer, E; Stevens, D A; Restrepo, A

    1992-01-01

    Conidia ingested by resident macrophages had an enhanced percentage of transformation to yeast cells compared with those in culture medium without macrophages. The yeast cells subsequently grew intracellularly by budding. Macrophages treated with cytokines from antigen-stimulated spleen cells from immunized mice significantly inhibited transformation of ingested conidia. PMID:1563800

  3. Fate of conidia of Paracoccidioides brasiliensis after ingestion by resident macrophages or cytokine-treated macrophages.

    PubMed

    Cano, L E; Brummer, E; Stevens, D A; Restrepo, A

    1992-05-01

    Conidia ingested by resident macrophages had an enhanced percentage of transformation to yeast cells compared with those in culture medium without macrophages. The yeast cells subsequently grew intracellularly by budding. Macrophages treated with cytokines from antigen-stimulated spleen cells from immunized mice significantly inhibited transformation of ingested conidia.

  4. Killing of Paracoccidioides brasiliensis conidia by pulmonary macrophages and the effect of cytokines.

    PubMed

    Cano, L E; Arango, R; Salazar, M E; Brummer, E; Stevens, D A; Restrepo, A

    1992-01-01

    The ability of conidia, the infectious form of the dimorphic fungus Paracoccidioides brasiliensis, to be killed in vitro by murine pulmonary macrophages was studied. Mice were immunized by intravenous injection of killed conidia, which resulted in cellular immunity demonstrated by delayed type hypersensitivity in vivo and macrophage migration inhibition factor production in vitro. Resident pulmonary macrophages from non-immune mice were able to significantly kill the conidia (28%). Such macrophages treated with supernatants (cytokines) from antigen-stimulated immune mononuclears had a markedly enhanced ability to kill conidia (73%). These results show that activated pulmonary macrophages are potent killers of conidia of P. brasiliensis and that immune mononuclears play a role in activation of macrophages. Activated macrophages may be important for pulmonary defense against the initial stages of infection with this fungus.

  5. Cell wall alpha1-3glucans induce the aggregation of germinating conidia of Aspergillus fumigatus.

    PubMed

    Fontaine, Thierry; Beauvais, Anne; Loussert, Céline; Thevenard, Benoît; Fulgsang, Claus C; Ohno, Naohito; Clavaud, Cécile; Prevost, Marie-Christine; Latgé, Jean-Paul

    2010-08-01

    The germination of Aspergillus fumigatus conidia can be divided into four stages: breaking of dormancy, isotropic swelling, establishment of cell polarity, and formation of a germ tube. Swelling of conidia is associated in liquid medium with a multi-cellular aggregation that produced large clumps of conidia. Conidial aggregation can be specifically prevented by the addition of alpha1-3glucanase. Swollen conidia specifically adhere to insoluble alpha1-3glucan chains. Electron microscopy studies showed that cell wall alpha1-3glucan chains became exposed at the cell surface during the swelling. These results demonstrate that cell wall alpha1-3glucans play an essential role in the aggregation between swollen conidia. Experiments with alpha1-3glucan coated latex beads show that alpha1-3glucan chains interacted between them without the requirement of any other cell wall component suggesting that biophysical properties of alpha1-3glucans are solely responsible for conidial aggregation.

  6. Hypocrea/Trichoderma: species with conidiophore elongations and green conidia.

    PubMed

    Chaverri, Priscila; Castlebury, Lisa A; Overton, Barrie E; Samuels, Gary J

    2003-01-01

    Species of Trichoderma and Hypocrea that have green conidia and sterile or fertile elongations of their conidiophores are described or redescribed and their phylogenetic position explored. The described species include T. crassum, T. fasciculatum, T. fertile, T. hamatum, T. longipile, T. oblongisporum, T. pubescens, T. spirale, T. strictipile, T. strigosum, T. stromaticum, T. tomentosum, Hypocrea aureoviridis f. macrospora, H. ceramica. and H. semiorbis. Trichoderma fasciculatum originally was described from cultures from ascospores of an unidentified Hypocrea specimen; it is considered to be a synonym of T. strictipile. The remaining species of Trichoderma considered here have not been linked to teleomorphs, and the Trichoderma anamorphs of H. aureoviridis f. macrospora and H. semiorbis have not been named. Five new species of Hypocrea are described, viz. H. cremea, H. cuneispora, H. estonica, H. strictipilosa and H. surrotunda. The phylogenetic relationships of these species were inferred based on partial RPB2 and EF-1α DNA sequence data and phenotypic characteristics, including teleomorph, anamorph, colony and growth rates. Trichoderma crassum was found to be a sister species to T. virens, based on molecular sequences and phenotypic data. Hypocrea surrotunda and H. cremea, H. cuneispora and T. longipile, T. fertile and T. oblongisporum, T. tomentosum and H. atrogelatinosa, and T. hamatum and T. pubescens, respectively, were found to be closely related phylogenetically, based on RPB2 and EF-1α gene genealogies. Anamorph and teleomorph phenotype, including conidiophore elongations, phialide morphology, conidial morphology, stroma anatomy and ascospore morphology are not useful predictors of relationships. Despite the shared phenotypic characters of these Trichoderma and Hypocrea species, they are distributed between two major clades of Trichoderma/Hypocrea. Redescriptions and a key to species of Hypocrea/Trichoderma with green conidia and conidiophore

  7. Physiological Response of Neurospora Conidia to Freezing in the Dehydrated, Hydrated, or Germinated State

    PubMed Central

    Leef, James L.; Mazur, Peter

    1978-01-01

    This study concerned the response to freezing of Neurospora crassa conidia in four different states: air-dry, hydrated in water, hydrated in Vogel medium lacking only sucrose, or hydrated in complete Vogel medium. All hydrated conidia were incubated in one of the above media for various times before freezing and were then washed and frozen in distilled water. Viability was estimated by three techniques, and the agreement among them was good. Hydration of air-dry conidia was found to be very rapid and, once hydrated, the conidia were much more sensitive to rapid freezing than they were before hydration. Rapidly cooled conidia survived freezing to a much higher extent when the warming rate was rapid than when it was slow; slowly cooled conidia showed little or no dependence on the warming rate. This sensitivity to rapid cooling and slow warming was attributed to the effects of intracellular ice. The sensitivity to freezing could be reversed by dehydrating the conidia in vacuo before freezing; thus, it was concluded that the presence or absence of water is the determining factor in the initial sensitivity due to freezing. In water, the sensitivity remained constant from 2 min to 15 days after hydration. Although conidia hydrated in growth medium lacking sucrose remained metabolically inactive, their sensitivity to rapid freezing decreased as a function of time in the medium before freezing. The reason for this decreased sensitivity is not understood. Conidia hydrated in complete growth medium (i.e., containing sucrose) became metabolically active and, after the initial sensitivity associated with hydration, became increasingly more sensitive to freezing as a function of their time in the medium. Drying itself was deleterious to metabolically active conidia, and those that survived dehydration did not exhibit a large absolute increase in resistance to subsequent freezing. The increase in sensitivity to freezing and to drying seems associated with the presence of

  8. Cadmium-induced formation of sulphide and cadmium sulphide particles in the aquatic hyphomycete Heliscus lugdunensis.

    PubMed

    Dobritzsch, Dirk; Ganz, Petra; Rother, Michael; Ehrman, James; Baumbach, Renate; Miersch, Jürgen

    2015-01-01

    Freshwater fungi which can survive under metal exposure receive increasing scientific attention. Enhanced synthesis of sulphide and glutathione but no phytochelatin synthesis in response to cadmium (up to 80 μM Cd(2+) in the medium) was measured in the aquatic hyphomycete Heliscus lugdunensis. Up to 25 μmol g(-1) dry mass the fungus formed sulphide in an exponentially Cd(2+)-concentration-dependent manner. Using light microscopy, precipitates were observed outside of the hyphae which could be determined as amorphous particles by X-ray diffraction (XRD). Energy dispersive X-ray spectroscopy (EDS) analysis indicated that these particles were mainly composed of Cd and S with an atomic ratio of 1:1, but some elements of the culture medium such as P and Cl were also present. Fungal cells exposed to Cd(2+) accumulated 12-28 μmol metal g(-1) dry mass over a period of 7-28 days. The results may indicate that sulphide could sequester excess Cd(2+) under oxygen deprived conditions and thereby reduce its toxicity via an additional avoidance mechanism of this fungus. Copyright © 2015 Elsevier GmbH. All rights reserved.

  9. Response of Aquatic Hyphomycete Communities to Changes in Heavy Metal Exposure

    NASA Astrophysics Data System (ADS)

    Sridhar, K. R.; Bärlocher, Felix; Krauss, Gerd-Joachim; Krauss, Gudrun

    2005-02-01

    Decomposition of Alnus glutinosa (alder) leaves was studied in a severely (site H4) and a moderately (site H8) heavy metal polluted stream in the former copper shale mining district of Mansfeld, Central Germany. Leaves at H8 had reduced fungal diversity and spore production but a high exponential decay rate (k = 0.065). No further mass loss of leaves occurred at H4 after 4-6 weeks, and fungal diversity and spore production were lower than in H8. Decay and sporulation rates gradually increased to values of H8 control leaves in leaves preincubated in H4 and then transferred to H8. These increases correlated with the invasion of transplanted leaves by Tetracladium marchalianum and Tricladium angulatum. In the reverse transplant experiment (H8 to H4), mass loss appeared to stop immediately. Sporulation rates also declined, but remained consistently above levels in H4 control leaves. Leaves precolonized in the laboratory by one of three aquatic hyphomycete species exhibited increased decay rates in both streams. Sporulation rates on these leaves were greater than those of control leaves in H4, but smaller than those of control leaves in H8.

  10. Identification and directed biosynthesis of efrapeptins in the fungusTolypocladium geodes gams (Deuteromycotina: Hyphomycetes).

    PubMed

    Krasnoff, S B; Gupta, S

    1991-10-01

    HPLC analysis of crude dichloromethane extracts of shaken liquid cultures of the hyphomycetous fungusTolypocladium geodes Gams revealed the presence of efrapeptins. These peptides, which have mitochondrial ATPase inhibitory activity as well as antifungal and insecticidal properties, are previously known only from the congeneric species,T. niveum Rostrup. The identity of efrapeptins was confirmed by fast atom bombardment mass spectrometry and by amino acid analysis. HPLC analyses of efrapeptins extracted from single isolates of bothT. geodes andT. niveum indicated that both species produced the same efrapeptins but the profile of relative abundance of the compounds produced was diagnostic for the isolates examined. Efrapeptin F was the major component of the mixture fromT. geodes with the order of abundance of the six efrapeptins detected being F >G>D∼E>H>C. Efrapeptin D was the major component fromT. niveum with the order of abundance of the six efrapeptins detected being D >E>F>C∼G>H. Efrapeptin F diifers from efrapeptin D by a single amino acid residue, efrapeptin F having an alanine where efrapeptin D has a glycine. Addition of alanine to the culture medium increased the relative abundance of efrapeptin F in the profile of both species. Conversely, addition of glycine increased the relative abundance of efrapeptin D in the profile of both species.

  11. Early events in macrophage killing of Aspergillus fumigatus conidia: new flow cytometric viability assay.

    PubMed

    Marr, K A; Koudadoust, M; Black, M; Balajee, S A

    2001-11-01

    Detailed investigations of macrophage phagocytosis and killing of Aspergillus fumigatus conidia have been limited by technical difficulties in quantifying fungal uptake and viability. In order to study early events in cell pathogen ingestion and killing, we developed a new flow cytometry assay that utilizes the fungus-specific viability dye FUN-1. Metabolically active A. fumigatus conidia accumulate orange fluorescence in vacuoles, while dormant or dead conidia stain green. After incubation within THP-1 cells, recovered conidia are costained with propidium iodide (PI) to discriminate between dormant and dead cells. Flow cytometric measurements of FUN-1 metabolism and PI uptake provide indicators of conidial viability, dormancy, and death. Conidial phagocytosis and killing are also assessed by measurement of green and orange FUN-1 fluorescence within the THP-1 cell population. Compared to previously described methods, this assay has less error introduced by membrane permeability changes and serial dilution of filamentous fungal forms. Results suggest that the THP-1 cells kill conidia rapidly (within 6 h) after exposure. Conidia that are preexposed to human serum are ingested and killed more quickly than are nonopsonized conidia.

  12. Differential Pathogenicity of Metarhizium Blastospores and Conidia Against Larvae of Three Mosquito Species.

    PubMed

    Alkhaibari, A M; Carolino, A T; Bull, J C; Samuels, R I; Butt, T M

    2017-05-01

    Biorational insecticides are being increasingly used in integrated pest management programs. In laboratory bioassays, the pathogenicity of blastospores and conidia of the entomopathogenic fungus Metarhizium brunneum ARSEF 4556 was evaluated against larvae of three mosquito species. Three propagule concentrations (1 × 106, 1 × 107, and 1 × 108 spores ml - 1) were used in the bioassays. Results showed that Aedes aegypti had lower survival rates when exposed to blastospores than when exposed to conidia, whereas the converse was true for Culex quinquefasciatus larvae. Anopheles stephensi larvae survival rates were similar when exposed to blastospores and conidia, except at the higher doses, where blastospores were more virulent. Several assays showed little difference in mortalities when using either 1 × 107 or 1 × 108 spores ml - 1, suggesting a threshold above which no higher control levels or economic benefit would be achieved. When tested at the lowest dose, the LT50 of Cx. quinquefasciatus using blastospores, wet conidia, and dry conidia was 3.2, 1.9, and 4.4 d, respectively. The LT50 of Ae. aegypti using blastospores, wet conidia, and dry conidia was 1.3, 3.3, and 6.2 d, respectively. The LT50 of An. stephensi using blastospores, wet conidia, and dry conidia was 2.0, 1.9, and 2.1 d, respectively. These observations suggest that for optimized control, two different formulations of the fungus may be needed when treating areas where there are mixed populations of Aedes, Anopheles, and Culex. © The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

  13. Ovularia puerariae Sawada is the hyphomycetous anamorph of a new Marasmius species on living leaves of kudzu (Pueraria montana, Fabaceae).

    PubMed

    Kirschner, Roland; Lee, I-Shu; Chen, Chee-Jen

    2013-01-01

    An arthroconidial hyphomycete on living leaves of kudzu (Pueraria montana, Fabaceae), originally described by Sawada in 1959 as Ovularia puerariae, was rediscovered. This anamorph is connected to an unknown Marasmius teleomorph belonging to section Globulares, which develops on the same living leaves. Ultrastructure and LSU rDNA sequence analysis of the anamorph confirm this connection. The fungus does not have only a unique biology among agarics, comparable only to Mycena citricolor, but also has the potential for application as a control agent of kudzu. During comparison with similar anamorph genera, Illosporium graminicola was found to be a synonym for Beniowskia sphaeroidea.

  14. Paecilomyces fumosoroseus (Deuteromycotina: Hyphomycetes) as a potential mycoparasite on Sphaerotheca fuliginea (Ascomycotina: Erysiphales).

    PubMed

    Kavková, Miloslava; Curn, Vladislav

    2005-01-01

    Hyphomycete Paecilomyces fumosoroseus that is well known as saprophytic and entomopatogenic fungus was investigated for its mycoparasitism on the cucumber powdery mildew pathogen. Mycoparasitism was documented by using standard bioassay and SEM. Effects of mycoparasitism were evaluated in three types of experiments. Paecilomyces fumosoroseus was applied in the form of graded suspensions into a colony of powdery mildew on a leaf segment. Interaction between both fungi was observed as the percentage of colonized area vs. experimental time. In the second experiment, young cucumber plants were sprayed with a suspension of Paecilomyces fumosoroseus 24 h before inoculation of Sphaerotheca fuliginea. Pre-treatment with P. fumosoroseus reduced development and spreading of powdery mildew infection significantly 15 days post-inoculation in contrast to pre-treatments with sulfur fungicide and distilled water. The development of pure culture powdery mildew under determined experimental conditions was observed and compared with treated variants. In the third experiment, mildewed plants were treated with a suspension of P. fumosoroseus. The control treatments with sulfur fungicide and distilled water were tested. Effects of P. fumosoroseus on the dispersion of powdery mildew during a 21-day period were observed. P. fumosoroseus suppressed the development and spread of cucumber powdery mildew significantly during the time of the experiment. The mechanical and physical damages and disruptions of vegetative and fruiting structures of powdery mildew were recorded under light microscopy and S.E.M. Results were concluded in pursuance to differences between the natural behaviour and development of S. fuliginea on cucumber plants treated with P. fumosoroseus and non-treated plants.

  15. Efrapeptin production byTolypocladium fungi (Deuteromycotina: Hyphomycetes): Intra- and interspecific variation.

    PubMed

    Krasnoff, S B; Gupta, S

    1992-10-01

    Production of the mitochondrial ATPase inhibitory peptides efrapeptins was evaluated by HPLC analysis in 44 strains of nine species of the fungal genusTolypocladium (Deuteromycotina: Hyphomycetes). Efrapeptin identification was confirmed by mass spectral data for the first time in the speciesT. cylindrosporum. HPLC retention time data indicated thatT. nubicola andT. tundrense, two species not previously known to produce efrapeptins, also produce the peptides. No efrapeptins were detected (<0.3 mg/100 ml broth) in single strains each ofT. balanoides, T. extinguens, T. parasiticum, andT. microsporum. Five strains ofT. geodes produced detectable amounts of efrapeptins and had compound F, or F and G, as the major component(s) in the mixture with the order of abundance being F> or ∼G > H ∼ D ∼ E > C. Eleven strains ofT. niveum produced detectable amounts of efrapeptins and had efrapeptins D and E as the primary and secondary components, respectively, in the mixture with the order of abundance being D > E > F > C ∼ G. A singleT. niveum strain had an efrapeptin profile similar to that of theT. geodes strains. Ten strains ofT. cylindrosporum had detectable amounts of efrapeptins. Of these, nine had F and one had G as the major component.T. cylindrosporum had higher ratios of E to D than didT. geodes. Efrapeptins were detected in one of twoT. nubicola strains analyzed (F > G > H) and one of threeT. tundrense strains (F > G > H > E).T. niveum strains could, in most cases, be identified to species on the basis of their efrapeptin profiles.

  16. Boric Acid Inhibition of Trichophyton rubrum Growth and Conidia Formation.

    PubMed

    Schmidt, Martin

    2017-04-08

    Trichophyton rubrum is a common human dermatophyte that is the causative agent of 80-93% of fungal infections of the skin and nails. While dermatophyte infections in healthy people are easily treatable with over-the-counter medications, such infections pose a higher risk for patients with compromised immune function and impaired regenerative potential. The efficacy of boric acid (BA) for the treatment of vaginal yeast infections prompted an investigation of the effect of BA on growth and morphology of T. rubrum. This is of particular interest since BA facilitates wound healing, raising the possibility that treating athlete's foot with BA, either alone or in combination with other antifungal drugs, would combine the benefits of antimicrobial activity and tissue regeneration to accelerate healing of infected skin. The data presented here show that BA represses T. rubrum growth at a concentration reported to be beneficial for host tissue regeneration. Oxygen exposure increases BA toxicity, and mycelia growing under BA stress avoid colonizing the surface of the growth surface, which leads to a suppression of aerial mycelium growth and surface conidia formation. BA penetrates into solid agar matrices, but the relative lack of oxygen below the substrate surface limits the effectiveness of BA in suppressing growth of embedded T. rubrum cells.

  17. Transformation of Epichloë typhina by electroporation of conidia

    PubMed Central

    2011-01-01

    Background Choke, caused by the endophytic fungus Epichloë typhina, is an important disease affecting orchardgrass (Dactylis glomerata L.) seed production in the Willamette Valley. Little is known concerning the conditions necessary for successful infection of orchardgrass by E. typhina. Detection of E. typhina in plants early in the disease cycle can be difficult due to the sparse distribution of hyphae in the plant. Therefore, a sensitive method to detect fungal infection in plants would provide an invaluable tool for elucidating the conditions for establishment of infection in orchardgrass. Utilization of a marker gene, such as the green fluorescent protein (GFP), transformed into Epichloë will facilitate characterization of the initial stages of infection and establishment of the fungus in plants. Findings We have developed a rapid, efficient, and reproducible transformation method using electroporation of germinating Epichloë conidia isolated from infected plants. Conclusions The GFP labelled E. typhina provides a valuable molecular tool to researchers studying conditions and mechanisms involved in the establishment of choke disease in orchardgrass. PMID:21375770

  18. Germination of fungal conidia after exposure to low concentration ozone atmospheres.

    USDA-ARS?s Scientific Manuscript database

    The germinability of conidia of Alternaria alternata, Aspergillus flavus, Aspergillus niger, Penicillium digitatum, Penicillium expansum, or Penicillium italicum was determined periodically during exposure for approximately 100 days to a humid atmosphere of air alone or air containing 150 ppb ozone ...

  19. THE POWER OF THE SMALL: THE EXAMPLE OF Paracoccidioides brasiliensis CONIDIA

    PubMed Central

    RESTREPO, Angela; CANO, Luz Elena; GONZALEZ, Ángel

    2015-01-01

    SUMMARY Research on Paracoccidioides brasiliensis has centered in the yeast cell probably because of the lack of distinctive features in the mycelium. In 1942 and for the first time, lateral conidia were noticed in the fungus' hyphae. Later on, Brazilian, Venezuelan and Argentinean researchers described "aleurias" when the fungus was grown in natural substrates. In 1970 authors became interested in the conidia and were able to obtain them in large numbers and treat them as individual units. Their shape and size were defined and the presence of all the elements of a competent eukaryotic cell were demonstrated. Conidia exhibited thermal dimorphism and, additionally, when given intranasally to BALB/c male mice, they converted into yeasts in the lungs and produce progressive pulmonary lesions with further dissemination to other organs. Studies on the phagocyte-conidia interaction were revealing and showed that these versatile structures allow a better understanding of the host- P. brasiliensis interactions. PMID:26465363

  20. A staining method for assessing the viability of Esteya vermicola conidia.

    PubMed

    Wang, Yunbo; Thang, NguyenTrong; Li, Zheng; Zhang, Yongan; Li, Jingjie; Xue, Jianjie; Gu, Lijuan; Hong, VuThuy; Mira, Lee; Sung, Changkeun

    2014-07-01

    The viability of conidia of Esteya vermicola, a potentially important biocontrol agent against the pinewood nematode Bursaphelenchus xylophilus, is usually determined by cultivation for 18-48 h in culture medium. As an alternative to this labor-intensive method, we have developed a rapid, simple, and low-cost staining method for assessing E vermicola conidia survival rates. A mixture of neutral red and methylene blue was found to be the most optimal among several stains that also included safranin O and Janus green B. This mixture stained nonviable conidia blue, in contrast to viable conidia, which were stained red in the cytoplasm and blue in the cell wall. This method may be particularly useful for traditional research laboratories, as it provides rapid results using common, relatively inexpensive laboratory equipment.

  1. Comparison of methodologies for conidia production by Alternaria alternata from citrus

    PubMed Central

    Carvalho, Daniel D. C.; Alves, Eduardo; Batista, Tereza R. S.; Camargos, Renato B.; Lopes, Eloísa A. G. L.

    2008-01-01

    Conidia production is a problem in the study of Alternaria alternata from citrus. Thus, this study aimed to compare existing methodologies for conidial production of A. alternata isolated from Ponkan tangerine (2 isolates), Cravo lemon (1 isolate), Pêra orange (2 isolates) and Murcott tangor (1 isolate). The methodologies used were conidia production with 12 and 24 hours under white fluorescent light, evaluation with 24 and 48 hours after applying fungal mycelium stress technique, cold stress followed by injury of mycelium and evaluation with 24 hours, using healthy vegetable tissue and the use of black fluorescent near ultraviolet (NUV) lamp. Satisfactory result was obtained with A. alternata isolate from Murcott tangor, with the production of 2.8 × 105 conidia mL-1, when fungal mycelium was stressed (Petri dish with 66.66% of fungi growth) and subsequently 24 h of growth. The use of white light (24 h) and black fluorescent NUV lamp also induced expressive conidia production by one isolate of Ponkan tangerine, which produced 17.2 × 105 and 10.1 × 105conidia mL-1 and another of Murcott tangor, which produced 13.9 × 105 and 10.1 × 105 conidia mL-1, respectively. The remaining methodologies analyzed in this study were not able to induce conidia production in satisfactory quantity. The use of both mycelium stress technique and white light (24 h) and black fluorescent NUV lamp allowed the production of enough quantities of conidia to be used in vitro (detection of fungitoxic substances) and in vivo (pathogenicity test) assays, respectively. PMID:24031309

  2. Comparison of methodologies for conidia production by Alternaria alternata from citrus.

    PubMed

    Carvalho, Daniel D C; Alves, Eduardo; Batista, Tereza R S; Camargos, Renato B; Lopes, Eloísa A G L

    2008-10-01

    Conidia production is a problem in the study of Alternaria alternata from citrus. Thus, this study aimed to compare existing methodologies for conidial production of A. alternata isolated from Ponkan tangerine (2 isolates), Cravo lemon (1 isolate), Pêra orange (2 isolates) and Murcott tangor (1 isolate). The methodologies used were conidia production with 12 and 24 hours under white fluorescent light, evaluation with 24 and 48 hours after applying fungal mycelium stress technique, cold stress followed by injury of mycelium and evaluation with 24 hours, using healthy vegetable tissue and the use of black fluorescent near ultraviolet (NUV) lamp. Satisfactory result was obtained with A. alternata isolate from Murcott tangor, with the production of 2.8 × 10(5) conidia mL(-1), when fungal mycelium was stressed (Petri dish with 66.66% of fungi growth) and subsequently 24 h of growth. The use of white light (24 h) and black fluorescent NUV lamp also induced expressive conidia production by one isolate of Ponkan tangerine, which produced 17.2 × 10(5) and 10.1 × 10(5)conidia mL(-1) and another of Murcott tangor, which produced 13.9 × 10(5) and 10.1 × 10(5) conidia mL(-1), respectively. The remaining methodologies analyzed in this study were not able to induce conidia production in satisfactory quantity. The use of both mycelium stress technique and white light (24 h) and black fluorescent NUV lamp allowed the production of enough quantities of conidia to be used in vitro (detection of fungitoxic substances) and in vivo (pathogenicity test) assays, respectively.

  3. Transcriptomic analysis of the exit from dormancy of Aspergillus fumigatus conidia

    PubMed Central

    Lamarre, Claude; Sokol, Sergueï; Debeaupuis, Jean-Paul; Henry, Christine; Lacroix, Céline; Glaser, Philippe; Coppée, Jean-Yves; François, Jean-Marie; Latgé, Jean-Paul

    2008-01-01

    Background Establishment of aspergillosis is depending upon the exit from dormancy and germination of the conidia of Aspergillus fumigatus in the lung. To gain an understanding of the molecular mechanisms underlying the early steps of conidial germination, we undertook a transcriptomic analysis using macroarrays constructed with PCR fragments from > 3,000 genes (around one third of the annotated A. fumigatus genome). Results Major results of this analysis are the following: (i) conidia stored pre-packaged mRNAs transcripts (27% of genes have transcripts in the resting conidia; (ii) incubation at 37°C in a nutritive medium induced up- and down-regulation of genes: 19% of the total number of genes deposited on the array were up-regulated whereas 22% of the genes with pre-packaged mRNA in the resting conidia were down-regulated; (iii) most modifications were seen during the first 30 min of germination whereas very little modification of gene expression occurred during the following hour; (iv) one-year old conidia and one-week old conidia behaved similarly at transcriptional level. Conclusion Transcriptomic data indicate that the exit from dormancy is associated with a shift from a fermentative metabolism to a respiratory metabolism as well as a trend toward immediate protein synthesis. PMID:18796135

  4. Interaction between Paracoccidioides brasiliensis conidia and the coagulation system: involvement of fibrinogen

    PubMed Central

    Tamayo, Diana; Hernández, Orville; Muñoz-Cadavid, Cesar; Cano, Luz Elena; González, Angel

    2013-01-01

    The infectious process starts with an initial contact between pathogen and host. We have previously demonstrated that Paracoccidioides brasiliensis conidia interact with plasma proteins including fibrinogen, which is considered the major component of the coagulation system. In this study, we evaluated the in vitro capacity of P. brasiliensis conidia to aggregate with plasma proteins and compounds involved in the coagulation system. We assessed the aggregation of P. brasiliensis conidia after incubation with human serum or plasma in the presence or absence of anticoagulants, extracellular matrix (ECM) proteins, metabolic and protein inhibitors, monosaccharides and other compounds. Additionally, prothrombin and partial thromboplastin times were determined after the interaction of P. brasiliensis conidia with human plasma. ECM proteins, monosaccharides and human plasma significantly induced P. brasiliensis conidial aggregation; however, anticoagulants and metabolic and protein inhibitors diminished the aggregation process. The extrinsic coagulation pathway was not affected by the interaction between P. brasiliensis conidia and plasma proteins, while the intrinsic pathway was markedly altered. These results indicate that P. brasiliensis conidia interact with proteins involved in the coagulation system. This interaction may play an important role in the initial inflammatory response, as well as fungal disease progression caused by P. brasiliensis dissemination. PMID:23827999

  5. Interaction between Paracoccidioides brasiliensis conidia and the coagulation system: involvement of fibrinogen.

    PubMed

    Tamayo, Diana; Hernández, Orville; Muñoz-Cadavid, Cesar; Cano, Luz Elena; González, Angel

    2013-06-01

    The infectious process starts with an initial contact between pathogen and host. We have previously demonstrated that Paracoccidioides brasiliensis conidia interact with plasma proteins including fibrinogen, which is considered the major component of the coagulation system. In this study, we evaluated the in vitro capacity of P. brasiliensis conidia to aggregate with plasma proteins and compounds involved in the coagulation system. We assessed the aggregation of P. brasiliensis conidia after incubation with human serum or plasma in the presence or absence of anticoagulants, extracellular matrix (ECM) proteins, metabolic and protein inhibitors, monosaccharides and other compounds. Additionally, prothrombin and partial thromboplastin times were determined after the interaction of P. brasiliensis conidia with human plasma. ECM proteins, monosaccharides and human plasma significantly induced P. brasiliensis conidial aggregation; however, anticoagulants and metabolic and protein inhibitors diminished the aggregation process. The extrinsic coagulation pathway was not affected by the interaction between P. brasiliensis conidia and plasma proteins, while the intrinsic pathway was markedly altered. These results indicate that P. brasiliensis conidia interact with proteins involved in the coagulation system. This interaction may play an important role in the initial inflammatory response, as well as fungal disease progression caused by P. brasiliensis dissemination.

  6. Melanin is required for the formation of the multi-cellular conidia in the endophytic fungus Pestalotiopsis microspora.

    PubMed

    Yu, Xi; Huo, Liang; Liu, Heng; Chen, Longfei; Wang, Yu; Zhu, Xudong

    2015-10-01

    Melanin plays an important role in regulating various biological processes in many fungi. However, its biological role in conidiation remains largely elusive. We report here that conidia production, morphogenesis, integrity, germination and their viability in Pestalotiopsis microspora require the polyketide-derived melanin. A polyketide synthase gene, pks1, was identified and demonstrated responsible for melanin biosynthesis in this fungus. A targeted deletion mutant strain Δpks1 displayed a defect in pigmentation of conidia and had an albino colonial phenotype. Interestingly, Δpks1 produced approximately 6-fold as many conidia as the wild type did, suggesting a negative modulation of melanin on conidia production in this fungus. Moreover, the conidia failed to develop into the normal five-cell morphology, rather the three main-body cells separated via constriction at the original septum position to generate three independent mutant conidia. This result suggests a novel role of melanin in the formation of the multi-cellular conidia. Germ tubes could develop from the three different types of mutant conidia and kept elongating, despite a significantly lower germination rate was observed for them. Still more, the unpigmented conidia became permeable to Calcofluor White and DAPI, suggesting the integrity of the conidia was impaired. Deliberate inhibition of melanin biosynthesis by a specific inhibitor, tricyclazole, led to a similar phenotypes. This work demonstrates a new function of fungal melanin in conidial development.

  7. Solid substrate production of Epicoccum nigrum conidia for biological control of brown rot on stone fruits.

    PubMed

    Larena, I; De Cal, A; Melgarejo, P

    2004-07-15

    Production of conidia of Epicoccum nigrum, a biocontrol agent of the fungal pathogen Monilinia laxa, was tested in liquid- and solid-state fermentation. Liquid fermentation was conducted in 250 ml Erlenmeyer flasks containing 50 ml of a mineral medium (containing per litre: 20 g lactose, 10 g NO3K, 1 g K2HPO4, 0.5 g MgSO4.7H2O, and 1 ml of a minor-element solution), inoculated with 2 x 10(5) E. nigrum conidia ml(-1), and incubated at 20-25 degrees C and 150 rpm for 7 days. Solid-state fermentation was carried out in specially designed plastic bags (600 cm3) (VALMIC) containing either 50 g of peat/vermiculite (1:1, w/w), or 50 g of peat/vermiculite/lentil meal (1:1:1, w/w/w) with 40% (v/w) initial moisture content. Substrate was inoculated with a conidial suspension of E. nigrum to give 10(5) conidia g(-1) substrate, and bags were incubated at 20-25 degrees C for 7 days in darkness. The amount of conidia of E. nigrum obtained in solid-state fermentation with substrate based on peat/vermiculite/lentil meal was 10-fold higher than with substrate based on peat/vermiculite or in liquid fermentation. Conidial production under these conditions was maintained in the range of 10(8) conidia g(-1) substrate from 10 to 150 days after inoculation. Germinability of these conidia was >90%. Addition of other nutrients than lentil meal to peat/vermiculite did not enhance production of conidia. Presence of peat in the substrate was necessary for good conidia production, but change in the kind of peat or vermiculite did not improve conidial production. Conidial production was similar when the substrate was inoculated with 10(5), 10(6) or 10(7) conidia g(-1) dry substrate. Incubation of bags in light conditions did not enhance conidial production. Fresh conidia produced in this solid-state fermentation system reduced the incidence and lesion diameter induced by M. laxa on peaches.

  8. Effects of Stachybotrys chartarum (atra) conidia and isolated toxin on lung surfactant production and homeostasis.

    PubMed

    Mason, C D; Rand, T G; Oulton, M; MacDonald, J M; Scott, J E

    1998-01-01

    This study evaluated the effects of Stachybotrys chartarum conidia and a trichothecene, isosatratoxin-F, on choline incorporation into DSPC by fetal rabbit alveolar type II cells and on alveolar surfactant subtypes in mice. Exposure of fetal rabbit type II cells to S. chartarum conidia at concentrations of 10(3) to 10(6) conidia ml(-1) significantly depressed [3H] choline incorporation after 24 h of exposure. Exposure of the rabbit cells to 10(5) to 10(6) conidia ml(-1) also resulted in significantly depressed [3H] choline uptake after 48 h. Additionally, fetal rabbit alveolar type II cells exposed to isosatratoxin-F in concentrations ranging from 10(-9) to 10(-4) M showed a significant reduction in [3H] choline incorporation into DSPC. Alveolar surfactant phospholipid concentrations in the different metabolic subfractions of lung lavage fluid of mice intratracheally exposed to either 50 microl of 10(7) ml(-1) S. chartarum conidia or 50 microl 10(-7) M isosatratoxin-F showed some significant changes at 12, 24, 48, and 72 h post-exposure, compared to the surfactant subfractions of control mice which were either untreated, exposed to saline or to 50 microl of 10(-7) ml(-1) Cladosporium cladosporioides conidia. In both the S. chartarum- and the isosatratoxin-F-treated mice, exposure significantly increased P10, P100, and S100 phospholipid concentrations, while the P60 phospholipid concentrations were depressed. In contrast, C. cladosporioides-treated mice showed only one significant change in subfraction phospholipid concentration: P60 was depressed at 48 h post-exposure. These results reveal that alveolar type II cells are sensitive to exposure to S. chartarum conidia and to isosatratoxin F. Sensitivity is manifest by alterations in the normal metabolic processing of alveolar surfactant. In exposed mice, this effect appears to involve a significant increase in newly secreted surfactant and an accumulation of the used surfactant forms.

  9. [Identification of genes associated with germination of conidia to form mycelia in the fungus Paracoccidioides brasiliensis].

    PubMed

    García, Ana María; Hernández, Orville; Aristizábal, Beatriz H; Cano, Luz Elena; Restrepo, Angela; McEwen, Juan G

    2009-09-01

    Paracoccidioides brasiliensis is a thermo-dimorphic fungus. At room temperature it grows as a mold that produces conidia, whereas in the vertebrate host it grows as a multiple-budding yeast. The molecular mechanisms involved in the germination from the conidia to the mycelia process remain unknown. The kinetics of conidia to mycelia germination process were studied in the dimorphic fungus P. brasiliensis. Gene expression during this process was evaluated by construction and analysis of an EST library. For the germination kinetics study, P. brasiliensis conidia were isolated as single cell units. Then, they were cultured at 18 degrees C in BHI (brain-heart infusion) broth for 24, 48, 72 and 96 hr. After each perion, they were examined by light microscopy. From conidia harvested at 96 hr, an EST library was constructed; at this stage the gene expression was presumed to be maximal for the germination process. During the conidia to the mycelia developmental process, the following germination rates were observed: at 24 hr, 11.7+/-1.2%; at 48 hr, 30+/-0.6%; at 72 hr, 43+/-1.3%; and at 96 hr, 66+/-2.4%. At the 96 hour stage, an EST library was constructed. It consisted of 129 sequences grouped in 4 contigs and 7 singlets for a total of 11 possible genes. Eight of the sequences had not been described previously in other EST libraries of this fungus. New genes were identified that were expressed during the conidia to the mycelia germination process and may represent genes specific to the germination process.

  10. [Influence of different drying temperatures for solid substrate after fermentation on conidia characteristics of the entomopathogenic fungus Beauveria bassiana].

    PubMed

    Li, Zunhua; Lin, Jianwen; Ma, Jincheng; Wu, Di; Zhang, Yongjun

    2008-07-01

    This study was to evaluate the effect of drying temperature for solid substrate after fermentation on conidia characteristics of entomopathogenic fungus Beauveria bassiana. Seven constant or varied temperatures between 28 degrees C and 35 degrees C were designed for drying the solid substrate and the quality of harvested conidia was analyzed. The results showed that the drying treatments at varied temperatures significantly decreased bacterial contamination in the harvested conidia powder. The conidia viability and germination speed were varied with different drying treatments. After drying at 35 degrees C for 5 h, there was no significant difference in viability between the dried and fresh conidia, while the median germination time (9.6 h) of the dried conidia was shortened by 9.3%. The tolerance of conidia to heat and UV radiation was increased by drying treatment. Compared to the drying treatment at a constant temperature at 28 degrees C or 35 degrees C, some varied temperature treatments were in favor of enhancing the stress tolerance of conidia. Drying treatments influenced accumulation of trehalose in harvested conidia, while neither heat resistance nor UV tolerance of conidia was obvious correlation with trehalose level. Optimizing drying temperature could increase the virulence of B. bassiana. After drying at 28 degrees C for 24 h and then 35 degrees C for 2 h or at 35 degrees C for 5 h, the LT(50S) to Myzus persicae at the dose of 370-450 conidia /mm2 were shortened by 10.6 h and 7.5 h, respectively. The results suggested that the drying temperature for post-fermentation solid substrate has an important influence on bacterial contamination in the harvested conidia powder, spore viability, stress tolerance and virulence in B. bassiana.

  11. Molecular and physiological effects of environmental UV radiation on fungal conidia.

    PubMed

    Braga, Gilberto U L; Rangel, Drauzio E N; Fernandes, Éverton K K; Flint, Stephan D; Roberts, Donald W

    2015-08-01

    Conidia are specialized structures produced at the end of the asexual life cycle of most filamentous fungi. They are responsible for fungal dispersal and environmental persistence. In pathogenic species, they are also involved in host recognition and infection. Conidial production, survival, dispersal, germination, pathogenicity and virulence can be strongly influenced by exposure to solar radiation, although its effects are diverse and often species dependent. UV radiation is the most harmful and mutagenic waveband of the solar spectrum. Direct exposure to solar radiation for a few hours can kill conidia of most fungal species. Conidia are killed both by solar UV-A and UV-B radiation. In addition to killing conidia, which limits the size of the fungal population and its dispersion, exposures to sublethal doses of UV radiation can reduce conidial germination speed and virulence. The focus of this review is to provide an overview of the effects of solar radiation on conidia and on the major systems involved in protection from and repair of damage induced by solar UV radiation. The efforts that have been made to obtain strains of fungi of interest such as entomopathogens more tolerant to solar radiation will also be reviewed.

  12. Interaction of Aspergillus fumigatus conidia with Acanthamoeba castellanii parallels macrophage-fungus interactions.

    PubMed

    Van Waeyenberghe, Lieven; Baré, Julie; Pasmans, Frank; Claeys, Myriam; Bert, Wim; Haesebrouck, Freddy; Houf, Kurt; Martel, An

    2013-12-01

    Aspergillus fumigatus and free-living amoebae are common inhabitants of soil. Mechanisms of A. fumigatus to circumvent the amoeba's digestion may facilitate overcoming the vertebrate macrophage defence mechanisms. We performed co-culture experiments using A. fumigatus conidia and the amoeba Acanthamoeba castellanii. Approximately 25% of the amoebae ingested A. fumigatus conidia after 1 h of contact. During intra-amoebal passage, part of the ingested conidia was able to escape the food vacuole and to germinate inside the cytoplasm of A. castellanii. Fungal release into the extra-protozoan environment by exocytosis of conidia or by germination was observed with light and transmission electron microscopy. These processes resulted in structural changes in A. castellanii, leading to amoebal permeabilization without cell lysis. In conclusion, A. castellanii internalizes A. fumigatus conidia, resulting in fungal intracellular germination and subsequent amoebal death. As such, this interaction highly resembles that of A. fumigatus with mammalian and avian macrophages. This suggests that A. fumigatus virulence mechanisms to evade macrophage killing may be acquired by co-evolutionary interactions among A. fumigatus and environmental amoebae.

  13. Production of Conidia by the Fungus Metarhizium anisopliae Using Solid-State Fermentation.

    PubMed

    Loera-Corral, Octavio; Porcayo-Loza, Javier; Montesinos-Matias, Roberto; Favela-Torres, Ernesto

    2016-01-01

    This chapter describes the production of conidia by Metarhizium anisopliae using solid-state fermentation. Before production of conidia, procedures for strains conservation, reactivation, and propagation are essential in order to provide genetic stability of the strains. The strain is conserved in freeze-dried vials and then reactivated through insect inoculation. Rice is used as a substrate for the conidia production in two different bioreactors: plastic bags and tubular bioreactor. The CO2 production in the tubular bioreactors is measured with a respirometer; this system allows calculating indirect growth parameters as lag time (tlag) (25-35 h), maximum rate of CO2 production (rCO2 max) (0.5-0.7 mg/gdm h), specific rate of CO2 production (μ) (0.10-0.15 1/h), and final CO2 production (CO2) (100-120 mg/gdm). Conidial yield per gram of dry substrate (gdm) should be above 1 × 10(9) conidia/gdm after 10 days of incubation. Germination and viability of conidia obtained after 10 days of incubation should be above 80 % and 75 %, respectively. Bioassays using of Tenebrio molitor as a host insect should yield a final mortality above 80 %.

  14. Maturation feeding and transmission of Bursaphelenchus xylophilus (Nematoda: Parasitaphelenchidae) by Monochamus alternatus (Coleoptera: Cerambycidae) inoculated with Beauveria bassiana (Deuteromycotina: Hyphomycetes).

    PubMed

    Maehara, Noritoshi; He, Xueyou; Shimazu, Mitsuaki

    2007-02-01

    We examined the amount of maturation feeding and transmission of pinewood nematodes, Bursaphelenchus xylophilus (Steiner et Buhrer) Nickle (Nematoda: Parasitaphelenchidae), to healthy pine (Pinus spp.) trees by pine sawyer Monochamus alternatus Hope (Coleoptera: Cerambycidae) adults infected with Beauveria bassiana (Balsamo) Vuill. (Deuteromycotina: Hyphomycetes). Inoculated beetles fed less than noninoculated beetles, probably because feeding by inoculated beetles began to decrease at about 4 d postinoculation and inoculated beetles ceased to feed for several days before their death. In inoculated beetles carrying >1,000 nematodes, some beetles died before nematode departure. The remaining heavily nematode-infested beetles lived until the beginning of nematode departure, but they had stopped feeding, preventing the nematodes from entering pine twigs. We suggest that microbial control of pine sawyer adults by B. bassiana may be effective in preventing transmission of pine wilt disease to healthy pine trees.

  15. Aquatic hyphomycete strains from metal-contaminated and reference streams might respond differently to future increase in temperature.

    PubMed

    Ferreira, Verónica; Gonçalves, Ana Lúcia; Canhoto, Cristina

    2012-01-01

    Aquatic hyphomycetes, a group of polyphyletic fungi, have been reported in streams contaminated with metals. This tolerance to metal contamination however can result in limited performance and limited ability to cope with additional environmental change. The predicted increase in water temperature, as a consequence of global warming, will have an additional effect on many streams. The sensitivity to temperature of strains of three aquatic hyphomycete species isolated from a metal-contaminated stream and an uncontaminated stream was assessed by determining their radial growth and activity (conidial production, oxygen consumption, mycelial biomass accumulation, fine particulate organic matter [FPOM] production, and microbial induced leaf mass loss) at 13 C (present water temperature in autumn) and at 18 C (predicted water temperature under global warming). Growth and reproductive activity generally were depressed for the strains isolated from the metal-contaminated stream when compared with those isolated from the unpolluted stream. These differences however were not translated into differences in FPOM production and leaf-litter mass loss, indicating that the strains isolated from the contaminated stream can decompose leaf litter similar to those of the reference stream. The 5 C increase in temperature stimulated fungal activity and litter decomposition, irrespective of species and strain. This might have strong effect on aquatic food-web and ecosystem functioning under global warming because increases in litter decomposition might lead to food shortage for higher trophic levels. The sensitivity to temperature depended on the response variable, species and strain. FPOM production was the variable most sensitive to temperature across strains and species and that for which temperature sensitivities differed most between strains. Fungal tolerance to metal contamination affects the extent to which its functions are stimulated by an increase in temperature, constituting

  16. A new species of Cladophialophora (hyphomycetes) from boreal and montane bryophytes.

    PubMed

    Davey, Marie L; Currah, Randolph S

    2007-01-01

    During a survey of bryophilous fungi from boreal and montane habitats in central Alberta, a hitherto undescribed species of Cladophialophora was recovered from Polytrichum juniperinum, Aulacomnium palustre, and Sphagnum fuscum. On potato dextrose agar (PDA) colonies grew slowly, attaining a diameter of 25 mm after 30 d, were dark grey, velvety, radially sulcate, and convolute and cracked at the centre. Micronematous conidiophores gave rise to branched chains of small (1-2 x 8-22 microm), cylindrical to fusiform conidia with truncate, swollen scars at each end. Phylogenies built on the ITS and ribosomal SSU regions indicate the isolates form a monophyletic clade within the family Herpotrichiellaceae (Chaetothyriales) that is composed of two geographically based groups, each with 99% within-group sequence similarity and 97-98% between-group sequence similarity. A teleomorph has not been found but would likely be similar to species of Capronia. In vitro inoculation of the isolates onto axenically grown P. juniperinum produced no discernible host symptoms, and host penetration could not be detected using light microscopy. The production of polyphenol oxidases by the fungus and the role of other Cladophialophora species as latent endophytes and saprobes suggest that a potential role for the fungus is the degradation of the polyphenol-rich cell walls of mosses. A dichotomous key to species of the genus Cladophialophora is provided.

  17. Susceptibility of biological stages of the horn fly, Haematobia irritans, to entomopathogenic fungi (Hyphomycetes)

    PubMed Central

    Angel-Sahagún, C. A.; Lezama-Gutiérrez, R.; Molina-Ochoa, J.; Galindo-Velasco, E.; López-Edwards, M.; Rebolledo-Domínguez, O.; Cruz-Vázquez, C.; Reyes-Velázquez, W. P.; Skoda, S. R.; Foster, J. E.

    2005-01-01

    The susceptibility of the egg, pupa, and adult of Haematobia irritans (L.) (Diptera: Muscidae) to isolates of the fungi Metarhizium anisopliae (Metsch.) Sor., Beauveria bassiana (Bals.) Vuill., and Paecilomyces fumosoroseus (Wize) Brown and Smith, was evaluated under laboratory conditions. Groups of 20 eggs than 4 h old, pupae less than 48h old and adults were sprayed with a conidial suspension of each isolate. Eggs, pupae and adults of horn fly were susceptible to these entomopathogenic fungi. For treated eggs, the isolates Ma3, Ma 15, Ma25, Pfr1, and Pfr8 reduced adult emergence to 3.8% to 6.3% in comparison with the control (72%). The mortality of pupae infected by the isolates Ma2, Ma25, and Pfr10 ranged between 50% and 71.3%. Mortality of adults after treatment with the isolates Ma6, Ma 10, Ma 14, Ma 15, Pfr 1, Pfr 9, Pfr 10, Pfr 11, and Pfr12 were higher than 90%. The isolate Ma6 produced the lowest LC50 against adult horn flies (8.08 × 102conidia/ml). These findings supported the hypotheses that isolates of M. anisopliae, and P. fumosoroseus are pathogenic against the different biological stages of horn flies by reducing adult emergence when applied on groups of eggs and pupae, and producing mortality when applied to adults. PMID:17119632

  18. Filter Paper Degrading Ability of a Trichoderma Strain With Multinucleate Conidia

    NASA Astrophysics Data System (ADS)

    Toyama, Hideo; Yano, Makiko; Hotta, Takeshi; Toyama, Nobuo

    The multinucleate conidia were produced from the green mature conidia of Trichoderma reesei Rut C-30 strain by colchicine treatment. The strain with higher Filter paper degrading ability was selected among those conidia using a double layer selection medium. The selected strain, JS-2 was able to collapse the filter paper within 15 min but the original strain took 25 min to collapse it completely. Moreover, the amount of reducing sugar in the L-type glass tube of the strain, JS-2, was greater than that of the original strain. The Avicel, CMC-Na, and Salicin hydrolyzing activity of the strain, JS-2, increased 2.1 times, 1.2 times, and 3.6 times higher than that of the original strain.

  19. Production of conidia by entomopathogenic fungi: from inoculants to final quality tests.

    PubMed

    Muñiz-Paredes, Facundo; Miranda-Hernández, Francisco; Loera, Octavio

    2017-03-01

    Demand for biopesticides is growing due to the increase of areas under integrated pest management worldwide. Conidia from entomopathogenic fungi play a major role as infective units in the current market of biopesticides. Success in a massive production of fungal conidia include the use of proper long-term conservation microbial methods, aimed at preserving the phenotypic traits of the strains. The development of suitable inoculants should also be considered since that favours a rapid germination and invasiveness of the substrate in solid state cultures (SSC). After the selection of a suitable fungal strain, proven optimization approaches for SSC mainly include the combination of substrates, moisture, texturizers, aeration and moderate stress to induce conidiation. Nonetheless, during storage and upon application in open fields, conidia either as free propagules or imbibed in formulations are subjected to stress due to abiotic factors, then quality should be preserved to resist such harsh conditions. All of these topics are analysed in this report.

  20. Early neutrophil recruitment and aggregation in the murine lung inhibit germination of Aspergillus fumigatus Conidia.

    PubMed

    Bonnett, Colin R; Cornish, E Jean; Harmsen, Allen G; Burritt, James B

    2006-12-01

    Several types of polymorphonuclear neutrophil (PMN) deficiency are a predisposing condition for fatal Aspergillus fumigatus infection. In order to study the defensive role of PMNs in the lungs, with particular reference to PMN recruitment and antimicrobial oxidant activity, responses to pulmonary instillation of A. fumigatus conidia were examined. Responses in BALB/c and C57BL/6 mice were compared with those in CXCR2(-/-) and gp91(phox-/-) mice, which are known to have delayed recruitment of PMN to the lungs in response to inflammatory stimuli and inactive NADPH oxidase, respectively. In BALB/c mice, PMNs were recruited to the lungs and formed oxidase-active aggregates with conidia, which inhibited germination. In C57BL/6, gp91(phox-/-), and CXCR2(-/-) mice, PMN recruitment was slower and there was increased germination compared to that in BALB/c mice at 6 and 12 h. In gp91(phox-/-) mice, germination was extensive in PMN aggregates but negligible in alveolar macrophages (AM). Lung sections taken at 6 and 48 h from BALB/c mice showed PMN accumulation at peribronchiolar sites but no germinating conidia. Those from C57BL/6 and CXCR2(-/-) mice showed germinating conidia at 6 h but not at 48 h and few inflammatory cells. In contrast, those from gp91(phox-/-) mice showed germination at 6 h with more-extensive hyphal proliferation and tissue invasion at 48 h. These results indicate that when the lungs are exposed to large numbers of conidia, in addition to the phagocytic activity of AM, early PMN recruitment and formation of oxidative-active aggregates are essential in preventing germination of A. fumigatus conidia.

  1. A Murine Inhalation Model to Characterize Pulmonary Exposure to Dry Aspergillus fumigatus Conidia

    PubMed Central

    Buskirk, Amanda D.; Green, Brett J.; Lemons, Angela R.; Nayak, Ajay P.; Goldsmith, W. Travis; Kashon, Michael L.; Anderson, Stacey E.; Hettick, Justin M.; Templeton, Steven P.; Germolec, Dori R.; Beezhold, Donald H.

    2014-01-01

    Most murine models of fungal exposure are based on the delivery of uncharacterized extracts or liquid conidia suspensions using aspiration or intranasal approaches. Studies that model exposure to dry fungal aerosols using whole body inhalation have only recently been described. In this study, we aimed to characterize pulmonary immune responses following repeated inhalation of conidia utilizing an acoustical generator to deliver dry fungal aerosols to mice housed in a nose only exposure chamber. Immunocompetent female BALB/cJ mice were exposed to conidia derived from Aspergillus fumigatus wild-type (WT) or a melanin-deficient (Δalb1) strain. Conidia were aerosolized and delivered to mice at an estimated deposition dose of 1×105 twice a week for 4 weeks (8 total). Histopathological and immunological endpoints were assessed 4, 24, 48, and 72 hours after the final exposure. Histopathological analysis showed that conidia derived from both strains induced lung inflammation, especially at 24 and 48 hour time points. Immunological endpoints evaluated in bronchoalveolar lavage fluid (BALF) and the mediastinal lymph nodes showed that exposure to WT conidia led to elevated numbers of macrophages, granulocytes, and lymphocytes. Importantly, CD8+ IL17+ (Tc17) cells were significantly higher in BALF and positively correlated with germination of A. fumigatus WT spores. Germination was associated with specific IgG to intracellular proteins while Δalb1 spores elicited antibodies to cell wall hydrophobin. These data suggest that inhalation exposures may provide a more representative analysis of immune responses following exposures to environmentally and occupationally prevalent fungal contaminants. PMID:25340353

  2. Morphological Transition of Paracoccidioides brasiliensis Conidia to Yeast Cells: In Vivo Inhibition in Females

    PubMed Central

    Aristizabal, Beatriz H.; Clemons, Karl V.; Stevens, David A.; Restrepo, Angela

    1998-01-01

    Clinical paracoccidioidomycosis is 13 times more common in men than in women. Estrogen inhibits the transition of mycelia or conidia (the saprophytic form of Paracoccidoides brasiliensis) to yeasts (the parasitic form) in vitro. Here, we show that, in male mice that were infected intranasally (mimicking natural infection) the transition of conidia in bronchoalveolar lavage fluids to intermediate forms and yeasts occurred over 24 to 96 h; CFU and yeasts (shown by histopathology) increased subsequently. In females, transition did not occur and infection cleared. These events in vivo are consistent with epidemiological and in vitro observations, suggesting that female hormones block transition and are responsible for resistance. PMID:9784579

  3. Recognition of laminin by Paracoccidioides brasiliensis conidia: a possible mechanism of adherence to human type II alveolar cells.

    PubMed

    Caro, Erika; Gonzalez, Angel; Muñoz, César; Urán, Marta E; Restrepo, Angela; John Hamilton, Andrew; Elena Cano, Luz

    2008-12-01

    This study addresses the recognition of laminin by Paracoccidioides brasiliensis conidia, as well as its possible role in the adherence of conidia to A549 cells. Adherence of conidia to immobilized laminin was shown to be specific, as anti-laminin antibodies, soluble laminin or the laminin-derived peptides IKVAV and CDPGYIGSR inhibited this interaction. RGD containing peptides and various monosaccharides had no effect on adherence, with the exception of N-acetylneuraminic acid. Pre-treatment of conidia with fibrinogen and fibronectin, but not with BSA, also resulted in significant inhibition, suggesting that P. brasiliensis conidia might cross-recognize host proteins involved in colonization. In assays using transmission electron microscopy, we observed internalization of conidia 30 min after exposition to A549 cells. Laminin present on the surface of A549 cells shown to serve as mediator of this interaction, with a significant decrease in fungal adherence when the epithelial cells were pre-treated with anti-laminin antibodies or when conidia were pre-incubated with either soluble laminin or the laminin-specific peptides. Together these results suggest that the recognition of laminin by P. brasiliensis conidia is a key process in the interaction with pulmonary epithelial cells, where this extracellular matrix protein acts as bridging molecule.

  4. Influence of Infection Court, Host Vigor, and Culture Filtrates on Canker Production by Botryodiplodia Theobromae Conidia in Sycamore

    Treesearch

    R. Lewis

    1978-01-01

    Some of the factors that influence canker development in American sycamores inoculated with Botryodiplodia theobromae conidia were determined. A combination of B. theobromae culture filtrates and conidia resulted in 100% canker production when introduced into stem wounds; however, a combination of Cephalosporium diospyri...

  5. Temperature, water activity and pH during conidia production affect the physiological state and germination time of Penicillium species.

    PubMed

    Nguyen Van Long, Nicolas; Vasseur, Valérie; Coroller, Louis; Dantigny, Philippe; Le Panse, Sophie; Weill, Amélie; Mounier, Jérôme; Rigalma, Karim

    2017-01-16

    Conidial germination and mycelial growth are generally studied with conidia produced under optimal conditions to increase conidial yield. Nonetheless, the physiological state of such conidia most likely differs from those involved in spoilage of naturally contaminated food. The present study aimed at investigating the impact of temperature, pH and water activity (aw) during production of conidia on the germination parameters and compatible solutes of conidia of Penicillium roqueforti and Penicillium expansum. Low temperature (5°C) and reduced aw (0.900 aw) during sporulation significantly reduced conidial germination times whereas the pH of the sporulation medium only had a slight effect at the tested values (2.5, 8.0). Conidia of P. roqueforti produced at 5°C germinated up to 45h earlier than those produced at 20°C. Conidia of P. roqueforti and P. expansum produced at 0.900 aw germinated respectively up to 8h and 3h earlier than conidia produced at 0.980 aw. Furthermore, trehalose and mannitol assessments suggested that earlier germination might be related to delayed conidial maturation even though no ultra-structural modifications were observed by transmission electron microscopy. Taken together, these results highlight the importance of considering environmental conditions during sporulation in mycological studies. The physiological state of fungal conidia should be taken into account to design challenge tests or predictive mycology studies. This knowledge may also be of interest to improve the germination capacity of fungal cultures commonly used in fermented foods.

  6. Optimization of compatible non-ionic surfactant for formulation development of hydrophobic conidia of entomopathogenic fungi, Beauveria bassiana (Hypocreales:Cordycipitaceae) and Metarhizium anisopliae Hypocreales:Clavicipita

    USDA-ARS?s Scientific Manuscript database

    Aerial conidia, especially dried conidia of entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae are hydrophobic, and therefore surfactants are needed for developing water-based formulations in laboratory studies, greenhouse bioassays, and field trials as well as commercial product ...

  7. Temperature during conidiation affects stress tolerance, pigmentation, and trypacidin accumulation in the conidia of the airborne pathogen Aspergillus fumigatus.

    PubMed

    Hagiwara, Daisuke; Sakai, Kanae; Suzuki, Satoshi; Umemura, Myco; Nogawa, Toshihiko; Kato, Naoki; Osada, Hiroyuki; Watanabe, Akira; Kawamoto, Susumu; Gonoi, Tohru; Kamei, Katsuhiko

    2017-01-01

    Asexual spores (conidia) are reproductive structures that play a crucial role in fungal distribution and survival. As fungal conidia are, in most cases, etiological agents of plant diseases and fungal lung disease, their stress resistance and interaction with their hosts have drawn increasing attention. In the present study, we investigated whether environmental temperature during conidiation affects the stress tolerance of the conidia of the human pathogenic fungus Aspergillus fumigatus. Conidia from a 25°C culture showed a lower tolerance to heat (60°C) and oxidative (H2O2) stresses and a marked resistance to ultraviolet radiation exposure, compared with those produced at 37 and 45°C. The accumulation of trehalose was lower in the conidia from the 25°C culture. Furthermore, the conidia from the 25°C culture showed darker pigmentation and increased transcripts of dihydroxynaphthalene (DHN)-melanin biosynthesis-related genes (i.e., pksP, arp1, and arp2). An RNA-sequencing analysis revealed that the transcription level of the trypacidin (tpc) gene cluster, which contains 13 genes, was sharply and coordinately activated in the conidia from the 25°C culture. Accordingly, trypacidin was abundant in the conidia from the 25°C culture, whereas there was little trypacidin in the conidia from the 37°C culture. Taken together, these data show that the environmental temperature during conidiation affects conidial properties such as stress tolerance, pigmentation, and mycotoxin accumulation. To enhance our knowledge, we further explored the temperature-dependent production of DHN-melanin and trypacidin in clinical A. fumigatus isolates. Some of the isolates showed temperature-independent production of DHN-melanin and/or trypacidin, indicating that the conidia-associated secondary metabolisms differed among the isolates.

  8. Temperature during conidiation affects stress tolerance, pigmentation, and trypacidin accumulation in the conidia of the airborne pathogen Aspergillus fumigatus

    PubMed Central

    Sakai, Kanae; Suzuki, Satoshi; Umemura, Myco; Nogawa, Toshihiko; Kato, Naoki; Osada, Hiroyuki; Watanabe, Akira; Kawamoto, Susumu; Gonoi, Tohru; Kamei, Katsuhiko

    2017-01-01

    Asexual spores (conidia) are reproductive structures that play a crucial role in fungal distribution and survival. As fungal conidia are, in most cases, etiological agents of plant diseases and fungal lung disease, their stress resistance and interaction with their hosts have drawn increasing attention. In the present study, we investigated whether environmental temperature during conidiation affects the stress tolerance of the conidia of the human pathogenic fungus Aspergillus fumigatus. Conidia from a 25°C culture showed a lower tolerance to heat (60°C) and oxidative (H2O2) stresses and a marked resistance to ultraviolet radiation exposure, compared with those produced at 37 and 45°C. The accumulation of trehalose was lower in the conidia from the 25°C culture. Furthermore, the conidia from the 25°C culture showed darker pigmentation and increased transcripts of dihydroxynaphthalene (DHN)-melanin biosynthesis-related genes (i.e., pksP, arp1, and arp2). An RNA-sequencing analysis revealed that the transcription level of the trypacidin (tpc) gene cluster, which contains 13 genes, was sharply and coordinately activated in the conidia from the 25°C culture. Accordingly, trypacidin was abundant in the conidia from the 25°C culture, whereas there was little trypacidin in the conidia from the 37°C culture. Taken together, these data show that the environmental temperature during conidiation affects conidial properties such as stress tolerance, pigmentation, and mycotoxin accumulation. To enhance our knowledge, we further explored the temperature-dependent production of DHN-melanin and trypacidin in clinical A. fumigatus isolates. Some of the isolates showed temperature-independent production of DHN-melanin and/or trypacidin, indicating that the conidia-associated secondary metabolisms differed among the isolates. PMID:28486558

  9. Density, Viability Conidia And Symptoms of Metarhizium anisopliae infection on Oryctes rhinoceros larvae

    NASA Astrophysics Data System (ADS)

    Indriyanti, D. R.; Putri, R. I. P.; Widiyaningrum, P.; Herlina, L.

    2017-04-01

    M. anisopliae is parasitic fungus on insect pests; it is used as a biocontrol agent. M. anisopliae can be propagated on maize or rice substrate. M. anisopliae is currently sold in the form of kaolin powder formulations. Before it is used to check the density, viability and pathogenicity of M. anisopliae. However the problem is the kaolin powder very soft, so it difficult to distinguish between kaolin and conidia. This article gives information on how to calculate conidia density, viability and symptoms of M. anisopliae infection on Oryctes rhinoceros larvae. The study was conducted in the laboratory to determine the density and viability. The pathogenicity testing was done using pots. The Pot is containing soil substrate mixed with M. Anispoliae and ten tails O. Rhinoceros larvae per pot. The results showed that the density of M. anisopliae conidia was 1.81 x 108 conidia mL-1 and the viability was 94% within 24 hours. The larval mortality began to emerge in the 1st week, and all larvae died at the sixth week. The symptom of M. anisopliae infection on Oryctes rhinoceros larvae, there was a black spot on the larval integument. The larvae movements become slow and poor appetite; it will die within 3-7 days. The larvae die hard, and the white hyphae grow on the body surface that turns green.

  10. Interactions of Aspergillus fumigatus Conidia with Airway Epithelial Cells: A Critical Review

    PubMed Central

    Croft, Carys A.; Culibrk, Luka; Moore, Margo M.; Tebbutt, Scott J.

    2016-01-01

    Aspergillus fumigatus is an environmental filamentous fungus that also acts as an opportunistic pathogen able to cause a variety of symptoms, from an allergic response to a life-threatening disseminated fungal infection. The infectious agents are inhaled conidia whose first point of contact is most likely to be an airway epithelial cell (AEC). The interaction between epithelial cells and conidia is multifaceted and complex, and has implications for later steps in pathogenesis. Increasing evidence has demonstrated a key role for the airway epithelium in the response to respiratory pathogens, particularly at early stages of infection; therefore, elucidating the early stages of interaction of conidia with AECs is essential to understand the establishment of infection in cohorts of at-risk patients. Here, we present a comprehensive review of the early interactions between A. fumigatus and AECs, including bronchial and alveolar epithelial cells. We describe mechanisms of adhesion, internalization of conidia by AECs, the immune response of AECs, as well as the role of fungal virulence factors, and patterns of fungal gene expression characteristic of early infection. A clear understanding of the mechanisms involved in the early establishment of infection by A. fumigatus could point to novel targets for therapy and prophylaxis. PMID:27092126

  11. Absorbing film assisted laser induced forward transfer of fungi (Trichoderma conidia)

    NASA Astrophysics Data System (ADS)

    Hopp, B.; Smausz, T.; Antal, Zs.; Kresz, N.; Bor, Zs.; Chrisey, D.

    2004-09-01

    We present an investigation on absorbing film assisted laser induced forward transfer (AFA-LIFT) of fungus (Trichoderma) conidia. A KrF excimer laser beam [λ =248nm,FWHM=30ns (FWHM, full width at half maximum)] was directed through a quartz plate and focused onto its silver coated surface where conidia of the Trichoderma strain were uniformly spread. The laser fluence was varied in the range of 0-2600mJ/cm2 and each laser pulse transferred a pixel of target material. The average irradiated area was 8×10-2mm2. After the transfer procedure, the yeast extract medium covered glass slide and the transferred conidia patterns were incubated for 20 h and then observed using an optical microscope. The transferred conidia pixels were germinated and the areas of the culture medium surfaces covered by the pixels were evaluated as a function of laser fluence. As the laser fluence was increased from 0 to 355mJ/cm2 the transferred and germinated pixel area increased from 0 to 0.25mm2. Further increase in fluence resulted in a drastic decrease down to an approximately constant value of 0.06mm2. The yield of successful transfer by AFA-LIFT and germination was as much as 75% at 355mJ/cm2. The results prove that AFA-LIFT can successfully be applied for the controlled transfer of biological objects.

  12. Evaluation of Soyscreen in an Oil-based Formulation for UV Protection of Beauveria bassiana Conidia

    USDA-ARS?s Scientific Manuscript database

    SoyScreen oil was studied as a formulation ingredient to protect Beauveria bassiana (Balsamo) Vuillemin conidia from UV degradation. Feruloylated soy glycerides, referred to as SoyScreen oil, are biobased ultraviolet-absorbing molecules made by combining molecules of soybean oil with ferulic acid. ...

  13. Metacridamides A and B, bioactive macrocycles from conidia of the entomopathogenic fungus Metarhizium acridum

    USDA-ARS?s Scientific Manuscript database

    Metarhizium acridum, an entomopathogenic fungus, has been commercialized and used successfully for biocontrol of grasshopper pests in Africa and Australia. Its conidia produce two novel 17-membered macrocycles, metacridamides A (1) and B (2), which consist of a Phe unit condensed with a nonaketide....

  14. Algorithm-based design of synthetic growth media stimulating virulence properties of Metarhizium anisopliae conidia.

    PubMed

    Hutwimmer, S; Wagner, S; Affenzeller, M; Burgstaller, W; Strasser, H

    2008-12-01

    Synthetic media should be designed for the production of Metarhizium anisopliae conidia with improved virulence properties. A genetic algorithm (GA), demonstrated to be suitable for the design of media for spore mass production (Hutwimmer et al. 2008), was utilized for a multi-objective medium design to improve conidia yield and three proposed virulence properties of conidia: C : N ratio, germination speed and amount of spore-bound Pr1 protease. After five iterative optimizations, 52 media were improved over Sabouraud dextrose agar (SDA). Four media exhibited medium performances (a factor derived from the four single optimization variables) of around 0.7; cf. SDA = 0.532; media with enhanced properties were reached for each single optimization variable; Bioassays against Tenebrio larvae indicated also a slight improvement in virulence of conidia from designed media. A degenerated phenotype of the same strain did not exhibit differences in colony appearance, spore characteristics and virulence if grown on designed media. The application of a problem-oriented GA is a practical and rapid method to design media for multi-objective purposes. The applicability of a GA for multi-objective medium design was demonstrated for the cultivation of anamorphic fungi on solid media.

  15. Genes Differentially Expressed in Conidia and Hyphae of Aspergillus fumigatus upon Exposure to Human Neutrophils

    PubMed Central

    Sugui, Janyce A.; Kim, H. Stanley; Zarember, Kol A.; Chang, Yun C.; Gallin, John I.; Nierman, Willian C.; Kwon-Chung, Kyung J.

    2008-01-01

    Background Aspergillus fumigatus is the most common etiologic agent of invasive aspergillosis in immunocompromised patients. Several studies have addressed the mechanism involved in host defense but only few have investigated the pathogen's response to attack by the host cells. To our knowledge, this is the first study that investigates the genes differentially expressed in conidia vs hyphae of A. fumigatus in response to neutrophils from healthy donors as well as from those with chronic granulomatous disease (CGD) which are defective in the production of reactive oxygen species. Methodology/Principal Findings Transcriptional profiles of conidia and hyphae exposed to neutrophils, either from normal donors or from CGD patients, were obtained by using the genome-wide microarray. Upon exposure to either normal or CGD neutrophils, 244 genes were up-regulated in conidia but not in hyphae. Several of these genes are involved in the degradation of fatty acids, peroxisome function and the glyoxylate cycle which suggests that conidia exposed to neutrophils reprogram their metabolism to adjust to the host environment. In addition, the mRNA levels of four genes encoding proteins putatively involved in iron/copper assimilation were found to be higher in conidia and hyphae exposed to normal neutrophils compared to those exposed to CGD neutrophils. Deletants in several of the differentially expressed genes showed phenotypes related to the proposed functions, i.e. deletants of genes involved in fatty acid catabolism showed defective growth on fatty acids and the deletants of iron/copper assimilation showed higher sensitivity to the oxidative agent menadione. None of these deletants, however, showed reduced resistance to neutrophil attack. Conclusion This work reveals the complex response of the fungus to leukocytes, one of the major host factors involved in antifungal defense, and identifies fungal genes that may be involved in establishing or prolonging infections in humans

  16. Growth under visible light increases conidia and mucilage production and tolerance to UV-B radiation in the plant pathogenic fungus Colletotrichum acutatum.

    PubMed

    de Menezes, Henrique D; Massola, Nelson S; Flint, Stephan D; Silva, Geraldo J; Bachmann, Luciano; Rangel, Drauzio E N; Braga, Gilberto U L

    2015-01-01

    Light conditions can influence fungal development. Some spectral wavebands can induce conidial production, whereas others can kill the conidia, reducing the population size and limiting dispersal. The plant pathogenic fungus Colletotrichum acutatum causes anthracnose in several crops. During the asexual stage on the host plant, Colletototrichum produces acervuli with abundant mucilage-embedded conidia. These conidia are responsible for fungal dispersal and host infection. This study examined the effect of visible light during C. acutatum growth on the production of conidia and mucilage and also on the UV tolerance of these conidia. Conidial tolerance to an environmentally realistic UV irradiance was determined both in conidia surrounded by mucilage on sporulating colonies and in conidial suspension. Exposures to visible light during fungal growth increased production of conidia and mucilage as well as conidial tolerance to UV. Colonies exposed to light produced 1.7 times more conidia than colonies grown in continuous darkness. The UV tolerances of conidia produced under light were at least two times higher than conidia produced in the dark. Conidia embedded in the mucilage on sporulating colonies were more tolerant of UV than conidia in suspension that were washed free of mucilage. Conidial tolerance to UV radiation varied among five selected isolates.

  17. An animal model for allergic penicilliosis induced by the intranasal instillation of viable Penicillium chrysogenum conidia

    PubMed Central

    Cooley, J; Wong, W; Jumper, C; Hutson, J; Williams, H; Schwab, C; Straus, D

    2000-01-01

    BACKGROUND—A study was undertaken to determine the consequences of long term intranasal instillation of Penicillium chrysogenum propagules in a mouse model.
METHODS—C57 Black/6 mice were inoculated intranasally each week for six weeks with 104 viable and non-viable P chrysogenum conidia. Cytokine levels and cellular responses in these animals were then measured.
RESULTS—Compared with controls, mice inoculated intranasally each week for six weeks with 104P chrysogenum conidia (average viability 25%) produced significantly more total serum IgE (mean difference 1823.11, lower and upper 95% confidence intervals (CI) 539.09 to 3107.13), peripheral eosinophils (mean difference 5.11, 95% CI 2.24 to 7.99), and airway eosinophilia (rank difference 11.33, 95% CI 9.0 to 20.0). With the exception of airway neutrophilia (mean difference 20.89, 95% CI 3.72 to 38.06), mice inoculated intranasally with 104 non-viable conidia did not show significant changes in total serum IgE, peripheral or airway eosinophils. However, when compared with controls, this group (104 non-viable) had a significant increase in total serum IgG2a (mean difference 1990.56, 95% CI 790.48 to 3190.63) and bronchoalveolar lavage (BAL) fluid levels of interferon (IFN)-γ (mean difference 274.72, 95% CI 245.26 to 304.19). In addition, lung lavages from mice inoculated intranasally with 104 viable P chrysogenum conidia had significantly increased levels of interleukin (IL)-4 (mean difference 285.28, 95% CI 108.73 to 461.82) and IL-5 (mean difference 16.61, 95% CI 11.23 to 21.99). The IgG2a/IgE ratio and the IFN-γ/IL-4 ratio was lower in the group of mice inoculated intranasally with 104 viable conidia than in the 104 non-viable conidia group and the controls. When proteins were extracted from P chrysogenum conidia, attached to microtitre plates and incubated with serum from the 104 viable group, significant increases in conidia-specific IgE and IgG1 were observed compared with controls, while serum

  18. Differential Regulation by Organic Compounds and Heavy Metals of Multiple Laccase Genes in the Aquatic Hyphomycete Clavariopsis aquatica

    PubMed Central

    Solé, Magali; Müller, Ines; Pecyna, Marek J.; Fetzer, Ingo; Harms, Hauke

    2012-01-01

    To advance the understanding of the molecular mechanisms controlling microbial activities involved in carbon cycling and mitigation of environmental pollution in freshwaters, the influence of heavy metals and natural as well as xenobiotic organic compounds on laccase gene expression was quantified using quantitative real-time PCR (qRT-PCR) in an exclusively aquatic fungus (the aquatic hyphomycete Clavariopsis aquatica) for the first time. Five putative laccase genes (lcc1 to lcc5) identified in C. aquatica were differentially expressed in response to the fungal growth stage and potential laccase inducers, with certain genes being upregulated by, e.g., the lignocellulose breakdown product vanillic acid, the endocrine disruptor technical nonylphenol, manganese, and zinc. lcc4 is inducible by vanillic acid and most likely encodes an extracellular laccase already excreted during the trophophase of the organism, suggesting a function during fungal substrate colonization. Surprisingly, unlike many laccases of terrestrial fungi, none of the C. aquatica laccase genes was found to be upregulated by copper. However, copper strongly increases extracellular laccase activity in C. aquatica, possibly due to stabilization of the copper-containing catalytic center of the enzyme. Copper was found to half-saturate laccase activity already at about 1.8 μM, in favor of a fungal adaptation to low copper concentrations of aquatic habitats. PMID:22544244

  19. Aquatic hyphomycete communities associated with decomposing alder leaf litter in reference headwater streams of the Basque Country (northern Spain).

    PubMed

    Pérez, Javier; Descals, Enrique; Pozo, Jesús

    2012-08-01

    The community of aquatic hyphomycetes associated with decomposing alder leaf litter was studied during autumn-winter in nine headwater reference streams of the Basque Country (northern Spain). In order to study the spatial variability in composition and community structure, three streams from each of three different river basins were compared. The colonization dynamics and community changes throughout the decomposition process were also followed in three of the rivers (one per basin). The taxonomic richness and community structure of these fungi varied among rivers, including similar streams of a given watershed. However, neither species diversity nor total abundance was statistically related to environmental variables. Only the conidial production of two of the species, Flagellospora curvula and Lunulospora curvula appeared to be enhanced by nitrate availability in the water. The taxonomic richness and the reproductive activity (sporulation rate) were positively related to the leaf litter decomposition rate. The changes in conidial production along the process were similar for all the streams and helped explain leaf litter quality dynamics.

  20. Inhibitory effect of deferoxamine or macrophage activation on transformation of Paracoccidioides brasiliensis conidia ingested by macrophages: reversal by holotransferrin.

    PubMed

    Cano, L E; Gomez, B; Brummer, E; Restrepo, A; Stevens, D A

    1994-04-01

    Conidia of P. brasiliensis ingested by murine macrophages at 37 degrees C showed enhanced transformation to yeast cells and further intracellular growth compared with conidia in culture medium alone. Treatment of macrophages with the iron chelator deferoxamine inhibited the intracellular conidium-to-yeast transformation. Cytokine-activated macrophages could also exert this inhibitory effect. Holotransferrin reversed the inhibitory effect of either deferoxamine or activated macrophages on intracellular conidium-to-yeast transformation. These results indicate that iron restriction is one of the mechanisms by which activated macrophages control the intracellular transformation of ingested conidia and growth of yeast cells.

  1. Inhibitory effect of deferoxamine or macrophage activation on transformation of Paracoccidioides brasiliensis conidia ingested by macrophages: reversal by holotransferrin.

    PubMed Central

    Cano, L E; Gomez, B; Brummer, E; Restrepo, A; Stevens, D A

    1994-01-01

    Conidia of P. brasiliensis ingested by murine macrophages at 37 degrees C showed enhanced transformation to yeast cells and further intracellular growth compared with conidia in culture medium alone. Treatment of macrophages with the iron chelator deferoxamine inhibited the intracellular conidium-to-yeast transformation. Cytokine-activated macrophages could also exert this inhibitory effect. Holotransferrin reversed the inhibitory effect of either deferoxamine or activated macrophages on intracellular conidium-to-yeast transformation. These results indicate that iron restriction is one of the mechanisms by which activated macrophages control the intracellular transformation of ingested conidia and growth of yeast cells. PMID:8132359

  2. Single tracheal inoculation of Aspergillus fumigatus conidia induced aspergillosis in juvenile falcons (Falco spp.).

    PubMed

    Fischer, Dominik; Van Waeyenberghe, Lieven; Failing, Klaus; Martel, An; Lierz, Michael

    2017-09-01

    Aspergillosis is a common and life-threatening respiratory disease in raptors with acute and chronic courses. Among raptors, gyrfalcons (Falco rusticolus) and their hybrids are often declared to be highly susceptible with juvenile individuals being the most susceptible. However, species- and age-specific experimental studies are lacking and minimal infective doses (IDs) for Aspergillus spp. conidia are unknown.Therefore, 8-week-old, healthy gyr-hybrid falcons (F. rusticolus X F. cherrug) (N = 18) were experimentally infected with Aspergillus fumigatus using a single intratracheal inoculation with varying dosages of conidia (10(2) to 10(7) conidia). Over 28 days, clinical signs were monitored as well as haematological and serological parameters. Following euthanasia, necropsy, histopathology, bacteriology, and mycology were performed. Re-isolated fungi were compared to the inoculum using microsatellite length polymorphisms. During the trial, clinical signs and dyspnoea correlated significantly with the ID. Necropsy revealed fungal lesions in the upper and lower airways of 10/18 inoculated falcons, but not in the control birds. In 9/18 inoculated falcons, fungal granulomas were confirmed in histopathology and A. fumigatus was re-isolated from these granulomas. Except one nasal isolate all re-isolated fungal strains were identical to the inoculum strain. Based on mycology and histopathology a minimal ID of 50% was calculated to be MID50% (±S.E.) = 10(4.52±0.67) for a single tracheal inoculation of A. fumigatus conidia. This study demonstrates for the first time that a single exposure is able to cause acute aspergillosis in juvenile falcons.

  3. Botrytis cinerea isolates collected from grapes present different requirements for conidia germination.

    PubMed

    Cotoras, Milena; García, Carolina; Mendoza, Leonora

    2009-01-01

    Botrytis cinerea presents high variability in several biological traits, which can be explained by the high degree of genotypic diversity among isolates. Because this genetic variability might be related to phenotypic differences the requirements for conidia germination of three natural isolates (G1, G5 and G11) obtained from grapes and belonging to the same genetic group were analyzed. The results showed that contact with a solid surface was a common requisite for conidia germination of the isolates but they differed in their nutritional requirements to germinate. Isolate G11 was able to germinate in the absence of a carbon or nitrogen source. G1 and G5 required the presence of a carbon source such as glucose, fructose or sucrose. In G11 and G5 isolates a much higher rate of germination was obtained in the presence of sucrose. It was shown with a pharmacological approach that the cAMP stimulated the germination only in those isolates requiring a carbon source. Conidia germination of G1 and G5 was inhibited by EGTA, a calcium chelator. Isolate G11 germinated in the presence of this compound. On the other hand the germination of three B. cinerea isolates required protein synthesis and did not require RNA synthesis. To explain the ability of isolate G11 to germinate in water the content of total and reducing sugars, mannitol/L-arabitol, trehalose, and proteins in the nongerminated conidia of the three isolates was compared. The isolates presented similar amounts of total and reducing sugars. In the three isolates the amount of mannitol/L-arabitol was higher than that of trehalose. In isolate G11 total protein content was twice higher than in the other isolates.

  4. In vitro photodynamic inactivation of conidia of the phytopathogenic fungus Colletotrichum graminicola with cationic porphyrins.

    PubMed

    Vandresen, Camila Chevonica; Gonçalves, Alan Guilherme; Ducatti, Diogo Ricardo Bazan; Murakami, Fabio Seigi; Noseda, Miguel Daniel; Duarte, Maria Eugenia Rabello; Barreira, Sandra Mara Woranovicz

    2016-05-11

    Photodynamic inactivation (PDI) is an efficient approach for the elimination of a series of microorganisms; however, PDI involving phytopathogenic filamentous fungi is scarce in the literature. In the present study, we have demonstrated the photoinactivating properties of five cationic meso-(1-methyl-4-pyridinio)porphyrins on conidia of the phytopathogen Colletotrichum graminicola. For this purpose, photophysical properties (photostability and (1)O2 singlet production) of the porphyrins under study were first evaluated. PDI assays were then performed with a fluence of 30, 60, 90 and 120 J cm(-2) and varying the porphyrin concentration from 1 to 25 μmol L(-1). Considering the lowest concentration that enabled the best photoinactivation, with the respective lowest effective irradiation time, the meso-(1-methyl-4-pyridinio)porphyrins herein studied could be ranked as follows: triple-charged 4 (1 μmol L(-1) with a fluence of 30 J cm(-2)) > double-charged-trans2 (1 μmol L(-1) with 60 J cm(-2)) > tetra-charged 5 (15 μmol L(-1) with 90 J cm(-2)) > mono-charged 1 (25 μmol L(-1) with 120 J cm(-2)). Double-charged-cis-porphyrin 3 inactivated C. graminicola conidia in the absence of light. Evaluation of the porphyrin binding to the conidia and fluorescence microscopic analysis were also performed, which were in agreement with the PDI results. In conclusion, the cationic porphyrins herein studied were considered efficient photosensitizers to inactivate C. graminicola conidia. The amount and position of positive charges are related to the compounds' amphiphilicity and therefore to their photodynamic activity.

  5. Lag time for germination of Penicillium chrysogenum conidia is induced by temperature shifts.

    PubMed

    Kalai, Safaa; Bensoussan, Maurice; Dantigny, Philippe

    2014-09-01

    In the environment, fungal conidia are subject to transient conditions. In particular, temperature is varying according to day/night periods. All predictive models for germination assume that fungal spores can adapt instantaneously to changes of temperature. The only study that supports this assumption (Gougouli and Koutsoumanis, 2012, Modelling germination of fungal spores at constant and fluctuating temperature conditions. International Journal of Food Microbiology, 152: 153-161) was carried out on Penicillium expansum and Aspergillus niger conidia that, in most cases, already produced germ tubes. In contrast, the present study focuses on temperature shifts applied during the first stages of germination (i.e., before the apparition of the germ tubes). Firstly, germination times were determined in steady state conditions at 10, 15, 20 and 25 °C. Secondly, temperature shifts (e.g., up-shifts and down-shifts) were applied at 1/4, 1/2, and 3/4 of germination times, with 5, 10 and 15 °C magnitudes. Experiments were carried out in triplicate on Penicillium chrysogenum conidia on Potato Dextrose Agar medium according to a full factorial design. Statistical analysis of the results clearly demonstrated that the assumption of instantaneous adaptation of the conidia should be rejected. Temperature shifts during germination led to an induced lag time or an extended germination time as compared to the experiments conducted ay steady state. The induced lag time was maximized when the amplitude of the shift was equal to 10 °C. Interaction between the instant and the direction of the shift was highlighted. A negative lag time was observed for a 15 °C down-shift applied at 1/4 of the germination time. This result suggested that at optimal temperature the rate of germination decreased with time, and that the variation of this rate with time depended on temperature.

  6. EVALUATION OF DIFFERENT METHODS FOR THE EXTRACTION OF DNA FROM FUNGAL CONIDIA BY QUANTITATIVE COMPETITIVE PCR ANALYSIS

    EPA Science Inventory

    Five different DNA extraction methods were evaluated for their effectiveness in recovering PCR templates from the conidia of a series of fungal species often encountered in indoor air. The test organisms were Aspergillus versicolor, Penicillium chrysogenum, Stachybotrys chartaru...

  7. EVALUATION OF DIFFERENT METHODS FOR THE EXTRACTION OF DNA FROM FUNGAL CONIDIA BY QUANTITATIVE COMPETITIVE PCR ANALYSIS

    EPA Science Inventory

    Five different DNA extraction methods were evaluated for their effectiveness in recovering PCR templates from the conidia of a series of fungal species often encountered in indoor air. The test organisms were Aspergillus versicolor, Penicillium chrysogenum, Stachybotrys chartaru...

  8. Location of Aryl Sulfatase in Conidia and Young Mycelia of Neurospora crassa

    PubMed Central

    Scott, Walter A.; Metzenberg, Robert L.

    1970-01-01

    Aryl sulfatase (arylsulfate sulfohydrolase, EC 3.1.6.1) was found to have multiple locations in Neurospora conidia. Some enzyme activity remained in the supernatant when a spore suspension was centrifuged or filtered. Part of the cell-bound activity could be detected by adding the assay ingredients to a suspension of intact spores (patent enzyme), and additional activity was only detectable when the spores were first treated to destroy their permeability barriers (cryptic enzyme). Such treatments include: disruption with an X-press, brief rinsing with chloroform or acetone, incubation at 60 C for 5 min, and incubation with phenethyl alcohol, nystatin, or ascosin. Part of the patent aryl sulfatase was inactivated by briefly acid treating the intact spores (no loss of conidial viability). This enzyme was considered to have a cell surface location. Some enzyme was acid-resistant in intact spores, but all of the enzyme was acid-sensitive in spores whose permeability barriers had been disrupted. The pH dependence, kinetic properties, and p-nitrophenyl sulfate uptake were investigated in acid-treated conidia. No aryl sulfatase was detected in ascospores. Young mycelia contained more aryl sulfatase than did conidia, but little, if any, was secreted into the growth medium. Cryptic activity was demonstrated in young mycelia by brief chloroform treatment or by rinsing the cells with 0.1 m acetate buffer. Enzyme activity in young mycelia was completely labile to acid treatment, as was cell viability. PMID:16559101

  9. Development of pulmonary fibrosis in mice during infection with Paracoccidioides brasiliensis conidia.

    PubMed

    Restrepo, S; Tobon, A; Trujillo, J; Restrepo, A

    1992-01-01

    Pulmonary fibrosis is a feared sequelae of paracoccidioidomycosis. We sought to determine if mice exposed to Paracoccidioides brasiliensis conidia would develop pulmonary fibrosis. BALB/c mice were infected intranasally with P. brasiliensis conidia and sacrificed at regular intervals. One lung was sectioned for histopathology and sections were stained with haematoxylin and eosin, trichromic and argentic stains; the other lung was homogenized and cultured to determine the viability of the fungus. One week post-challenge, only small peribronchial foci were apparent. After 4 weeks, reticular fibres appeared disorganized and disrupted. Six to 8 weeks later peribronchial infiltrates were larger and appeared surrounded by reticular fibres; thick collagen I fibres were noticed in the infiltrated areas at this time. On weeks 10-12, infiltrates were confluent and reticular fibres were concentrated around the inflammatory foci; collagenization was apparent. Observations up to 16 weeks revealed diffuse involvement of the lung parenchyma with extensive collagenization. Lung cultures were always positive. We suggest that inhalation of P. brasiliensis conidia induces adverse lung responses leading to changes in the proportion of collagen fibres I and III.

  10. Effects of fluctuating moisture and temperature regimes on the persistence of quiescent conidia of Isaria fumosorosea.

    PubMed

    Bouamama, N; Vidal, C; Fargues, J

    2010-10-01

    Conidia of Isaria fumosorosea were submitted to three regimes of temperature and moisture to simulate microclimatic conditions which prevail in temperate (43% RH and 28 degrees C to 98% RH and 15 degrees C), subtropical (75% RH and 35 degrees C to 98% RH and 25 degrees C), and arid areas (13% RH and 40 degrees C to 33% RH and 15 degrees C) with daily fluctuating cycles. Germination, conidial viability, and virulence to Spodoptera frugiperda larvae were less affected after 20 days exposure under temperate cycling conditions than under arid and subtropical conditions. Exposure of conidia for 20 days to constant nocturnal simulated conditions of any tested regime weakly affected conidial persistence, whereas diurnal conditions exerted the most detrimental effects of high temperatures. However, when tested at both 45 degrees C and 50 degrees C at 33% RH for 160 h, the persistence of I. fumosorosea conidia was relatively higher than expected. These results emphasize that climatic conditions prevailing in environments and ecological fitness of fungal isolates have to be taken into account for assessing microbial control strategies.

  11. Ecological study of Paracoccidioides brasiliensis in soil: growth ability, conidia production and molecular detection

    PubMed Central

    Terçarioli, Gisela Ramos; Bagagli, Eduardo; Reis, Gabriela Martins; Theodoro, Raquel Cordeiro; Bosco, Sandra De Moraes Gimenes; Macoris, Severino Assis da Graça; Richini-Pereira, Virgínia Bodelão

    2007-01-01

    Background Paracoccidioides brasiliensis ecology is not completely understood, although several pieces of evidence point to the soil as its most probable habitat. The present study aimed to investigate the fungal growth, conidia production and molecular pathogen detection in different soil conditions. Methods Soils samples of clayey, sandy and medium textures were collected from ground surface and the interior of armadillo burrows in a hyperendemic area of Paracoccidioidomycosis. P. brasiliensis was inoculated in soil with controlled humidity and in culture medium containing soil extracts. The molecular detection was carried out by Nested PCR, using panfungal and species specific primers from the ITS-5.8S rDNA region. Results The soil texture does not affect fungus development and the growth is more abundant on/in soil saturated with water. Some soil samples inhibited the development of P. brasiliensis, especially those that contain high values of Exchangeable Aluminum (H+Al) in their composition. Some isolates produced a large number of conidia, mainly in soil-extract agar medium. The molecular detection was positive only in samples collected from armadillo burrows, both in sandy and clayey soil. Conclusion P. brasiliensis may grow and produce the infectious conidia in sandy and clayey soil, containing high water content, mainly in wild animal burrows, but without high values of H+Al. PMID:17953742

  12. Fungal tyrosine betaine, a novel secondary metabolite from conidia of entomopathogenic Metarhizium spp. fungi.

    PubMed

    Carollo, Carlos A; Calil, Ana Luiza A; Schiave, Letícia A; Guaratini, Thais; Roberts, Donald W; Lopes, Norberto P; Braga, Gilberto U L

    2010-01-01

    Fungi, including the entomopathogenic deuteromycete Metarhizium anisopliae, produce a wide diversity of secondary metabolites that either can be secreted or stored in specific developmental structures, e.g., conidia. Some secondary metabolites, such as pigments, polyols and mycosporines, are associated with pathogenicity and/or fungal tolerance to several stress-inducing environmental factors, including temperature and solar radiation extremes. Extracts of M. anisopliae var. anisopliae (strain ESALQ-1037) conidia were purified by chromatographic procedures and the isolated compounds analyzed by ¹H and ¹³C nuclear magnetic resonance spectroscopy and high-resolution mass spectrometry. LC-MS analyses were carried out to search for mycosporines (the initial targets), but no compounds of this class were detected. A molecule whose natural occurrence was previously undescribed was identified. It consists of betaine conjugated with tyrosine, and the structure was identified as 2-{[1-carboxy-2-(4-hydroxyphenyl)ethyl]amino}-N,N,N-trimethyl-2-oxoethanammonium. Mannitol was the predominant compound in the alcoholic conidial extract, but no amino acids other than tyrosine were found to be conjugated with betaine in conidia. The fungal tyrosine betaine was detected also in conidial extracts of three other M. anisopliae var. anisopliae (ARSEF 1095, 5626 and 5749) and three M. anisopliae var. acridum isolates (ARSEF 324, 3391 and 7486), but it was not detected in Aspergillus nidulans conidial extract (ATCC 10074).

  13. Production and quality of conidia by Metarhizium anisopliae var. lepidiotum: critical oxygen level and period of mycelium competence.

    PubMed

    Garcia-Ortiz, Nohemi; Tlecuitl-Beristain, Saúl; Favela-Torres, Ernesto; Loera, Octavio

    2015-03-01

    Mycoinsecticides application within Integral Pest Management requires high quantities of conidia, with the proper quality and resistance against environmental conditions. Metarhizium anisopliae var. lepidiotum conidia were produced in normal atmospheric conditions (21 % O2) and different concentrations of oxygen pulses (16, 26, 30, and 40 %); conidia obtained under hypoxic conditions showed significantly lower viability, hydrophobicity, and virulence against Tenebrio molitor larvae or mealworm, compared with those obtained under normal atmospheric conditions. Higher concentrations of oxygen (26 and 30 %) improved conidial production. However, when a 30 % oxygen concentration was applied, maximal conidial yields were obtained at earlier times (132 h) relative to 26 % oxygen pulses (156 h); additionally, with 30 % oxygen pulses, conidia thermotolerance was improved, maintaining viability, hydrophobicity, and virulence. Although conidial production was not affected when 40 % oxygen pulses were applied, viability and virulence were diminished in those conidia. In order to find a critical time for mycelia competence to respond to these oxidant conditions, oxygen pulses were first applied either at 36, 48, 60, and 72 h. A critical time of 60 h was determined to be the best time for the M. anisopliae var. lepidiotum mycelia to respond to oxygen pulses in order to increase conidial production and also to maintain the quality features. Therefore, oxygen-enriched (30 %) pulses starting at 60 h are recommended for a high production without the impairment of quality of M. anisopliae var. lepidiotum conidia.

  14. Germination of Aspergillus niger conidia is triggered by nitrogen compounds related to L-amino acids.

    PubMed

    Hayer, Kimran; Stratford, Malcolm; Archer, David B

    2014-10-01

    Conidial germination is fundamentally important to the growth and dissemination of most fungi. It has been previously shown (K. Hayer, M. Stratford, and D. B. Archer, Appl. Environ. Microbiol. 79:6924-6931, 2013, http://dx.doi.org/10.1128/AEM.02061-13), using sugar analogs, that germination is a 2-stage process involving triggering of germination and then nutrient uptake for hyphal outgrowth. In the present study, we tested this 2-stage germination process using a series of nitrogen-containing compounds for the ability to trigger the breaking of dormancy of Aspergillus niger conidia and then to support the formation of hyphae by acting as nitrogen sources. Triggering and germination were also compared between A. niger and Aspergillus nidulans using 2-deoxy-D-glucose (trigger), D-galactose (nontrigger in A. niger but trigger in A. nidulans), and an N source (required in A. niger but not in A. nidulans). Although most of the nitrogen compounds studied served as nitrogen sources for growth, only some nitrogen compounds could trigger germination of A. niger conidia, and all were related to L-amino acids. Using L-amino acid analogs without either the amine or the carboxylic acid group revealed that both the amine and carboxylic acid groups were essential for an L-amino acid to serve as a trigger molecule. Generally, conidia were able to sense and recognize nitrogen compounds that fitted into a specific size range. There was no evidence of uptake of either triggering or nontriggering compounds over the first 90 min of A. niger conidial germination, suggesting that the germination trigger sensors are not located within the spore.

  15. Detection of Airborne Stachybotrys chartarum Macrocyclic Trichothecene Mycotoxins on Particulates Smaller than Conidia

    PubMed Central

    Brasel, T. L.; Douglas, D. R.; Wilson, S. C.; Straus, D. C.

    2005-01-01

    Highly respirable particles (diameter, <1 μm) constitute the majority of particulate matter found in indoor air. It is hypothesized that these particles serve as carriers for toxic compounds, specifically the compounds produced by molds in water-damaged buildings. The presence of airborne Stachybotrys chartarum trichothecene mycotoxins on particles smaller than conidia (e.g., fungal fragments) was therefore investigated. Cellulose ceiling tiles with confluent Stachybotrys growth were placed in gas-drying containers through which filtered air was passed. Exiting particulates were collected by using a series of polycarbonate membrane filters with decreasing pore sizes. Scanning electron microscopy was employed to determine the presence of conidia on the filters. A competitive enzyme-linked immunosorbent assay (ELISA) specific for macrocyclic trichothecenes was used to analyze filter extracts. Cross-reactivity to various mycotoxins was examined to confirm the specificity. Statistically significant (P < 0.05) ELISA binding was observed primarily for macrocyclic trichothecenes at concentrations of 50 and 5 ng/ml and 500 pg/ml (58.4 to 83.5% inhibition). Of the remaining toxins tested, only verrucarol and diacetylverrucarol (nonmacrocyclic trichothecenes) demonstrated significant binding (18.2 and 51.7% inhibition, respectively) and then only at high concentrations. The results showed that extracts from conidium-free filters demonstrated statistically significant (P < 0.05) antibody binding that increased with sampling time (38.4 to 71.9% inhibition, representing a range of 0.5 to 4.0 ng/ml). High-performance liquid chromatography analysis suggested the presence of satratoxin H in conidium-free filter extracts. These data show that S. chartarum trichothecene mycotoxins can become airborne in association with intact conidia or smaller particles. These findings may have important implications for indoor air quality assessment. PMID:15640178

  16. Detection of airborne Stachybotrys chartarum macrocyclic trichothecene mycotoxins on particulates smaller than conidia.

    PubMed

    Brasel, T L; Douglas, D R; Wilson, S C; Straus, D C

    2005-01-01

    Highly respirable particles (diameter, <1 microm) constitute the majority of particulate matter found in indoor air. It is hypothesized that these particles serve as carriers for toxic compounds, specifically the compounds produced by molds in water-damaged buildings. The presence of airborne Stachybotrys chartarum trichothecene mycotoxins on particles smaller than conidia (e.g., fungal fragments) was therefore investigated. Cellulose ceiling tiles with confluent Stachybotrys growth were placed in gas-drying containers through which filtered air was passed. Exiting particulates were collected by using a series of polycarbonate membrane filters with decreasing pore sizes. Scanning electron microscopy was employed to determine the presence of conidia on the filters. A competitive enzyme-linked immunosorbent assay (ELISA) specific for macrocyclic trichothecenes was used to analyze filter extracts. Cross-reactivity to various mycotoxins was examined to confirm the specificity. Statistically significant (P < 0.05) ELISA binding was observed primarily for macrocyclic trichothecenes at concentrations of 50 and 5 ng/ml and 500 pg/ml (58.4 to 83.5% inhibition). Of the remaining toxins tested, only verrucarol and diacetylverrucarol (nonmacrocyclic trichothecenes) demonstrated significant binding (18.2 and 51.7% inhibition, respectively) and then only at high concentrations. The results showed that extracts from conidium-free filters demonstrated statistically significant (P < 0.05) antibody binding that increased with sampling time (38.4 to 71.9% inhibition, representing a range of 0.5 to 4.0 ng/ml). High-performance liquid chromatography analysis suggested the presence of satratoxin H in conidium-free filter extracts. These data show that S. chartarum trichothecene mycotoxins can become airborne in association with intact conidia or smaller particles. These findings may have important implications for indoor air quality assessment.

  17. Vacuolar system of ungerminated Colletotrichum graminicola conidia: convergence of autophagic and endocytic pathways.

    PubMed

    Schadeck, Ruth Janice Guse; Randi, Marco Antonio Ferreira; de Freitas Buchi, Dorly; Leite, Breno

    2003-01-28

    Vacuoles of ungerminated Colletotrichum graminicola conidia engulf cytoplasmic structures by a process analogous to microautophagy, demonstrated by using a vacuolar membrane acid phosphatase marker. Fusion of vesicles with vacuoles, without deposition of the acid phosphatase reaction product has been observed, suggesting other pathways of material delivery to vacuoles than microautophagy. Plasma membrane invaginations, multivesicular bodies and retention of neutral red into small vesicles, which were internalized by the vacuole, were verified. These results provided evidence for endocytosis and an active endosomal system. Together, our findings with C. graminicola demonstrated that vacuoles are very dynamic compartments, playing roles in autophagy and endocytic processes.

  18. Methods for reducing particle concentrations of Aspergillus fumigatus conidia and mouldy hay dust.

    PubMed

    Edwards, J H; Trotman, D M; Mason, O F

    1985-08-01

    The effectiveness of commercially available domestic air purifiers to reduce airborne Aspergillus fumigatus conidia and mouldy hay dust was investigated. It was found that the rate of particle clearance is a function of the volume of air passing through the purifiers but that the low throughflow of air makes their use of little value in clearing particles from a normal sized room. Vacuum cleaners were more effective than air purifiers because of their higher air throughput, so too were high volume fan systems in conjunction with simple filtration units. Ionizers had no effect but steam condensation was very efficient at clearing airborne particles.

  19. Environmental Factors Affecting Production, Release, and Field Populations of Conidia of Alternaria alternata, the Cause of Brown Spot of Citrus.

    PubMed

    Timmer, L W; Solel, Z; Gottwald, T R; Ibañez, A M; Zitko, S E

    1998-11-01

    ABSTRACT Alternaria brown spot, caused by Alternaria alternata pv. citri, affects many tangerines and their hybrids, causing loss of immature leaves and fruit and reducing the marketability of the remaining fruit. Conidial production of A. alternata was greatest on mature leaves moistened and maintained at near 100% relative humidity (RH) for 24 h, whereas leaves that had been soaked or maintained at moderate RH produced few conidia. Conidial release from filter paper cultures and infected leaves was studied in a computer-controlled environmental chamber. Release of large numbers of conidia was triggered from both substrates by sudden drops in RH or by simulated rainfall events. Vibration induced release of low numbers of conidia, but red/infrared irradiation had no effect. In field studies from 1994 to 1996, air sampling with a 7-day recording volumetric spore trap indicated that conidia were present throughout the year with periodic large peaks. The number of conidia captured was not closely related to rainfall amounts or average wind speed, but was weakly related to the duration of leaf wetness. Likewise, disease severity on trap plants placed in the field weekly during 1995 to 1996 was not closely related to conidial numbers or rainfall amounts, but was weakly related to leaf wetness duration. Sufficient inoculum appears to be available to allow infection to occur throughout the year whenever susceptible host tissue and moisture are available.

  20. Cyclic AMP-dependent protein kinase A negatively regulates conidia formation by the tangerine pathotype of Alternaria alternata.

    PubMed

    Tsai, Hsieh-Chin; Yang, Siwy Ling; Chung, Kuang-Ren

    2013-02-01

    The necrotrophic fungal pathogen Alternaria alternata causes brown spot diseases in many citrus cultivars. The FUS3 and SLT2 mitogen-activated protein kinases (MAPK)-mediated signaling pathways have been shown to be required for conidiation. Exogenous application of cAMP to this fungal pathogen decreased conidia formation considerably. This study determined whether a cAMP-activated protein kinase A (PKA) is required for conidiation. Using loss-of-function mutations in PKA catalytic and regulatory subunit-coding genes, we demonstrated that PKA negatively regulates conidiation. Fungal mutants lacking PKA catalytic subunit gene (PKA ( cat )) reduced growth, lacked detectable PKA activity, and produced higher amounts of conidia compared to wild-type. Introduction of a functional copy of PKA ( cat ) into a null mutant partially restored PKA activity and produced wild-type level of conidia. In contrast, fungi lacking PKA regulatory subunit gene (PKA ( reg )) produced detectable PKA activity, exhibited severe growth reduction, formed swelling hyphal segments, and produced no mature conidia. Introduction of the PKA ( reg ) gene to a regulatory subunit mutant restored all phenotypes to wild type. PKA ( reg )-null mutants induced fewer necrotic lesions on citrus compared to wild-type, whereas PKA ( cat ) mutant displayed wild-type virulence. Overall, our studies indicate that PKA and FUS3-mediated signaling pathways apparently have very different roles in the regulation of conidia production and A. alternata pathogenesis in citrus.

  1. EST analysis of cDNA libraries from the entomopathogenic fungus Beauveria (Cordyceps) bassiana. I. Evidence for stage-specific gene expression in aerial conidia, in vitro blastospores and submerged conidia.

    PubMed

    Cho, Eun-Min; Liu, Li; Farmerie, William; Keyhani, Nemat O

    2006-09-01

    The entomopathogenic fungus Beauveria (Cordyceps) bassiana holds much promise as a pest biological control agent. B. bassiana produces at least three in vitro single cell infectious propagules, including aerial conidia, vegetative cells termed blastospores and submerged conidia, that display different morphological, biochemical and virulence properties. Populations of aerial conidia, blastospores and submerged conidia were produced on agar plates, rich liquid broth cultures and under conditions of nutrient limitation in submerged cultures, respectively. cDNA libraries were generated from mRNA isolated from each B. bassiana cell type and approximately 2,500 5' end sequences were determined from each library. Sequences derived from aerial conidia clustered into 284 contigs and 963 singlets, with those derived from blastospores and submerged conidia forming 327 contigs with 788 singlets, and 303 contigs and 1,079 contigs, respectively. Almost half (40-45 %) of the sequences in each library displayed either no significant similarity (e value >10(-4)) or similarity to hypothetical proteins found in the NCBI database. The expressed sequence tag dataset also included sequences representing a significant portion of proteins in cellular metabolism, information storage and processing, transport and cell processes, including cell division and posttranslational modifications. Transcripts encoding a diverse array of pathogenicity-related genes, including proteases, lipases, esterases, phosphatases and enzymes producing toxic secondary metabolites, were also identified. Comparative analysis between the libraries identified 2,416 unique sequences, of which 20-30 % were unique to each library, and only approximately 6 % of the sequences were shared between all three libraries. The unique and divergent representation of the B. bassiana transcriptome in the cDNA libraries from each cell type suggests robust differential gene expression profiles in response to environmental

  2. A Non-canonical Melanin Biosynthesis Pathway Protects Aspergillus terreus Conidia from Environmental Stress.

    PubMed

    Geib, Elena; Gressler, Markus; Viediernikova, Iuliia; Hillmann, Falk; Jacobsen, Ilse D; Nietzsche, Sandor; Hertweck, Christian; Brock, Matthias

    2016-05-19

    Melanins are ubiquitous pigments found in all kingdoms of life. Most organisms use them for protection from environmental stress, although some fungi employ melanins as virulence determinants. The human pathogenic fungus Aspergillus fumigatus and related Ascomycetes produce dihydroxynaphthalene- (DHN) melanin in their spores, the conidia, and use it to inhibit phagolysosome acidification. However, biosynthetic origin of melanin in a related fungus, Aspergillus terreus, has remained a mystery because A. terreus lacks genes for synthesis of DHN-melanin. Here we identify genes coding for an unusual NRPS-like enzyme (MelA) and a tyrosinase (TyrP) that A. terreus expressed under conidiation conditions. We demonstrate that MelA produces aspulvinone E, which is activated for polymerization by TyrP. Functional studies reveal that this new pigment, Asp-melanin, confers resistance against UV light and hampers phagocytosis by soil amoeba. Unexpectedly, Asp-melanin does not inhibit acidification of phagolysosomes, thus likely contributing specifically to survival of A. terreus conidia in acidic environments. Copyright © 2016 Elsevier Ltd. All rights reserved.

  3. Comparison between gene expression of conidia and germinating phase in Trichophyton rubrum.

    PubMed

    Yang, Li; Wang, LingLing; Peng, JunPing; Yu, Lu; Liu, Tao; Leng, WenChuan; Yang, Jian; Chen, LiHong; Zhang, WenLiang; Zhang, Qian; Qi, YiPeng; Jin, Qi

    2007-06-01

    Trichophyton rubrum is a dominating superficial dermatophyte, whose conidial germination is correlated to pathopoiesis and a highly important developmental process. To investigate the changes of physiology, biochemistry and cytology during the germination, we selected 3364 function identified ESTs from T. rubrum cDNA library to construct cDNA microarrays, and compared the gene expression levels of conidia and germinating phase. Data analysis indicated that 335 genes were up-regulated during the germination, which mainly encoded translated, modified proteins and structural proteins. The constituents of cell wall and cell membrane were synthetized abundantly, suggesting that they are the foundation of cell morphogenesis. The ingredients of the two-component signal transduction system were up-regulated, presuming that they were important for the conidial germination. Genes of various metabolic pathways were expressed prosperously, especially the genes that participated in glycolysis and oxidative phosphorylation were up-regulated on the whole, demonstrating that in the environment with sufficient oxygen and glucose, conidia obtained energy through aerobic respiration. This paper provides important clues which are helpful to understanding the changes in gene expression, signal conduction and metabolism characteristics during T. rubrum conidial germination, and possess significant meaning to the study of other superficial dermatophytes.

  4. Conidia of the insect pathogenic fungus, Metarhizium anisopliae, fail to adhere to mosquito larval cuticle

    PubMed Central

    Greenfield, Bethany P. J.; Lord, Alex M.; Dudley, Ed; Butt, Tariq M.

    2014-01-01

    Adhesion of conidia of the insect pathogenic fungus, Metarhizium anisopliae, to the arthropod host cuticle initially involves hydrophobic forces followed by consolidation facilitated by the action of extracellular enzymes and secretion of mucilage. Gene expression analysis and atomic force microscopy were used to directly quantify recognition and adhesion between single conidia of M. anisopliae and the cuticle of the aquatic larval stage of Aedes aegypti and a representative terrestrial host, Tenebrio molitor. Gene expression data indicated recognition by the pathogen of both hosts; however, the forces for adhesion to the mosquito were approximately five times lower than those observed for Tenebrio. Although weak forces were recorded in response to Aedes, Metarhizium was unable to consolidate firm attachment. An analysis of the cuticular composition revealed an absence of long-chain hydrocarbons in Aedes larvae which are thought to be required for fungal development on host cuticle. This study provides, to our knowledge, the first evidence that Metarhizium does not form firm attachment to Ae. aegypti larvae in situ, therefore preventing the normal route of invasion and pathogenesis from occuring. PMID:26064542

  5. Automated quantification of the phagocytosis of Aspergillus fumigatus conidia by a novel image analysis algorithm

    PubMed Central

    Kraibooj, Kaswara; Schoeler, Hanno; Svensson, Carl-Magnus; Brakhage, Axel A.; Figge, Marc Thilo

    2015-01-01

    Studying the pathobiology of the fungus Aspergillus fumigatus has gained a lot of attention in recent years. This is due to the fact that this fungus is a human pathogen that can cause severe diseases, like invasive pulmonary aspergillosis in immunocompromised patients. Because alveolar macrophages belong to the first line of defense against the fungus, here, we conduct an image-based study on the host-pathogen interaction between murine alveolar macrophages and A. fumigatus. This is achieved by an automated image analysis approach that uses a combination of thresholding, watershed segmentation and feature-based object classification. In contrast to previous approaches, our algorithm allows for the segmentation of individual macrophages in the images and this enables us to compute the distribution of phagocytosed and macrophage-adherent conidia over all macrophages. The novel automated image-based analysis provides access to all cell-cell interactions in the assay and thereby represents a framework that enables comprehensive computation of diverse characteristic parameters and comparative investigation for different strains. We here apply automated image analysis to confocal laser scanning microscopy images of the two wild-type strains ATCC 46645 and CEA10 of A. fumigatus and investigate the ability of macrophages to phagocytose the respective conidia. It is found that the CEA10 strain triggers a stronger response of the macrophages as revealed by a higher phagocytosis ratio and a larger portion of the macrophages being active in the phagocytosis process. PMID:26106370

  6. Effects of Single-Drop Impactions and Natural and Simulated Rains on the Dispersal of Botryosphaeria dothidea Conidia.

    PubMed

    Ahimera, Neph; Gisler, Simon; Morgan, David P; Michailides, Themis J

    2004-11-01

    ABSTRACT Laboratory and field experiments were conducted to study the dispersal of Botryosphaeria dothidea conidia using single-drop impactions and natural and simulated precipitations. For laboratory studies, 200 single drops were released from a height of 1 m on infected pistachio nuts. On pieces of photographic film, 50% of the droplets were collected within 20 mm (average droplet travel distance) of the target area, and the droplets ranged from 0.041 to 3.19 mm in diameter, with an average of 0.3 mm. Each droplet carried an average of 23 B. dothidea conidia. In 3 years of field experiments, rainwater was collected in funnels connected to bottles positioned at different heights inside the tree canopy and at different distances away from the edge of tree canopy in three commercial pistachio orchards in San Joaquin, Yolo, and Glenn counties in California. Numbers of conidia in rainwater varied among and within sampling seasons by sampling dates and orchards. Up to 67,000 conidia/ml were obtained in rainwater samples collected from an orchard in Yolo County. Rainwater from orchards in Yolo and Glenn counties contained a consistently higher number of conidia than rainwater collected from the orchard in San Joaquin County. Variation in numbers of conidia also existed among heights where bottles were located. There were significantly more conidia in rainwater collected inside than outside tree canopies. Inside tree canopies, bottles located at 100 and 150 cm above ground collected more B. dothidea conidia than those placed at 50 and 200 cm. Conidia were collected as far as 1 m from the tree canopy edge. Based on data from the Glenn County orchard, a linear relationship between number of conidia (Y) and rainfall amount (X) in millimeters was determined as Y = 240X - 3,867, with r(2) = 0.91, which meant that a minimum of 16.1 mm of rain was needed to disperse conidia of B. dothidea. The power law model best described the dispersal gradients of B. dothidea propagules in

  7. Toxicity of ozone gas to conidia of Penicillium digitatum, P. italicum, and Botrytis cinerea and control of gray mold on table grapes

    USDA-ARS?s Scientific Manuscript database

    Penicillium digitatum, P. italicum, and Botrytis cinerea attack fresh fruit and cause significant postharvest decay losses and the toxicity of ozone (O3) gas at different relative humidities to control their conidia was determined. Conidia were exposed to an atmosphere containing 200 to 350 µL L-1 o...

  8. A Quantitative Dynamic Simulation of Bremia lactucae Airborne Conidia Concentration above a Lettuce Canopy.

    PubMed

    Fall, Mamadou Lamine; Van der Heyden, Hervé; Carisse, Odile

    2016-01-01

    Lettuce downy mildew, caused by the oomycete Bremia lactucae Regel, is a major threat to lettuce production worldwide. Lettuce downy mildew is a polycyclic disease driven by airborne spores. A weather-based dynamic simulation model for B. lactucae airborne spores was developed to simulate the aerobiological characteristics of the pathogen. The model was built using the STELLA platform by following the system dynamics methodology. The model was developed using published equations describing disease subprocesses (e.g., sporulation) and assembled knowledge of the interactions among pathogen, host, and weather. The model was evaluated with four years of independent data by comparing model simulations with observations of hourly and daily airborne spore concentrations. The results show an accurate simulation of the trend and shape of B. lactucae temporal dynamics of airborne spore concentration. The model simulated hourly and daily peaks in airborne spore concentrations. More than 95% of the simulation runs, the daily-simulated airborne conidia concentration was 0 when airborne conidia were not observed. Also, the relationship between the simulated and the observed airborne spores was linear. In more than 94% of the simulation runs, the proportion of the linear variation in the hourly-observed values explained by the variation in the hourly-simulated values was greater than 0.7 in all years except one. Most of the errors came from the deviation from the 1:1 line, and the proportion of errors due to the model bias was low. This model is the only dynamic model developed to mimic the dynamics of airborne inoculum and represents an initial step towards improved lettuce downy mildew understanding, forecasting and management.

  9. A Quantitative Dynamic Simulation of Bremia lactucae Airborne Conidia Concentration above a Lettuce Canopy

    PubMed Central

    Fall, Mamadou Lamine; Van der Heyden, Hervé; Carisse, Odile

    2016-01-01

    Lettuce downy mildew, caused by the oomycete Bremia lactucae Regel, is a major threat to lettuce production worldwide. Lettuce downy mildew is a polycyclic disease driven by airborne spores. A weather-based dynamic simulation model for B. lactucae airborne spores was developed to simulate the aerobiological characteristics of the pathogen. The model was built using the STELLA platform by following the system dynamics methodology. The model was developed using published equations describing disease subprocesses (e.g., sporulation) and assembled knowledge of the interactions among pathogen, host, and weather. The model was evaluated with four years of independent data by comparing model simulations with observations of hourly and daily airborne spore concentrations. The results show an accurate simulation of the trend and shape of B. lactucae temporal dynamics of airborne spore concentration. The model simulated hourly and daily peaks in airborne spore concentrations. More than 95% of the simulation runs, the daily-simulated airborne conidia concentration was 0 when airborne conidia were not observed. Also, the relationship between the simulated and the observed airborne spores was linear. In more than 94% of the simulation runs, the proportion of the linear variation in the hourly-observed values explained by the variation in the hourly-simulated values was greater than 0.7 in all years except one. Most of the errors came from the deviation from the 1:1 line, and the proportion of errors due to the model bias was low. This model is the only dynamic model developed to mimic the dynamics of airborne inoculum and represents an initial step towards improved lettuce downy mildew understanding, forecasting and management. PMID:26953691

  10. [Compatibility of two fungal biocontrol agents conidia with commercial chemical acaricides].

    PubMed

    Jiang, Yu; Feng, Mingguang

    2006-07-01

    In this paper, the biological compatibility of fungal biocontrol agents Beauveria bassiana and Paecilomyces fumosoroseus conidia with 10 commercial chemical acaricides were assayed, based on the conidial germination rates in nutritional liquid and on SDAY plate. The results showed that nutritional liquid was more available than SDAY plate in the assay. So far as the 24 h conidial viability concerned, there were significant differences among the test acaricides with the concentrations of recommended for field spray and 5 and 10 fold dilutions, as well as between the two fungal agents. Since acaricides azocyclotin, liuyangmycin, dicofol and avermectin had strong inhibitory effects on the 24 h germination rate of both fungal agents conidia, their combined application with fungal agents was unsuitable for mite control. However, the combined application of pyridaben, propargite, chlorpyrifos, hexythiazox or amitraz with either B. bassiana or P. fumosoroseus was practical, because of their short-term compatibility. When the mixtures of oil-based B. bassiana formulation with the three concentrations of pyridaben, propargite and chlorpyrifos were stored at 4 degrees C or at ambient temperature for 12 months, none of the three chemicals was considered to be good enough for a combined formulation due to the great variability in long-term compatibility. Nevertheless, chlorpyrifos exhibited an encouraging long-term compatibility with B. bassiana, because its low concentration in the fungal formulation did not affect the conidial viability during a 6.5-month period of storage at ambient temperature. When stored at 4 degrees C in dark, the B. bassiana formulation containing low or medium concentration of chlorpyrifos retained the conidial viability of > 90% for up to 12 months.

  11. Effects of a Cover Crop on Splash Dispersal of Colletotrichum acutatum Conidia.

    PubMed

    Ntahimpera, N; Ellis, M A; Wilson, L L; Madden, L V

    1998-06-01

    ABSTRACT A rain simulator, with generated rains of 11 and 30 mm/h, was used to determine the effect of a cover crop or intercrop on the splash dispersal of Colletotrichum acutatum conidia. Dispersal through sudangrass, which can be used as a 'living mulch', was tested at two planting densities (140 or 280 kg/ha) and two heights (5 and 20 cm) and compared with a control consisting of a bare soil. Dispersal of C. acutatum conidia was assessed by counting colonies formed from spore-bearing splash droplets deposited in sheltered petri plates containing a selective medium. Both a cover crop and rain intensity significantly affected splash dispersal as measured by the interpolated total number of colonies (denoted by Sigma) from 0 to 72 cm from the inoculum source and in a time span of 61 min of generated rain (P < 0.001). However, there was no significant interaction of cover crop and intensity (P > 0.90). Dispersal with a 30-mm/h rain was higher than dispersal with a 11-mm/h rain, and presence of a cover crop significantly reduced dispersal compared with bare soil (P < 0.001). Of the treatments with sudangrass, cover crop planting density did not affect dispersal overall, but there was greater spore dispersal with the taller sudangrass at the higher planting density, due in part to the higher rate of water splashing with the tall grass compared with the short grass. Spore deposition in the petri plates could be functionally related to distance and time using a diffusion-type model, and parameter estimates could be used to explain the effects of cover crop on Sigma. Although the relationship between cover crop properties and splash dispersal is complex, results show the potential beneficial effects of the cover crop on disease management.

  12. Thermal inactivation of Botrytis cinerea conidia in synthetic medium and strawberry puree.

    PubMed

    Villa-Rojas, R; Sosa-Morales, M E; López-Malo, A; Tang, J

    2012-04-16

    Botrytis cinerea is one of the most important post-harvest molds that cause quality deterioration of strawberries and other fruits even during refrigeration storage. This research studied the effects of thermal inactivation of B. cinerea in synthetic medium and strawberry puree using hot water baths at different temperatures. These media were studied in order to determine if results obtained in a solution with the major components of the fruit (synthetic media), are comparable to the ones obtained in fruit purees. The results demonstrated that B. cinerea spores can be inactivated by heat treatments using relatively low temperatures (42-46 °C). Inactivation curves were well described by first order kinetics (R² 0.91-0.99). B. cinerea conidia inoculated in synthetic medium required less time to achieve one log reduction in population than those inoculated in the fruit puree. D values were 22, 8.5, 4 and 1.4 min at 42, 44, 46 and 48 °C, respectively, in synthetic medium; while D values in strawberry puree were 44.9, 13.8, 4.7 and 1.4 min at 42, 44, 46 and 48 °C, respectively. The z values obtained were 4.15 and 5.08 °C for the strawberry puree and synthetic medium respectively, showing higher sensitivity of B. cinerea in fruit purees than in the synthetic medium. Thus, a change in the medium composition had a marked difference in the heat inactivation of B. cinerea conidia, and the results obtained in synthetic medium are not accurate to describe the behavior of the microorganism in the fruit.

  13. Two Novel Genes Induced by Hard-Surface Contact of Colletotrichum gloeosporioides Conidia

    PubMed Central

    Kim, Yeon-Ki; Liu, Zhi-Mei; Li, Daoxin; Kolattukudy, Pappachan E.

    2000-01-01

    Germinating conidia of many phytopathogenic fungi must differentiate into an infection structure called the appressorium in order to penetrate into their hosts. This differentiation is known to require contact with a hard surface. However, the molecular basis for this requirement is not known. Induction of this differentiation in the avocado pathogen, Colletotrichum gloeosporioides, by chemical signals such as the host's surface wax or the fruit-ripening hormone, ethylene, requires contact of the conidia with a hard surface for about 2 h. To study molecular events triggered by hard-surface contact, we isolated several genes expressed during the early stage of hard-surface treatment by a differential-display method. The genes that encode Colletotrichum hard-surface induced proteins are designated chip genes. In this study, we report the characterization of CHIP2 and CHIP3 genes that would encode proteins with molecular masses of 65 and 64 kDa, respectively, that have no homology to any known proteins. The CHIP2 product would contain a putative nuclear localization signal, a leucine zipper motif, and a heptad repeat region which might dimerize into coiled-coil structure. The CHIP3 product would be a nine-transmembrane-domain-containing protein. RNA blots showed that CHIP2 and CHIP3 are induced by a 2-h hard-surface contact. However, disruption of these genes did not affect the appressorium-forming ability and did not cause a significant decrease in virulence on avocado or tomato fruits suggesting that C. gloeosporioides might have genes functionally redundant to CHIP2 and CHIP3 or that these genes induced by hard-surface contact control processes not directly involved in pathogenesis. PMID:10940006

  14. Importance of complement 3 and mannose receptors in phagocytosis of Paracoccidioides brasiliensis conidia by Nramp1 congenic macrophages lines.

    PubMed

    Jiménez, María del P; Restrepo, Angela; Radzioch, Danuta; Cano, Luz E; García, Luis F

    2006-06-01

    Genetic factors influence susceptibility to Paracoccidioidomycosis, a Latin American endemic mycosis. The pattern of susceptibility of congenic mouse strains infected with Paracoccidioides brasiliensis resembles the pattern of the Nramp1 gene. Thus, congenic murine bone-marrow-derived macrophage lines B10R (Nramp1rGly169) and B10S (null Nramp1 protein expression, Nramp1sAsp169) were infected with P. brasiliensis conidia and compared, under opsonic and nonopsonic conditions. Opsonization increased the percentage of phagocytosis by both cell lines. B10R macrophages exhibited a higher percentage of cells with associated conidia and higher number of conidia per macrophage than B10S. Heat-inactivation and EDTA treatment of serum used for opsonization, and treatment of macrophages with anti-complement receptor 3 (CR3) decreased phagocytosis by both cell lines. alpha-methyl-d-mannoside reduced phagocytosis by B10R macrophages, suggesting that the mannose receptor participates in phagocytosis by these cells. The CR3 expression was similar on both cell lines and B10R expressed more mannose receptors, but neither cell line expressed CR1. IFNgamma decreased the conversion of conidia to the yeast form of P. brasiliensis in B10R, but not in B10S macrophages.

  15. Arthrornyces and Blastosporella, two new genera of conidia-producing lyophylloid agarics (Agaricales, Basidiornycota) from the neotropics

    Treesearch

    Timothy J. Baroni; Ana Esperanza Franco-molano; D. Jean Lodge; Daniel L. Lindner; Egon Horak; Valerie Hofstetter

    2007-01-01

    Two new genera encompassing three new species of lyophylloid agarics that produce conidia on the basidiomata are described. Arthromyces is a genus comprised of two very different arthrospore-producing mushroom species found in the Greater Antilles and Central America. Blastosporella is a monotypic genus with spherical balls of...

  16. Arthromyces and Blastosporella, two new genera of conidia-producing lyophylloid agarics (Agaricales, Basidiomycota) from the neotropics

    Treesearch

    Timothy J. Baroni; Ana Esperanza Franco-Molano; D. Jean Lodge; Daniel L. Lindner; Egon Horak; Valerie Hofstetter

    2007-01-01

    Two new genera encompassing three new species of lyophylloid agarics that produce conidia on the basidiomata are described. Arthromyces is a genus comprised of two very different arthrospore-producing mushroom species found in the Greater Antilles and Central America. Blastosporella is a monotypic genus with spherical balls of blastospores covering the pileus surface...

  17. QUANTIFICATION OF STACHYBOTRYS CHARTARUM CONIDIA IN INDOOR DUST USING REAL TIME, FLUORESCENT PROBE-BASED DETECTION OF PCR PRODUCTS

    EPA Science Inventory

    Analyses of fungal spores or conidia in indoor dust samples can be useful for determining the contamination status of building interiors and in signaling instances where potentially harmful exposures of building occupants to these organisms may exist. A recently developed method ...

  18. A proteomic approach to identifying proteins differentially expressed in conidia and mycelium of the entomopathogenic fungus Metarhizium acridum.

    PubMed

    Barros, Bruno H R; da Silva, Sérgio H; dos ReisMarques, Everaldo Dos Reis; Rosa, José C; Yatsuda, Ana Patrícia; Roberts, Donald W; Braga, Gilberto U L

    2010-07-01

    Metarhizium spp. is an important worldwide group of entomopathogenic fungi used as an interesting alternative to chemical insecticides in programs of agricultural pest and disease vector control. Metarhizium conidia are important in fungal propagation and also are responsible for host infection. Despite their importance, several aspects of conidial biology, including their proteome, are still unknown. We have established conidial and mycelial proteome reference maps for Metarhizium acridum using two-dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF MS). In all, 1130±102 and 1200±97 protein spots were detected in ungerminated conidia and fast-growing mycelia, respectively. Comparison of the two protein-expression profiles reveled that only 35% of the protein spots were common to both developmental stages. Out of 94 2-DE protein spots (65 from conidia, 25 from mycelia and two common to both) analyzed using mass spectrometry, seven proteins from conidia, 15 from mycelia and one common to both stages were identified. The identified protein spots exclusive to conidia contained sequences similar to known fungal stress-protector proteins (such as heat shock proteins (HSP) and 6-phosphogluconate dehydrogenase) plus the fungal allergen Alt a 7, actin and the enzyme cobalamin-independent methionine synthase. The identified protein spots exclusive to mycelia included proteins involved in several cell housekeeping biological processes. Three proteins (HSP 90, 6-phosphogluconate dehydrogenase and allergen Alt a 7) were present in spots in conidial and mycelial gels, but they differed in their locations on the two gels.

  19. A novel approach of utilization of the fungal conidia biomass to remove heavy metals from the aqueous solution through immobilization

    NASA Astrophysics Data System (ADS)

    Cai, Chun-Xiang; Xu, Jian; Deng, Nian-Fang; Dong, Xue-Wei; Tang, Hao; Liang, Yu; Fan, Xian-Wei; Li, You-Zhi

    2016-11-01

    The biomass of filamentous fungi is an important cost-effective biomass for heavy metal biosorption. However, use of free fungal cells can cause difficulties in the separation of biomass from the effluent. In this study, we immobilized the living conidia of the heavy metal-resistant Penicillium janthinillum strain GXCR by polyvinyl alcohol (PVA)-sodium alginate (SA) beads to remove heavy metals from an aqueous solution containing a low concentration (70 mg/L) of Cu, Pb, and Cd. The PVA-SA-conidia beads showed perfect characters of appropriate mechanical strength suitable for metal removal from the dynamic wastewater environment, an ideal settleability, easy separation from the solution, and a high metal biosorption and removal rate even after four cycles of successive sorption-desorption of the beads, overcoming disadvantages when fungal biomasses alone are used for heavy metal removal from wastewater. We also discuss the major biosorption-affecting factors, biosorption models, and biosorption mechanisms.

  20. A novel approach of utilization of the fungal conidia biomass to remove heavy metals from the aqueous solution through immobilization.

    PubMed

    Cai, Chun-Xiang; Xu, Jian; Deng, Nian-Fang; Dong, Xue-Wei; Tang, Hao; Liang, Yu; Fan, Xian-Wei; Li, You-Zhi

    2016-11-16

    The biomass of filamentous fungi is an important cost-effective biomass for heavy metal biosorption. However, use of free fungal cells can cause difficulties in the separation of biomass from the effluent. In this study, we immobilized the living conidia of the heavy metal-resistant Penicillium janthinillum strain GXCR by polyvinyl alcohol (PVA)-sodium alginate (SA) beads to remove heavy metals from an aqueous solution containing a low concentration (70 mg/L) of Cu, Pb, and Cd. The PVA-SA-conidia beads showed perfect characters of appropriate mechanical strength suitable for metal removal from the dynamic wastewater environment, an ideal settleability, easy separation from the solution, and a high metal biosorption and removal rate even after four cycles of successive sorption-desorption of the beads, overcoming disadvantages when fungal biomasses alone are used for heavy metal removal from wastewater. We also discuss the major biosorption-affecting factors, biosorption models, and biosorption mechanisms.

  1. A novel approach of utilization of the fungal conidia biomass to remove heavy metals from the aqueous solution through immobilization

    PubMed Central

    Cai, Chun-Xiang; Xu, Jian; Deng, Nian-Fang; Dong, Xue-Wei; Tang, Hao; Liang, Yu; Fan, Xian-Wei; Li, You-Zhi

    2016-01-01

    The biomass of filamentous fungi is an important cost-effective biomass for heavy metal biosorption. However, use of free fungal cells can cause difficulties in the separation of biomass from the effluent. In this study, we immobilized the living conidia of the heavy metal-resistant Penicillium janthinillum strain GXCR by polyvinyl alcohol (PVA)-sodium alginate (SA) beads to remove heavy metals from an aqueous solution containing a low concentration (70 mg/L) of Cu, Pb, and Cd. The PVA-SA-conidia beads showed perfect characters of appropriate mechanical strength suitable for metal removal from the dynamic wastewater environment, an ideal settleability, easy separation from the solution, and a high metal biosorption and removal rate even after four cycles of successive sorption-desorption of the beads, overcoming disadvantages when fungal biomasses alone are used for heavy metal removal from wastewater. We also discuss the major biosorption-affecting factors, biosorption models, and biosorption mechanisms. PMID:27848987

  2. Localization of melanin in conidia of Alternaria alternata using phage display antibodies.

    PubMed

    Carzaniga, Raffaella; Fiocco, Daniela; Bowyer, Paul; O'Connell, Richard J

    2002-03-01

    Melanins derived from 1,8-dihydroxynaphthalene (DHN) are important for the pathogenicity and survival of fungi causing disease in both plants and animals. However, precise information on their location within fungal cell walls is lacking. To obtain antibodies for the immunocytochemical localization of melanin, 83 phage antibodies binding to 1,8-DHN were selected from a naive semisynthetic single-chain Fv (scFv) phage display library. Sequence analysis of the heavy chain binding domains of 17 antibodies showed a high frequency of positively charged amino acids. One antibody, designated M1, was characterized in detail. M1 bound specifically to 1,8-DHN in competitive inhibition enzyme-linked immunosorbent assays, showing no cross-reaction with nine structurally related phenolic compounds. Epitope recognition required two hydroxyl groups in a 1,8 configuration. M1 also bound to naturally occurring melanin isolated from mycelia of Alternaria alternata, suggesting that epitopes remain accessible in polymerized melanin. Transmission electron microscopy-immunogold labeling, using M1 in the form of soluble scFv fragments, showed that melanin was located in the septa and outer (primary) walls of wild-type A. alternata conidia, but not those of an albino mutant, AKT88-1. The M1 antibody provides a new tool for detecting melanized pathogens in plant and animal tissues and for precisely mapping the distribution of the polymer within spores, appressoria, and hyphae.

  3. IL-1 signaling inhibits Trichophyton rubrum conidia development and modulates the IL-17 response in vivo.

    PubMed

    Yoshikawa, Fábio Seiti Yamada; Ferreira, Lucas Gonçalves; de Almeida, Sandro Rogério

    2015-01-01

    Dermatophytosis are one of the most common fungal infections in the world. They compromise keratinized tissues and the main etiological agent is Trichophyton rubrum. Macrophages are key cells in innate immunity and prominent sources of IL-1β, a potent inflammatory cytokine whose main production pathway is by the activation of inflammasomes and caspase-1. However, the role of inflammasomes and IL-1 signaling against T.rubrum has not been reported. In this work, we observed that bone marrow-derived macrophages produce IL-1β in response to T.rubrum conidia in a NLRP3-, ASC- and caspase-1-dependent fashion. Curiously, lack of IL-1 signaling promoted hyphae development, uncovering a protective role for IL-1β in macrophages. In addition, mice lacking IL-1R showed reduced IL-17 production, a key cytokine in the antifungal defense, in response to T.rubrum. Our findings point to a prominent role of IL-1 signaling in the immune response to T.rubrum, opening the venue for the study of this pathway in other fungal infections.

  4. Influence of mechanical stress and surface interaction on the aggregation of Aspergillus niger conidia.

    PubMed

    Grimm, L H; Kelly, S; Völkerding, I I; Krull, R; Hempel, D C

    2005-12-30

    Productivity of fungal cultures is closely linked with their morphologic development. Morphogenesis of coagulating filamentous fungi, like Aspergillus niger, starts with aggregation of conidia, also denominated as spores. Several parameters are presumed to control this event, but little is known about their mode of action. Rational process optimization requires models that mirror the underlying reaction mechanisms. An approach in this regard is suggested and supported by experimental data. Aggregation kinetics was examined for the first 15 h of cultivation under different cultivation conditions. Mechanical stress was considered as well as pH-dependent surface interaction. Deliberations were based on a two-step aggregation mechanism. The first aggregation step is only affected by the pH-value, not by the fluid dynamic conditions in the bioreactor. The second aggregation step, in contrast, depends on the pH-value as well as on agitation and aeration induced power input. For the given experimental set-up, agitation had much more influence than aeration. In addition, hyphal growth rate was determined to be the driving force for the second aggregation step.

  5. Nitric Oxide Participation in the Fungicidal Mechanism of Gamma Interferon-Activated Murine Macrophages against Paracoccidioides brasiliensis Conidia

    PubMed Central

    Gonzalez, Angel; de Gregori, Waldemar; Velez, Diana; Restrepo, Angela; Cano, Luz E.

    2000-01-01

    Paracoccidioidomycosis, a systemic mycosis restricted to Latin America and produced by the dimorphic fungus Paracoccidioides brasiliensis, is probably acquired by inhalation of conidia produced by the mycelial form. The macrophage (Mφ) represents the major cell defense against this pathogen; when activated with gamma interferon (IFN-γ), murine Mφs kill the fungus by an oxygen-independent mechanism. Our goal was to determine the role of nitric oxide in the fungicidal effect of Mφs on P. brasiliensis conidia. The results revealed that IFN-γ-activated murine Mφs inhibited the conidium-to-yeast transformation process in a dose-dependent manner; maximal inhibition was observed in Mφs activated with 50 U/ml and incubated for 96 h at 37°C. When Mφs were activated with 150 to 200 U of cytokine per ml, the number of CFU was 70% lower than in nonactivated controls, indicating that there was a fungicidal effect. The inhibitory effect was reversed by the addition of anti-IFN-γ monoclonal antibodies. Activation by IFN-γ also enhanced Mφ nitric oxide production, as revealed by increasing NO2 values (8 ± 3 μM in nonactivated Mφs versus 43 ± 13 μM in activated Mφs). The neutralization of IFN-γ also reversed nitric oxide production at basal levels (8 ± 5 μM). Additionally, we found that there was a significant inverse correlation (r = −0.8975) between NO2− concentration and transformation of P. brasiliensis conidia. Additionally, treatment with any of the three different nitric oxide inhibitors used (arginase, NG-monomethyl-l-arginine, and aminoguanidine), reverted the inhibition of the transformation process with 40 to 70% of intracellular yeast and significantly reduced nitric oxide production. These results show that IFN-γ-activated murine Mφs kill P. brasiliensis conidia through the l-arginine–nitric oxide pathway. PMID:10768942

  6. Arthromyces and Blastosporella, two new genera of conidia-producing lyophylloid agarics (Agaricales, Basidiomycota) from the neotropics.

    PubMed

    Baroni, Timothy J; Franco-Molano, Ana Esperanza; Lodge, D Jean; Lindner, Daniel L; Horak, Egon; Hofstetter, Valerie

    2007-05-01

    Two new genera encompassing three new species of lyophylloid agarics that produce conidia on the basidiomata are described. Arthromyces is a genus comprised of two very different arthrospore-producing mushroom species found in the Greater Antilles and Central America. Blastosporella is a monotypic genus with spherical balls of blastospores covering the pileus surface with age and is known from Hispaniola and Colombia. A key to the species of Arthromyces is included.

  7. A proteomic and genetic analysis of the Neurospora crassa conidia cell wall proteins identifies two glycosyl hydrolases involved in cell wall remodeling

    PubMed Central

    Ao, Jie; Aldabbous, Mash’el; Notaro, Marysa J.; Lojacono, Mark; Free, Stephen J.

    2016-01-01

    A proteomic analysis of the conidial cell wall identified 35 cell wall proteins. A comparison with the proteome of the vegetative hyphae showed that 16 cell wall proteins were shared, and that these shared cell wall proteins were cell wall biosynthetic proteins or cell wall structural proteins. Deletion mutants for 34 of the genes were analyzed for phenotypes indicative of conidial cell wall defects. Mutants for two cell wall glycosyl hydrolases, the CGL-1 β-1,3-glucanase (NCU07523) and the NAG-1 exochitinase (NCU10852), were found to have a conidial separation phenotype. These two enzymes function in remodeling the cell wall between adjacent conidia to facilitate conidia formation and dissemination. Using promoter::RFP and promoter::GFP constructs, we demonstrated that the promoters for 15 of the conidia-specific cell wall genes, including cgl-1 and nag-1, provided for conidia-specific gene expression or for a significant increase in their expression during conidiation. PMID:27381444

  8. Surface structure characterization of Aspergillus fumigatus conidia mutated in the melanin synthesis pathway and their human cellular immune response.

    PubMed

    Bayry, Jagadeesh; Beaussart, Audrey; Dufrêne, Yves F; Sharma, Meenu; Bansal, Kushagra; Kniemeyer, Olaf; Aimanianda, Vishukumar; Brakhage, Axel A; Kaveri, Srini V; Kwon-Chung, Kyung J; Latgé, Jean-Paul; Beauvais, Anne

    2014-08-01

    In Aspergillus fumigatus, the conidial surface contains dihydroxynaphthalene (DHN)-melanin. Six-clustered gene products have been identified that mediate sequential catalysis of DHN-melanin biosynthesis. Melanin thus produced is known to be a virulence factor, protecting the fungus from the host defense mechanisms. In the present study, individual deletion of the genes involved in the initial three steps of melanin biosynthesis resulted in an altered conidial surface with masked surface rodlet layer, leaky cell wall allowing the deposition of proteins on the cell surface and exposing the otherwise-masked cell wall polysaccharides at the surface. Melanin as such was immunologically inert; however, deletion mutant conidia with modified surfaces could activate human dendritic cells and the subsequent cytokine production in contrast to the wild-type conidia. Cell surface defects were rectified in the conidia mutated in downstream melanin biosynthetic pathway, and maximum immune inertness was observed upon synthesis of vermelone onward. These observations suggest that although melanin as such is an immunologically inert material, it confers virulence by facilitating proper formation of the A. fumigatus conidial surface.

  9. The pbrB gene encodes a laccase required for DHN-melanin synthesis in conidia of Talaromyces (Penicillium) marneffei.

    PubMed

    Sapmak, Ariya; Boyce, Kylie J; Andrianopoulos, Alex; Vanittanakom, Nongnuch

    2015-01-01

    Talaromyces marneffei (Basionym: Penicillium marneffei) is a significant opportunistic fungal pathogen in patients infected with human immunodeficiency virus in Southeast Asia. T. marneffei cells have been shown to become melanized in vivo. Melanins are pigment biopolymers which act as a non-specific protectant against various stressors and which play an important role during virulence in fungi. The synthesis of the two most commonly found melanins in fungi, the eumelanin DOPA-melanin and the allomelanin DHN-melanin, requires the action of laccase enzymes. The T. marneffei genome encodes a number of laccases and this study describes the characterization of one of these, pbrB, during growth and development. A strain carrying a PbrB-GFP fusion shows that pbrB is expressed at high levels during asexual development (conidiation) but not in cells growing vegetatively. The pbrB gene is required for the synthesis of DHN-melanin in conidia and when deleted results in brown pigmented conidia, in contrast to the green conidia of the wild type.

  10. Modulation of conidia production and expression of the gene bbrgs1 from Beauveria bassiana by oxygen pulses and light.

    PubMed

    Rodriguez-Gomez, Divanery; Marcial-Quino, Jaime; Loera, Octavio

    2015-09-01

    Light and oxidant states affect the conidiation in diverse fungi, although the response has not been described when both stimuli are applied simultaneously. Conidial production and quality in Beauveria bassiana were analysed under four conditions for a wild-type (wt) strain and a previously isolated mutant (mt): normal atmosphere (21% O2; NA) or oxygen-enriched pulses (26% O2; OEP), with either light (L) or darkness (D). The response was complemented by following the expression of the bbrgs1 gene, encoding a regulator of the G-protein signal associated to conidia production. Conidiation was not significantly affected in the mutant strain by any condition (highest value with NA-L: 2.7×10(8)concm(-2)). Relative to maximal levels under NA (NA-D: 4×10(7)concm(2)), the wt strain diminished conidiation by 34-fold under OEP. The expression of bbrgs1 was higher (up to 188 times) in the mutant strain in every condition relative to the wt strain, in fact expression levels were consistent with the conidiation yields between strains. Viability and hydrophobicity were less affected by culture conditions, although pathogenicity parameters improved in conidia from OEP. The response to OEP, either with light or darkness, was strain-dependent for conidial production, viability, hydrophobicity and infectivity of conidia, then these parameters could be modulated in mass production processes. Copyright © 2015 Elsevier Inc. All rights reserved.

  11. Surface Structure Characterization of Aspergillus fumigatus Conidia Mutated in the Melanin Synthesis Pathway and Their Human Cellular Immune Response

    PubMed Central

    Bayry, Jagadeesh; Beaussart, Audrey; Dufrêne, Yves F.; Sharma, Meenu; Bansal, Kushagra; Kniemeyer, Olaf; Aimanianda, Vishukumar; Brakhage, Axel A.; Kaveri, Srini V.; Kwon-Chung, Kyung J.

    2014-01-01

    In Aspergillus fumigatus, the conidial surface contains dihydroxynaphthalene (DHN)-melanin. Six-clustered gene products have been identified that mediate sequential catalysis of DHN-melanin biosynthesis. Melanin thus produced is known to be a virulence factor, protecting the fungus from the host defense mechanisms. In the present study, individual deletion of the genes involved in the initial three steps of melanin biosynthesis resulted in an altered conidial surface with masked surface rodlet layer, leaky cell wall allowing the deposition of proteins on the cell surface and exposing the otherwise-masked cell wall polysaccharides at the surface. Melanin as such was immunologically inert; however, deletion mutant conidia with modified surfaces could activate human dendritic cells and the subsequent cytokine production in contrast to the wild-type conidia. Cell surface defects were rectified in the conidia mutated in downstream melanin biosynthetic pathway, and maximum immune inertness was observed upon synthesis of vermelone onward. These observations suggest that although melanin as such is an immunologically inert material, it confers virulence by facilitating proper formation of the A. fumigatus conidial surface. PMID:24818666

  12. The pbrB Gene Encodes a Laccase Required for DHN-Melanin Synthesis in Conidia of Talaromyces (Penicillium) marneffei

    PubMed Central

    Sapmak, Ariya; Boyce, Kylie J.; Andrianopoulos, Alex; Vanittanakom, Nongnuch

    2015-01-01

    Talaromyces marneffei (Basionym: Penicillium marneffei) is a significant opportunistic fungal pathogen in patients infected with human immunodeficiency virus in Southeast Asia. T. marneffei cells have been shown to become melanized in vivo. Melanins are pigment biopolymers which act as a non-specific protectant against various stressors and which play an important role during virulence in fungi. The synthesis of the two most commonly found melanins in fungi, the eumelanin DOPA-melanin and the allomelanin DHN-melanin, requires the action of laccase enzymes. The T. marneffei genome encodes a number of laccases and this study describes the characterization of one of these, pbrB, during growth and development. A strain carrying a PbrB-GFP fusion shows that pbrB is expressed at high levels during asexual development (conidiation) but not in cells growing vegetatively. The pbrB gene is required for the synthesis of DHN-melanin in conidia and when deleted results in brown pigmented conidia, in contrast to the green conidia of the wild type. PMID:25866870

  13. Taxonomy, nomenclature and phylogeny of three cladosporium-like hyphomycetes, Sorocybe resinae, Seifertia azaleae and the Hormoconis anamorph of Amorphotheca resinae

    PubMed Central

    Seifert, K.A.; Hughes, S.J.; Boulay, H.; Louis-Seize, G.

    2007-01-01

    Using morphological characters, cultural characters, large subunit and internal transcribed spacer rDNA (ITS) sequences, and provisions of the International Code of Botanical Nomenclature, this paper attempts to resolve the taxonomic and nomenclatural confusion surrounding three species of cladosporium-like hyphomycetes. The type specimen of Hormodendrum resinae, the basis for the use of the epithet resinae for the creosote fungus {either as Hormoconis resinae or Cladosporium resinae) represents the mononematous synanamorph of the synnematous, resinicolous fungus Sorocybe resinae. The phylogenetic relationships of the creosote fungus, which is the anamorph of Amorphotheca resinae, are with the family Myxotrichaceae, whereas S. resinae is related to Capronia (Chaetothyriales, Herpotrichiellaceae). Our data support the segregation of Pycnostysanus azaleae, the cause of bud blast of rhododendrons, in the recently described anamorph genus Seifertia, distinct from Sorocybe; this species is related to the Dothideomycetes but its exact phylogenetic placement is uncertain. To formally stabilize the name of the anamorph of the creosote fungus, conservation of Hormodendrum resinae with a new holotype should be considered. The paraphyly of the family Myxotrichaceae with the Amorphothecaceae suggested by ITS sequences should be confirmed with additional genes. PMID:18491002

  14. Aspergillus fumigatus germ tube growth and not conidia ingestion induces expression of inflammatory mediator genes in the human lung epithelial cell line A549.

    PubMed

    Bellanger, Anne-Pauline; Millon, Laurence; Khoufache, Khaled; Rivollet, Danièle; Bièche, Ivan; Laurendeau, Ingrid; Vidaud, Michel; Botterel, Françoise; Bretagne, Stéphane

    2009-02-01

    Inhalation of conidia is the main cause of invasive pulmonary aspergillosis (IPA) and the respiratory epithelium is the first line of defence. To explore the triggering factor for the inflammatory response to Aspergillus fumigatus, the species mainly responsible for IPA, this study analysed the differential expression of three inflammatory genes in A549 cells after challenge with live and killed conidia. The influence of steroids, one of the main risk factors for developing IPA, was also investigated. Quantification of mRNAs of the inflammatory mediator genes encoding interleukin (IL)-8, tumour necrosis factor (TNF)-alpha and granulocyte-monocyte colony-stimulating factor (GM-CSF) was carried out using real-time PCR. Ingestion rates were studied for the conidia of A. fumigatus and Penicillium chrysogenum using a fluorescence brightener. Similar results were obtained for both species, with ingestion rates ranging from 35 to 40 %. Exposure of A549 cells to live A. fumigatus conidia only induced a four- to fivefold increase in the mRNA levels of the three genes, starting 8 h after the initial contact. Both inactivation of live A. fumigatus conidia and treatment by dexamethasone (10(-7) M) prevented the overexpression of TNF-alpha, IL-8 and GM-CSF. Fungal growth, rather than conidia ingestion, appears to be the main stimulus for the production of inflammatory mediators by epithelial cells, and this production is inhibited by steroid therapy. These results underline the role that the epithelium plays in the innate response against IPA.

  15. Intact Cell/Spore Mass Spectrometry of Fusarium Macro Conidia for Fast Isolate and Species Differentiation

    NASA Astrophysics Data System (ADS)

    Dong, Hongjuan; Marchetti-Deschmann, Martina; Winkler, Wolfgang; Lohninger, Hans; Allmaier, Guenter

    The focus of this paper is the development of an approach called intact cell mass spectrometry (ICMS) or intact spore mass spectrometry (ISMS) based on the technique matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) for the rapid differentiation and identification of Fusarium species. Several parameters, which are known to affect the quality of IC mass spectra, have been investigated in detail by varying the MALDI matrix as well as the solvent system, in which the matrix has been dissolved, the solvent system for sample purification and the type of sample/MALDI matrix deposition technique. In the end characteristic as well as highly reproducible IC or IS mass spectra or peptide/protein fingerprints of three Fusarium species (F. cerealis, F. graminearum and F. poae) including 16 Fusarium isolates derived from different hosts and geographical locations have been obtained. Unscaled hierarchical cluster analysis based on ICMS data of eight selected Fusarium isolates of two species F. graminearum and F. poae revealed significant difference among the peptide/protein pattern of them. The results of the applied cluster analysis proved that, ICMS is a powerful approach for the rapid differentiation of Fusarium species. In addition, an on-target tryptic digestion was applied to Fusarium macro conidia spores to identify proteins using MALDI post source decay (PSD) fragment ion analysis. Two kinds of trypsin, namely bead-immobilized - to favor cleavage of surface-associated proteins - and non-immobilized trypsin were applied and compared. The results showed that the latter is more suitable for generating sequence tags by PSD fragment ion analysis.

  16. Protein translation inhibition by Stachybotrys chartarum conidia with and without the mycotoxin containing polysaccharide matrix.

    PubMed

    Karunasena, Enusha; Cooley, J Danny; Straus, Douglas; Straus, David C

    2004-07-01

    Recent studies have correlated the presence of Stachybotrys chartarum in structures with SBS. S. chartarum produces mycotoxins that are thought to produce some of the symptoms reported in sick-building syndrome (SBS). The conidia (spores) produced by Stachybotrys species are not commonly found in the air of buildings that have been found to contain significant interior growth of this organism. This could be due in part to the large size of the Stachybotrys spores, or the organism growing in hidden areas such as wall cavities. However, individuals in buildings with significant Stachybotrys growth frequently display symptoms that may be attributed to exposure to the organism's mycotoxins. In addition, Stachybotrys colonies produce a "slime" or polysaccharide (carbohydrate) matrix that coats the hyphae and the spores. The intent of this project was to determine whether the carbohydrate matrix and the mycotoxins embedded in it could be removed from the spores by repeated washings with either aqueous or organic solvents. The results demonstrated that the process of spore washing removed compounds that were toxic in a protein translation assay as compared to spores that were washed with an organic solution, however a correlation between carbohydrate removal during the washing process and the removal of mycotoxins from the spore surface was not observed. These data demonstrated that mycotoxins are not likely to be found exclusively in the carbohydrate matrix of the spores. Therefore, mycotoxin removal from the spore surface can occur without significant loss of polysaccharide. We also showed that toxic substances may be removed from the spore surface with an aqueous solution. These results suggest that satratoxins are soluble in aqueous solutions without being bound to water-soluble moieties, such as the carbohydrate slime matrix.

  17. [Variation in the viability and the metabolism of internally reserved nutrients of Beauveria bassiana conidia during storage at different regimes of water content and temperature].

    PubMed

    Feng, Mingguang; Ying, Shenghua

    2002-04-01

    While the fresh powder (with water content of 58.9 +/- 1.6%) and vacuum-freeze dried (VFD) one (with water content of 7.4 +/- 0.9%) of Beauveria bassiana SGBB8702 conidia were stored at 4 degrees C and 20 degrees C, their viability and contents of internally reserved saccharides and proteins were examined at 5 days intervals. It was found during one-month storage that the water content and temperature interactively affected the levels of conidia viability and the contents of the reserved saccharides and proteins, which in turn correlated considerably well to the viability. Stored at 4 degrees C and 20 degrees C, the VFD conidia lost content of saccharides by 13.4% and 14.1%, content of proteins by 39.2% and 38.2%, and viability by 32.0% and 55.8% when germinating in water only and 6.7% and 10.4% when germinating in 2% glucose solution plus 0.5% peptone, respectively. In contrast, the four estimates of the fresh conidia stored at 4 degrees C and 20 degrees C decreased by 42.4% and 43.2%, 66.3% and 65.4%, 96.4% and 99.4%, and 9.9% and 98.4%, respectively. Thus, water content of B. bassiana conidia affected variation ranges of the viability and the internally reserved nutrients whereas storage temperature influenced their variation rates. However, depletion of the internally reserved nutrients did not necessarily inactivate the conidia. Instead, such conidia may germinate at relatively high level during a period of time as long as being supplied with sufficient nutrients. In another experiment, the powder of VFD conidia with water content being reduced to 4.0 +/- 0.9% were half-monthly examined for viability during one-year storage. The viability only decreased slightly from 99.0% to 90.2% at 4 degrees C by the end of one year. Stored at 20 degrees C, the viability also declined slowly during the first 165 days but dramatically decreased afterwards with little being detected on day 240. A modeling analysis showed that the time for the VFD conidia to lose 50% viability

  18. Development of a TaqMan real-time PCR assay for quantification of airborne conidia of Botrytis squamosa and management of botrytis leaf blight of onion.

    PubMed

    Carisse, O; Tremblay, D M; Lévesque, C A; Gindro, K; Ward, P; Houde, A

    2009-11-01

    The use of a DNA-based method for quantifying airborne inoculum of Botrytis squamosa, a damaging pathogen of onion, was investigated. A method for purifying DNA from conidia collected using rotating-arm samplers and quantifying it using a TaqMan real-time quantitative polymerase chain reaction (qPCR) assay is described. The sensitivity of the qPCR assay was high, with a detection limit of 2 conidia/rod. A linear relationship between numbers of conidia counted with a compound microscope and those determined with the qPCR assay was obtained. Receiver operating characteristic curve analysis was used to evaluate the reliability of the two methods of conidia quantification (microscope examination and qPCR assay) to predict the risk of disease being below or above a damage threshold (D(th)). In total, 142 field samples from commercial onion fields were analyzed. At damage thresholds of 5 or 10 lesions/leaf, conidia quantification with the qPCR assay was more reliable at predicting disease risk than conidia quantification based on microscope counts. The proportion of decisions where the disease was present and predicted was higher for the qPCR assay than for the microscope counts, with values of 0.95 and 0.89 compared with 0.79 and 0.81 for D(th) of 5 and 10 lesions/leaf, respectively. The proportion of decisions where the disease was present but not predicted was lower for the qPCR assay than for microscope counts, with values of 0.05 and 0.11 compared with 0.20 and 0.19 for D(th) of 5 and 10 lesions/leaf, respectively. The results demonstrated that this new qPCR assay was reliable for quantifying B. squamosa airborne inoculum in commercial onion fields and that molecular conidia quantification could be used as a component of a risk management system for Botrytis leaf blight.

  19. Field trials using the fungal pathogen, Metarhizium anisopliae (Deuteromycetes: Hyphomycetes) to control the ectoparasitic mite, Varroa destructor (Acari: Varroidae) in honey bee, Apis mellifera (Hymenoptera: Apidae) colonies.

    PubMed

    Kanga, Lambert Houssou Ble; Jones, Walker A; James, Rosalind R

    2003-08-01

    The potential for Metarhizium anisopliae (Metschinkoff) to control the parasitic mite, Varroa destructor (Anderson and Trueman) in honey bee colonies was evaluated in field trials against the miticide, tau-fluvalinate (Apistan). Peak mortality of V. destructor occurred 3-4 d after the conidia were applied; however, the mites were still infected 42 d posttreatments. Two application methods were tested: dusts and strips coated with the fungal conidia, and both methods resulted in successful control of mite populations. The fungal treatments were as effective as the Apistan, at the end of the 42-d period of the experiment. The data suggested that optimum mite control could be achieved when no brood is being produced, or when brood production is low, such as in the early spring or late fall. M. anisopliae was harmless to the honey bees (adult bees, or brood) and colony development was not affected. Mite mortality was highly correlated with mycosis in dead mites collected from sticky traps, indicating that the fungus was infecting and killing the mites. Because workers and drones drift between hives, the adult bees were able to spread the fungus between honey bee colonies in the apiary, a situation that could be beneficial to beekeepers.

  20. Photodynamic inactivation of conidia of the fungus Colletotrichum abscissum on Citrus sinensis plants with methylene blue under solar radiation.

    PubMed

    Gonzales, Júlia C; Brancini, Guilherme T P; Rodrigues, Gabriela B; Silva-Junior, Geraldo José; Bachmann, Luciano; Wainwright, Mark; Braga, Gilberto Ú L

    2017-09-14

    Antimicrobial photodynamic treatment (APDT) is a promising light based approach to control diseases caused by plant-pathogenic fungi. In the present study, we evaluated the effects of APDT with the phenothiazinium photosensitizer methylene blue (MB) under solar radiation on the germination and viability of conidia of the pathogenic fungus Colletotricum abscissum (former Colletotrichum acutatum sensu lato). Experiments were performed both on petals and leaves of sweet orange (Citrus sinensis) in different seasons and weather conditions. Conidial suspensions were deposited on the leaves and petals surface, treated with the PS (25 or 50μM) and exposed to solar radiation for only 30min. The effects of APDT on conidia were evaluated by counting the colony forming units recovered from leaves and petals and by direct evaluating conidial germination on the surface of these plant organs after the treatment. To better understand the mechanistic of conidial photodynamic inactivation, the effect of APDT on the permeability of the conidial plasma membrane was assessed using the fluorescent probe propidium iodide (PI) together with flow cytometry and fluorescence microscopy. APDT with MB and solar exposure killed C. abscissum conidia and prevented their germination on both leaves and petals of citrus. Reduction of conidial viability was up to three orders of magnitude and a complete photodynamic inactivation was achieved in some of the treatments. APDT damaged the conidial plasma membrane and increased its permeability to PI. No damage to sweet orange flowers or leaves was observed after APDT. The demonstration of the efficacy of APDT on the plant host represents a further step towards the use of the method for control phytopathogens in the field. Copyright © 2017 Elsevier B.V. All rights reserved.

  1. Hypocrea rufa/Trichoderma viride: a reassessment, and description of five closely related species with and without warted conidia

    PubMed Central

    Jaklitsch, Walter M.; Samuels, Gary J.; Dodd, Sarah L.; Lu, Bing-Sheng; Druzhinina, Irina S.

    2006-01-01

    The type species of the genus Hypocrea (Hypocreaceae, Hypocreales, Ascomycota, Fungi), H. rufa, is re-defined and epitypified using a combination of phenotype (morphology of teleomorphs and anamorphs, and characteristics in culture) and phylogenetic analyses of the translation-elongation factor 1α gene. Its anamorph, T. viride, the type species of Trichoderma, is re-described and epitypified. Eidamia viridescens is combined as Trichoderma viridescens and is recognised as one of the most morphologically and phylogenetically similar relatives of T. viride. Its teleomorph is newly described as Hypocrea viridescens. Contrary to frequent citations of H. rufa and T. viride in the literature, this species is relatively rare. Although both T. viride and T. viridescens have a wide geographic distribution, their greatest genetic diversity appears to be in Europe and North America. Hypocrea vinosa is characterised and its anamorph, T. vinosum sp. nov., is described. Conidia of T. vinosum are subglobose and warted. The new species T. gamsii is proposed. It shares eidamia-like morphology of conidiophores with T. viridescens, but it has smooth, ellipsoidal conidia that have the longest L/W ratio that we have seen in Trichoderma. Trichoderma scalesiae, an endophyte of trunks of Scalesia pedunculata in the Galapagos Islands, is described as new. It only produces conidia on a low-nutrient agar to which filter paper has been added. Additional phylogenetically distinct clades are recognised and provisionally delimited from the species here described. Trichoderma neokoningii, a T. koningii-like species, is described from a collection made in Peru on a fruit of Theobroma cacao infected with Moniliophthora roreri. PMID:18490991

  2. Dark Period Following UV-C Treatment Enhances Killing of Botrytis cinerea Conidia and Controls Gray Mold of Strawberries.

    PubMed

    Janisiewicz, Wojciech J; Takeda, Fumiomi; Glenn, D Michael; Camp, Mary J; Jurick, Wayne M

    2016-04-01

    Strawberries are available throughout the year either from production in the field or from high and low tunnel culture. Diversity of production conditions results in new challenges in controlling diseases before and after harvest. Fungicides have traditionally been used to control these diseases; however, their limitations necessitate a search for new approaches. We found that UV-C irradiation of Botrytis cinerea, a major pathogen of strawberry, can effectively kill this fungus if a dark period follows the treatment. The inclusion of a 4-h dark period resulted in almost complete kill of B. cinerea conidia on agar media at a dose of 12.36 J/m2. The UV-C dose did not cause a reduction in photosynthesis in strawberry leaves or discoloration of sepals, even after exposing plants repeatedly (twice a week) for 7 weeks. Although irradiation of dry conidia of B. cinerea with this dose resulted in some survival, the conidia were not infective and not able to cause decay even when inoculated onto a highly susceptible mature apple fruit. Irradiation of strawberry pollen at 12.36 J/m2 did not affect pollen germination, tube growth and length in vitro, or germination and tube growth in the style of hand-pollinated emasculated strawberry flowers. No negative effect of the UV-C treatment was observed on fruit yield and quality in high tunnel culture. In the fruit and flower petal inoculation tests, the UV-C treatment was highly effective in reducing fruit decay and petal infection. This UV-C treatment with an exposure time of 60 s may be useful in controlling gray mold in tunnel production of strawberries and may also have the potential for use in intensive field and indoor production of other fruits and vegetables providing that a 4-h dark period follows the irradiation.

  3. Binding of live conidia of Aspergillus fumigatus activates in vitro-generated human Langerhans cells via a lectin of galactomannan specificity

    PubMed Central

    PERSAT, F; NOIREY, N; DIANA, J; GARIAZZO, M-J; SCHMITT, D; PICOT, S; VINCENT, C

    2003-01-01

    Aspergillus fumigatus is the most common aetiological fungus responsible for human pulmonary aspergilloses. This study investigated the primary contact between Langerhans cells (LC), corresponding to dendritic cells present in pulmonary mucosa and live conidia of A. fumigatus. LC play a key role in antigen presentation for initiation of the primary T cell response. In vitro-generated LC (iLC) were differentiated from cultured human cord blood CD34+ cells and incubated at 4°C or 37°C with fluorescein-isothiocyanate (FITC)-stained conidia or control latex beads. In vitro, conidia were shown by microscopy and cytometry to adhere to iLC in a dose- and time-dependent manner. This adhesion was not limited to iLC because interstitial dendritic and other cells also fluoresced in the presence of conidia-FITC. A lectin other than mannose receptor-type lectin was demonstrated to be responsible of conidial binding. Inhibition of binding was observed with heterologous galactomannan and EDTA, indicating a C-lectin-like receptor with galactomannan structure specificity. After binding only a few conidia were internalized in acidic vesicles, as indicated by the cessation of conidial fluorescence. Conidial binding was followed by activation and maturation of iLC, suggesting that LC present in the lung may play a role in cellular host defence against aspergilloses. PMID:12930363

  4. Impact of urban air pollution on the allergenicity of Aspergillus fumigatus conidia: Outdoor exposure study supported by laboratory experiments.

    PubMed

    Lang-Yona, Naama; Shuster-Meiseles, Timor; Mazar, Yinon; Yarden, Oded; Rudich, Yinon

    2016-01-15

    Understanding the chemical interactions of common allergens in urban environments may help to decipher the general increase in susceptibility to allergies observed in recent decades. In this study, asexual conidia of the allergenic mold Aspergillus fumigatus were exposed to air pollution under natural (ambient) and controlled (laboratory) conditions. The allergenic activity was measured using two immunoassays and supported by a protein mass spectrometry analysis. The allergenicity of the conidia was found to increase by 2-5 fold compared to the control for short exposure times of up to 12h (accumulated exposure of about 50 ppb NO2 and 750 ppb O3), possibly due to nitration. At higher exposure times, the allergenicity increase lessened due to protein deamidation. These results indicate that during the first 12h of exposure, the allergenic potency of the fungal allergen A. fumigatus in polluted urban environments is expected to increase. Additional work is needed in order to determine if this behavior occurs for other allergens. Copyright © 2015 Elsevier B.V. All rights reserved.

  5. Verruculogen associated with Aspergillus fumigatus hyphae and conidia modifies the electrophysiological properties of human nasal epithelial cells.

    PubMed

    Khoufache, Khaled; Puel, Olivier; Loiseau, Nicolas; Delaforge, Marcel; Rivollet, Danièle; Coste, André; Cordonnier, Catherine; Escudier, Estelle; Botterel, Françoise; Bretagne, Stéphane

    2007-01-23

    The role of Aspergillus fumigatus mycotoxins in the colonization of the respiratory tract by conidia has not been studied extensively, even though patients at risk from invasive aspergillosis frequently exhibit respiratory epithelium damage. In a previous study, we found that filtrates of A. fumigatus cultures can specifically alter the electrophysiological properties of human nasal epithelial cells (HNEC) compared to those of non pathogenic moulds. We fractionated the organic phase of filtrate from 3-day old A. fumigatus cultures using high-performance liquid chromatography. The different fractions were tested for their ability to modify the electrophysiological properties of HNEC in an in vitro primary culture model. The fraction collected between 20 and 30 min mimicked the effects of the whole filtrate, i.e. decrease of transepithelial resistance and increase of potential differences, and contained secondary metabolites such as helvolic acid, fumagillin, and verruculogen. Only verruculogen (10(-8) M) had effects similar to the whole filtrate. We verified that verruculogen was produced by a collection of 67 human, animal, plant and environmental A. fumigatus isolates. Using MS-MS analysis, we found that verruculogen was associated with both mycelium and conidia extracts. Verruculogen is a secondary metabolite that modifies the electrophysiological properties of HNEC. The role of these modifications in the colonization and invasion of the respiratory epithelium by A. fumigatus on first contact with the epithelium remains to be determined.

  6. Verruculogen associated with Aspergillus fumigatus hyphae and conidia modifies the electrophysiological properties of human nasal epithelial cells

    PubMed Central

    Khoufache, Khaled; Puel, Olivier; Loiseau, Nicolas; Delaforge, Marcel; Rivollet, Danièle; Coste, André; Cordonnier, Catherine; Escudier, Estelle; Botterel, Françoise; Bretagne, Stéphane

    2007-01-01

    Background The role of Aspergillus fumigatus mycotoxins in the colonization of the respiratory tract by conidia has not been studied extensively, even though patients at risk from invasive aspergillosis frequently exhibit respiratory epithelium damage. In a previous study, we found that filtrates of A. fumigatus cultures can specifically alter the electrophysiological properties of human nasal epithelial cells (HNEC) compared to those of non pathogenic moulds. Results We fractionated the organic phase of filtrate from 3-day old A. fumigatus cultures using high-performance liquid chromatography. The different fractions were tested for their ability to modify the electrophysiological properties of HNEC in an in vitro primary culture model. The fraction collected between 20 and 30 min mimicked the effects of the whole filtrate, i.e. decrease of transepithelial resistance and increase of potential differences, and contained secondary metabolites such as helvolic acid, fumagillin, and verruculogen. Only verruculogen (10-8 M) had effects similar to the whole filtrate. We verified that verruculogen was produced by a collection of 67 human, animal, plant and environmental A. fumigatus isolates. Using MS-MS analysis, we found that verruculogen was associated with both mycelium and conidia extracts. Conclusion Verruculogen is a secondary metabolite that modifies the electrophysiological properties of HNEC. The role of these modifications in the colonization and invasion of the respiratory epithelium by A. fumigatus on first contact with the epithelium remains to be determined. PMID:17244350

  7. Expression and arrangement of extracellular matrix proteins in the lungs of mice infected with Paracoccidioides brasiliensis conidia

    PubMed Central

    González, Angel; Lenzi, Henrique Leonel; Motta, Ester Maria; Caputo, Luzia; Restrepo, Angela; Cano, Luz Elena

    2008-01-01

    Extracellular matrix (ECM) proteins are important modulators of migration, differentiation and proliferation for the various cell types present in the lungs; they influence the immune response as well as participate in the adherence of several fungi including Paracoccidioides brasiliensis. The expression, deposition and arrangement of ECM proteins such as laminin, fibronectin, fibrinogen, collagen and proteoglycans in the lungs of mice infected with P. brasiliensis conidia has been evaluated in this study, together with the elastic fibre system. Lungs of BALB/c mice infected with P. brasiliensis conidia were analysed for the different ECM proteins by histological and immunohistochemical procedures at different times of infection. In addition, laser scanning confocal microscopy and scanning electron microscopy were used. During the early periods, the lungs of infected animals showed an inflammatory infiltrate composed mainly of polymorphonuclear neutrophils (PMNs) and macrophages, while during the later periods, mice presented a chronic inflammatory response with granuloma formation. Re-arrangement and increased expression of all ECM proteins tested were observed throughout all studied periods, especially during the occurrence of inflammatory infiltration and formation of the granuloma. The elastic fibre system showed an elastolysis process in all experiments. In conclusion, this study provides new details of pulmonary ECM distribution during the course of paracoccidioidomycosis. PMID:18336528

  8. Effects of resveratrol on the ultrastructure of Botrytis cinerea conidia and biological significance in plant/pathogen interactions.

    PubMed

    Adrian, Marielle; Jeandet, Philippe

    2012-12-01

    Many roles have been ascribed to stilbenes, namely as antimicrobial, deterrent or repellent compounds in plants, protecting them from attacks by fungi, bacteria, nematodes or herbivores, acting both as constitutive and active defense (phytoalexin) compounds. More recently, stilbenes (especially resveratrol and its derivatives) were acclaimed for their wondrous effects and wide range of purported healing and preventive powers as cardioprotective, antitumor, neuroprotective and antioxidant agents. Although there is a huge number of works concerning the role of resveratrol in human health, reports on the antifungal activity of this compound are still scarce. This study was thus conducted in order to investigate the toxicity of resveratrol at an ultra- structural level to dormant conidia of Botrytis cinerea, the causal microorganism for gray mold. In grapevine particularly, this disease can affect all the green organs but is particularly damaging for ripening berries. Observations using transmission electron microscopy showed the occurrence of damages on conidia treated with sub-lethal doses, that is, 60μg/mL (2.6×10(-4)M) of resveratrol, a concentration usually reached in grapevine leaves and grape berries challenged by this pathogen. These results provide further data about the overall mode of action of this phytoalexin and its role in the B. cinerea/grapevine interaction.

  9. Conidia immobilization of T-DNA inserted Trichoderma atroviride mutant AMT-28 with dichlorvos degradation ability and exploration of biodegradation mechanism.

    PubMed

    Sun, Wenliang; Chen, Yunpeng; Liu, Lixing; Tang, Jun; Chen, Jie; Liu, Peng

    2010-12-01

    An immobilizing conidia approach was used to study the degradation ability of dichlorvos in Trichoderma atroviride T-DNA insertional mutant AMT-28. Beads with 10(7) immobilized conidia per 100 mL of Na-alginate solution exhibited the highest degradation abilities. The immobilized conidia showed enhanced degradation abilities compared with immobilized or freely suspended mycelia. The immobilized cells kept good storage capacity and reusability. Dichlorvos was confirmed to be completely removed by mycelia of AMT-28 within 7 days using HPLC analysis. The dichlorvos degradation rates in auxotrophic Burk media varied and were significantly affected by nitrogen sources. There was no detectable biosorption and the removal of dichlorvos in AMT-28 was primarily attributed to a kind of Biomineralization process.

  10. Effect of exposure method to Beauveria bassiana and conidia concentration on mortality, mycosis, and sporulation in cadavers of Chilo partellus (Lepidoptera: Pyralidae).

    PubMed

    Tefera, Tadele; Pringle, K L

    2003-10-01

    The effects of exposure methods, conidial concentrations, and temperature on mortality, mycosis and sporulation in second instar Chilo partellus cadavers infected by Beauveria bassiana was investigated in laboratory studies. Larvae directly sprayed with conidia, exposed to conidia-treated leaves, and dipped into conidial suspension resulted in high mortality (98-100%). The longest LT50 (3.5 days) and days to mortality (2.6 days) were observed in the treated-leaves exposure method. The shortest LT50 (1 day) and days to mortality (1 day) were recorded for the dipping method. With increasing conidial concentrations, there were decreasing LT50 and days to mortality. Larvae exposed to treated leaves and larvae directly sprayed with conidial suspensions produced high mycoses in cadavers. Exposure of larvae to treated-leaves resulted in high sporulation. At lower concentrations of conidia, both mycosis and sporulation in cadavers were high. The optimum temperature for mycosis was 20 and 15 degrees C for sporulation.

  11. Role of iron in the nitric oxide-mediated fungicidal mechanism of IFN-gamma-activated murine macrophages against Paracoccidioides brasiliensis conidia.

    PubMed

    Gonzalez, Angel; Restrepo, Angela; Cano, Luz E

    2007-01-01

    Iron is an essential growth element of virtually all microorganisms and its restriction is one of the mechanisms used by macrophages to control microbial multiplication. Paracoccidioides brasiliensis, the agent of paracoccidioidomycosis, an important systemic mycosis in Latin America, is inhibited in its conidia-to-yeast conversion in the absence of iron. We studied the participation of iron in the nitric oxide (NO)-mediated fungicidal mechanism against conidia. Peritoneal murine macrophages activated with 50 U/mL of IFN-gamma or treated with 35 microM Deferoxamine (DEX) and infected with P. brasiliensis conidia, were co-cultured and incubated for 96 h in the presence of different concentrations of holotransferrin (HOLO) and FeS04. The supernatants were withdrawn in order to assess NO2 production by the Griess method. The monolayers were fixed, stained and observed microscopically. The percentage of the conidia-to-yeast transition was estimated by counting 200 intracellular propagules. IFN-gamma-activated or DEX-treated Mthetas presented marked inhibition of the conidia-to-yeast conversion (19 and 56%, respectively) in comparison with non-activated or untreated Mthetas (80%). IFN-gamma-activated macrophages produced high NO levels in comparison with the controls. Additionally, when the activated or treated-macrophages were supplemented with iron donors (HOLO or FeSO4), the inhibitory action was reversed, although NO production remained intact. These results suggest that the NO-mediated fungicidal mechanism exerted by IFN-gamma-activated macrophages against P. brasiliensis conidia, is dependent of an iron interaction.

  12. Copulation Activity, Sperm Production and Conidia Transfer in Aedes aegypti Males Contaminated by Metarhizium anisopliae: A Biological Control Prospect.

    PubMed

    Garza-Hernández, Javier A; Reyes-Villanueva, Filiberto; Russell, Tanya L; Braks, Marieta A H; Garcia-Munguia, Alberto M; Rodríguez-Pérez, Mario A

    2015-01-01

    Dengue is the most prevalent arboviral disease transmitted by Aedes aegypti worldwide, whose chemical control is difficult, expensive, and of inconsistent efficacy. Releases of Metarhizium anisopliae--exposed Ae. aegypti males to disseminate conidia among female mosquitoes by mating represents a promising biological control approach against this important vector. A better understanding of fungus virulence and impact on reproductive parameters of Ae. aegypti, is need before testing auto-dissemination strategies. Mortality, mating competitiveness, sperm production, and the capacity to auto-disseminate the fungus to females up to the 5 th copulation, were compared between Aedes aegypti males exposed to 5.96 x 10(7) conidia per cm2 of M. anisopliae and uninfected males. Half (50%) of fungus-exposed males (FEMs) died within the first 4 days post-exposure (PE). FEMs required 34% more time to successively copulate with 5 females (165 ± 3 minutes) than uninfected males (109 ± 3 minutes). Additionally, fungus infection reduced the sperm production by 87% at 5 days PE. Some beneficial impacts were observed, FEMs were able to successfully compete with uninfected males in cages, inseminating an equivalent number of females (about 25%). Under semi-field conditions, the ability of FEMs to search for and inseminate females was also equivalent to uninfected males (both inseminating about 40% females); but for the remaining females that were not inseminated, evidence of tarsal contact (transfer of fluorescent dust) was significantly greater in FEMs compared to controls. The estimated conidia load of a female exposed on the 5th copulation was 5,200 mL(-1) which was sufficient to cause mortality. Our study is the first to demonstrate auto-dissemination of M. anisopliae through transfer of fungus from males to female Ae. aegypti during mating under semi-field conditions. Our results suggest that auto-dissemination studies using releases of FEMs inside households could successfully

  13. Copulation Activity, Sperm Production and Conidia Transfer in Aedes aegypti Males Contaminated by Metarhizium anisopliae: A Biological Control Prospect

    PubMed Central

    Russell, Tanya L.; Braks, Marieta A. H.

    2015-01-01

    Background Dengue is the most prevalent arboviral disease transmitted by Aedes aegypti worldwide, whose chemical control is difficult, expensive, and of inconsistent efficacy. Releases of Metarhizium anisopliae—exposed Ae. aegypti males to disseminate conidia among female mosquitoes by mating represents a promising biological control approach against this important vector. A better understanding of fungus virulence and impact on reproductive parameters of Ae. aegypti, is need before testing auto-dissemination strategies. Methodology/Principal Findings Mortality, mating competitiveness, sperm production, and the capacity to auto-disseminate the fungus to females up to the 5thcopulation, were compared between Aedes aegypti males exposed to 5.96 x 107 conidia per cm2 of M. anisopliae and uninfected males. Half (50%) of fungus-exposed males (FEMs) died within the first 4 days post-exposure (PE). FEMs required 34% more time to successively copulate with 5 females (165 ± 3 minutes) than uninfected males (109 ± 3 minutes). Additionally, fungus infection reduced the sperm production by 87% at 5 days PE. Some beneficial impacts were observed, FEMs were able to successfully compete with uninfected males in cages, inseminating an equivalent number of females (about 25%). Under semi-field conditions, the ability of FEMs to search for and inseminate females was also equivalent to uninfected males (both inseminating about 40% females); but for the remaining females that were not inseminated, evidence of tarsal contact (transfer of fluorescent dust) was significantly greater in FEMs compared to controls. The estimated conidia load of a female exposed on the 5th copulation was 5,200 mL-1 which was sufficient to cause mortality. Conclusion/Significance Our study is the first to demonstrate auto-dissemination of M. anisopliae through transfer of fungus from males to female Ae. aegypti during mating under semi-field conditions. Our results suggest that auto-dissemination studies

  14. Yield and cold storage of Trichoderma conidia is influenced by substrate pH and storage temperature.

    PubMed

    Steyaert, Johanna M; Chomic, Anastasia; Nieto-Jacobo, Maria; Mendoza-Mendoza, Artemio; Hay, Amanda J; Braithwaite, Mark; Stewart, Alison

    2017-05-01

    In this study we examined the influence of the ambient pH during morphogenesis on conidial yield of Trichoderma sp. "atroviride B" LU132 and T. hamatum LU593 and storage at low temperatures. The ambient pH of the growth media had a dramatic influence on the level of Trichoderma conidiation and this was dependent on the strain and growth media. On malt-extract agar, LU593 yield decreased with increasing pH (3-6), whereas yield increased with increasing pH for LU132. During solid substrate production the reverse was true for LU132 whereby yield decreased with increasing pH. The germination potential of the conidia decreased significantly over time in cold storage and the rate of decline was a factor of the strain, pH during morphogenesis, growth media, and storage temperature. © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  15. A soluble fucose-specific lectin from Aspergillus fumigatus conidia--structure, specificity and possible role in fungal pathogenicity.

    PubMed

    Houser, Josef; Komarek, Jan; Kostlanova, Nikola; Cioci, Gianluca; Varrot, Annabelle; Kerr, Sheena C; Lahmann, Martina; Balloy, Viviane; Fahy, John V; Chignard, Michel; Imberty, Anne; Wimmerova, Michaela

    2013-01-01

    Aspergillus fumigatus is an important allergen and opportunistic pathogen. Similarly to many other pathogens, it is able to produce lectins that may be involved in the host-pathogen interaction. We focused on the lectin AFL, which was prepared in recombinant form and characterized. Its binding properties were studied using hemagglutination and glycan array analysis. We determined the specificity of the lectin towards l-fucose and fucosylated oligosaccharides, including α1-6 linked core-fucose, which is an important marker for cancerogenesis. Other biologically relevant saccharides such as sialic acid, d-mannose or d-galactose were not bound. Blood group epitopes of the ABH and Lewis systems were recognized, Le(Y) being the preferred ligand among others. To provide a correlation between the observed functional characteristics and structural basis, AFL was crystallized in a complex with methyl-α,L-selenofucoside and its structure was solved using the SAD method. Six binding sites, each with different compositions, were identified per monomer and significant differences from the homologous AAL lectin were found. Structure-derived peptides were utilized to prepare anti-AFL polyclonal antibodies, which suggested the presence of AFL on the Aspergillus' conidia, confirming its expression in vivo. Stimulation of human bronchial cells by AFL led to IL-8 production in a dose-dependent manner. AFL thus probably contributes to the inflammatory response observed upon the exposure of a patient to A. fumigatus. The combination of affinity to human epithelial epitopes, production by conidia and pro-inflammatory activity is remarkable and shows that AFL might be an important virulence factor involved in an early stage of A. fumigatus infection.

  16. Interactions of Chemical Additives, pH, and Temperature on Conidia Germination and Virulence of Colletotrichum truncatum, a Bioherbicide of Sesbania exaltata

    USDA-ARS?s Scientific Manuscript database

    We studied several factors that influence conidial germination and pathogenicity of the fungus Colletotrichum truncatum, which is a potential bioherbicide for hemp sesbania. The conidia germinated on 2% water agar over the pH range of 3.5 to 9.0, and the optimal pH for germination depended upon the ...

  17. Application of hydrophilic-lypophilic balance (HLB) number to optimize a compatible non-ionic surfactant for dried aerial conidia of Beauveria bassiana

    USDA-ARS?s Scientific Manuscript database

    The hydrophilic-lipophilic balance (HLB) number system was used to optimize a compatible non-ionic surfactant, TDA(polyoxyethylene tridecyl ether) in formulations for two Beauveria bassiana strains, NI8 and GHA. The optimal HLB number for TDA was determined on the basis of wetting times for conidia...

  18. A proposed role for the cuticular fatty amides of Liposcelis bostrychophila (Psocoptera: Liposcelidae) in preventing adhesion of entomopathogenic fungi with dry-conidia.

    PubMed

    Lord, Jeffrey C; Howard, Ralph W

    2004-08-01

    Maximum challenge exposure of Liposcelis bostrychophila to Beauveria bassiana, Paecilomyces fumosoroseus, Aspergillus parasiticus or Metarhizium anisopliae resulted in no more than 16% mortality. We investigated several of L. bostrychophila's cuticular lipids for possible contributions to its tolerance for entomopathogenic fungi. Saturated C14 and C16 fatty acids did not reduce the germination rates of B. bassiana or M. anisopliae conidia. Saturated C6 to C12 fatty acids that have not been identified in L. bostrychophila cuticular extracts significantly reduced germination, but the reduction was mitigated by the presence of stearamide. Cis-6-hexadecenal did not affect germination rates. Mycelial growth of either fungal species did not occur in the presence of caprylic acid, was reduced by the presence of lauric acid, and was not significantly affected by palmitic acid. Liposcelis bostrychophila is the only insect for which fatty acid amides have been identified as cuticular components. Stearamide, its major fatty amide, did not reduce germination of B. bassiana or M. anisopliae conidia or growth of their mycelia. Adhesion of conidia to stearamide preparations did not differ significantly from adhesion to the cuticle of L. bostrychophila. Pretreatment of a beetle known to be fungus-susceptible, larval Oryzaephilus surinamensis, with stearamide significantly decreased adhesion of B. bassiana or M. anisopliae conidia to their cuticles. This evidence indicates that cuticular fatty amides may contribute to L. bostrychophila's tolerance for entomopathogenic fungi by decreasing hydrophobicity and static charge, thereby reducing conidial adhesion.

  19. Transcription profile of Trichophyton rubrum conidia grown on keratin reveals the induction of an adhesin-like protein gene with a tandem repeat pattern.

    PubMed

    Bitencourt, Tamires Aparecida; Macedo, Claudia; Franco, Matheus Eloy; Assis, Amanda Freire; Komoto, Tatiana Takahasi; Stehling, Eliana Guedes; Beleboni, Rene Oliveira; Malavazi, Iran; Marins, Mozart; Fachin, Ana Lúcia

    2016-03-18

    Trichophyton rubrum is a cosmopolitan filamentous fungus that can infect human keratinized tissue (skin, nails and, rarely, hair) and is the major agent of all chronic and recurrent dermatophytoses. The dermatophyte infection process is initiated through the release of arthroconidial adhesin, which binds to the host stratum corneum. The conidia then germinate, and fungal hyphae invade keratinized skin structures through the secretion of proteases. Although arthroconidia play a central role in pathogenesis, little is known about the dormancy and germination of T. rubrum conidia and the initiation of infection. The objective of this study was to evaluate the transcriptional gene expression profile of T. rubrum conidia during growth on keratin- or elastin-containing medium, mimicking superficial and deep dermatophytosis, respectively. A transcriptional profiling analysis was conducted using a custom oligonucleotide-based microarray by comparing T. rubrum conidia grown on elastin and keratin substrates. This comparison shows differences according to protein source used, but consisted of a very small set of genes, which could be attributed to the quiescent status of conidia. The modulated genes were related to the dormancy, survival and germination of conidia, including genes involved in the respiratory chain, signal transduction and lipid metabolism. However, an induction of a great number of proteases occurred when T. rubrum was grown in the presence of keratin such as the subtilisin family of proteases (Sub 1 and Sub 3) and leucine aminopeptidase (Lap 1 and Lap 2). Interestingly, keratin also promoted the up-regulation of a gene encoding an adhesin-like protein with a tandem repeat sequence. In silico analysis showed that the protein contains a domain related to adhesin that may play a role in host-pathogen interactions. The expression of this adhesin-like gene was also induced during the co-culture of T. rubrum with a human keratinocyte cell line, confirming its

  20. Aspergillus flavus Conidia-derived Carbon/Sulfur Composite as a Cathode Material for High Performance Lithium–Sulfur Battery

    NASA Astrophysics Data System (ADS)

    Xu, Maowen; Jia, Min; Mao, Cuiping; Liu, Sangui; Bao, Shujuan; Jiang, Jian; Liu, Yang; Lu, Zhisong

    2016-01-01

    A novel approach was developed to prepare porous carbon materials with an extremely high surface area of 2459.6 m2g‑1 by using Aspergillus flavus conidia as precursors. The porous carbon serves as a superior cathode material to anchor sulfur due to its uniform and tortuous morphology, enabling high capacity and good cycle lifetime in lithium sulfur-batteries. Under a current rate of 0.2 C, the carbon-sulfur composites with 56.7 wt% sulfur loading deliver an initial capacity of 1625 mAh g‑1, which is almost equal to the theoretical capacity of sulfur. The good performance may be ascribed to excellent electronic networks constructed by the high-surface-area carbon species. Moreover, the semi-closed architecture of derived carbons can effectively retard the polysulfides dissolution during charge/discharge, resulting in a capacity of 940 mAh g‑1 after 120 charge/discharge cycles.

  1. Use of a granular bioplastic formulation for carrying conidia of a non-aflatoxigenic strain of Aspergillus flavus.

    PubMed

    Accinelli, Cesare; Saccà, M Ludovica; Abbas, Hamed K; Zablotowicz, Robert M; Wilkinson, Jeffery R

    2009-09-01

    Previous research demonstrated that aflatoxin contamination in corn is reduced by field application of wheat grains pre-inoculated with the non-aflatoxigenic Aspergillus flavus strain NRRL 30797. To facilitate field applications of this biocontrol isolate, a series of laboratory studies were conducted on the reliability and efficiency of replacing wheat grains with the novel bioplastic formulation Mater-Bi to serve as a carrier matrix to formulate this fungus. Mater-Bi granules were inoculated with a conidial suspension of NRRL 30797 to achieve a final cell density of approximately log 7 conidia/granule. Incubation of 20-g soil samples receiving a single Mater-Bi granule for 60-days resulted in log 4.2-5.3 propagules of A. flavus/g soil in microbiologically active and sterilized soil, respectively. Increasing the number of granules had no effect on the degree of soil colonization by the biocontrol fungus. In addition to the maintenance of rapid vegetative growth and colonization of soil samples, the bioplastic formulation was highly stable, indicating that Mater-Bi is a suitable substitute for biocontrol applications of A. flavus NRRL 30797.

  2. Aspergillus flavus Conidia-derived Carbon/Sulfur Composite as a Cathode Material for High Performance Lithium–Sulfur Battery

    PubMed Central

    Xu, Maowen; Jia, Min; Mao, Cuiping; Liu, Sangui; Bao, Shujuan; Jiang, Jian; Liu, Yang; Lu, Zhisong

    2016-01-01

    A novel approach was developed to prepare porous carbon materials with an extremely high surface area of 2459.6 m2g−1 by using Aspergillus flavus conidia as precursors. The porous carbon serves as a superior cathode material to anchor sulfur due to its uniform and tortuous morphology, enabling high capacity and good cycle lifetime in lithium sulfur-batteries. Under a current rate of 0.2 C, the carbon-sulfur composites with 56.7 wt% sulfur loading deliver an initial capacity of 1625 mAh g−1, which is almost equal to the theoretical capacity of sulfur. The good performance may be ascribed to excellent electronic networks constructed by the high-surface-area carbon species. Moreover, the semi-closed architecture of derived carbons can effectively retard the polysulfides dissolution during charge/discharge, resulting in a capacity of 940 mAh g−1 after 120 charge/discharge cycles. PMID:26732547

  3. Evaluation of Conidia-Dust Formulation of the Entomopathogenic Fungus, Metarhizium anisopliae to Biocontrol the Brown-Banded Cockroach, Supella longipalpa F.

    PubMed Central

    Sharififard, Mona; Mossadegh, Mohammad Saeed; Vazirianzadeh, Babak; Latifi, Seyed Mahmood

    2014-01-01

    Background: The brown-banded cockroach Supella longipalpa (F.) as a mechanical vector of pathogens and source of allergens has recently become widespread in the city of Ahvaz, southwestern Iran. Objectives: This research was done to evaluate the efficacy of a dust-formulation of Metarhizium anisopliae isolate IRAN 437C, as a common entomopathogenous fungus, against S. longipalpa. Materials and Methods: Conidia dust-formulations of M. anisopliae were prepared in proportions of 1%, 5%, 10%, 25%, 50% and 100% with bad wheat flour as the carrier. Cockroaches were exposed to surfaces treated with 1.5 mg/cm2 of the formulations under laboratory and semi-field conditions. Results: Cockroach mortality rates increased and survival times (ST50) decreased with an increased proportion of conidia from 1% to 100% but records taken for mortality and survival time from proportions of 25%, 50% and 100% were not significantly different. The mortality rates reached 100% and 90-100% in adults and nymphs, respectively on the seventh day. The lowest ST50 was related to the proportion of 100% (3 days). Probit analysis indicated LD50 and LD90 values of 1.7 × 106 and 1.7 × 107 conidia/cm2 for adults and these values changed to 4.5 × 106 and 2.9 × 107 for third and fourth instar nymphs at three days post exposure. Proportion of 25% caused mortality rates of 87%, 81% and 73% in adult, adult & nymph and nymph populations, respectively at four days after exposure under room conditions. Conclusions: Conidia dust-formulation of M. anospliae isolate IRAN 437C could present a promising alternative to control the brown-banded cockroach. PMID:25371804

  4. Modelling the effect of temperature, pH, water activity, and organic acids on the germination time of Penicillium camemberti and Penicillium roqueforti conidia.

    PubMed

    Kalai, Safaa; Anzala, Lexane; Bensoussan, Maurice; Dantigny, Philippe

    2017-01-02

    In this study, the influence of environmental factors on the germination time of Penicillium camemberti and Penicillium roqueforti conidia was evaluated. To do so, the effects of i/temperature, pH, water activity, and ii/organic acids were determined using models based on i/cardinal values, and ii/minimum inhibitory concentration (MIC) respectively. Cardinal values for germination of conidia were not observed to be species dependent. Minimum temperatures were estimated to be below the freezing point, with an optimum of 26.9°C, and a maximum of 33.5°C. For both species, minimal and optimal aw values were found to be 0.83 and 0.99, respectively, while for pH these values corresponded to 2.9, and 5.6. MIC values could not be determined for lactic acid because conidia of both species germinated in up to 1M concentrations, the highest concentration tested. At pH5.6, P. camemberti (MIC=0.197M) was more sensitive to propionic acid than P. roqueforti (MIC=0.796M).

  5. Repellent activity of desiccant dusts and conidia of the entomopathogenic fungus Beauveria bassiana when tested against poultry red mites (Dermanyssus gallinae) in laboratory experiments.

    PubMed

    Kilpinen, Ole; Steenberg, Tove

    2016-11-01

    Desiccant dusts and entomopathogenic fungi have previously been found to hold potential against the poultry red mite, which is an important pest in egg production and notoriously difficult to control. Both control agents may cause repellence in other arthropods and potentially also influence control levels adversely when used against the poultry red mite. Five desiccant dust products with good efficacy against the poultry red mite Dermanyssus gallinae caused avoidance behavior in mites when tested in bioassays. The repellent activity was correlated with efficacy, which was found to depend on both dose and relative humidity (RH). However, one desiccant dust was significantly less repellent compared to other dusts with similar levels of efficacy. Further, dry conidia of the fungus Beauveria bassiana were also shown to be repellent to poultry red mites, both when applied on its own and when admixed with a low dose of the desiccant dust Diamol. The pick-up of desiccant dust particles and fungus conidia from treated surfaces by mites did not differ depending on RH, whereas the overall efficacy of the two control agents were significantly higher at 75 than at 85 % RH. In addition, the combined effect of the two substances was synergistic when tested in a bioassay where mites could choose whether to cross a treated surface. This is the first time a member of Acari has been shown to be repelled by desiccant dusts and by conidia of an entomopathogenic fungus.

  6. The physiological effects of multi-walled carbon nanotubes (MWCNTs) on conidia and the development of the entomopathogenic fungus, Metarhizium anisopliae (Metsch.) Sorok.

    PubMed

    Gorczyca, Anna; Kasprowicz, Marek J; Lemek, Tadeusz

    2014-01-01

    The aim of the study was an in vitro evaluation of the effect of MWCNTs on the conidia of two strains of entomopathogenic fungus, Metarhizium anisopliae. The study made use of water suspensions of MWCNTs (concentration ∼ 3 mg·mL(-1)) made from commercial nanotubes and centrifuged. The conidia were placed in contact with nanotubes for 240 h. An assessment of MWCNT influence on conidia was performed after 1, 24, 72 and 240 h and focused on the linear growth of vegetative mycelium derived from these conidia, mycelium sporulation in subcultures and pathogenicity. Using TEM imaging, it was demonstrated that carbon nanotubes are able to damage cell membranes of the examined fungi conidia. However, the absence was noted of a significantly fungistatic effect of both MWCNT suspensions on the examined strains with respect to the physiological features in question. The increase in vegetative mycelium effected by spores after contact with MWCNTs was characterized by a slight modification in relation to the control. There was no strong trend (inhibition - stimulation), in relation to the effect of the tested suspension of carbon nanotubes, on the development of the vegetative mycelium in in vitro culture. Sporulation of the mycelium after completion of the culture only occurred in one case (strain Ma73F and culture of spores after 24-h contact with MWCNTs) significantly more intensely than in the controls. With respect to pathogenicity for test insects compared to the control strain, Ma73F spores grown from the longest contact with nanotubes suspensions performed significantly better. On the basis of the calculated of mycelium index growth rates and the time of death of the test insects (LT50), it was found that the adverse effects of water suspension MWCNTs on the spores of M. anisopliae were applied after a short contact with biological material. This indicates unfavorable physical rather than chemical effects on the tested cell. Over time, nanotube aggregation in water

  7. Survival of Penicillium spp. conidia during deep-frying and baking steps of frozen chicken nuggets processing.

    PubMed

    Wigmann, Évelin Francine; Moreira, Rafael Chelala; Alvarenga, Verônica Ortiz; Sant'Ana, Anderson S; Copetti, Marina Venturini

    2016-05-01

    This study aimed at determining whether Penicillium spp. strains could survive through the heat treatment applied during the processing of frozen chicken nuggets. Firstly, it was found that the conidia of Penicillium were not able to survive the heat shock in phosphate buffer at pH 7.2 in thermal death tubes (TDT) at 80 °C/30 min. Subsequently, each Penicillium strain was inoculated in frozen chicken nuggets, which were subjected to the following treatments: i) only deep frying (frying oil at 195-200 °C), ii) only baking (120-130 °C until the internal temperature reached 70 °C) and iii) deep frying followed by baking (frying oil temperature of 195-200 °C and baking temperature of 120-130 °C, until the internal temperature reached 70 °C). The results indicated that Penicillium polonicum NGT 23/12, Penicillium commune NGT 16/12, Penicillium solitum NGT 30/12 and Penicillium crustosum NGT 51/12 were able to survive after the combined treatment (deep frying followed by baking) when inoculated in chicken nuggets. P. polonicum NGT 23/12 was the most resistant strain to the combined treatment (deep frying and baking), as its population was reduced by 3 log cycles CFU/g, when the internal temperature reached 78 °C after 10 min and 30 s of baking. The present data show that if Penicillium spp. is present in high numbers in raw materials, such as breading flours, it will survive the thermal processing applied during chicken nuggets production.

  8. Effects of Airflow and Changing Humidity on the Aerosolization of Respirable Fungal Fragments and Conidia of Botrytis cinerea

    PubMed Central

    2012-01-01

    The purpose of this study was to investigate the aerosolization of particles (micro- and macroconidia and fragments) from Botrytis cinerea cultures in relation to potential human inhalation in indoor environments. The influence of the following factors on the aerosolization of B. cinerea particles was studied: exposure to airflow, relative humidity (rh), changing rh, and plant or building materials. The aerodynamic diameter (da) and the respirable fraction of the aerosolized particles were determined. Conidia and fragments of B. cinerea were not aerosolized as a response to a decrease in the rh. In contrast, both micro- and macroconidia and fungal fragments were aerosolized when exposed to an airflow of 1.5 m s−1 or 0.5 m s−1. Significantly more particles of microconidial size and fragment size were aerosolized at a low rh (18 to 40% rh) than at a higher rh (60 to 80% rh) when cultures were exposed to airflow. The size of the respirable fraction of the aerosolized particles was dependent on the rh but not on the growth material. At high rh, about 30% of the aerosolized particles were of respirable size, while at low rh, about 70% were of respirable size. During low rh, more fungal (1→3)-β-d-glucan and chitinase were aerosolized than during high rh. In conclusion, exposure to external physical forces such as airflow is necessary for the aerosolization of particles from B. cinerea. The amount and size distribution are highly affected by the rh, and more particles of respirable sizes were aerosolized at low rh than at high rh. PMID:22447608

  9. Structural and Topographic Dynamics of Pulmonary Histopathology and Local Cytokine Profiles in Paracoccidioides brasiliensis Conidia-Infected Mice

    PubMed Central

    Cruz, Oswaldo G.; Restrepo, Angela; Cano, Luz Elena; Lenzi, Henrique Leonel

    2011-01-01

    Background Paracoccidioidomycosis (PCM), an endemic systemic mycosis caused by the fungus Paracoccidioides brasiliensis (Pb), usually results in severe lung damage in patients. Methods and Findings Considering the difficulties to sequentially study the infection in humans, this work was done in mice inoculated intranasally with infective Pb-conidia. Lungs of control and Pb-infected mice were studied after 2-hours, 4, 8, 12 and 16-weeks post-infection (p.i) in order to define histopathologic patterns of pulmonary lesions, multiplex-cytokine profiles and their dynamics during the course of this mycosis. Besides the nodular/granulomatous lesions previously informed, results revealed additional non-formerly described lung abnormalities, such as periarterial sheath inflammation and pseudotumoral masses. The following chronologic stages occurring during the course of the experimental infection were defined: Stage one (2-hours p.i): mild septal infiltration composed by neutrophils and macrophages accompanied by an intense “cytokine burst” represented by significant increases in IL-1α, IL-1β, IL-4, IL-5, IL-6, IL-10, IL12p70, IL-13, IL-17, Eotaxin, G-CSF, MCP1, MIP1α, GM-CSF, IFN-γ, MIP1β and TNFα levels. Stage two (4-weeks p.i): presence of nodules, evidence of incipient periarterial- and intense but disperse parenchymal- inflammation, abnormalities that continued to be accompanied by hyper-secretion of those cytokines and chemokines mentioned in the first stage of infection. Stages three and four (8 and 12-weeks p.i.): fungal proliferation, inflammation and collagenesis reached their highest intensity with particular involvement of the periarterial space. Paradoxically, lung cytokines and chemokines were down-regulated with significant decreases in IL-2,IL-3,IL-5,IL-9,IL-13,IL-15,GM-CSF,IFN-γ,MIP1β and TNFα. Stage five (16-weeks p.i.): inflammation decreased becoming limited to the pseudotumoral masses and was accompanied by a “silent” cytokine response

  10. Liquid culture production of microsclerotia and submerged conidia by Trichoderma harzianum active against damping-off disease caused by Rhizoctonia solani.

    PubMed

    Kobori, Nilce N; Mascarin, Gabriel M; Jackson, Mark A; Schisler, David A

    2015-04-01

    Media and culturing protocols were identified that supported the formation of submerged conidia and microsclerotia (MS) by Trichoderma harzianum Rifai strain T-22 using liquid culture fermentation. Liquid media with a higher carbon concentration (36 g L(-1)) promoted MS formation at all C:N ratios tested. Hyphae aggregated to form MS after 2 d growth and after 7 d MS were fully melanized. This is the first report of MS formation by T. harzianum or any species of Trichoderma. Furthermore, submerged conidia formation was induced by liquid culture media, but yields, desiccation tolerance, and storage stability varied with C:N ratio and carbon rate. Air-dried MS granules (<4% moisture) retained excellent shelf life under cool and unrefrigerated storage conditions with no loss in conidial production. A low-cost complex nitrogen source based on cottonseed flour effectively supported high MS yields. Amending potting mix with dried MS formulations reduced or eliminated damping-off of melon seedlings caused by Rhizoctonia solani. Together, the results provide insights into the liquid culture production, stabilization process, and bioefficacy of the hitherto unreported MS of T. harzianum as a potential biofungicide for use in integrated management programs against soilborne diseases.

  11. Identification of a gene product induced by hard-surface contact of Colletotrichum gloeosporioides conidia as a ubiquitin-conjugating enzyme by yeast complementation.

    PubMed

    Liu, Z M; Kolattukudy, P E

    1998-07-01

    The germinating conidia of many phytopathogenic fungi on hosts must differentiate into an infection structure called the appressorium in order to penetrate their hosts. Chemical signals, such as the host's surface wax or fruit ripening hormone, ethylene, trigger germination and appressorium formation of the avocado pathogen Colletotrichum gloeosporioides only after the conidia are in contact with a hard surface. What role this contact plays is unknown. Here, we describe isolation of genes expressed during the early stage of hard-surface treatment by a differential-display method and report characterization of one of these cloned genes, chip1 (Colletotrichum hard-surface induced protein 1 gene), which encodes a ubiquitin-conjugating enzyme. RNA blots clearly showed that it is induced by hard-surface contact and that ethylene treatment enhanced this induction. The predicted open reading frame (ubc1Cg) would encode a 16.2-kDa ubiquitin-conjugating enzyme, which shows 82% identity to the Saccharomyces cerevisiae UBC4-UBC5 E2 enzyme, comprising a major part of total ubiquitin-conjugating activity in stressed yeast cells. UBC1Cg can complement the proteolysis deficiency of the S. cerevisiae ubc4 ubc5 mutant, indicating that ubiquitin-dependent protein degradation is involved in conidial germination and appressorial differentiation.

  12. Environmental Scanning Electron Microscopy characterization of the adhesion of conidia from Penicillium expansum to cedar wood substrata at different pH values.

    PubMed

    El Abed, Soumya; Ibnsouda, Saad Koraichi; Latrache, Hassan; Meftah, Hasna; Tahri, Nezha Joutey; Hamadi, Fatima

    2012-04-01

    Initial microbial adhesion to surfaces is a complicated process that is affected by a number of factors. An important property of a solution that may influence adhesion is pH. The surface properties of the cedar wood were characterized by the sessile drop technique. Moreover, the interfacial free energy of surface adhesion to the cedar wood was determined under pH values (2, 3, 5, 7, 9 and 11). The results showed that cedar wood examined at different pH levels could be considered hydrophobic ranged from Giwi = -13.1 mJ/m(2) to Giwi = -75 mJ/m(2). We noted that the electron-donor character of cedar wood was important at both basic and limit acidic conditions (pH 11 and pH 3) and it decreased at intermediate pH (pH 5). The cedar wood substratum presents a weak electron acceptor under various pH's. In addition, the adhesion of conidia from Penicilllium expansum to the cedar wood surfaces at different pH values (2, 3, 5, 7, 9 and 11) was investigated using Environmental Scanning Electron Microscopy and image analysis was assessed with the Mathlab(®) program. The data analysis showed that the conidia from P. expansum were strongly influenced by the pH. The maximum adhesion occurs in the pH 11 and pH 3 and decreased to 24% at pH 5.

  13. Combination of driselase and lysing enzyme in one molar potassium chloride is effective for the production of protoplasts from germinated conidia of Fusarium verticillioides.

    PubMed

    Ramamoorthy, Vellaisamy; Govindaraj, Lavanya; Dhanasekaran, Madhumitha; Vetrivel, Sharmilee; Kumar, Krish K; Ebenezar, Edward

    2015-04-01

    Various cell wall degrading enzymes and the protoplasting media were evaluated for the production of protoplast in Fusarium verticillioides. Among the various enzymes tested, driselase at 12.5 mg/ml in 1 M KCl protoplasting medium produced the maximum number of protoplast. Next to driselase, lysing enzyme at 10 mg/ml in 1.2 M MgSO4 protoplasting medium was found to be the second best enzyme for the production of protoplast. More interestingly, the combined use of driselase @ 12.5 mg/ml and lysing enzyme @ 10 mg/ml in 1 M KCl exhibited the additive effect on protoplast formation. Germinated conidia of F. verticillioides are the most susceptible fungal material for protoplast production. The use of sucrose at 1.2 M in the regeneration medium supported the maximum regeneration of protoplast. From the present study, we recommend driselase (12.5 mg/ml) and lysing enzyme (10 mg/ml) in 1 M KCl protoplasting medium and germinated conidia of F. verticillioides for the maximum production of protoplasts and 1.2 M sucrose is the best osmoticum for the regeneration of protoplasts. Copyright © 2015 Elsevier B.V. All rights reserved.

  14. Increased rDNA synthesis in germinated conidia of Neurospora crassa is caused by RNA primer molecules found in its culture medium

    SciTech Connect

    Dutta, S.K.; Beljanski, M.

    1984-01-01

    Purine rich small primer RNA molecules (10-15 nucleotides) were isolated from growth medium of germinated (3 hr sprout) conidia of N. crassa. These RNA-primer molecules strongly stimulated in vitro DNA synthesis in N. crassa 74A wild type, as well as in DNAs from mice spleen and lung, and quail testis. These increases of in vitro DNA synthesis was dependent on the concentration of these RNA primer molecules. In contrast, such molecules were not found in 1 or 10 hour sprouts, nor in the culture medium of mycelia (24 hr). These RNA-primer molecules could be hydrolyzed by T1 RNAse but not by pancreatic RNase. Dutta et al. reported increased (250) copies of rRNA genes in germinated conidia (3 hr sprouts) compared to 100 copies of rRNA genes in mycelial cells grown for 24 hours. These observations suggest excessive transcription of rDNAs in the germinated conidial cells which undergo cleavages by nucleates after 3-4 hours of cell growth. Some degradation products were excreted into the culture medium and acted as RNA-primers.

  15. New collection of Acrogenospora spharerocephala from freshwater habitats of Northern Thailand

    NASA Astrophysics Data System (ADS)

    Liu, X. J.; Li, F. P.; Liu, W.; Huang, X. L.; Zhang, H.

    2017-01-01

    During investigations of freshwater fungi in Thailand, we collected dematiaceous hyphomycetes Acrogenospora spharerocephalan in lotic freshwater habitat. The description and morphological photographs of new collection is illustrated. Acrogenospora spharerocephala was characterized by unbranched conidiophores and producing solitary non-septate conidia acrogenously from recurrently proliferating conidiogenous cells. The conidia are dark brown to black when mature, paler at young, spherical to subspherical.

  16. Inhibition of spoilage mould conidia by acetic acid and sorbic acid involves different modes of action, requiring modification of the classical weak-acid theory.

    PubMed

    Stratford, Malcolm; Plumridge, Andrew; Nebe-von-Caron, Gerhardt; Archer, David B

    2009-11-30

    Fungal spoilage of many foods is prevented by weak-acid preservatives such as sorbic acid or acetic acid. We show that sorbic and acetic acids do not both inhibit cells by lowering of internal pH alone and that the "classical weak-acid theory" must be revised. The "classical weak-acid theory" suggests that all lipophilic acids with identical pK(a) values are equally effective as preservatives, causing inhibition by diffusion of molecular acids into the cell, dissociation, and subsequent acidification of the cytoplasm. Using a number of spoilage fungi from different genera, we have shown that sorbic acid was far more toxic than acetic acid, and no correlation existed between resistance to acetic acid and resistance to sorbic acid. The molar ratio of minimum inhibitory concentrations (MICs) (acetic: sorbic) was 58 for Paecilomyces variotii and 14 for Aspergillus phoenicis. Using flow cytometry on germinating conidia of Aspergillusniger, acetic acid at pH 4.0 caused an immediate decline in the mean cytoplasmic pH (pH(i)) falling from neutrality to approximately pH 4.7 at the MIC (80 mM). Sorbic acid also caused a rapid but far smaller drop in pH(i), at the MIC (4.5 mM); the pH remained above pH 6.3. Over 0-5 mM, a number of other weak acids caused a similar fall in cytoplasmic pH. It was concluded that while acetic acid inhibition of A. niger conidia was due to cytoplasmic acidification, inhibition by sorbic acid was not. A possible membrane-mediated mode of action of sorbic acid is discussed.

  17. Low pressure mode combined with OsO4 vapor fixation and sputter-coating for the preservation of delicate aerial hyphae and conidia in the ESEM.

    PubMed

    Coetzee, Stephan H; Jordaan, Anine; Mpuchane, Sisai F

    2005-08-01

    The newer generation of environmental scanning electron microscopes (ESEMs) allows samples to be viewed under a range of different vacuum conditions. No specific sample preparation protocols are required with the ESEMs, as fresh, unfixed samples are used and discarded later. We have worked out a method that preserves aerial hyphae on biltong that closely resemble fresh specimens and may be stored for viewing at a later date. Another advantage is that fixed samples are more resilient to the variable vacuum encountered in the ESEM. When biltong samples with fungal growth were first studied, we observed that vacuum-related artifacts were induced unless vacuum conditions and changes in pressure were carefully controlled. Damage readily occured in conidia and its delicate hyphae. Fresh, unfixed samples are very vulnerable to these artifacts. In addition, biltong proved to be a problematic study sample because of its high salt content, its hygroscopic nature as well as being laden with spices. To eliminate these artifacts, the preservation of specimens by OsO4 vapor fixation combined with a special sputter-coating technique is described. Previous studies confirmed that OsO4 vapor fixation is superior to traditional immersion fixation methods for the examination of hyphae and conidia of various fungi in a conventional SEM. However, both preparation methods induce sample shrinkage. We observed that OsO4 vapor fixation followed by Au coating under strictly controlled vacuum conditions induced fewer artifacts and gave the best images with minimum distortion in low pressure (LP) mode. The proposed method allows samples to be viewed both in ESEM and LP mode. There are however some disadvantages inherent to ESEM mode. Even when viewing fixed, coated samples, care should be taken to maintain a pressure of not lower than 0.2 Torr in the specimen chamber. It is critical that different samples have the same vacuum exposure history, as shrinkage and collapse were found to be

  18. Role of ozone in UV-C disinfection, demonstrated by comparison between wild-type and mutant conidia of Aspergillus niger.

    PubMed

    Liu, Jing; Zhou, Lin; Chen, Ji-Hong; Mao, Wang; Li, Wen-Jian; Hu, Wei; Wang, Shu-Yang; Wang, Chun-Ming

    2014-01-01

    This study aimed to investigate the tolerance of a melanized wild-type strain of Aspergillus niger (CON1) and its light-colored mutant (MUT1) to UV-C light and the concomitantly generated ozone. Treatments were segregated into four groups based on whether UV irradiation was used and the presence or absence of ozone: (-UV, -O3), (-UV, +O3), (+UV, -O3) and (+UV, +O3). The survival of CON1 and MUT1 conidia under +UV decreased as the exposure time increased, with CON1 showing greater resistance to UV irradiation than MUT1. Ozone induced CON1 conidium inactivation only under conditions of UV radiation exposure. While, the inactivation effect of ozone on MUT1 was always detectable regardless of the presence of UV irradiation. Furthermore, the CON1 conidial suspension showed lower UV light transmission than MUT1 when examined at the same concentration. Compared with the pigment in MUT1, the melanin in CON1 exhibited more potent radical-scavenging activity and stronger UV absorbance. These results suggested that melanin protected A. niger against UV disinfection via UV screening and free radical scavenging. The process by which UV-C disinfection induces a continual decrease in conidial survival suggests that UV irradiation and ozone exert a synergistic fungicidal effect on A. niger prior to reaching a plateau.

  19. Pathogenicity of conidia-based preparations of entomopathogenic fungi against the greenhouse pest aphids Myzus persicae, Aphis gossypii, and Aulacorthum solani (Hemiptera: Aphididae).

    PubMed

    Jandricic, S E; Filotas, M; Sanderson, J P; Wraight, S P

    2014-05-01

    Seeking new isolates of entomopathogenic fungi with greater virulence against greenhouse aphid pests than those currently registered in North America for control of these insects, single-dose screening assays of 44 selected fungal isolates and 4 commercially available strains were conducted against first-instar nymphs of Myzus persicae and Aphis gossypii. The assays identified a number of Beauveria and Metarhizium isolates with virulence equal to or greater than that of the commercial strains against the nymphal aphids, but none exhibited exceptionally high virulence. Virulence of Isaria isolates was unexpectedly low (<31% mortality at doses>1000conidia/mm(2)). In dose-response assays, Beauveria ARSEF 5493 proved most virulent against M. persicae and A. gossypii; however, LC50s of this isolate did not differ significantly from those of B. bassiana commercial strain JW-1. Dose-response assays were also conducted with Aulacorthum solani, the first reported evaluations of Beauveria and Metarhizium against this pest. The novel isolate Metarhizium 5471 showed virulence⩾that of Beauveria 5493 in terms of LC25 and LC50, but 5493 produced a steeper dose response (slope). Additional tests showed that adult aphids are more susceptible than nymphs to fungal infection but confirmed that infection has a limited pre-mortem effect on aphid reproduction. Effects of assay techniques and the potential of fungal pathogens as aphid-control agents are discussed. Copyright © 2014 Elsevier Inc. All rights reserved.

  20. Visual expression analysis of the responses of the alternative oxidase gene (aox1) to heat shock, oxidative, and osmotic stresses in conidia of citric acid-producing Aspergillus niger.

    PubMed

    Honda, Yuki; Hattori, Takasumi; Kirimura, Kohtaro

    2012-03-01

    The citric acid-producing filamentous fungus Aspergillus niger WU-2223L shows cyanide-insensitive respiration catalyzed by alternative oxidase in addition to the cytochrome pathway. Sequence analysis of the 5' flanking region of the alternative oxidase gene (aox1) revealed a potential heat shock element (HSE) and a stress response element (STRE). We have previously confirmed aox1 expression in conidia. In this study, to confirm whether the upstream region of aox1 responds to various stresses, we used a visual expression analysis system for single-cell conidia of the A. niger strain AOXEGFP-1. This strain harbored a fusion gene comprising aox1 and egfp, which encodes the enhanced green fluorescent protein (EGFP). The fluorescence intensity of EGFP increased in conidia of A. niger AOXEGFP-1 that were subjected to heat shock at 35-45 °C, oxidative stress by exposure to 5mM paraquat or 1 mM t-butylhydroperoxide, or osmotic stresses by exposure to 0.5 M KCl or 1.0 M mannitol. These results indicate that the putative HSE and STRE in the upstream region of aox1 directly or indirectly respond to heat shock, oxidative, and osmotic stresses.

  1. Nomenclatural notes on some cercosporoid hyphomycetes

    USDA-ARS?s Scientific Manuscript database

    Cercospora haleniae is validated, Cercospora tragiicola sp. nov. and Passalora halesiicola sp. nov. (Cercospora halesiae nom. inval., Passalora halesiae comb. inval.) are described and the combination Pseudocercospora nepheloides is introduced....

  2. Characterization of NpgA, a 4'-phosphopantetheinyl transferase of Aspergillus nidulans, and evidence of its involvement in fungal growth and formation of conidia and cleistothecia for development.

    PubMed

    Kim, Jung-Mi; Song, Ha-Yeon; Choi, Hyo-Jin; So, Kum-Kang; Kim, Dae-Hyuk; Chae, Keon-Sang; Han, Dong-Min; Jahng, Kwang-Yeop

    2015-01-01

    The null pigmentation mutant (npgA1) in Aspergillus nidulans results in a phenotype with colorless organs, decreased branching growth, delayed of asexual spore development, and aberrant cell wall structure. The npgA gene was isolated from A. nidulans to investigate these pleiomorphic phenomena of npgA1 mutant. Sequencing analysis of the complementing gene indicated that it contained a 4'-phosphopantetheinyl transferase (PPTase) superfamily domain. Enzymatic assay of the PPTase, encoded by the npgA gene, was implemented in vivo and in vitro. Loss-of-function of LYS5, which encoded a PPTase in Saccharomyces cerevisiae, was functionally complemented by NpgA, and Escherichia coli-derived NpgA revealed phosphopantetheinylation activity with the elaboration of 3'5'-ADP. Deletion of the npgA gene caused perfectly a lethal phenotype and the absence of asexual/sexual sporulation and secondary metabolites such as pigments in A. nidulans. However, a cross feeding effect with A. nidulans wild type allowed recovery from deletion defects, and phased-culture filtrate from the wild type were used to verify that the npgA gene was essential for formation of metabolites needed for development as well as growth. In addition, forced expression of npgA promoted the formation of conidia and cleistothecia as well as growth. These results indicate that the npgA gene is involved in the phosphopantetheinylation required for primary biological processes such as growth, asexual/sexual development, and the synthesis of secondary metabolites in A. nidulans.

  3. The pmr gene, encoding a Ca2+-ATPase, is required for calcium and manganese homeostasis and normal development of hyphae and conidia in Neurospora crassa.

    PubMed

    Bowman, Barry J; Abreu, Stephen; Johl, Jessica K; Bowman, Emma Jean

    2012-11-01

    The pmr gene is predicted to encode a Ca(2+)-ATPase in the secretory pathway. We examined two strains of Neurospora crassa that lacked PMR: the Δpmr strain, in which pmr was completely deleted, and pmr(RIP), in which the gene was extensively mutated. Both strains had identical, complex phenotypes. Compared to the wild type, these strains required high concentrations of calcium or manganese for optimal growth and had highly branched, slow-growing hyphae. They conidiated poorly, and the shape and size of the conidia were abnormal. Calcium accumulated in the Δpmr strains to only 20% of the wild-type level. High concentrations of MnCl(2) (1 to 5 mM) in growth medium partially suppressed the morphological defects but did not alter the defect in calcium accumulation. The Δpmr Δnca-2 double mutant (nca-2 encodes a Ca(2+)-ATPase in the plasma membrane) accumulated 8-fold more calcium than the wild type, and the morphology of the hyphae was more similar to that of wild-type hyphae. Previous experiments failed to show a function for nca-1, which encodes a SERCA-type Ca(2+)-ATPase in the endoplasmic reticulum (B. J. Bowman, S. Abreu, E. Margolles-Clark, M. Draskovic, and E. J. Bowman, Eukaryot. Cell 10:654-661, 2011). The pmr(RIP) Δnca-1 double mutant accumulated small amounts of calcium, like the Δpmr strain, but exhibited even more extreme morphological defects. Thus, PMR can apparently replace NCA-1 in the endoplasmic reticulum, but NCA-1 cannot replace PMR. The morphological defects in the Δpmr strain are likely caused, in part, by insufficient concentrations of calcium and manganese in the Golgi compartment; however, PMR is also needed to accumulate normal levels of calcium in the whole cell.

  4. Proteomic analysis of conidia germination in Fusarium oxysporum f. sp. cubense tropical race 4 reveals new targets in ergosterol biosynthesis pathway for controlling Fusarium wilt of banana.

    PubMed

    Deng, Gui-Ming; Yang, Qiao-Song; He, Wei-Di; Li, Chun-Yu; Yang, Jing; Zuo, Cun-Wu; Gao, Jie; Sheng, Ou; Lu, Shao-Yun; Zhang, Sheng; Yi, Gan-Jun

    2015-09-01

    Conidial germination is a crucial step of the soilborne fungus Fusarium oxysporum f. sp. cubense tropical race 4 (Foc TR4), a most important lethal disease of banana. In this study, a total of 3659 proteins were identified by isobaric tags for relative and absolute quantitation (iTRAQ)-based comparative proteomic approach, of which 1009 were differentially expressed during conidial germination of the fungus at 0, 3, 7, and 11 h. Functional classification and bioinformatics analysis revealed that the majority of the differentially expressed proteins are involved in six metabolic pathways. Particularly, all differential proteins involved in the ergosterol biosynthesis pathway were significantly upregulated, indicating the importance of the ergosterol biosynthesis pathway to the conidial germination of Foc TR4. Quantitative RT-PCR, western blotting, and in vitro growth inhibition assay by several categories of fungicides on the Foc TR4 were used to validate the proteomics results. Four enzymes, C-24 sterol methyltransferase (ERG6), cytochrome P450 lanosterol C-14α-demethylase (EGR11), hydroxymethylglutaryl-CoA synthase (ERG13), and C-4 sterol methyl oxidase (ERG25), in the ergosterol biosynthesis pathway were identified and verified, and they hold great promise as new targets for effective inhibition of Foc TR4 early growth in controlling Fusarium wilt of banana. To the best of our knowledge, this report represents the first comprehensive study on proteomics profiling of conidia germination in Foc TR4. It provides new insights into a better understanding of the developmental processes of Foc TR4 spores. More importantly, by host plant-induced gene silencing (HIGS) technology, the new targets reported in this work allow us to develop novel transgenic banana leading to high protection from Fusarium wilt and to explore more effective antifungal drugs against either individual or multiple target proteins of Foc TR4.

  5. Formation of 1-octen-3-ol from Aspergillus flavus conidia is accelerated after disruption of cells independently of Ppo oxygenases, and is not a main cause of inhibition of germination.

    PubMed

    Miyamoto, Kana; Murakami, Tomoko; Kakumyan, Pattana; Keller, Nancy P; Matsui, Kenji

    2014-01-01

    Eight-carbon (C8) volatiles, such as 1-octen-3-ol, are ubiquitous among fungi. They are the volatiles critical for aroma and flavor of fungi, and assumed to be signals controlling germination of several fungi. In this study, we found that intact Aspergillus flavus conidia scarcely synthesized C8 volatiles but repeated freeze-thaw treatment that made the cell membrane permeable promoted (R)-1-octen-3-ol formation. Loss or down regulation of any one of five fatty acid oxygenases (PpoA, PpoB, PpoC, PpoD or lipoxygenase) hypothesized contribute to 1-octen-3-ol formation had little impact on production of this volatile. This suggested that none of the oxygenases were directly involved in the formation of 1-octen-3-ol or that compensatory pathways exist in the fungus. Germination of the conidia was markedly inhibited at high density (1.0 × 10(9)spores mL(-1)). It has been postulated that 1-octen-3-ol is an autoinhibitor suppressing conidia germination at high density. 1-Octen-3-ol at concentration of no less than 10 mM was needed to suppress the germination while the concentration of 1-octen-3-ol in the suspension at 1.0 × 10(9) mL(-1) was under the detection limit (<1 µM). Thus, 1-octen-3-ol was not the principal component responsible for inhibition of germination. Instead, it was evident that the other heat-labile factor(s) suppressed conidial germination.

  6. Formation of 1-octen-3-ol from Aspergillus flavus conidia is accelerated after disruption of cells independently of Ppo oxygenases, and is not a main cause of inhibition of germination

    PubMed Central

    Miyamoto, Kana; Murakami, Tomoko; Kakumyan, Pattana; Keller, Nancy P.

    2014-01-01

    Eight-carbon (C8) volatiles, such as 1-octen-3-ol, are ubiquitous among fungi. They are the volatiles critical for aroma and flavor of fungi, and assumed to be signals controlling germination of several fungi. In this study, we found that intact Aspergillus flavus conidia scarcely synthesized C8 volatiles but repeated freeze-thaw treatment that made the cell membrane permeable promoted (R)-1-octen-3-ol formation. Loss or down regulation of any one of five fatty acid oxygenases (PpoA, PpoB, PpoC, PpoD or lipoxygenase) hypothesized contribute to 1-octen-3-ol formation had little impact on production of this volatile. This suggested that none of the oxygenases were directly involved in the formation of 1-octen-3-ol or that compensatory pathways exist in the fungus. Germination of the conidia was markedly inhibited at high density (1.0 × 109spores mL−1). It has been postulated that 1-octen-3-ol is an autoinhibitor suppressing conidia germination at high density. 1-Octen-3-ol at concentration of no less than 10 mM was needed to suppress the germination while the concentration of 1-octen-3-ol in the suspension at 1.0 × 109 mL−1 was under the detection limit (<1 µM). Thus, 1-octen-3-ol was not the principal component responsible for inhibition of germination. Instead, it was evident that the other heat-labile factor(s) suppressed conidial germination. PMID:24883255

  7. Influence of plant root exudates, germ tube orientation and passive conidia transport on biological control of fusarium wilt by strains of nonpathogenic Fusarium oxysporum.

    PubMed

    Mandeel, Qaher A

    2006-03-01

    reached at 96 h. The presented data confirm the previous findings that attributes important for nonpathogenic fusaria to induce resistant are: rapid spore germination and orientation in response to root exudate; active root penetration and passive conidia transport in stem to initiate defence reaction without pathogenicity and enough lag period between induction and challenge inoculation. Strain C14 possesses all these qualifications and hence its ability to enhance host resistance is superior than strain C5.

  8. Species and ecological diversity within the Cladosporium cladosporioides complex (Davidiellaceae, Capnodiales)

    USDA-ARS?s Scientific Manuscript database

    The genus Cladosporium is one of the largest genera of dematiaceous hyphomycetes, and is characterised by a coronate scar structure, conidia in acropetal chains and Davidiella teleomorphs. Based on morphology and DNA phylogeny, the species complexes of C. herbarum and C. sphaerospermum have been res...

  9. John Webster: a personal perspective.

    PubMed

    Fisher, Peter J

    2005-05-01

    A personal perspective of John Webster, with an emphasis of studies of aero-aquatic hyphomycetes and the techniques pioneered to enable them to be studied, illustrated by examples particularly from the genera with helicoid conidia, Helicodendron and Helicoon, showing flotation devices.

  10. Filipin is a reliable in situ marker of ergosterol in the plasma membrane of germinating conidia (spores) of Penicillium discolor and stains intensively at the site of germ tube formation.

    PubMed

    Van Leeuwen, M R; Smant, W; de Boer, W; Dijksterhuis, J

    2008-08-01

    Filipin, a widely used fluorescent sterol marker is also a potent antibiotic. In this study we address the reliability of filipin as a monitor of ergosterol in fungal cells. A revised staining protocol was developed to minimize any biological effect of the compound. Germinating conidia of Penicillium discolor stained with filipin, displayed a fluorescent cap at the location of germ tube appearance and formation. During germ tube emergence, the fluorescent intensity of the cap increased. This was confirmed by HPLC as an increase of the net cellular ergosterol content. Filipin staining is absent during early germination, while FM dyes, similar molecules, stain the plasma membrane after 1 h. This indicates that the conidial cell wall is no barrier for filipin. To evaluate if filipin does bind ergosterol in situ, natamycin, more specific to ergosterol, was added before filipin staining. This resulted in a marked decrease in fluorescence indicating high ergosterol levels. This was characterized further in ergDelta-mutant cells of Saccharomyces cerevisiae containing altered sterols. Here ergosterol containing cells showed a high fluorescence decrease. Taken together, these data suggest that filipin monitors an ergosterol-enriched cap in germinating conidia at the site of germ tube formation. Furthermore, the sterol-rich cap decreases and reappears after a period of actin disruption. Myriocin that affects sphingolipid synthesis results in an increase of cellular ergosterol and overall filipin fluorescence, but not at the ergosterol cap, where fluorescence is significantly lowered. In conclusion, in this work we have demonstrated an effective revised method for ergosterol staining with filipin and demonstrated its specificity in both Penicillium and Saccharomyces.

  11. SPM and fungal spores in the ambient air of west Korea during the Asian dust (Yellow sand) period

    NASA Astrophysics Data System (ADS)

    Yeo, Hwan-Goo; Kim, Jong-Ho

    The relationship between suspended particulate matter (SPM) and fungal spore was investigated in Seosan, a rural county along the west coast of Korea, in the spring of 2000. SPM concentrations in the air were 199.8 μg m -3 in the first Asian dust period (23-24 March), 249.4 μg m -3 in the second Asian dust period (7-9 April) and 98.9 μg m -3 in the non-Asian dust period (12-16 May), respectively. The majority of the total SPM were composed of coarse particles sized about 5 μm during the two Asian dust periods. Four molds genera grown from airborne fungal spores were identified in colonies grown from SPM samples taken during the Asian dust periods. All the genera found, Fusarium, Aspergillus, Penicillium and Basipetospora, are hyphomycetes in the division Deuteromycota. Morphologically, more diversified mycelia of hyphomycetes were grown on the sample captured from 1.1 to 2.1 μm sized SPM than on the other sized samples gathered in the dust periods. On the other hand, no mold was observed on the sample of 1.1-2.1 μm sized SPM in the non-Asian dust period. From these results, it seems evident that several sorts of fine sized fungal spores were suspended in the atmospheric environment of this study area during Asian dust periods.

  12. The impact of biotechnology on hyphomycetous fungal insect biocontrol agents.

    PubMed

    Hegedus, D D; Khachatourians, G G

    1995-01-01

    The potential for the control of insect pests by entomopathogenic fungi has been touted for decades, if not centuries. Only recently have advances in biotechnology provided the tools for indepth analysis of the mechanisms involved in pathogenesis and host death at the molecular level. This review outlines the current state of knowledge regarding the mode of infection and targets several key components that are amenable to improvement via biotechnology. Realization of the considerable economic potential of fungal bioinsecticides can occur only through a combined and coordinated effort involving fundamental science, formulation technology and field applications.

  13. Quantitative real-time PCR as a promising tool for the detection and quantification of leaf-associated fungal species – A proof-of-concept using Alatospora pulchella

    PubMed Central

    Feckler, Alexander; Schrimpf, Anne; Bundschuh, Mirco; Bärlocher, Felix; Baudy, Patrick; Cornut, Julien; Schulz, Ralf

    2017-01-01

    Traditional methods to identify aquatic hyphomycetes rely on the morphology of released conidia, which can lead to misidentifications or underestimates of species richness due to convergent morphological evolution and the presence of non-sporulating mycelia. Molecular methods allow fungal identification irrespective of the presence of conidia or their morphology. As a proof-of-concept, we established a quantitative real-time polymerase chain reaction (qPCR) assay to accurately quantify the amount of DNA as a proxy for the biomass of an aquatic hyphomycete species (Alatospora pulchella). Our study showed discrimination even among genetically closely-related species, with a high sensitivity and a reliable quantification down to 9.9 fg DNA (3 PCR forming units; LoD) and 155.0 fg DNA (47 PCR forming units; LoQ), respectively. The assay’s specificity was validated for environmental samples that harboured diverse microbial communities and likely contained PCR-inhibiting substances. This makes qPCR a promising tool to gain deeper insights into the ecological roles of aquatic hyphomycetes and other microorganisms. PMID:28384166

  14. Transformation of Epichloë typhina by electroporation of conidia

    USDA-ARS?s Scientific Manuscript database

    Choke, caused by the endophytic fungus Epichloë typhina is an important disease of orchardgrass (Dactylis glomerata L.) grown for seed. Very little is known concerning the conditions necessary for successful infection of orchardgrass by E. typhina. Detection of E. typhina early in the disease cycle ...

  15. Environmental factors impact Passalora sequoiae conidia counts from Leyland Cypress

    USDA-ARS?s Scientific Manuscript database

    Needle blight disease, caused by Passalora sequoiae, results in a progressive lost of leaf tissue on Leyland cypress (×Cupressocyparis leylandii) within container and field tree nurseries, in the landscape, and on Christmas tree farms. Fungicide schedules were developed in response to seasonal sympt...

  16. Ramophialophora humicola and Fibulochlamys chilensis, two new microfungi from soil.

    PubMed

    Madrid, H; Cano, J; Stchigel, A; Gené, J; Guarro, J

    2010-01-01

    In a study on soil microfungi from different countries two new hyphomycetes were found. The first one, Ramophialophora humicola, isolated from a soil sample collected in Ronda (Spain), is characterized by producing profusely branched conidiophores ending in sterile, sometimes swollen apices, and subhyaline, dacryoid conidia borne from both integrated and discrete phialides with conspicuous collaretes. ITS sequence data reveal its relationships with members of the Sordariales and its genetic differences with other fungi morphologically close, such as Cladorrhinum spp. The second species, Fibulochlamys chilensis, isolated from a soil sample collected in LaJunta (Chile), is characterized by micronematous, clamped, mostly branched conidiophores producing thallic, one-celled, thick-walled conidia that exhibit strongly wrinkled surfaces in age. The analysis of partial sequences of the ITS region and 28S rRNA gene reveal that this fungus is close to members of the gilled Agaricales.

  17. The hyphomycete genus Piricauda, with the description of a new species.

    PubMed

    Mercado Sierra, Angel; Guarro, Josep; Heredia, Gabriela

    2005-06-01

    The genus Piricauda comprises eight species characterized by monotretic conidiogenous cells and micronematous arched conidiophores: P. cochinensis, P. cubensis, P. longispora, P. mexicana, P. paraguayensis, P. pseudarthriae, P. taiwanensis, and P. vulcanensis sp. nov. P. caribensis is transferred to Acrodictys. Species usually colonize dead petioles and rachides of palms in tropical and subtropical areas, but can also be found on branches and leaves of different trees. The species are illustrated and briefly described, and a key to them is provided.

  18. CELL-WALL DEGRADING ENZYMES OF AQUATIC HYPHOMYCETES: A REVIEW. (U915444)

    EPA Science Inventory

    The perspectives, information and conclusions conveyed in research project abstracts, progress reports, final reports, journal abstracts and journal publications convey the viewpoints of the principal investigator and may not represent the views and policies of ORD and EPA. Concl...

  19. Virulence of Metarhizium anisopliae (Deuteromycotina: Hyphomycetes) on Anastrepha ludens (Diptera: Tephritidae): laboratory and field trials.

    PubMed

    Lezama-Gutiérrez, R; Trujillo-de la Luz, A; Molina-Ochoa, J; Rebolledo-Dominguez, O; Pescador, A R; López-Edwards, M; Aluja, M

    2000-08-01

    Twenty isolates of the fungus Metarhizium anisopliae (Metsch). Sorkin (Ma) were evaluated to determine their virulence against last instar and adult emergence of Mexican fruit fly, Anastrepha ludens (Loew). Larvae were exposed by immersion in a conidial suspension at a concentration of 10(8) UFC/ml under laboratory conditions. Larvae and pupae cumulative mortality rates ranged from 37.9 to 98.75%. Thirteen isolates caused mortality rates > 83.7%, and their LT50 values ranged from 1.8 to 6.2 d. The Ma2, Ma8, and Ma16 isolates were evaluated at seven different concentrations ranging from 10(1) to 10(7) UFC/ml, showing LC50 values from 3.7 to 4.8 x 10(5) UFC/ml. In a field-cage experiment, 200 ml of a conidial suspension of Ma2, at a concentration of 2.5 x 10(6) UFC/ml, was applied on 2,500 cm2 soil surface (2 x 10(5) UFC/cm2). The fungus reduced adult emergence, 22% fewer adults emerging in a sandy loam soil, and 43% fewer in loam soil, compared with the controls. M. anisopliae may offer a preferable alternative to chemicals as a biological control agent against A. ludens.

  20. Carbon requirements of some nematophagous, entomopathogenic and mycoparasitic hyphomycetes as fungal biocontrol agents.

    PubMed

    Sun, ManHong; Liu, XingZhong

    2006-05-01

    Thirty-three carbon sources were evaluated for their effects on spore germination, hyphal growth and sporulation of 11 fungal biocontrol agents, i.e. the nematophagous fungi Paecilomyces lilacinus, Pochonia chlamydosporia, Hirsutella rhossiliensis, H. minnesotensis and Arkansas Fungus 18, the entomopathogenic fungi Lecanicillium lecanii, Beauveria bassiana and Metarhizium anisopliae, and the mycoparasitic fungus Trichoderma viride. Variations in carbon requirements were found among the fungal species or strains tested. All strains studied except for T. viride grew on most carbon sources, although B. bassiana had more fastidious requirements for spore germination. Monosaccharides and disaccharides were suitable for fungal growth. For most isolates, D-glucose, D-mannose, sucrose and trehalose were superior to pectin and soluble starch among the polysaccharides and lactic acid among the organic acids. Both ethanol and methanol could accelerate growth of most isolates but not biomass. D-mannose, D-fructose and D-xylose were excellent carbon sources for sporulation, while D-glucose, sucrose, cellobiose, trehalose, chitin, dextrin, gelatin and lactic acid were better for some isolates. Neither sorbic acid nor linoleic acid could be utilized as a single carbon source. These findings provided a better understanding of the nutritional requirements of different fungal biocontrol agents that can benefit the mass production process.

  1. The Role of Antennae in Removing Entomopathogenic Fungi from Cuticle of the Termite, Coptotermes formosanus

    PubMed Central

    Yanagawa, Aya; Yokohari, Fumio; Shimizu, Susumu

    2009-01-01

    Our previous research has shown that the termite, Coptotermes formosanus Shiraki (Isoptera: Rhinotermitidae), protects itself from entomopathogenic fungi by mutual grooming behavior. The termite removes and discards foreign organisms, such as fungal conidia, from the body surface of its nestmates by mutual grooming behavior. The role of the antennae in detecting the condia was examind here. Three entomopathogenic fungi were used, Beauveria brongniartii 782 (Saccardo) (Hypocreales), Paecilomyces fumosoroseus K3 (Wize) (Hyphomycetes), and Metarhizium anisopliae 455 Sorokin (Hyphomycetes). Termites with antennae removed conidia more efficiently than termites without antennae. There were differences between termites with and without antennae in selection of sites to be groomed on nestmates, in the length of grooming and in occurrence of grooming. Electroantennogram (EAG) responses were recorded from termite antennae and the waveforms were rather specific to the kinds of fungi used as odor sources. Termites were able to distinguish between the tested fungi in feeding tests. These results show that the antennae play important roles in the mutual grooming behavior of the termite. PMID:19611249

  2. Seasonal and substrate preferences of fungi colonizing leaves in streams: traditional versus molecular evidence.

    PubMed

    Nikolcheva, Liliya G; Bärlocher, Felix

    2005-02-01

    Aquatic hyphomycetes are the main fungal decomposers of plant litter in streams. We compared the importance of substrate (three leaf species, wood) and season on fungal colonization. Substrates were exposed for 12 4-week periods. After recovery, mass loss, fungal biomass and release of conidia by aquatic hyphomycetes were measured. Fungal communities were characterized by counting and identifying released conidia and by extracting and amplifying fungal DNA (ITS2), which was subdivided into phylotypes by denaturing gradient gel electrophoresis (DGGE) and terminal-restriction fragment length polymorphism (T-RFLP). Mass loss, fungal biomass and reproduction were positively correlated with stream temperature. Conidial diversity was highest between May and September. Numbers of different phylotypes were more stable. Principal coordinate analyses (PCO) and canonical analyses of principal coordinates (CAP) of presence/absence data (DGGE bands, T-RFLP peaks and conidial species) showed a clear seasonal trend (Por=0.88). Season was also a significant factor when proportional similarities of conidial communities or relative intensities of DGGE bands were evaluated (P

  3. Whole-stream nitrate addition affects litter decomposition and associated fungi but not invertebrates.

    PubMed

    Ferreira, Verónica; Gulis, Vladislav; Graça, Manuel A S

    2006-10-01

    We assessed the effect of whole-stream nitrate enrichment on decomposition of three substrates differing in nutrient quality (alder and oak leaves and balsa veneers) and associated fungi and invertebrates. During the 3-month nitrate enrichment of a headwater stream in central Portugal, litter was incubated in the reference site (mean NO3-N 82 microg l-1) and four enriched sites along the nitrate gradient (214-983 microg NO3-N l-1). A similar decomposition experiment was also carried out in the same sites at ambient nutrient conditions the following year (33-104 microg NO3-N l-1). Decomposition rates and sporulation of aquatic hyphomycetes associated with litter were determined in both experiments, whereas N and P content of litter, associated fungal biomass and invertebrates were followed only during the nitrate addition experiment. Nitrate enrichment stimulated decomposition of oak leaves and balsa veneers, fungal biomass accrual on alder leaves and balsa veneers and sporulation of aquatic hyphomycetes on all substrates. Nitrate concentration in stream water showed a strong asymptotic relationship (Michaelis-Menten-type saturation model) with temperature-adjusted decomposition rates and percentage initial litter mass converted into aquatic hyphomycete conidia for all substrates. Fungal communities did not differ significantly among sites but some species showed substrate preferences. Nevertheless, certain species were sensitive to nitrogen concentration in water by increasing or decreasing their sporulation rate accordingly. N and P content of litter and abundances or richness of litter-associated invertebrates were not affected by nitrate addition. It appears that microbial nitrogen demands can be met at relatively low levels of dissolved nitrate, suggesting that even minor increases in nitrogen in streams due to, e.g., anthropogenic eutrophication may lead to significant shifts in microbial dynamics and ecosystem functioning.

  4. Orbilia querci sp. nov. and its knob-forming nematophagous anamorph.

    PubMed

    Liu, Bin; Liu, Xing-Zhong; Zhuang, Wen-Ying

    2005-04-01

    Orbilia querci, a new nematode-trapping fungus, was found on rotten wood of Quercus sp. in Huai-rou County, Beijing, China. It is characterized by having a tear-shaped spore body in the cylindrical ascospore. Pure culture was obtained from the ascospores. Conidiophores were simple or occasionally branched, bearing a single conidium on the tip. Conidia were spindle-shaped, mostly with 3-septa. Nematodes were captured by means of adhesive stalked knobs. The adhesive knobs were produced frequently on nutritional agar plates even in the absence of challenging nematodes. Its anamorph is placed in Dactylellina and named as D. querci. The sequence divergence of the ITS1 region between the fungus and the other knob-forming species tested was 23.8-33.4%, supporting O. querci as a distinct species. This is the first report of the connection between a knob-forming nematophagous hyphomycete and an Orbilia teleomorph.

  5. Preillumination of rice blast conidia induces tolerance to subsequent oxidative stress

    USDA-ARS?s Scientific Manuscript database

    Many environmental factors, alone or combined, affect organisms by changing a pro-/antioxidant balance. Here we tested rice blast fungus (Magnaporthe oryzae) for possible cross-adaptations caused by relatively intense light and protecting from artificially formed reactive oxygen species (ROS) and RO...

  6. Commercial detergents effective against conidia and chlamydospores of Fusarium oxysporum f. sp. vasinfectum

    USDA-ARS?s Scientific Manuscript database

    Current containment recommendations for limiting the spread of race 4 of Fusarium oxysporum f. sp. vasinfectum in California lack non-corrosive yet effective alternatives to bleach for sanitizing equipment used in farming operations. To find an equivalent to Farmcleanse, an Australian product recomm...

  7. Production of Macrophomina phaseolina conidia by multiple soybean isolates in culture

    USDA-ARS?s Scientific Manuscript database

    Macrophomina phaseolina (Tassi) Goid is the cause of charcoal rot of soybean, Glycine max (L.) (Merr.). Soybean resistance to M. phaseolina in commercial cultivars is not common, but is especially needed in locations where the disease is chronic and severe. The objective of this study was to develop...

  8. Effect of storage temperature and sphacellum age on C. africana conidia survival

    USDA-ARS?s Scientific Manuscript database

    Sorghum ergot, caused by Claviceps africana Frederickson, Mantle & de Milliano, is a disease that poses a serious treat to sorghum, especially in hybrid seed production. The initial sign of the disease is called a sphacelium. Sphacelia containing macroconidia could play a role in the survival of t...

  9. Storage conditions affect speed of germination in Beauveria bassiana and Metarhizium anisopliae conidia

    USDA-ARS?s Scientific Manuscript database

    The retention of high viability during storage is essential for effectiveness and thus market acceptance of fungus-based biopesticides. The length of adequate shelf-lives for mycoinsecticides is controversial, with proposed requirements varying from a few weeks to 18 months. Shelf-life determination...

  10. Shelf-life of Beauveria bassiana conidia in gaseous atmospheres under high temperature regimes

    USDA-ARS?s Scientific Manuscript database

    The retention of high viability during storage is essential for effectiveness and thus market acceptance of fungus-based biopesticides. The length of adequate shelf-lives for mycoinsecticides is controversial, with proposed requirements varying from a few weeks to 18 months. Shelf-life determination...

  11. Beauveria bassiana, Metarhizium anisopliae, and Metarhizium anisopliae var. acridum conidia: tolerance to imbibitional damage

    USDA-ARS?s Scientific Manuscript database

    When dry fungal cells are immersed in water, rapid imbibition (water uptake) may compromise the plasma membrane, killing the cell. This study investigated the impact of imbibitional damage (measured in terms of reduced viability) on Beauveria bassiana (Bb), Metarhizium anisopliae (Ma) and M. anisop...

  12. Imbibitional damage in conidia of the entomopathogenic fungi Beauveria bassiana, Metarhizium anisopliae, and Metarhizium acridum

    USDA-ARS?s Scientific Manuscript database

    When dried organisms are immersed in water, rapid imbibition may cause severe damage to plasma membranes; in unicellular organisms, such damage is usually lethal. This study investigated effects of pre-immersion moisture levels and immersion temperature on imbibitional damage in three insect pathoge...

  13. [Aerobiological study of Alternaria and Cladosporium conidia in the atmosphere of Almeria (SE Spain)].

    PubMed

    Sabariego Ruiz, Silvia; Díaz de la Guardia Guerrero, Consuelo; Alba Sánchez, Francisca

    2004-09-01

    This study analysed the seasonal and intradiurnal behaviour of fungal spores from Alternaria and Cladosporium on air samples collected in the city of Almeria (SE Spain), as well as the influence that meteorological parameters have on the concentration of these type of spores. Aerobiological sampling was made during four years (1998-2001) using a Hirst-type volumetric spore trap. Spores of Alternaria and Cladosporium were detected throughout the year, reaching the highest concentrations from May to October. The diurnal patterns of these taxons reflected a presence similar of spores during a 24 h period, with values horary close to 4% of total sampling daily. The correlations show a positive association with temperature, hours of sunshine and accumulate rainfall, but negative with daily rainfall.

  14. Confirming resistance in bottle gourd germplasm by quantifying powdery mildew conidia using a cellometer

    USDA-ARS?s Scientific Manuscript database

    Powdery mildew (PM) caused by Podosphaera xanthii, an important foliar disease affecting cucurbit crops grown in the United States, commonly occurs on foliage, petioles, and stems. We have developed two highly resistant bottle gourd (Lagenaria siceraria) germplasm (USVL351 and USVL482) for use in o...

  15. Inactivation of conidia of Penicillium chrysogenum, P. digitatum and P. italicum by ethanol solutions and vapours.

    PubMed

    Dao, Thien; Bensoussan, Maurice; Gervais, Patrick; Dantigny, Philippe

    2008-02-29

    A fractional factorial design, 2(5-1) experiments, was used for assessing the influence of 5 factors: water activity, aw [0.7, 0.9], temperature, T ( degrees C) [10, 30], mode of application, A [liquid, vapour], ethanol concentration, E (% w/w) [5, 10] and time, t (d) [1, 4] on the inactivation of spores of Penicillium chrysogenum, P. digitatum and P. italicum. Survival was determined by germination at optimal conditions within 3d. The experimental response was log (N 0/Nt), where N 0 and Nt (spore ml(-1)) the concentrations of viable spores at t=0 and t respectively. By a decreasing order of sensitivity to ethanol, moulds were ranked as followed: P. digitatum, P. italicum and P. chrysogenum. A greater inactivation for P. digitatum, P. italicum, that were the most sensitive moulds to ethanol, was obtained by applying vapour rather than ethanol solution. The order of significance of the main factors depended upon the mould. The key factor for explaining inactivation of P. chrysogenum was water activity. But, temperature was the main factor for explaining inactivation of P. digitatum and P. italicum. In the more drastic conditions, (i.e., 0.7 aw, 30 degrees C, 10% w/w ethanol), all spores were inactivated by applying liquid solution for 4d.

  16. Evaluation of disinfectants to control contamination of citrus degreening rooms with conidia, 2008

    USDA-ARS?s Scientific Manuscript database

    Early season oranges are harvested and treated with ethylene gas for two or more days in humidified ‘degreening’ rooms at 20C to accelerate the degradation of chlorophyll to enhance the orange color of the fruit rind. Decay of the fruit during this process by P. digitatum is common and between cycle...

  17. Beauveria bassiana yeast phase on agar medium and its pathogenicity against Diatraea saccharalis (Lepidoptera: Crambidae) and Tetranychus urticae (Acari: Tetranychidae).

    PubMed

    Alves, Sérgio Batista; Rossi, Luciana Savoi; Lopes, Rogério Biaggioni; Tamai, Marco Antonio; Pereira, Roberto M

    2002-10-01

    Beauveria bassiana colonizes insect hosts initially through a yeast phase, which is common in some artificial liquid cultures, but not reported on artificial solid media. We describe a yeast-like phase for B. bassiana isolate 447 (ATCC 20872) on MacConkey agar and its virulence toward Diatraea saccharalis and Tetranychus urticae. The yeast-like cells of B. bassiana developed by budding from germinating conidia after 24-h incubation. Cells were typically 5-10 microm and fungal colonies were initially circular and mucoid, but later were covered with mycelia and conidia. Ability to produce yeast-like cells on MacConkey medium was relatively common among different B. bassiana isolates, but growth rate and timing of yeast-like cell production also varied. Metarhizium anisopliae and Paecilomyces spp. isolates did not grow as yeast-like cells on MacConkey medium. Yeast-like cells of B. bassiana 447 were more virulent against D. saccharalis than conidia when 10(7)cells/ml were used. At 10(8)cells/ml, the estimated mean survival time was 5.4 days for the yeast suspension and 7.7 days for the conidial suspension, perhaps due to faster germination. The LC(50) was also lower for yeast than conidial suspensions. Yeast-like cells and conidia had similar virulence against T. urticae; the average mortalities with yeast-like cells and conidia were, respectively, 42.8 and 45.0%, with 10(7)cells/ml, and 77.8 and 74.4%, with 10(8)cells/ml. The estimated mean survival times were 3.6 and 3.9 for yeast and conidial suspensions, respectively. The bioassay results demonstrate the yeast-like structures produced on MacConkey agar are effective as inoculum for B. bassiana applications against arthropod pests, and possibly superior to conidia against some species. Obtaining well-defined yeast phase cultures of entomopathogenic hyphomycetes may be an important step in studies of the biology and nutrition, pathogenesis, and the genetic manipulation of these fungi.

  18. Pseudosigmoidea ibarakiensis sp. nov., a Dark Septate Endophytic Fungus from a Cedar Forest in Ibaraki, Japan

    PubMed Central

    Diene, Ousmane; Wang, Wei; Narisawa, Kazuhiko

    2013-01-01

    A dark septate fungus of Pseudosigmoidea, Hyphomycetes, was recovered from forest soil in Ibaraki prefecture, Japan. The isolate is characterized by pale to brown conidia with up to 8 septa measuring 68–132 × 4–7.9 μm. It is also unique in producing conidia borne by long conidogenious cells in agar medium with or without water, compared to P. cranei, which must be immersed in water to sporulate. Morphological analysis indicated that the isolate is distinct from P. cranei and is described as a new species, P. ibarakiensis sp. nov. Pathogenicity tests of Chinese cabbage and cucumber seedlings indicated that the fungus grows as an endophyte and colonizes, inter and intracellularly, the root epidermal and cortical layers without causing apparent disease symptoms in the host. This endophyte showed the ability to support cucumber plant growth under conditions where NaNO3 was replaced by organic nitrogen but also conferred to Chinese cabbage the ability to grow at low pH. It also became successfully established in six other plants, including the Brassicae, Solanaceae, Poaceae, and Liliacea families, suggesting its adaptability to a broad range of host plants. PMID:24005845

  19. Determining Diversity of Freshwater Fungi on Decaying Leaves: Comparison of Traditional and Molecular Approaches

    PubMed Central

    Nikolcheva, Liliya G.; Cockshutt, Amanda M.; Bärlocher, Felix

    2003-01-01

    Traditional microscope-based estimates of species richness of aquatic hyphomycetes depend upon the ability of the species in the community to sporulate. Molecular techniques which detect DNA from all stages of the life cycle could potentially circumvent the problems associated with traditional methods. Leaf disks from red maple, alder, linden, beech, and oak as well as birch wood sticks were submerged in a stream in southeastern Canada for 7, 14, and 28 days. Fungal biomass, estimated by the amount of ergosterol present, increased with time on all substrates. Alder, linden, and maple leaves were colonized earlier and accumulated the highest fungal biomass. Counts and identifications of released conidia suggested that fungal species richness increased, while community evenness decreased, with time (up to 11 species on day 28). Conidia of Articulospora tetracladia dominated. Modifications of two molecular methods—denaturing gradient gel electrophoresis (DGGE) and terminal restriction fragment length polymorphism (T-RFLP) analysis—suggested that both species richness and community evenness decreased with time. The dominant ribotype matched that of A. tetracladia. Species richness estimates based on DGGE were consistently higher than those based on T-RFLP analysis and exceeded those based on spore identification on days 7 and 14. Since traditional and molecular techniques assess different aspects of the fungal organism, both are essential for a balanced view of fungal succession on leaves decaying in streams. PMID:12732520

  20. Pseudosigmoidea ibarakiensis sp. nov., a dark septate endophytic fungus from a cedar forest in Ibaraki, Japan.

    PubMed

    Diene, Ousmane; Wang, Wei; Narisawa, Kazuhiko

    2013-01-01

    A dark septate fungus of Pseudosigmoidea, Hyphomycetes, was recovered from forest soil in Ibaraki prefecture, Japan. The isolate is characterized by pale to brown conidia with up to 8 septa measuring 68-132 × 4-7.9 mm. It is also unique in producing conidia borne by long conidogenious cells in agar medium with or without water, compared to P. cranei, which must be immersed in water to sporulate. Morphological analysis indicated that the isolate is distinct from P. cranei and is described as a new species, P. ibarakiensis sp. nov. Pathogenicity tests of Chinese cabbage and cucumber seedlings indicated that the fungus grows as an endophyte and colonizes, inter and intracellularly, the root epidermal and cortical layers without causing apparent disease symptoms in the host. This endophyte showed the ability to support cucumber plant growth under conditions where NaNO3 was replaced by organic nitrogen but also conferred to Chinese cabbage the ability to grow at low pH. It also became successfully established in six other plants, including the Brassicae, Solanaceae, Poaceae, and Liliacea families, suggesting its adaptability to a broad range of host plants.

  1. Septoria-like pathogens causing leaf and fruit spot of pistachio.

    PubMed

    Crous, Pedro W; Quaedvlieg, William; Sarpkaya, Kamil; Can, Canan; Erkılıç, Ali

    2013-12-01

    Several species of Septoria are associated with leaf and fruit spot of pistachio (Pistacia vera), though their identity has always been confused, making identification problematic. The present study elucidates the taxonomy of the Septoria spp. associated with pistachio, and distinguishes four species associated with this host genus. Partial nucleotide sequence data for five gene loci, ITS, LSU, EF-1α, RPB2 and Btub were generated for a subset of isolates. Cylindroseptoria pistaciae, which is associated with leaf spots of Pistacia lentiscus in Spain, is characterised by pycnidial conidiomata that give rise to cylindrical, aseptate conidia. Two species of Septoria s. str. are also recognised on pistachio, S. pistaciarum, and S. pistaciae. The latter is part of the S. protearum species complex, and appears to be a wide host range pathogen occurring on hosts in several different plant families. Septoria pistacina, a major pathogen of pistachio in Turkey, is shown to belong to Pseudocercospora, and not Septoria as earlier suspected. Other than for its pycnidial conidiomata, it is a typical species of Pseudocercospora based on its smooth, pigmented conidiogenous cells and septate conidia. This phenomenon has also been observed in Pallidocercospora, and seriously questions the value of conidiomatal structure at generic level, which has traditionally been used to separate hyphomycetous from coelomycetous ascomycetes. Other than DNA barcodes to facilitate the molecular identification of these taxa occurring on pistachio, a key is also provided to distinguish species based on morphology.

  2. A Beauveria phylogeny inferred from nuclear ITS and EF1-alpha sequences: evidence for cryptic diversification and links to Cordyceps teleomorphs.

    PubMed

    Rehner, Stephen A; Buckley, Ellen

    2005-01-01

    Beauveria is a globally distributed genus of soil-borne entomopathogenic hyphomycetes of interest as a model system for the study of entomopathogenesis and the biological control of pest insects. Species recognition in Beauveria is difficult due to a lack of taxonomically informative morphology. This has impeded assessment of species diversity in this genus and investigation of their natural history. A gene-genealogical approach was used to investigate molecular phylogenetic diversity of Beauveria and several presumptively related Cordyceps species. Analyses were based on nuclear ribosomal internal transcribed spacer (ITS) and elongation factor 1-alpha (EF1-alpha) sequences for 86 exemplar isolates from diverse geographic origins, habitats and insect hosts. Phylogenetic trees were inferred using maximum parsimony and Bayesian likelihood methods. Six well supported clades within Beauveria, provisionally designated A-F, were resolved in the EF1-alpha and combined gene phylogenies. Beauveria bassiana, a ubiquitous species that is characterized morphologically by globose to subglobose conidia, was determined to be non-monophyletic and consists of two unrelated lineages, clades A and C. Clade A is globally distributed and includes the Asian teleomorph Cordyceps staphylinidaecola and its probable synonym C. bassiana. All isolates contained in Clade C are anamorphic and originate from Europe and North America. Clade B includes isolates of B. brongniartii, a Eurasian species complex characterized by ellipsoidal conidia. Clade D includes B. caledonica and B. vermiconia, which produce cylindrical and comma-shaped conidia, respectively. Clade E, from Asia, includes Beauveria anamorphs and a Cordyceps teleomorph that both produce ellipsoidal conidia. Clade F, the basal branch in the Beauveria phylogeny includes the South American species B. amorpha, which produces cylindrical conidia. Lineage diversity detected within clades A, B and C suggests that prevailing morphological

  3. Potential of topic applications, leaf residues and soil drenches of the fungus Paecilomyces fumosoroseus (Deuteromycotina: Hyphomycetes) for management of the Diaprepes root weevil: laboratory and greenhouse investigations

    USDA-ARS?s Scientific Manuscript database

    Diaprepes root weevil, (DRW) Diaprepes abbreviatus is a key pest of citrus and ornamental plants in Florida and Texas. DRW larvae burrow through the soil feeding on roots which when girdled causes secondary infection of the structural roots or root crown by Phytopthora spp. wherein mature citrus tre...

  4. Influence of shredder feeding and nutrients on fungal activity and community structure in headwater streams

    Treesearch

    Namil Chung; Keller. Suberkropp

    2008-01-01

    In stream detrital food webs, interactions occur between aquatic hyphomycetes associated with decomposing leaves and shredders consuming those leaves. However, few studies have examined how the feeding activity of shredders affects aquatic hyphomycetes. We examined the effect of shredder feeding on aquatic hyphomycete communities associated with submerged leaves in two...

  5. Graphene oxide exhibits broad-spectrum antimicrobial activity against bacterial phytopathogens and fungal conidia by intertwining and membrane perturbation.

    PubMed

    Chen, Juanni; Peng, Hui; Wang, Xiuping; Shao, Feng; Yuan, Zhaodong; Han, Heyou

    2014-01-01

    To understand the interaction mechanism between graphene oxide (GO) and typical phytopathogens, a particular investigation was conducted about the antimicrobial activity of GO against two bacterial pathogens (P. syringae and X. campestris pv. undulosa) and two fungal pathogens (F. graminearum and F. oxysporum). The results showed that GO had a powerful effect on the reproduction of all four pathogens (killed nearly 90% of the bacteria and repressed 80% macroconidia germination along with partial cell swelling and lysis at 500 μg mL(-1)). A mutual mechanism is proposed in this work that GO intertwinds the bacteria and fungal spores with a wide range of aggregated graphene oxide sheets, resulting in the local perturbation of their cell membrane and inducing the decrease of the bacterial membrane potential and the leakage of electrolytes of fungal spores. It is likely that GO interacts with the pathogens by mechanically wrapping and locally damaging the cell membrane and finally causing cell lysis, which may be one of the major toxicity actions of GO against phytopathogens. The antibacterial mode proposed in this study suggests that the GO may possess antibacterial activity against more multi-resistant bacterial and fungal phytopathogens, and provides useful information about the application of GO in resisting crop diseases.

  6. Effect of a heterotrimeric G protein alpha subunit on conidia germination, stress response, and roquefortine C production in Penicillium roqueforti.

    PubMed

    García-Rico, Ramón O; Chávez, Renato; Fierro, Francisco; Martín, Juan F

    2009-06-01

    Heterotrimeric G protein signaling regulates many processes in fungi, such as development, pathogenicity, and secondary metabolite biosynthesis. For example, the Galpha subunit Pga1 from Penicillium chrysogenum regulates conidiation and secondary metabolite production in this fungus. The dominant activating allele, pga1G42R, encoding a constitutively active Pga1 Galpha subunit, was introduced in Penicillium roqueforti by transformation, resulting in a phenotype characterized by low sporulation and slow growth. In this work, the effect of the constitutively active Pga1G42R Galpha subunit on conidial germination, stress tolerance, and roquefortine C production of P. roqueforti was studied. Pga1G42R triggered germination in the absence of a carbon source, in addition to negatively regulating thermal and osmotic stress tolerance. The presence of the Pga1G42R Galpha subunit also had an important effect on roquefortine C biosynthesis, increasing production and maintaining high levels of the mycotoxin throughout a culture period of 30 days. Together, the results suggest that G protein-mediated signaling participates in the regulation of these three processes in P. roqueforti.

  7. rDNA-targeted PCR primers and FISH probe in the detection of Ophiocordyceps sinensis hyphae and conidia.

    PubMed

    Zhong, Xin; Peng, Qingyun; Qi, Lili; Lei, Wei; Liu, Xin

    2010-11-01

    Ophiocordyceps sinensis (Berk.) Sung, Sung, Hywel-Jones & Spatafora (syn. Cordyceps sinensis) one of the entomopathogenic fungi, is a rare Traditional Chinese Medicine (TCM) found in the Qinghai-Tibetan Plateau. Polymerase Chain Reaction (PCR) and Fluorescence in situ hybridization (FISH) methods are necessary to identify the mycelia or spores of O. sinensis from its habitat and to monitor its dispersal, colonization and infectivity. To develop both primers and probe specific to O. sinensis, ribosomal DNA (rDNA) amplified with universal primers from O. sinensis genomic DNA and seven closely related fungi were sequenced. According to these sequences, the upper and lower primers (OsT-F and OsT-R) were designed within internal transcribed spacer region 1 (ITS1) and ITS2 and flanked by universal primers ITS5 and ITS4, respectively. The designed primers were used for general PCR, touchdown PCR, or both together with the universal primers for nested-touchdown PCR. The results showed that only the extracted DNA of O. sinensis was specifically amplified. The sensitivity of nested-touchdown PCR with extracted DNA of O. sinensis is as low as 10(-14)g (10 fg) and at least 1000 times higher than the other PCR methods. In addition, Cy5-labeled probe (OsLSU) for cytoplasmic LSU rRNA was hybridized with the ascospores of O. sinensis. It showed a strong red fluorescence throughout the whole cell but did not cross-react with other entomopathogenic fungi. Taken together, these methods were useful for studying the biology and ecology of O. sinensis.

  8. Use of a Granular Bioplastic Formulation for Carrying Conidia of a Non-aflatoxigenic Strain of Aspergillus flavus

    USDA-ARS?s Scientific Manuscript database

    Previous research demonstrated that aflatoxin contamination in corn grown in Mississippi is reduced by field application of wheat grains pre-inoculated with the non-aflatoxigenic Aspergillus flavus strain NRRL 30797. To facilitate field applications of the biocontrol isolate, a series of laboratory ...

  9. Using Soyscreen in an Oil-Based Biopesticide Formulation to Protect Beauveria bassiana Conidia from Degradation by Ultraviolet Light Energy

    USDA-ARS?s Scientific Manuscript database

    Improving the efficacy of biological control treatments for pest control in crops will help to reduce dependence on chemical pesticide applications. Although effective for controlling insects, entomopathogenic microbes degrade rapidly when applied to field crops because of the adverse effects of su...

  10. Graphene oxide exhibits broad-spectrum antimicrobial activity against bacterial phytopathogens and fungal conidia by intertwining and membrane perturbation

    NASA Astrophysics Data System (ADS)

    Chen, Juanni; Peng, Hui; Wang, Xiuping; Shao, Feng; Yuan, Zhaodong; Han, Heyou

    2014-01-01

    To understand the interaction mechanism between graphene oxide (GO) and typical phytopathogens, a particular investigation was conducted about the antimicrobial activity of GO against two bacterial pathogens (P. syringae and X. campestris pv. undulosa) and two fungal pathogens (F. graminearum and F. oxysporum). The results showed that GO had a powerful effect on the reproduction of all four pathogens (killed nearly 90% of the bacteria and repressed 80% macroconidia germination along with partial cell swelling and lysis at 500 μg mL-1). A mutual mechanism is proposed in this work that GO intertwinds the bacteria and fungal spores with a wide range of aggregated graphene oxide sheets, resulting in the local perturbation of their cell membrane and inducing the decrease of the bacterial membrane potential and the leakage of electrolytes of fungal spores. It is likely that GO interacts with the pathogens by mechanically wrapping and locally damaging the cell membrane and finally causing cell lysis, which may be one of the major toxicity actions of GO against phytopathogens. The antibacterial mode proposed in this study suggests that the GO may possess antibacterial activity against more multi-resistant bacterial and fungal phytopathogens, and provides useful information about the application of GO in resisting crop diseases.To understand the interaction mechanism between graphene oxide (GO) and typical phytopathogens, a particular investigation was conducted about the antimicrobial activity of GO against two bacterial pathogens (P. syringae and X. campestris pv. undulosa) and two fungal pathogens (F. graminearum and F. oxysporum). The results showed that GO had a powerful effect on the reproduction of all four pathogens (killed nearly 90% of the bacteria and repressed 80% macroconidia germination along with partial cell swelling and lysis at 500 μg mL-1). A mutual mechanism is proposed in this work that GO intertwinds the bacteria and fungal spores with a wide range of aggregated graphene oxide sheets, resulting in the local perturbation of their cell membrane and inducing the decrease of the bacterial membrane potential and the leakage of electrolytes of fungal spores. It is likely that GO interacts with the pathogens by mechanically wrapping and locally damaging the cell membrane and finally causing cell lysis, which may be one of the major toxicity actions of GO against phytopathogens. The antibacterial mode proposed in this study suggests that the GO may possess antibacterial activity against more multi-resistant bacterial and fungal phytopathogens, and provides useful information about the application of GO in resisting crop diseases. Electronic supplementary information (ESI) available. See DOI: 10.1039/c3nr04941h

  11. QUANTITATIVE MEASUREMENT OF STACHYBOTRYS CHARTARUM CONIDIA USING REAL TIME DETECTION OF PCR PRODUCTS WITH THE TAQMAN TM FLUOROGENIC PROBE SYSTEM

    EPA Science Inventory

    The occurence of Stachybotrys chartarum in indoor environments has been associated with a number of human health concerns, including fatal pulmonary haemosiderosis in infants. Currently used culture-based and microscopic methods of fungal species identification are poorly suited ...

  12. Recovery of viability and radiation resistance by heat-injured conidia of Penicillium expansum Lk. ex Thom.

    PubMed

    Baldy, R W; Sommer, N F; Buckley, P M

    1970-05-01

    Spores heated in water at 54 C for up to 1 hr were plated on nutrient agar immediately or held for 3 days in aerated water at 23 C and then plated. Under these conditions, holding was optimal for recovery, increasing survival percentage up to 20-fold over values for immediate plating. Recovery was prevented partially or completely, however, when spores were held in any of the following solutions: glucose, potassium phosphate, ammonium or sodium acetate, sodium azide, or 2,4-dinitrophenol, or in the sodium or potassium salts of pyruvate, and tricarboxylic acid cycle acids. Both anaerobiosis and incubation at 0 C prevented recovery. Survivors of a heat treatment were more sensitive to gamma radiation than were unheated spores. Conditions which affected the recovery of viability had the same effect on restoration of radiation resistance. Thus, many of the processes for restoration of radiation resistance seem involved also in recovery of viability after heating. After a 99% inactivating treatment (about 30 min at 54 C), heated spores respired as fast as unheated spores, or faster. Malate, citrate, succinate, and acetate stimulated respiration in unheated spores and inhibited it in heated spores.

  13. Species and ecological diversity within the Cladosporium cladosporioides complex (Davidiellaceae, Capnodiales)

    PubMed Central

    Bensch, K.; Groenewald, J.Z.; Dijksterhuis, J.; Starink-Willemse, M.; Andersen, B.; Summerell, B.A.; Shin, H.-D.; Dugan, F.M.; Schroers, H.-J.; Braun, U.; Crous, P.W.

    2010-01-01

    The genus Cladosporium is one of the largest genera of dematiaceous hyphomycetes, and is characterised by a coronate scar structure, conidia in acropetal chains and Davidiella teleomorphs. Based on morphology and DNA phylogeny, the species complexes of C. herbarum and C. sphaerospermum have been resolved, resulting in the elucidation of numerous new taxa. In the present study, more than 200 isolates belonging to the C. cladosporioides complex were examined and phylogenetically analysed on the basis of DNA sequences of the nuclear ribosomal RNA gene operon, including the internal transcribed spacer regions ITS1 and ITS2, the 5.8S nrDNA, as well as partial actin and translation elongation factor 1-α gene sequences. For the saprobic, widely distributed species Cladosporium cladosporioides, both a neotype and epitype are designated in order to specify a well established circumscription and concept of this species. Cladosporium tenuissimum and C. oxysporum, two saprobes abundant in the tropics, are epitypified and shown to be allied to, but distinct from C. cladosporioides. Twenty-two species are newly described on the basis of phylogenetic characters and cryptic morphological differences. The most important phenotypic characters for distinguishing species within the C. cladosporioides complex, which represents a monophyletic subclade within the genus, are shape, width, length, septation and surface ornamentation of conidia and conidiophores; length and branching patterns of conidial chains and hyphal shape, width and arrangement. Many of the treated species, e.g., C. acalyphae, C. angustisporum, C. australiense, C. basiinflatum, C. chalastosporoides, C. colocasiae, C. cucumerinum, C. exasperatum, C. exile, C. flabelliforme, C. gamsianum, and C. globisporum are currently known only from specific hosts, or have a restricted geographical distribution. A key to all species recognised within the C. cladosporioides complex is provided. PMID:20877444

  14. Potential of two entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae (Coleoptera: Scarabaeidae), as biological control agents against the June beetle

    PubMed Central

    Erler, Fedai; Ates, A. Ozgur

    2015-01-01

    The aim of this study was to evaluate the effectiveness of the entomopathogenic fungi (EPF), Beauveria bassiana (Bals.) Vuill. (Deuteromycotina: Hyphomycetes) strain PPRI 5339 [BroadBand, an emulsifiable spore concentrate (EC) formulation] and Metarhizium anisopliae (Metsch.) Sorokin (Hypocreales: Clavicipitaceae) strain F52 [Met52, both EC and granular (GR) formulations] against the larvae of Polyphylla fullo (L.) (Coleoptera: Scarabaeidae). Larvicidal bioassays were performed in foam boxes (100 by 75 by 50 cm; length by width by height), containing moist soil medium with some humus and potato tubers as food. Although the B. bassiana product (min. 4 × 109 conidia/ml) was applied at 100, 150, and 200 ml/100 l water; M. anisopliae strain F52 was applied at 500, 1,000, and 1,500 g/m3 of moist soil medium for GR (9 × 108 cfu/g) and 75, 100, and 125 ml/100 l water for EC (5.5 × 109 conidia/ml) formulation. Both fungi were pathogenic to larvae of the pest; however, young larvae (1st and 2nd instars) were more susceptible to infection than older ones (3rd instar). Mortality rates of young and older larvae varied with conidial concentration of both fungi and elapsed time after application. The B. bassiana product was more effective than both of the formulations of the M. anisopliae product, causing mortalities up to 79.8 and 71.6% in young and older larvae, respectively. The highest mortality rates of young and older larvae caused by the M. anisopliae product were 74.1 and 67.6% for the GR formulation, 70.2 and 61.8% for the EC formulation, respectively. These results may suggest that both fungi have potential to be used for management of P. fullo. PMID:25881632

  15. Potential of two entomopathogenic fungi, Beauveria bassiana and Metarhizium anisopliae (Coleoptera: Scarabaeidae), as biological control agents against the June beetle.

    PubMed

    Erler, Fedai; Ates, A Ozgur

    2015-01-01

    The aim of this study was to evaluate the effectiveness of the entomopathogenic fungi (EPF), Beauveria bassiana (Bals.) Vuill. (Deuteromycotina: Hyphomycetes) strain PPRI 5339 [BroadBand, an emulsifiable spore concentrate (EC) formulation] and Metarhizium anisopliae (Metsch.) Sorokin (Hypocreales: Clavicipitaceae) strain F52 [Met52, both EC and granular (GR) formulations] against the larvae of Polyphylla fullo (L.) (Coleoptera: Scarabaeidae). Larvicidal bioassays were performed in foam boxes (100 by 75 by 50 cm; length by width by height), containing moist soil medium with some humus and potato tubers as food. Although the B. bassiana product (min. 4 × 10(9) conidia/ml) was applied at 100, 150, and 200 ml/100 l water; M. anisopliae strain F52 was applied at 500, 1,000, and 1,500 g/m(3) of moist soil medium for GR (9 × 10(8) cfu/g) and 75, 100, and 125 ml/100 l water for EC (5.5 × 10(9) conidia/ml) formulation. Both fungi were pathogenic to larvae of the pest; however, young larvae (1st and 2nd instars) were more susceptible to infection than older ones (3rd instar). Mortality rates of young and older larvae varied with conidial concentration of both fungi and elapsed time after application. The B. bassiana product was more effective than both of the formulations of the M. anisopliae product, causing mortalities up to 79.8 and 71.6% in young and older larvae, respectively. The highest mortality rates of young and older larvae caused by the M. anisopliae product were 74.1 and 67.6% for the GR formulation, 70.2 and 61.8% for the EC formulation, respectively. These results may suggest that both fungi have potential to be used for management of P. fullo.

  16. Common but different: The expanding realm of Cladosporium.

    PubMed

    Bensch, K; Groenewald, J Z; Braun, U; Dijksterhuis, J; de Jesús Yáñez-Morales, M; Crous, P W

    2015-09-01

    The genus Cladosporium (Cladosporiaceae, Dothideomycetes), which represents one of the largest genera of dematiaceous hyphomycetes, has been intensively investigated during the past decade. In the process, three major species complexes (C. cladosporioides, C. herbarum and C. sphaerospermum) were resolved based on morphology and DNA phylogeny, and a monographic revision of the genus (s. lat.) published reflecting the current taxonomic status quo. In the present study a further 19 new species are described based on phylogenetic characters (nuclear ribosomal RNA gene operon, including the internal transcribed spacer regions ITS1 and ITS2, as well as partial actin and translation elongation factor 1-α gene sequences) and morphological differences. For a selection of the species with ornamented conidia, scanning electron microscopic photos were prepared to illustrate the different types of surface ornamentation. Surprisingly, during this study Cladosporium ramotenellum was found to be a quite common saprobic species, being widely distributed and occurring on various substrates. Therefore, an emended species description is provided. Furthermore, the host range and distribution data for several previously described species are also expanded.

  17. Pathogenicity of the Fungus, Aspergillus clavatus, Isolated from the Locust, Oedaleus senegalensis, Against Larvae of the Mosquitoes Aedes aegypti, Anopheles gambiae and Culex quinquefasciatus

    PubMed Central

    Seye, Fawrou; Faye, Oumar; Ndiaye, Mady; Njie, Ebrima; Marie Afoutou, José

    2009-01-01

    The use of insect pathogenic fungi is a promising alternative to chemical control against mosquitoes. Among the Hyphomycetes isolated from insects for mosquito control, the genus Aspergillus remains the least studied. In September 2005, four fungi were isolated from the Senegalese locust, Oedaleus senegalensis Kraus (Orthoptera: Acrididae), collected in Dakar, Senegal. One of these fungi, identified as Aspergillus clavatus, Desmazières (Eurotiales: Trichocomaceae) was highly pathogenic against larvae of the mosquitoes Aedes aegypti L., Anopheles gambiae s.l. Giles and Culex quinquefasciatus Say (Diptera: Culicidae). An application of 1.2 mg/ml dry conidia yielded 100% mortality after 24 hours against both Ae. aegypti and Cx. quinquefasciatus while with An. gambiae it was 95%. With unidentified species in the genus Aspergillus, mortality after 24 h was <5% against all the larval species. Application of A. clavatus produced in a wheat powder medium using doses ranging between 4.3 to 21×107 spores/ml, caused 11 to 68% mortality against Cx. quinquefasciatus at 24h, and 37 to 100% against Ae. aegypti. Microscopic observations showed fungal germination on both Ae. aegypti and Cx. quinquefasciatus larvae. Histological studies revealed that A. clavatus penetrated the cuticle, invaded the gut and disintegrated its cells. Some Cx. quinquefasciatus larvae, treated with A. clavatus reached the pupal stage and produced infected adults. However, the infection was mainly located on the extremity of their abdomen. These results suggest that A. clavatus could be an effective tool to manage mosquito proliferation. PMID:20050773

  18. Pathogenicity of the Fungus, Aspergillus clavatus, isolated from the locust, Oedaleus senegalensis, against larvae of the mosquitoes Aedes aegypti, Anopheles gambiae and Culex quinquefasciatus.

    PubMed

    Seye, Fawrou; Faye, Oumar; Ndiaye, Mady; Njie, Ebrima; Marie Afoutou, José

    2009-01-01

    The use of insect pathogenic fungi is a promising alternative to chemical control against mosquitoes. Among the Hyphomycetes isolated from insects for mosquito control, the genus Aspergillus remains the least studied. In September 2005, four fungi were isolated from the Senegalese locust, Oedaleus senegalensis Kraus (Orthoptera: Acrididae), collected in Dakar, Senegal. One of these fungi, identified as Aspergillus clavatus, Desmazières (Eurotiales: Trichocomaceae) was highly pathogenic against larvae of the mosquitoes Aedes aegypti L., Anopheles gambiae s.l. Giles and Culex quinquefasciatus Say (Diptera: Culicidae). An application of 1.2 mg/ml dry conidia yielded 100% mortality after 24 hours against both Ae. aegypti and Cx. quinquefasciatus while with An. gambiae it was 95%. With unidentified species in the genus Aspergillus, mortality after 24 h was <5% against all the larval species. Application of A. clavatus produced in a wheat powder medium using doses ranging between 4.3 to 21x107 spores/ml, caused 11 to 68% mortality against Cx. quinquefasciatus at 24h, and 37 to 100% against Ae. aegypti. Microscopic observations showed fungal germination on both Ae. aegypti and Cx. quinquefasciatus larvae. Histological studies revealed that A. clavatus penetrated the cuticle, invaded the gut and disintegrated its cells. Some Cx. quinquefasciatus larvae, treated with A. clavatus reached the pupal stage and produced infected adults. However, the infection was mainly located on the extremity of their abdomen. These results suggest that A. clavatus could be an effective tool to manage mosquito proliferation.

  19. Diversity of endophytic fungi in the roots of mangrove species on the west coast of India.

    PubMed

    Ananda, K; Sridhar, K R

    2002-10-01

    Because mangrove plant species are a valuable source of useful metabolites, their endophytes have gained more importance. Randomly sampled surface-sterilized whole root segments of four mangrove plant species, Acanthus ilicifolius, Avicennia officinalis, Rhizophora mucronata, and Sonneratia caseolaris from the mangroves of Udyavara (Karnataka) on the west coast of India, were characterized for fungal communities by direct plating, damp chamber, and bubbling chamber incubation methods. The richness of endophytic fungal species from whole root segments after direct plating and damp chamber incubation was greatest for R. mucronata than for other plants (18 vs. 8-13). Incubation of whole root segments in bubbling chambers yielded conidia of two freshwater hyphomycetes: Mycocentrospora acerina (in Avicennia officinalis) and Triscelophorus acuminatus (in R. mucronata and in S. caseolaris). Surface-sterilized whole root and root bark segments of R. mucronata sampled from the mid-tide level on direct plating yielded more fungi than that of the root segments sampled from low-tide and high-tide levels. The greatest number of isolates, species richness, and diversity of fungi were shown by the whole root segments of R. mucronata from the mid-tide level. Rarefaction indices also revealed the highest expected number of species out of 150 random isolations from the mid-tide level samples of whole root and root bark segments of R. mucronata. The present study showed that fungi in mangrove roots are composed of a consortium of soil, marine, and freshwater fungi.

  20. Nutrient enrichment in water more than in leaves affects aquatic microbial litter processing.

    PubMed

    Biasi, Cristiane; Graça, Manuel A S; Santos, Sandro; Ferreira, Verónica

    2017-06-01

    Nutrient enrichment of soils and water will intensify in the future and has the potential to alter fundamental ecosystem processes, such as litter decomposition. We tested the direct (via water nutrient enrichment) and indirect (via changes in leaf chemistry) effects of nutrient enrichment on microbial activity and decomposability of Quercus robur L. (oak) leaves in laboratory microcosms simulating streams. Senescent leaves of oak trees grown without and with fertilization were incubated under ambient and elevated water nutrient [nitrogen (N) and phosphorus (P)] concentrations for 60 days. Soil fertilization led to an increase in leaf (3.4×) and leaf litter (2.3×) N concentration. Increased water-dissolved nutrients concentrations stimulated microbial activity (N uptake, microbial respiration, fungal biomass buildup and conidia production by aquatic hyphomycetes) that translated into accelerated litter decomposition (2.1× for unfertilized and 1.6× for fertilized trees). Leaves from fertilized trees had higher microbial activity and decomposition rates than leaves from unfertilized trees only at low dissolved nutrient availability. When both litter and water nutrients concentration increased, microbial activity and leaf decomposition were stimulated, but the effects were additive and direct effects from increased dissolved nutrient availability were stronger than those mediated by increases in litter N concentration (indirect effects). Our results suggest that increases in water nutrient availability (within the range used in this study) may exert a stronger control on microbial activity and litter decomposition than litter nutrient enrichment.

  1. Taxa-area relationship of aquatic fungi on deciduous leaves

    PubMed Central

    Cássio, Fernanda; Pascoal, Cláudia; Bärlocher, Felix

    2017-01-01

    One of the fundamental patterns in macroecology is the increase in the number of observed taxa with size of sampled area. For microbes, the shape of this relationship remains less clear. The current study assessed the diversity of aquatic fungi, by the traditional approach based on conidial morphology (captures reproducing aquatic hyphomycetes) and next generation sequencing (NGS; captures other fungi as well), on graded sizes of alder leaves (0.6 to 13.6 cm2). Leaves were submerged in two streams in geographically distant locations: the Oliveira Stream in Portugal and the Boss Brook in Canada. Decay rates of alder leaves and fungal sporulation rates did not differ between streams. Fungal biomass was higher in Boss Brook than in Oliveira Stream, and in both streams almost 100% of the reads belonged to active fungal taxa. In general, larger leaf areas tended to harbour more fungi, but these findings were not consistent between techniques. Morphospecies-based diversity increased with leaf area in Boss Brook, but not in Oliveira Stream; metabarcoding data showed an opposite trend. The higher resolution of metabarcoding resulted in steeper taxa-accumulation curves than morphospecies-based assessments (fungal conidia morphology). Fungal communities assessed by metabarcoding were spatially structured by leaf area in both streams. Metabarcoding promises greater resolution to assess biodiversity patterns in aquatic fungi and may be more accurate for assessing taxa-area relationships and local to global diversity ratios. PMID:28719634

  2. Studies on mould growth and biomass production using waste banana peel.

    PubMed

    Essien, J P; Akpan, E J; Essien, E P

    2005-09-01

    Hyphomycetous (Aspergillus fumigatus) and Phycomycetous (Mucor hiemalis) moulds were cultivated in vitro at room temperature (28 + 20 degrees C) to examined their growth and biomass production on waste banana peel agar (BPA) and broth (BPB) using commercial malt extract agar (MEA) and broth (MEB) as control. The moulds grew comparatively well on banana peel substrates. No significant difference (p > 0.05) in radial growth rates was observed between moulds cultivated on PBA and MEA, although growth rates on MEA were slightly better. Slight variations in sizes of asexual spores and reproductive hyphae were also observed between moulds grown on MEA and BPA. Smaller conidia and sporangiospores, and shorter aerial hyphae (conidiophores and sporangiophores) were noticed in moulds grown on BPA than on MEA. The biomass weight of the test moulds obtained after one month of incubation with BPB were only about 1.8 mg and 1.4 mg less than values recorded for A. fumigatus and M. hiemalis respectively, grown on MEB. The impressive performance of the moulds on banana peel substrate may be attributed to the rich nutrient (particularly the crude protein 7.8% and crude fat 11.6% contents) composition of banana peels. The value of this agricultural waste can therefore be increased by its use not only in the manufacture of mycological medium but also in the production of valuable microfungal biomass which is rich in protein and fatty acids.

  3. [Isolation, identification and anticancer activity of an endophytic fungi from Juglans mandshurica].

    PubMed

    Li, Meiya; Wu, Yunwei; Jiang, Fusheng; Yu, Xiangli; Tang, Kexuan; Miao, Zhiqi

    2009-07-01

    The endophytic fungus named FSN006 was isolated from the inner bark of Juglans mandshurica. It grew quickly and formed circular colony on PDA plate. The upper side of the colony was white, while the lower side of the colony and the conditioned medium were light yellow as a result of significant yellow pigment substances were produced and secreted by the fungi. Green elliptic conidia appeared when cultured on CMX plate. Based on the morphology identification and ITS sequence, it was clear that this fungus belonged to the Deuteromycotina, HyPhomycetes, Moniliales, Trichoderma longibrachiatum. The conditioned medium of FSN006 showed a high anti-tumor ability against liver cancer cell-HepG2, and reached its IC50 concentration after being diluted 20 times, while the IC50 concentration of curcumine was(11.49 +/- 0.12) mg x L(-1). In addition, there was preeminent selective inhibiting effect against the normal liver cell strain HL-7702 and its caner counter strain HepG2. The inhibiting effect against strain HL-7702 was only one quarter of that against HepG2 at the concentration of IC50. Therefore, the fermentation of FSN006 may provide a possible way to produce anticancer drug with higher efficiency and lower toxicity.

  4. Common but different: The expanding realm of Cladosporium

    PubMed Central

    Bensch, K.; Groenewald, J.Z.; Braun, U.; Dijksterhuis, J.; de Jesús Yáñez-Morales, M.; Crous, P.W.

    2015-01-01

    The genus Cladosporium (Cladosporiaceae, Dothideomycetes), which represents one of the largest genera of dematiaceous hyphomycetes, has been intensively investigated during the past decade. In the process, three major species complexes (C. cladosporioides, C. herbarum and C. sphaerospermum) were resolved based on morphology and DNA phylogeny, and a monographic revision of the genus (s. lat.) published reflecting the current taxonomic status quo. In the present study a further 19 new species are described based on phylogenetic characters (nuclear ribosomal RNA gene operon, including the internal transcribed spacer regions ITS1 and ITS2, as well as partial actin and translation elongation factor 1-α gene sequences) and morphological differences. For a selection of the species with ornamented conidia, scanning electron microscopic photos were prepared to illustrate the different types of surface ornamentation. Surprisingly, during this study Cladosporium ramotenellum was found to be a quite common saprobic species, being widely distributed and occurring on various substrates. Therefore, an emended species description is provided. Furthermore, the host range and distribution data for several previously described species are also expanded. PMID:26955200

  5. [Infectivity of the entomopathogenic fungus Verticillium lecanii in mice and guinea pigs].

    PubMed

    Mier, T; Rivera, F; Rodríguez-Ponce, M P; Carrillo-Farga, J; Toriello, C

    1994-01-01

    The resistance of plague insects to chemical insecticides as well as the importance of a healthy environment demands an alternative for agricultural plagues. Among others, biological control seems an alternate strategy with fungal entomopathogens playing a relevant role. The hyphomycete Verticillium lecanii is a natural bioregulator of aphids, scales and white-flies that attack different agricultural plantations. Its use in biological control programs must be assessed previously by safety procedures such as its innocuity in mammals and useful animals and plants. The aim of this study was pointed at demonstrating the innocuity of V. lecanii in mice and guinea pigs. Two strains of the fungus were injected intraperitoneally (10(8) conidia/kg of animal weight) to 130 mice and 66 guinea pigs. Two control groups were included, one injected with heat-killed fungi and the other with sterile physiological saline. The animals were killed at 8, 30 and 70 days after infection, and mycological and histopathological studies performed in their organs. Negative results obtained with the live fungus in the same manner as the two control groups, suggest the innocuity of V. lecanii in mice and guinea pigs.

  6. [Cultural Properties and Taxonomic Position of Helminthosporium-Like Fungal Isolates from the White Sea].

    PubMed

    Kireev, Ya V; Konovalova, O P; Myuge, N S; Shnyreva, A V; Bubnova, E N

    2015-01-01

    Morphological, cultural and physiological characteristics of 19 Helminthosporium-like hyphomycetes isolated from the White Sea were studied. Taxonomic status of the isolates was verified using molecular genetics techniques. One of the isolates was identified as Alternatia sp., while the rest of marine isolates belonged to the species Paradendriphiella salina (G.K. Sutherl.) Woudenb. & Crous. The specific features of the isolates studied were characterized as adaptive. Optimum salinity for their growth was 1-2% NaCl, which is lower than the value for the known open ocean isolates. This is probably due to relatively low salinity of White Sea (22-24 per thousand) as compared with the ocean water (35 per thousand). While the temperature optimum for growth was 22 per thousand, growth and sporulation occurred at 6 degrees C, which has not been reported for marine fungi isolated from warmer seawater. All isolates studied grew and sporulated efficiently on the medium supplied with the Fucus algae extract and in the sea water layer. Conidia of the isolates submerged in the sea water were propagated efficiently, unlike the soil-born fungi. Holoblastic conidiogenesis was demonstrated by light and scanning electron microscopy, confirming the separation of P. salina from the genus Scolecobasidium.

  7. Ultrastructural and cytochemical characterization of brown soft scale Coccus hesperidum (Hemiptera: Coccidae) infected by the Lecanicillium lecanii (Ascomycota: Hypocreales).

    PubMed

    Liu, Weimin; Xie, Yingping; Xue, Jiaoliang; Zhang, Yanfeng; Zhang, Xiaomin

    2011-01-01

    The ultrastructural and cytochemical characterization of the brown soft scale, Coccus hesperidum L. (Hemiptera: Coccidae) infected by the hyphomycete Lecanicillium lecanii (Zimmermann) Gams & Zare, belonging to the phylum Ascomycota and order Hypocreales, was investigated by light, scanning and transmission electron microscopy. Gold cytochemistry was used to label chitin in the cuticle of the scale insect. The results revealed that the pathogenic fungus, L. lecanii generally infected by penetrating the integument, especially at anus, vulva, spiracles, stigmatic furrow, body margin, and the areas of cuticle with grooves, fissures and rugoses areas. The conidia became attached to the host body surface and germinated into hyphae that established colonies by branching repeatedly. Hyphae penetrated the integument by means of their penetration pegs using mechanical force and extracellular enzymes. During integument penetration, the hyphae extended vertically or parallel along the cuticle. Labeling with the WGA/Ovo-G complex showed disruption of the parallel sheets of chitin and a decrease in the density of the gold particles surrounding the penetrated hyphae. Hyphal invasion also separated the cuticle and epidermis from each other. Once in the haemocoele, blastospores of the fungus infected the haemocytes and internal organs. After some time, the nutritive value of the haemocoele decreased and the insect's internal organs disappeared. The hyphae then produced conidiophores and released them through the cuticle of the scale insect cadaver.

  8. Taxa-area relationship of aquatic fungi on deciduous leaves.

    PubMed

    Duarte, Sofia; Cássio, Fernanda; Pascoal, Cláudia; Bärlocher, Felix

    2017-01-01

    One of the fundamental patterns in macroecology is the increase in the number of observed taxa with size of sampled area. For microbes, the shape of this relationship remains less clear. The current study assessed the diversity of aquatic fungi, by the traditional approach based on conidial morphology (captures reproducing aquatic hyphomycetes) and next generation sequencing (NGS; captures other fungi as well), on graded sizes of alder leaves (0.6 to 13.6 cm2). Leaves were submerged in two streams in geographically distant locations: the Oliveira Stream in Portugal and the Boss Brook in Canada. Decay rates of alder leaves and fungal sporulation rates did not differ between streams. Fungal biomass was higher in Boss Brook than in Oliveira Stream, and in both streams almost 100% of the reads belonged to active fungal taxa. In general, larger leaf areas tended to harbour more fungi, but these findings were not consistent between techniques. Morphospecies-based diversity increased with leaf area in Boss Brook, but not in Oliveira Stream; metabarcoding data showed an opposite trend. The higher resolution of metabarcoding resulted in steeper taxa-accumulation curves than morphospecies-based assessments (fungal conidia morphology). Fungal communities assessed by metabarcoding were spatially structured by leaf area in both streams. Metabarcoding promises greater resolution to assess biodiversity patterns in aquatic fungi and may be more accurate for assessing taxa-area relationships and local to global diversity ratios.

  9. Selection of Wetting Agent for Dried Conidia of two Beauveria bassiana Strains from Different Production Systems to Control Tarnished Plant Bugs

    USDA-ARS?s Scientific Manuscript database

    The hydrophilic–lipophilic balance (HLB) number was used to select surfactant, TDA (polyoxy-ethylene tridecyl ether) with a range of HLB numbers in formulation development of two Beauveria bassiana strains, NI8 and GHA produced from different systems. The results indicated that optimal HLB number of...

  10. A RAPID METHOD FOR THE EXTRACTION OF FUNGAL DNA FROM ENVIRONMENTAL SAMPLES: EVALUATION IN THE QUANTITATIVE ANALYSIS OF MEMNONIELLA ECHINATA CONIDIA USING REAL TIME DETECTION OF PCR PRODUCTS

    EPA Science Inventory

    New technologies are creating the potential for using nucleic acid sequence detection to perform routine microbiological analyses of environmental samples. Our laboratory has recently reported on the development of a method for the quantitative detection of Stachybotrys chartarum...

  11. Application of modified atmosphere packaging (gas flushing and active packaging) for extending the shelf life of Beauveria bassiana conidia at high temperatures

    USDA-ARS?s Scientific Manuscript database

    Limited shelf life has long been a major constraint to the development of fungus-based bioinsecticides (mycoinsecticides). Fungal spores comprising the active ingredients of most products typically lose viability within a few months when stored in conventional packaging at temperatures >30 deg C. Me...

  12. Virulence of Mexican isolates of entomopathogenic fungi upon Rhipicephalus-Boophilus microplus larvae and the efficacy of conidia formulations to reduce larval tick density under field conditions

    USDA-ARS?s Scientific Manuscript database

    The first objective was laboratory evaluation of the virulence of 53 Mexican isolates of fungi against larvae of Rhipicephalus (Boophilus) microplus. Thirty three isolates of Metarhizium anisopliae var. anisopliae (Metschnickoff) Sorokin (Hypocreales: Clavicipitaceae) and 20 isolates of Isaria (Paec...

  13. Loss of msnA, a Putative Stress Regulatory Gene, in Aspergillus parasiticus and Aspergillus flavus Increased Production of Conidia, Aflatoxins and Kojic Acid

    PubMed Central

    Chang, Perng-Kuang; Scharfenstein, Leslie L.; Luo, Meng; Mahoney, Noreen; Molyneux, Russell J.; Yu, Jiujiang; Brown, Robert L.; Campbell, Bruce C.

    2011-01-01

    Production of the harmful carcinogenic aflatoxins by Aspergillus parasiticus and Aspergillus flavus has been postulated to be a mechanism to relieve oxidative stress. The msnA gene of A. parasiticus and A. flavus is the ortholog of Saccharomyces cerevisiae MSN2 that is associated with multi-stress response. Compared to wild type strains, the msnA deletion (∆msnA) strains of A. parasiticus and A. flavus exhibited retarded colony growth with increased conidiation. The ∆msnA strains also produced slightly higher amounts of aflatoxins and elevated amounts of kojic acid on mixed cereal medium. Microarray assays showed that expression of genes encoding oxidative stress defense enzymes, i.e., superoxide dismutase, catalase, and cytochrome c peroxidase in A. parasiticus ∆msnA, and the catalase A gene in A. flavus ∆msnA, was up-regulated. Both A. parasiticus and A. flavus ∆msnA strains produced higher levels of reactive oxygen species (ROS), and ROS production of A. flavus msnA addback strains was decreased to levels comparable to that of the wild type A. flavus. The msnA gene appears to be required for the maintenance of the normal oxidative state. The impairment of msnA resulted in the aforementioned changes, which might be used to combat the increased oxidative stress in the cells. PMID:22069691

  14. A RAPID METHOD FOR THE EXTRACTION OF FUNGAL DNA FROM ENVIRONMENTAL SAMPLES: EVALUATION IN THE QUANTITATIVE ANALYSIS OF MEMNONIELLA ECHINATA CONIDIA USING REAL TIME DETECTION OF PCR PRODUCTS

    EPA Science Inventory

    New technologies are creating the potential for using nucleic acid sequence detection to perform routine microbiological analyses of environmental samples. Our laboratory has recently reported on the development of a method for the quantitative detection of Stachybotrys chartarum...

  15. Interactions between stream fungi and bacteria associated with decomposing leaf litter at different levels of nutrient availability

    Treesearch

    Vladislav Gulis; Keller Suberkropp

    2003-01-01

    We examined the potential for interactions between aquatic hyphomycetes and bacteria isolated from leaves decaying in a headwater stream. In agar plate assays, culture filtrates of each of 28 aquatic hyphomycete isolates tested (5 species) inhibited bacterial growth (16 Gram-negative bacterial isolates belonging to 6 colony morphotypes were tested). Inhibition of...

  16. Assessment of Fungal Pathogens as Biocontrol Agents of Myriophyllum Spicatum.

    DTIC Science & Technology

    1996-01-01

    Gliocladium roseum, 2 Indeterminate Hyphomycete (producing only chlamydospores), Acremoniurn sp, Cylindrocarpon destructans, Embeilisia nr. telituster, Fusarium ... solani , Geotrchum candidum Coniothyrium fuckelli, Cryptosporiopsis sp, Glomerella cingulata and an Indeterminate Coelomycete.

  17. Evaluation of Metarhizium anisopliae, Beauveria bassiana and Paecilomyces fumosoroseus as entomopathogens of the cactus moth, Cactoblastis cactorum (Lepidoptera:Pyralidae)

    USDA-ARS?s Scientific Manuscript database

    The fungal pathogens Metarhizium anisopliae (Metchnikoff) Sorokin (Hypocreales: Clavicipitaceae), Paecilomyces fumosoroseus (Wize) Brown & Smith (Deuteromycotina: Hyphomycetes), and Beauveria bassiana (Bals.-Criv.) Vuill. (Hypocreales: Clavicipitaceae) were evaluated as potential biological control ...

  18. Fungal propagules and DNA in feces of two detritus-feeding amphipods.

    PubMed

    Sridhar, Kandikere Ramaiah; Beaton, Margaret; Bärlocher, Felix

    2011-01-01

    Aquatic shredders (leaf-eating invertebrates) preferentially ingest and digest leaves colonized by aquatic hyphomycetes (fungi). This activity destroys leaf-associated fungal biomass and detritial resources in streams. Fungal counter-adaptations may include the ability to survive passage through the invertebrate's digestive tract. When fecal pellets of Gammarus tigrinus and Hyalella azteca were incubated with sterile leaves, spores of nine (G. tigrinus) and seven (H. azteca) aquatic hyphomycete species were subsequently released from the leaves, indicating the presence of viable fungal structures in the feces. Extraction, amplification, and sequencing of DNA from feces revealed numerous fungal phylotypes, two of which could be assigned unequivocally to an aquatic hyphomycete. The estimated contributions of major fungal groups varied depending on whether 18S or ITS sequences were amplified and cloned. We conclude that a variable proportion of fungal DNA in the feces of detritivores may originate from aquatic hyphomycetes. Amplified DNA may be associated with metabolically active, dormant, or dead fungal cells.

  19. Do Invertebrate Activity and Current Velocity Affect Fungal Assemblage Structure in Leaves?

    NASA Astrophysics Data System (ADS)

    Ferreira, Verónica; Graça, Manuel A. S.

    2006-02-01

    In this study we assessed the effect of current velocity and shredder presence, manipulated in artificial channels, on the structure of the fungal assemblage colonizing alder (Alnus glutinosa (L.) Gaertner) leaves incubated in coarse and fine mesh bags. Fungal sporulation rates, cumulative conidial production and number of species of aquatic hyphomycetes were higher in leaves exposed to high rather than to low current velocity. The opposite was observed regarding Simpson's index (D) on the fungal assemblage. Some species of aquatic hyphomycetes were consistently stimulated in high current channels. No effect of shredders or of mesh type was observed.

  20. Susceptibility of larvae of Galleria mellonella to infection by Aspergillus fumigatus is dependent upon stage of conidial germination.

    PubMed

    Renwick, Julie; Daly, Paul; Reeves, Emer P; Kavanagh, Kevin

    2006-06-01

    The ability of conidia of the human pathogenic fungus Aspergillus fumigatus to kill larvae of the insect Galleria mellonella was investigated. Conidia at different stages of the germination process displayed variations in their virulence as measured using the Galleria infection model. Non-germinating ('resting') conidia were avirulent except when an inoculation density of 1 x 10(7) conidia per insect was used. Conidia that had been induced to commence the germination process by pre-culturing in growth medium for 3 h were capable of killing larvae at densities of 1 x 10(6) and 1 x 10(7) per insect. An inoculation density of 1 x 10(5) conidia per insect remained avirulent. Conidia in the outgrowth phase of germination (characterised as the formation of a germ tube) were the most virulent and were capable of killing 100% of larvae after 5 or 24 h when 1 x 10(7) or 1 x 10(6) conidia, that had been allowed to germinate for 24 h, were used. Examination of the response of insect haemocytes to conidia at different stages of the germination process established that haemocytes could engulf non-germinating conidia and those in the early stages of the germination process but that conidia, which had reached the outgrowth stages of germination were not phagocytosed. The results presented here indicate that haemocytes of G. mellonella are capable of phagocytosing A. fumigatus conidia less than 3.0 microm in diameter but that conidia greater than this are too large to be engulfed. The virulence of A. fumigatus in G. mellonella larvae can be ascertained within 60-90 h if infection densities of 1 x 10(6) or 1 x 10(7) activated conidia (pre-incubated for 2-3 h) per insect are employed.

  1. Microcycle Conidiation in Cercospora zeae-maydis.

    PubMed

    Lapaire, Carrie L; Dunkle, Larry D

    2003-02-01

    ABSTRACT Conidia of Cercospora zeae-maydis are the primary inoculum causing gray leaf spot of maize. On nutrient-deficient substrates, but not on water on the leaf surface, conidia germinate and develop secondary conidia on conidiophores produced from germ tubes or conidial cells. A population of conidia increases its numbers more than twofold by 2 days on the surface of a water droplet and by fourfold on trichomes. This microcycle conidiation is suppressed by hydrogen peroxide and ammonium compounds but not by nitrate compounds, amino acids, or simple sugars. Microcycle conidiation is sensitive to alpha-amanitin and cycloheximide, suggesting that new RNA and proteins must be synthesized. Upon transfer from a humid to a dry atmosphere, secondary conidia and conidiophores dehydrate and collapse. Mature, dehydrated, secondary conidia are liberated by wind speeds approximately one-third those required to liberate hydrated conidia. The dispersed secondary conidia can rehydrate and germinate normally. Because this microcycle conidiation occurs at the expense of endogenous reserves, the ability to produce secondary conidia is lost after four successive cycles without a period of growth on nutrient media. This alternative method of maintaining inoculum potential during periods of fluctuating relative humidity may have epidemiological consequences when primary conidia fail to infect.

  2. Potassium influences forage bermudagrass yield and fungal leaf disease severity in Mississippi

    USDA-ARS?s Scientific Manuscript database

    Leaf spot diseases are associated with K deficiency in forage bermudagrass. In 2010, a natural disease epiphytotic caused by six species of Bipolaris, Curvularia, and Exserohilum (dematiaceous hyphomycetes) was evaluated in 56 plots of ‘Tifton 44' bermudagrass in Mississippi. Pathogen occurrence, di...

  3. Two new species of Stemphylium from Northwest China

    USDA-ARS?s Scientific Manuscript database

    During surveys of hyphomycetes from the Northwest of China, two undescribed taxa of Stemphylium were discovered from diseased leaves of Malus sieversii and Pyrus sinkiangensis in Sinkiang Provence, which are described as S. mali and S. pyrinum. The two species differ from similar species morphologi...

  4. Stachbotrys Root Rot

    USDA-ARS?s Scientific Manuscript database

    Stachybotrys root rot is caused by Stachybotrys chartarum, a cellulytic saprophytic hyphomycete fungus. The pathogen produces mycotoxins including a host of immunosupressant compounds for human and is one of the causes of the "sick building syndrome." Although S. chartarum is rarely known as a plan...

  5. Biological control of Spreading Dayflower (Commelina diffusa) with the fungal pathogen Phoma commelinicola

    USDA-ARS?s Scientific Manuscript database

    Greenhouse and field experiments showed that conidia of the fungal pathogen, Phoma commelinicola exhibited bioherbicidal activity on spreading dayflower (Commelina diffusa) seedlings when applied at concentrations of 106 to 109 conidia ml-1. Greenhouse tests determined an optimal temperature for co...

  6. Presence of adhesive vesicles in the mycoherbicide Alternaria helianthi

    USDA-ARS?s Scientific Manuscript database

    Alternaria helianthi conidia have been shown to cause disease on common cocklebur. Conidia were applied to slides made hydrophobic by coating with dimethyldicholorosilane (mimics leaf surface), then rinsed and treated with FITC-Con A to stain the adhesive material. Alternaria helianthi coni...

  7. Colonization by Cladosporium spp. of painted metal surfaces associated with heating and air conditioning systems

    NASA Technical Reports Server (NTRS)

    Ahearn, D. G.; Simmons, R. B.; Switzer, K. F.; Ajello, L.; Pierson, D. L.

    1991-01-01

    Cladosporium cladosporioides and C. hebarum colonized painted metal surfaces of covering panels and register vents of heating, air conditioning and ventilation systems. Hyphae penetrated the paint film and developed characteristic conidiophores and conidia. The colonies were tightly appressed to the metal surface and conidia were not readily detectable via standard air sampling procedures.

  8. FleA Expression in Aspergillus fumigatus Is Recognized by Fucosylated Structures on Mucins and Macrophages to Prevent Lung Infection

    PubMed Central

    Sinha, Meenal; McCabe, Orla; Palmer, Jonathan M.; Choera, Tsokyi; Yun Lim, Fang; Wimmerova, Michaela; Carrington, Stephen D.; Yuan, Shaopeng; Lowell, Clifford A.; Oscarson, Stefan; Keller, Nancy P.; Fahy, John V.

    2016-01-01

    The immune mechanisms that recognize inhaled Aspergillus fumigatus conidia to promote their elimination from the lungs are incompletely understood. FleA is a lectin expressed by Aspergillus fumigatus that has twelve binding sites for fucosylated structures that are abundant in the glycan coats of multiple plant and animal proteins. The role of FleA is unknown: it could bind fucose in decomposed plant matter to allow Aspergillus fumigatus to thrive in soil, or it may be a virulence factor that binds fucose in lung glycoproteins to cause Aspergillus fumigatus pneumonia. Our studies show that FleA protein and Aspergillus fumigatus conidia bind avidly to purified lung mucin glycoproteins in a fucose-dependent manner. In addition, FleA binds strongly to macrophage cell surface proteins, and macrophages bind and phagocytose fleA-deficient (∆fleA) conidia much less efficiently than wild type (WT) conidia. Furthermore, a potent fucopyranoside glycomimetic inhibitor of FleA inhibits binding and phagocytosis of WT conidia by macrophages, confirming the specific role of fucose binding in macrophage recognition of WT conidia. Finally, mice infected with ΔfleA conidia had more severe pneumonia and invasive aspergillosis than mice infected with WT conidia. These findings demonstrate that FleA is not a virulence factor for Aspergillus fumigatus. Instead, host recognition of FleA is a critical step in mechanisms of mucin binding, mucociliary clearance, and macrophage killing that prevent Aspergillus fumigatus pneumonia. PMID:27058347

  9. Morphology and molecular taxonomy of Evlachovaea-like fungi, and the status of this unusual conidial genus

    USDA-ARS?s Scientific Manuscript database

    The entomopathogenic anamorphic genus Evlachovaea was described to differ from other fungi in forming its conidia obliquely to the axis of the conidiogenous cell and with successive conidia having alternate orientations with a zipper- or chevron-like arrangement resulting in flat, ribbon-like chains...

  10. Ulocladium cantlous sp. nov. isolated from Northwest China: morphology and molecular phylogenetic position

    USDA-ARS?s Scientific Manuscript database

    A new species of Ulocladium was isolated from diseased leaves from two Cucumis spp. growing in Sinkiang and Gansu Provinces of China. Conidia were isolated from diseased leaf areas, grown in single-spore pure culture, and conidia harvested 1, 3, and 7 days after incubation for morphological comparis...

  11. Comparative efficacy of emulsifiable-oil, wettable-powder, and unformulated-powder preparations of Beauveria bassiana against the melon aphid Aphis gossypii

    USDA-ARS?s Scientific Manuscript database

    Aphicidal efficacy of two commercial formulations of Beauveria bassiana strain GHA conidia, an emulsifiable-paraffinic oil dispersion (OD) and a clay-based wettable powder (WP), were compared to unformulated conidia in laboratory tests with adult melon aphids (Aphis gossypii). For the initial 24 hou...

  12. Adhesion of the entomopathogenic fungus Beauveria (Cordyceps) bassiana to substrata.

    PubMed

    Holder, Diane J; Keyhani, Nemat O

    2005-09-01

    The entomopathogenic fungus Beauveria bassiana produces at least three distinct single-cell propagules, aerial conidia, vegetative cells termed blastospores, and submerged conidia, which can be isolated from agar plates, from rich broth liquid cultures, and under nutrient limitation conditions in submerged cultures, respectively. Fluorescently labeled fungal cells were used to quantify the kinetics of adhesion of these cell types to surfaces having various hydrophobic or hydrophilic properties. Aerial conidia adhered poorly to weakly polar surfaces and rapidly to both hydrophobic and hydrophilic surfaces but could be readily washed off the latter surfaces. In contrast, blastospores bound poorly to hydrophobic surfaces, forming small aggregates, bound rapidly to hydrophilic surfaces, and required a longer incubation time to bind to weakly polar surfaces than to hydrophilic surfaces. Submerged conidia displayed the broadest binding specificity, adhering to hydrophobic, weakly polar, and hydrophilic surfaces. The adhesion of the B. bassiana cell types also differed in sensitivity to glycosidase and protease treatments, pH, and addition of various carbohydrate competitors and detergents. The outer cell wall layer of aerial conidia contained sodium dodecyl sulfate-insoluble, trifluoroacetic acid-soluble proteins (presumably hydrophobins) that were not present on either blastospores or submerged conidia. The variations in the cell surface properties leading to the different adhesion qualities of B. bassiana aerial conidia, blastospores, and submerged conidia could lead to rational design decisions for improving the efficacy and possibly the specificity of entomopathogenic fungi for host targets.

  13. Conidial vigor vs. viability as predictors of virulence of entomopathogenic fungi

    USDA-ARS?s Scientific Manuscript database

    We tested the hypothesis that debilitated conidia exhibiting slow-germination (requiring greater than 16 hours to germinate) are less virulent than vigorous conidia exhibiting fast germination (requiring less than or equal to 16 hours to germinate). Preparations of Beauveria bassiana s.l. strain CG ...

  14. Adhesion of the Entomopathogenic Fungus Beauveria (Cordyceps) bassiana to Substrata†

    PubMed Central

    Holder, Diane J.; Keyhani, Nemat O.

    2005-01-01

    The entomopathogenic fungus Beauveria bassiana produces at least three distinct single-cell propagules, aerial conidia, vegetative cells termed blastospores, and submerged conidia, which can be isolated from agar plates, from rich broth liquid cultures, and under nutrient limitation conditions in submerged cultures, respectively. Fluorescently labeled fungal cells were used to quantify the kinetics of adhesion of these cell types to surfaces having various hydrophobic or hydrophilic properties. Aerial conidia adhered poorly to weakly polar surfaces and rapidly to both hydrophobic and hydrophilic surfaces but could be readily washed off the latter surfaces. In contrast, blastospores bound poorly to hydrophobic surfaces, forming small aggregates, bound rapidly to hydrophilic surfaces, and required a longer incubation time to bind to weakly polar surfaces than to hydrophilic surfaces. Submerged conidia displayed the broadest binding specificity, adhering to hydrophobic, weakly polar, and hydrophilic surfaces. The adhesion of the B. bassiana cell types also differed in sensitivity to glycosidase and protease treatments, pH, and addition of various carbohydrate competitors and detergents. The outer cell wall layer of aerial conidia contained sodium dodecyl sulfate-insoluble, trifluoroacetic acid-soluble proteins (presumably hydrophobins) that were not present on either blastospores or submerged conidia. The variations in the cell surface properties leading to the different adhesion qualities of B. bassiana aerial conidia, blastospores, and submerged conidia could lead to rational design decisions for improving the efficacy and possibly the specificity of entomopathogenic fungi for host targets. PMID:16151112

  15. Colonization by Cladosporium spp. of painted metal surfaces associated with heating and air conditioning systems

    NASA Technical Reports Server (NTRS)

    Ahearn, D. G.; Simmons, R. B.; Switzer, K. F.; Ajello, L.; Pierson, D. L.

    1991-01-01

    Cladosporium cladosporioides and C. hebarum colonized painted metal surfaces of covering panels and register vents of heating, air conditioning and ventilation systems. Hyphae penetrated the paint film and developed characteristic conidiophores and conidia. The colonies were tightly appressed to the metal surface and conidia were not readily detectable via standard air sampling procedures.

  16. Interaction of Aceria mangiferae with Fusarium mangiferae, the causal agent of mango malformation disease

    USDA-ARS?s Scientific Manuscript database

    The study examines the role of the mango bud mite, Aceria mangiferae, in carrying Fusarium mangiferae’s conidia, vectoring them into the penetration sites and assisting fungal penetration and dissemination. Conidia that were exposed to a green fluorescent protein (gfp)-marked isolate of F. mangifer...

  17. Variabilities of two Drechslerella dactyloides isolates in Korea and high predacity against Bursaphelenchus xylophilus.

    PubMed

    Wang, Zhen; Wang, Chun-Yan; Gu, Li-Juan; Sun, Bai-Shen; Zhang, Dong-Liang; Liu, Lei; Lee, Mi-Ra; Wang, Chun-Ling; Li, Zheng; Mo, Eun-Kyung; Sung, Chang-Keun

    2011-02-01

    Drechslerella dactyloides generates elongate ellipsoid conidia and three-celled rings. Recently, Drechslerella dactyloides CNU 091025 and CNU 091026 were isolated in Korea, which generated elongate ellipsoid, Y-shaped and reaphook-shaped conidia, three-celled rings and fishhook-shaped traps. Therefore, molecular phylogenetic analysis, morphological variability, and nematode capturing ability were tested in this study. CNU 091025 generated two-celled or three-celled Y-shaped conidia, reaphook-shaped conidia, and elongate ellipsoid conidia, 17.2, 40.9, and 41.9%, respectively. Some fishhook-shaped rings connecting together formed two-dimensional web. Both fungi showed high trap-forming and nematode-capturing ability; especially CNU 091026 captured 100% Bursaphelenchus xylophilus within 24 h after inoculation.

  18. Soil mycoflora from the Dead Sea Oases of Ein Gedi and Einot Zuqim (Israel).

    PubMed

    Steiman, R; Guiraud, P; Sage, L; Seigle-Murandi, F

    1997-10-01

    Samples were taken from the top 10 cm of soils from 24 points in the Ein Gedi area. Among 329 isolates, 142 species were identified: 11 genera of ascomycetes, one genus of coelomycetes, 28 genera of hyphomycetes, 7 genera of zygomycetes and one yeast, in addition to some unidentified basidiomycetes. The hyphomycetes were represented by 17 dematiaceous, 9 mucedinaceous and two tuberculariaceous. Melanconiaceous and stilbellaceous genera were not found. Two new varieties of Microascus recently described were reisolated. No strict thermophiles or halophiles were obtained. There is apparently no very characteristic or specific fungal flora of the Dead Sea Oases although it was different from that found in the desert soil surrounding this area.

  19. Influence of leaf quality in microbial decomposition in a headwater stream in the Brazilian cerrado: a 1-year study.

    PubMed

    Sales, M A; Gonçalves, J F; Dahora, J S; Medeiros, A O

    2015-01-01

    The temporal dynamics of leaf decomposition in a tropical stream were evaluated by measuring the input of leaves from riparian vegetation. The resulting mixture of leaves was processed by decomposer microorganisms and, in particular, by aquatic hyphomycetes, the most important microorganisms associated with leaf litter decomposition. Leaf decomposition and the characteristics and activity of the microbial community were investigated over the course of 1 year; these parameters depended on the composition and availability of leaves in the stream. We assessed the biomass, activity, and community structure of the fungi associated with the decomposing leaf mixtures monthly in a tropical stream with little seasonal variation in water characteristics. The leaf material included 27 riparian species; the amount of leaves of each contributing species varied from month to month, with higher overall values in the rainy season. Leaf decomposition was slow, and leaf mass loss did not occur monthly; however, leaf mass loss was significantly lower in May and September. The values of ATP, the ergosterol concentration, and the sporulation of aquatic hyphomycetes varied during the experiment, and only the ergosterol concentration (fungal biomass) was correlated with leaf mass loss. The sporulation rates of aquatic hyphomycetes peaked at the beginning of the rainy season (October), when only three hyphomycete species were present out of the total of seven that were found during the year. In this tropical stream, where the physical and chemical characteristics of the stream water varied little during the year, we hypothesize that changes related to leaf chemical composition could be the cause of changes in decomposition and the abundance and composition of the fungal assemblages. No significant differences were found for the parameters analyzed (lignin, cellulose, fiber, phosphorous, and polyphenols).

  20. Dimilin effects on leaf-decomposing aquatic fungi on the Fernow Experimental Forest, West Virginia

    Treesearch

    T. Dubey; S. L. Stephenson; P. J. Edwards

    1995-01-01

    Dimilin was applied to two watersheds on the Fernow Experimental Forest on May 16,1992, as part of a study to evaluate its effect on non-target organisms. Data were obtained on the occurrence, conidial production, and leaf litter colonization of aquatic hyphomycetes 5 days prior to and 2, 10, 25, and 55 days following application in the two treated watersheds and two...

  1. Temperature affects leaf litter decomposition in low-order forest streams: field and microcosm approaches.

    PubMed

    Martínez, Aingeru; Larrañaga, Aitor; Pérez, Javier; Descals, Enrique; Pozo, Jesús

    2014-01-01

    Despite predicted global warming, the temperature effects on headwater stream functioning are poorly understood. We studied these effects on microbial-mediated leaf decomposition and the performance of associated aquatic hyphomycete assemblages. Alder leaves were incubated in three streams differing in winter water temperature. Simultaneously, in laboratory, leaf discs conditioned in these streams were incubated at 5, 10 and 15 °C. We determined mass loss, leaf N and sporulation rate and diversity of aquatic hyphomycete communities. In the field, decomposition rate correlated positively with temperature. Decomposition rate and leaf N presented a positive trend with dissolved nutrients, suggesting that temperature was not the only factor determining the process velocity. Under controlled conditions, it was confirmed that decomposition rate and leaf N were positively correlated with temperature, leaves from the coldest stream responding most clearly. Sporulation rate correlated positively with temperature after 9 days of incubation, but negatively after 18 and 27 days. Temperature rise affected negatively the sporulating fungi richness and diversity only in the material from the coldest stream. Our results suggest that temperature is an important factor determining leaf processing and aquatic hyphomycete assemblages and that composition and activity of fungal communities adapted to cold environments could be more affected by temperature rises. Highlight: Leaf decomposition rate and associated fungal communities respond to temperature shifts in headwater streams.

  2. Effects of metals on growth and sporulation of aquatic fungi.

    PubMed

    Azevedo, Maria M; Cássio, Fernanda

    2010-07-01

    Aquatic hyphomycetes are a relevant group of fungi that play a crucial role as intermediaries between plant detritus and invertebrates in clean or metal-polluted streams. In this study, we investigated the effects of Zn, Cu, Ni, and Cd on the growth and sporulation of several aquatic hyphomycete species. Effects of these metals on growth were assessed in solid and liquid media with different compositions [1% malt extract (ME) and a mineral medium supplemented with vitamins and 2% glucose (MK)], and fungal sensitivity to metals was compared. The exposure to Zn or Cd inhibited the sporulation of Heliscus submersus and Tricladium chaetocladium, with these effects being stronger in the latter species. In solid medium, mydelial growth was linear, and, in most cases, metals negatively affected fungal growth. The sensitivity of aquatic hyphomycetes to metals, assessed as the metal concentration inhibiting biomass production in 50% (EC(50)), showed that Ypsilina graminea and Varicosporium elodeae were the most resistant species to Zn, while Alatospora acuminata, H. submersus, and Flagellospora curta appeared to be the most resistant fungus to Cu. Generally, lower toxicity of Zn or Cu than Ni or Cd was found. However, EC(50) values were about 20 times higher in solid than in liquid medium. Changes in nutrient supplies to fungi affected metal toxicity, as shown by higher EC(50) values in MK than ME. Complementarily, fungal tolerance to metals varied with fungal species as well as metal type.

  3. Leaf Barriers to Fungal Colonization and Shredders (Tipula lateralis) Consumption of Decomposing Eucalyptus globulus.

    PubMed

    Canhoto; Graça

    1999-04-01

    > Abstract Herein we assess the importance of leaf cuticle, polyphenolic, and essential oils contents of Eucalyptus globulus leaves to hyphomycete colonization and shredder consumption. Optical and electron microscopy revealed that, at least during the first 5 weeks of conditioning, the cuticle remains virtually intact. Stomata provide the main access for hyphae to internal leaf tissues and, eventually, for spore release. We suggest that in E. globulus leaves, fungal decomposition progresses predominantly in and from the eucalyptus leaf mesophyll to the outside. Malt extract agar media supplemented with either eucalyptus essential oils or tannic acid completely inhibited (Articulospora tetracladia, Lemonniera aquatica, and Tricladium gracile) or depressed (Heliscus lugdunensis, Lunulospora curvula, and Tricladium angulatum) aquatic hyphomycetes growth. The transference of both secondary compounds to alder leaves induced similar and significant reduction in Tipula lateralis larval consumption. Results consistently indicate that eucalyptus oils are stronger deterrents than polyphenols. The waxy cuticle of E. globulus appears to be a key physical factor delaying fungal colonization during decomposition. We hypothesize that the relative influence of leaf phenols and essential oils to aquatic hyphomycetes and shredders may be related to three main factors: (a) initial distribution of such compounds in the leaves; (b) possibility of their decrease through decomposition; and (c) consumption strategies of detritivores.

  4. Abundant respirable ergot alkaloids from the common airborne fungus Aspergillus fumigatus.

    PubMed

    Panaccione, Daniel G; Coyle, Christine M

    2005-06-01

    Ergot alkaloids are mycotoxins that interact with several monoamine receptors, negatively affecting cardiovascular, nervous, reproductive, and immune systems of exposed humans and animals. Aspergillus fumigatus, a common airborne fungus and opportunistic human pathogen, can produce ergot alkaloids in broth culture. The objectives of this study were to determine if A. fumigatus accumulates ergot alkaloids in a respirable form in or on its conidia, to quantify ergot alkaloids associated with conidia produced on several different substrates, and to measure relevant physical properties of the conidia. We found at least four ergot alkaloids, fumigaclavine C, festuclavine, fumigaclavine A, and fumigaclavine B (in order of abundance), associated with conidia of A. fumigatus. Under environmentally relevant conditions, the total mass of ergot alkaloids often constituted >1% of the mass of the conidium. Ergot alkaloids were extracted from conidia produced on all media tested, and the greatest quantities were observed when the fungus was cultured on latex paint or cultured maize seedlings. The values for physical properties of conidia likely to affect their respirability (i.e., diameter, mass, and specific gravity) were significantly lower for A. fumigatus than for Aspergillus nidulans, Aspergillus niger, and Stachybotrys chartarum. The demonstration of relatively high concentrations of ergot alkaloids associated with conidia of A. fumigatus presents opportunities for investigations of potential contributions of the toxins to adverse health effects associated with the fungus and to aspects of the biology of the fungus that contribute to its success.

  5. Interactions of Aspergillus fumigatus with endothelial cells: internalization, injury, and stimulation of tissue factor activity.

    PubMed

    Lopes Bezerra, Leila M; Filler, Scott G

    2004-03-15

    Invasive aspergillosis causes significant mortality among patients with hematologic malignancies. This infection is characterized by vascular invasion and thrombosis. To study the pathogenesis of invasive aspergillosis, we investigated the interactions of Aspergillus fumigatus conidia and hyphae with endothelial cells in vitro. We found that both forms of the organism induced endothelial cell microfilament rearrangement and subsequent endocytosis. Conidia were endocytosed 2-fold more avidly than hyphae, and endocytosis was independent of fungal viability. Endocytosed conidia and hyphae caused progressive endothelial cell injury after 4 hours of infection. Live conidia induced more endothelial cell injury than did live hyphae. However, endothelial cell injury caused by conidia was dependent on fungal viability, whereas injury caused by hyphae was not, indicating that conidia and hyphae injure endothelial cells by different mechanisms. Neither live nor killed conidia increased tissue factor activity of endothelial cells. In contrast, both live and killed hyphae stimulated significant endothelial cell tissue factor activity, as well as the expression of tissue factor antigen on the endothelial cell surface. These results suggest that angioinvasion and thrombosis caused by A fumigatus hyphae in vivo may be due in part to endothelial cell invasion, induction of injury, and stimulation of tissue factor activity.

  6. Conidial germination in Scedosporium apiospermum, S. aurantiacum, S. minutisporum and Lomentospora prolificans: influence of growth conditions and antifungal susceptibility profiles

    PubMed Central

    de Mello, Thaís Pereira; Aor, Ana Carolina; de Oliveira, Simone Santiago Carvalho; Branquinha, Marta Helena; dos Santos, André Luis Souza

    2016-01-01

    In the present study, we have investigated some growth conditions capable of inducing the conidial germination in Scedosporium apiospermum, S. aurantiacum, S. minutisporum and Lomentospora prolificans. Germination in Sabouraud medium (pH 7.0, 37ºC, 5% CO2) showed to be a typically time-dependent event, reaching ~75% in S. minutisporum and > 90% in S. apiospermum, S. aurantiacum and L. prolificans after 4 h. Similar germination rate was observed when conidia were incubated under different media and pHs. Contrarily, temperature and CO2 tension modulated the germination. The isotropic conidial growth (swelling) and germ tube-like projection were evidenced by microscopy and cytometry. Morphometric parameters augmented in a time-dependent fashion, evidencing changes in size and granularity of fungal cells compared with dormant 0 h conidia. In parallel, a clear increase in the mitochondrial activity was measured during the transformation of conidia-into-germinated conidia. Susceptibility profiles to itraconazole, fluconazole, voriconazole, amphotericin B and caspofungin varied regarding each morphotype and each fungal species. Overall, the minimal inhibitory concentrations for hyphae were higher than conidia and germinated conidia, except for caspofungin. Collectively, our study add new data about the conidia-into-hyphae transformation in Scedosporium and Lomentospora species, which is a relevant biological process of these molds directly connected to their antifungal resistance and pathogenicity mechanisms. PMID:27355215

  7. Aspergillus fumigatus spore proteomics and genetics reveal that VeA represses DefA-mediated DNA damage response.

    PubMed

    Shin, Kwang-Soo; Park, Hee-Soo; Kim, Young; Heo, In-Beom; Kim, Young Hwan; Yu, Jae-Hyuk

    2016-10-04

    Aspergillus fumigatus reproduces and infects host by forming a high number of small asexual spores (conidia). The velvet proteins are global transcriptional regulators governing the complex process of conidiogenesis in this fungus. Here, to further understand the velvet-mediated regulation, we carried out comparative proteomic analyses of conidia of wild type (WT) and three velvet mutants (ΔveA, ΔvelB and ΔvosA). Cluster analysis of 184 protein spots showing at least 1.5-fold differential accumulation between WT and mutants reveal the clustering of WT- ΔveA and ΔvelB-ΔvosA. Among 43 proteins identified by Nano-LC-ESI-MS/MS, 23 including several heat shock proteins showed more than two-fold reduction in both the ∆velB and ∆vosA conidia. On the contrary, three proteins exhibited more than five-fold increase in ∆veA only, including the putative RNA polymerase II degradation factor DefA. The deletion of defA resulted in a reduced number of conidia and restricted colony growth. In addition, the defA deletion mutant conidia showed hypersensitivity against the DNA damaging agents NQO and MMS, while the ΔveA mutant conidia were more resistant against to NQO. Taken together, we propose that VeA controls protein level of DefA in conidia, which are dormant and equipped with multiple layers of protection against environmental cues.

  8. Mutually facilitated dispersal between the nonmotile fungus Aspergillus fumigatus and the swarming bacterium Paenibacillus vortex

    PubMed Central

    Ingham, Colin J.; Kalisman, Oren; Finkelshtein, Alin; Ben-Jacob, Eshel

    2011-01-01

    In the heterogeneous environment surrounding plant roots (the rhizosphere), microorganisms both compete and cooperate. Here, we show that two very different inhabitants of the rhizosphere, the nonmotile fungus Aspergillus fumigatus and the swarming bacterium Paenibacillus vortex, can facilitate each other's dispersal. A. fumigatus conidia (nonmotile asexual fungal spores) can be transported by P. vortex swarms over distances of at least 30 cm and at rates of up to 10.8 mm h−1. Moreover, conidia can be rescued and transported by P. vortex from niches of adverse growth conditions. Potential benefit to the bacteria may be in crossing otherwise impenetrable barriers in the soil: fungal mycelia seem to act as bridges to allow P. vortex to cross air gaps in agar plates. Transport of conidia was inhibited by proteolytic treatment of conidia or the addition of purified P. vortex flagella, suggesting specific contacts between flagella and proteins on the conidial surface. Conidia were transported by P. vortex into locations where antibiotics inhibited bacteria growth, and therefore, growth and sporulation of A. fumigatus were not limited by bacterial competition. Conidia from other fungi, similar in size to those fungi from A. fumigatus, were not transported as efficiently by P. vortex. Conidia from a range of fungi were not transported by another closely related rhizosphere bacterium, Paenibacillus polymyxa, or the more distantly related Proteus mirabilis, despite both being efficient swarmers. PMID:22106274

  9. Effects of successive subculturing on stability, virulence, conidial yield, germination and shelf-life of entomopathogenic fungi.

    PubMed

    Ansari, M A; Butt, T M

    2011-06-01

    To determine the stability and conidial yield of two strains of the entomopathogenic fungus Metarhizium anisopliae and one strain of M. brunneum, being developed for the control of insect pests. The conidial yields and the shelf-life of the conidia of two commercially viable strains of M. anisopliae V275 (=F52) and ARSEF 4556 and one strain of M. brunneum (ARSEF 3297) were determined after harvesting conidia from in vitro subcultures on Sabouraud dextrose agar (SDA) and broken basmati rice. The strains were stable and showed no decline in virulence against Tenebrio molitor, even when subcultured successively 12 times on SDA. Conidia-bound Pr1 protease activity decreased in conidia harvested from SDA and mycosed cadavers after the 1st subculture, but increased in conidia produced on rice. The C:N ratio of conidia from mycosed cadavers was lower than that of conidia from rice or SDA. Irrespective of the number of subcultures, strain ARSEF 4556 produced significantly higher conidial yields than ARSEF 3297 and V275. The 12th subculture of V275 and ARSEF 3297 produced the lowest conidial yield. Shelf-life studies showed that conidia of strain ARSEF 4556 had a higher conidial viability than V275 and ARSEF 3297 after 4 months, stored at 4°C. The current study shows that determining strain stability and conidial yield through successive subculturing is an essential component for selecting the best strain for commercial purposes.   This is the first study to compare quality control parameters in the production of conidia on rice, and it shows that the level of Pr1 is comparatively high for inoculum produced on rice. © 2011 The Authors. Journal of Applied Microbiology © 2011 The Society for Applied Microbiology.

  10. [Production of infectious units of Isaria fumosorosea (Hypocreales: Cordycipitaceae) from different indigenous isolates of northeastern Mexico using 3 propagation strategies].

    PubMed

    Gandarilla-Pacheco, Fatima L; Morales-Ramos, Lilia H; Pereyra-Alférez, Benito; Elías-Santos, Myriam; Quintero-Zapata, Isela

    2017-09-26

    The aim of this study was to evaluate the production of blastospores and conidia of different native isolates and a strain of Isaria fumosorosea using different propagation techniques. Two liquid culture media of casamino acids and peptone as nitrogen sources and glucose as carbon source for both media cultures were respectively used in the production of blastospores, while for the production of conidia, the fungi were grown in potato dextrose agar; from these cultures, solutions of conidia to a concentration of 1×10(6) per milliliter were prepared to inoculate flasks with Sabouraud dextrose broth for the liquid phase of the biphasic culture, also known as preculture. Subsequently, rice grain bags were inoculated with the preculture and the conidia solutions, which were incubated for 14 days for solid fermentation and biphasic culture, respectively. The HIB-23 isolate recorded a concentration of 4.90×10(8) blastospores/ml in the casamino acid medium, while a concentration of 2.15×10(8) blastospores/ml was obtained in the peptone collagen medium. For the Pfr-612 strain, the conidia production in solid-state fermentation was 1.58×10(9) conidia/g, and for HIB-30 in the biphasic culture of 9.00×10(6) conidia/g. Solid-state fermentation proved to be the most effective method with an average of 1.09×10(9) conidia/g, whereas the biphasic culture was the least effective method with 2.76×10(6) conidia/g; no significant difference was reported for the submerged production media. Copyright © 2017 Asociación Argentina de Microbiología. Publicado por Elsevier España, S.L.U. All rights reserved.

  11. Comparison between superficial and solid-state cultures of Isaria fumosorosea: conidial yields, quality and sensitivity to oxidant conditions.

    PubMed

    Muñiz-Paredes, Facundo; Garza-López, Paul Misael; Viniegra-González, Gustavo; Loera, Octavio

    2016-07-01

    Conidia production and quality from mycoinsecticides in solid-state cultures (SSC) are frequently inferred from superficial culture (SC) results. Both parameters were evaluated for two Isaria fumosorosea strains (ARSEF 3302 and CNRCB1), in SC and SSC, using culture media with the same chemical composition. For both strains, conidia production was higher in SC than SSC in terms of conidia per gram of dry substrate. Germination in both strains did not show significant differences between SC and SSC (>90 %). Similarly, conidia viability in ARSEF 3302 strain did not show differences at early stages between SC and SSC, but was higher in SC compared to SSC in the late stage of culture; in contrast, conidia from CNRCB1 strain did not differ between both culture systems. Some infectivity parameters improved in conidia from SSC, compared to SC at the early stages, but these differences disappeared at the final stage, independently of the strain. Both strains showed decreased conidia production when 26 % O2 pulses were applied; nevertheless, conidiation in SSC was two orders of magnitude more sensitive to oxidant pulses. In SC with 26 % O2 pulses, conidia viability for both strains at early stages, was higher than in normal atmospheric conditions. Infectivity towards Galleria mellonella larvae was similar between conidia from normal atmosphere and oxidant conditions; notably, for the strain ARSEF 3302 infectivity decreased at the final stage. This study shows the intrinsic differences between SC and SSC, which should be considered when using SC as a model to design production processes in SSC.

  12. Insight into the feeding behavior of predatory mites on Beauveria bassiana, an arthropod pathogen

    PubMed Central

    Wu, Shengyong; Zhang, Ye; Xu, Xuenong; Lei, Zhongren

    2016-01-01

    Interactions between fungal entomopathogens and pest predators are particularly relevant in control of agricultural insect pests. In a laboratory study, we confirmed that the predatory mite, Neoseiulus barkeri, exhibited feeding behavior on the entomopathogenic fungus Beauveria bassiana conidia through DNA extracts. Using transmission electron microscopy, we determined that the majority of conidia found in the mite gut tended to dissolve within 24 h post ingestion, suggesting that the conidia had probably lost their viability. To our knowledge this is the first report of feeding behavior of phytoseiid mites on entomopathogenic fungus. The findings expand our knowledge of fungus–predator interactions. PMID:27041703

  13. Insight into the feeding behavior of predatory mites on Beauveria bassiana, an arthropod pathogen.

    PubMed

    Wu, Shengyong; Zhang, Ye; Xu, Xuenong; Lei, Zhongren

    2016-04-04

    Interactions between fungal entomopathogens and pest predators are particularly relevant in control of agricultural insect pests. In a laboratory study, we confirmed that the predatory mite, Neoseiulus barkeri, exhibited feeding behavior on the entomopathogenic fungus Beauveria bassiana conidia through DNA extracts. Using transmission electron microscopy, we determined that the majority of conidia found in the mite gut tended to dissolve within 24 h post ingestion, suggesting that the conidia had probably lost their viability. To our knowledge this is the first report of feeding behavior of phytoseiid mites on entomopathogenic fungus. The findings expand our knowledge of fungus-predator interactions.

  14. Morphology, cultural characteristics, and pathogenicity of Rhizosphaera kalkhoffii on Picea spp. in northern Minnesota and Wisconsin

    Treesearch

    Jennifer Juzwik

    1993-01-01

    Morphology, cultural characteristics, and pathogenicity of Rhizosphaera kalkhoffii from spruce (Picea spp.) showing premature needle loss in northern Minnesota and Wisconsin were investigated. Pycnidiospores from needles, conidia of the Hormonema-like synanamorph, and pycnidiospores produced in culture were...

  15. Differential allergy responses to Metarhizium anisopliae fungal component extracts in BALB/c mice

    EPA Science Inventory

    Intratracheal aspiration (IA) exposure to Metarhizium anisopliae crude antigen (MACA), which is composed of equal protein amounts of mycelium (MYC), conidia (CON) and inducible proteases/chitinases (IND) extracts/filtrates, has resulted in responses characteristic of human allerg...

  16. DIFFERENTIAL ALLERGIC AND NEUROTROPHIN RESPONSES TO FUNGAL COMPONENT EXTRACTS IN BALB/C MICE

    EPA Science Inventory

    Metarhizium anisopliae mycelium (MYC), conidia (CON) and inducible protease (IND) extracts were combined to produce the antigen MACA to screen for allergenic potential. Involuntary aspiration (IA) exposure to MACA in BALB/c mice has caused immune, inflammatory and physiological ...

  17. Scanning electron microscopy and energy-dispersive X-ray microanalysis of Penicillium brevicompactum treated with cobalt.

    PubMed

    Farrag, Rasha M; Mohamadein, Mohamed M; Mekawy, Amal A

    2008-01-01

    Scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDX) were used to study the morphology and elemental composition of the conidia, phialids and hyphae of Penicillium brevicompactum grown in the presence of cobalt concentrations of 0, 50, 200, 500, 800 and 1000 ppm (mg/l). Cobalt uptake was through the hyphae, phialids and the conidia with maximum uptake being by the conidia at a concentration of 1000 ppm. EDX revealed the increase in the percentage of calcium and magnesium in the hyphae, conidia and phialids, compared to corresponding controls, accompanying the increase in cobalt uptake. Alternatively a decrease in sulfur percentage was observed. This study might reflect the possibility of using SEM-EDX as a new technique in understanding the mechanism of tolerance.

  18. The genetic activity of 6-N-hydroxylaminopurine in Aspergillus nidulans.

    PubMed

    Babudri, N; Salvini, D; Pimpinelli, S; Morpurgo, G

    1994-04-01

    The activity of a base analog (6-N-hydroxylaminopurine, HAP) has been tested on Aspergillus nidulans. In germinating haploid conidia HAP is a strong mutagen, while it does not have any activity in resting conidia. Moreover, HAP does not increase the frequency of recombination in germinating conidia. The mutagenic activity of this base analog has also been tested in diploid conidia of A. nidulans; in fact, it has been shown (Pavlov et al., 1991) that the HAP-induced frequency of heteroallelic recessive mutations in diploid cells of the yeast S. cerevisiae is higher than expected. In A. nidulans, we did not observe any increase in the frequency of recessive homozygous fpaA/fpaA (p-fluorophenylalanine-resistant) mutants over the expected one, which has been calculated on the basis of the observed mutation frequency in the haploid strain.

  19. Differential allergy responses to Metarhizium anisopliae fungal component extracts in BALB/c mice

    EPA Science Inventory

    Intratracheal aspiration (IA) exposure to Metarhizium anisopliae crude antigen (MACA), which is composed of equal protein amounts of mycelium (MYC), conidia (CON) and inducible proteases/chitinases (IND) extracts/filtrates, has resulted in responses characteristic of human allerg...

  20. DIFFERENTIAL ALLERGIC AND NEUROTROPHIN RESPONSES TO FUNGAL COMPONENT EXTRACTS IN BALB/C MICE

    EPA Science Inventory

    Metarhizium anisopliae mycelium (MYC), conidia (CON) and inducible protease (IND) extracts were combined to produce the antigen MACA to screen for allergenic potential. Involuntary aspiration (IA) exposure to MACA in BALB/c mice has caused immune, inflammatory and physiological ...

  1. [Inhibition of Bacillus pumilus AR03 on Alternaria alternata and Erysiphe cichoracearum on tobacco].

    PubMed

    Wang, Jing; Tian, Hua; Kong, Fan-yu; Wang, Yi-hong; Zhang, Cheng-sheng; Feng, Chao

    2015-10-01

    This study aimed to evaluate the efficiency of Bacillus pumilus AR03 against Altenaria alternata and Erysiphe cichoracearum. The antagonistic activities were studied in the way of co-culture on plate, inhibition of pathogen conidia germination on concave slides and LB agar medium. In the investigation, the water suspension of living cells of strain AR03, at 3 x 10(8) cfu . mL-1 had a remarkable inhibition effect on hyphae growth and conidia germination of A. alternata and caused hyphae deformation, shorter and swollen nodes, winding hyphae accumulation, abnormal tubes with tips expanded or deformed. Conidia did not germinate and the tissues of compartment became swollen or formed a round spherical bubble. In addition, the inhibition rate of conidia germination of E. cichoracearum was 91. 3% and 69. 3%, respectively when treated with strain AR03 at 1 x 10(7) cfu . mL-1 and 1.5 x 10(6) cfu . mL-1. Conidia treated by living cells of AR03 became swollen, deformed, the protoplasm of conidia shrinked, disintegrated gradually and separated from the conidia wall. And some conidia were hollow because the protoplasm leaked out from inside. Greenhouse results revealed that the effects of living cells of AR03 with different concentrations were significantly different. Bacterial suspension of AR03 at 3 x 10(8) cfu . mL-1 was strongly antagonistic to E. cichoracearum with the control efficiencies 7 days and 15 days after treatment of two spays being 83.8% and 90.3%, respectively, while the control efficiencies of AR03 at 3 x 10(6) cfu . mL-1 were 70.0% and 73.3%, respectively. Strain AR03 had a long persistence against powdery mildew more than 30 days.

  2. Germination-defective mutant of Neurospora crassa that responds to siderophores

    NASA Technical Reports Server (NTRS)

    Charlang, G.; Williams, N. P.

    1977-01-01

    A conditionally germination-defective mutant of Neurospora crassa has been found to be partially curable by ferricrocin and other siderophores. The mutant conidia rapidly lose their membrane-bound siderophores when suspended in buffer or growth media. Germination is consequently delayed unless large numbers of conidia are present (positive population effect). This indicates that the mutant has a membrane defect involving the siderophore attachment site.

  3. Horizontal Transmission of Metarhizium anisopliae in Fruit Flies and Effect of Fungal Infection on Egg Laying and Fertility

    PubMed Central

    Dimbi, Susan; Maniania, Nguya K.; Ekesi, Sunday

    2013-01-01

    Fly-to-fly transmission of conidia of the entomopathogenic fungus Metarhizium anisopliae and the effect of fungal infection on the reproductive potential of females surviving infection were investigated in three fruit fly species, Ceratitis cosyra, C. fasciventris, and C. capitata. The number of conidia picked up by a single fruit fly was determined in C. cosyra. The initial uptake (Day 0) of conidia by a single fly was approx. 1.1 × 106 conidia after exposure to the treated substrate. However, the number of conidia dropped from 7.2 × 105 to 4.1 × 105 conidia after 2 and 8 h post-exposure, respectively. The number of conidia picked up by a single fungus-treated fly (“donor”) varied between 3.8 × 105 and 1.0 × 106 in the three fruit fly species, resulting in 100% mortality 5–6 days post-exposure. When fungus-free flies of both sexes (“recipient” flies) were allowed to mate with “donor” flies, the number of conidia picked up by a single fly varied between 1.0 × 105 and 2.5 × 105, resulting in a mortality of 83–100% in C. capitata, 72–85% in C. cosyra and 71–93% in C. fasciventris 10–15 days post-inoculation. There was an effect of fungal infection on female egg laying in the three species of fruit flies as control flies laid more eggs than fungus-treated females. The percentage reduction in fecundity in flies infected with M. anisopliae was 82, 73 and 37% in C. capitata, C. fasciventris and C. cosyra, respectively. The results are discussed with regard to application in autodissemination techniques. PMID:26464386

  4. Conidial Dihydroxynaphthalene Melanin of the Human Pathogenic Fungus Aspergillus fumigatus Interferes with the Host Endocytosis Pathway.

    PubMed

    Thywißen, Andreas; Heinekamp, Thorsten; Dahse, Hans-Martin; Schmaler-Ripcke, Jeannette; Nietzsche, Sandor; Zipfel, Peter F; Brakhage, Axel A

    2011-01-01

    Aspergillus fumigatus is the most important air-borne fungal pathogen of humans. The interaction of the pathogen with the host's immune system represents a key process to understand pathogenicity. For elimination of invading microorganisms, they need to be efficiently phagocytosed and located in acidified phagolysosomes. However, as shown previously, A. fumigatus is able to manipulate the formation of functional phagolysosomes. Here, we demonstrate that in contrast to pigmentless pksP mutant conidia of A. fumigatus, the gray-green wild-type conidia inhibit the acidification of phagolysosomes of alveolar macrophages, monocyte-derived macrophages, and human neutrophil granulocytes. Therefore, this inhibition is independent of the cell type and applies to the major immune effector cells required for defense against A. fumigatus. Studies with melanin ghosts indicate that the inhibitory effect of wild-type conidia is due to their dihydroxynaphthalene (DHN)-melanin covering the conidia, whereas the hydrophobin RodA rodlet layer plays no role in this process. This is also supported by the observation that pksP conidia still exhibit the RodA hydrophobin layer, as shown by scanning electron microscopy. Mutants defective in different steps of the DHN-melanin biosynthesis showed stronger inhibition than pksP mutant conidia but lower inhibition than wild-type conidia. Moreover, A. fumigatus and A. flavus led to a stronger inhibition of phagolysosomal acidification than A. nidulans and A. terreus. These data indicate that a certain type of DHN-melanin that is different in the various Aspergillus species, is required for maximal inhibition of phagolysosomal acidification. Finally, we identified the vacuolar ATPase (vATPase) as potential target for A. fumigatus based on the finding that addition of bafilomycin which inhibits vATPase, led to complete inhibition of the acidification whereas the fusion of phagosomes containing wild-type conidia and lysosomes was not affected.

  5. Conidial Dihydroxynaphthalene Melanin of the Human Pathogenic Fungus Aspergillus fumigatus Interferes with the Host Endocytosis Pathway

    PubMed Central

    Thywißen, Andreas; Heinekamp, Thorsten; Dahse, Hans-Martin; Schmaler-Ripcke, Jeannette; Nietzsche, Sandor; Zipfel, Peter F.; Brakhage, Axel A.

    2011-01-01

    Aspergillus fumigatus is the most important air-borne fungal pathogen of humans. The interaction of the pathogen with the host's immune system represents a key process to understand pathogenicity. For elimination of invading microorganisms, they need to be efficiently phagocytosed and located in acidified phagolysosomes. However, as shown previously, A. fumigatus is able to manipulate the formation of functional phagolysosomes. Here, we demonstrate that in contrast to pigmentless pksP mutant conidia of A. fumigatus, the gray-green wild-type conidia inhibit the acidification of phagolysosomes of alveolar macrophages, monocyte-derived macrophages, and human neutrophil granulocytes. Therefore, this inhibition is independent of the cell type and applies to the major immune effector cells required for defense against A. fumigatus. Studies with melanin ghosts indicate that the inhibitory effect of wild-type conidia is due to their dihydroxynaphthalene (DHN)-melanin covering the conidia, whereas the hydrophobin RodA rodlet layer plays no role in this process. This is also supported by the observation that pksP conidia still exhibit the RodA hydrophobin layer, as shown by scanning electron microscopy. Mutants defective in different steps of the DHN-melanin biosynthesis showed stronger inhibition than pksP mutant conidia but lower inhibition than wild-type conidia. Moreover, A. fumigatus and A. flavus led to a stronger inhibition of phagolysosomal acidification than A. nidulans and A. terreus. These data indicate that a certain type of DHN-melanin that is different in the various Aspergillus species, is required for maximal inhibition of phagolysosomal acidification. Finally, we identified the vacuolar ATPase (vATPase) as potential target for A. fumigatus based on the finding that addition of bafilomycin which inhibits vATPase, led to complete inhibition of the acidification whereas the fusion of phagosomes containing wild-type conidia and lysosomes was not affected. PMID

  6. Germination-defective mutant of Neurospora crassa that responds to siderophores

    NASA Technical Reports Server (NTRS)

    Charlang, G.; Williams, N. P.

    1977-01-01

    A conditionally germination-defective mutant of Neurospora crassa has been found to be partially curable by ferricrocin and other siderophores. The mutant conidia rapidly lose their membrane-bound siderophores when suspended in buffer or growth media. Germination is consequently delayed unless large numbers of conidia are present (positive population effect). This indicates that the mutant has a membrane defect involving the siderophore attachment site.

  7. Heat-stressed Metarhizium anisopliae: viability (in vitro) and virulence (in vivo) assessments against the tick Rhipicephalus sanguineus.

    PubMed

    Alves, Fabrício M; Bernardo, Cíntia C; Paixão, Flávia R S; Barreto, Lucas P; Luz, Christian; Humber, Richard A; Fernandes, Éverton K K

    2017-01-01

    The current study investigated the thermotolerance of Metarhizium anisopliae s.l. conidia from the commercial products Metarril® SP Organic and Metarril® WP. The efficacy of these M. anisopliae formulations against the tick Rhipicephalus sanguineus s.l. was studied in laboratory under optimum or heat-stress conditions. The products were prepared in water [Tween® 80, 0.01 % (v/v)] or pure mineral oil. Conidia from Metarril® SP Organic suspended in water presented markedly delayed germination after heating to constant 40 °C (for 2, 4, or 6 h) compared to conidia suspended in mineral oil. Metarril® SP Organic suspended in oil and exposed to daily cycles of heat-stress (40 °C for 4 h and 25 °C for 19 h for 5 consecutive days) presented relative germination of conidia ranging from 92.8 to 87.2 % from day 1 to day 5, respectively. Conversely, germination of conidia prepared in water ranged from 79.3 to 39.1 % from day 1 to day 5, respectively. Culturability of Metarril® WP decreased from 96 % when conidia were cultured for 30 min prior to heat exposure (40 °C for 4 h) to 9 % when conidia were cultured for 8 h. Tick percent control was distinctly higher when engorged females were treated with oil suspensions rather than water suspensions, even when treated ticks were exposed to heat-stress regimen. Oil-based applications protected fungal conidia against heat-stress. Although Metarril® is not registered for tick control, it may be useful for controlling R. sanguineus, especially if it is prepared in mineral oil.

  8. Entomopathogenic Fungus as a Biological Control for an Important Vector of Livestock Disease: The Culicoides Biting Midge

    PubMed Central

    Ansari, Minshad Ali; Pope, Edward C.; Carpenter, Simon; Scholte, Ernst-Jan; Butt, Tariq M.

    2011-01-01

    Background The recent outbreak of bluetongue virus in northern Europe has led to an urgent need to identify control measures for the Culicoides (Diptera: Ceratopogonidae) biting midges that transmit it. Following successful use of the entomopathogenic fungus Metarhizium anisopliae against larval stages of biting midge Culicoides nubeculosus Meigen, we investigated the efficacy of this strain and other fungi (Beauveria bassiana, Isaria fumosorosea and Lecanicillium longisporum) as biocontrol agents against adult C. nubeculosus in laboratory and greenhouse studies. Methodology/Findings Exposure of midges to ‘dry’ conidia of all fungal isolates caused significant reductions in survival compared to untreated controls. Metarhizium anisopliae strain V275 was the most virulent, causing a significantly decrease in midge survival compared to all other fungal strains tested. The LT50 value for strain V275 was 1.42 days compared to 2.21–3.22 days for the other isolates. The virulence of this strain was then further evaluated by exposing C. nubeculosus to varying doses (108–1011 conidia m−2) using different substrates (horse manure, damp peat, leaf litter) as a resting site. All exposed adults were found to be infected with the strain V275 four days after exposure. A further study exposed C. nubeculosus adults to ‘dry’ conidia and ‘wet’ conidia (conidia suspended in 0.03% aq. Tween 80) of strain V275 applied to damp peat and leaf litter in cages within a greenhouse. ‘Dry’ conidia were more effective than ‘wet’ conidia, causing 100% mortality after 5 days. Conclusion/Significance This is the first study to demonstrate that entomopathogenic fungi are potential biocontrol agents against adult Culicoides, through the application of ‘dry’ conidia on surfaces (e.g., manure, leaf litter, livestock) where the midges tend to rest. Subsequent conidial transmission between males and females may cause an increased level of fungi-induced mortality in midges

  9. Entomopathogenic fungus as a biological control for an important vector of livestock disease: the Culicoides biting midge.

    PubMed

    Ansari, Minshad Ali; Pope, Edward C; Carpenter, Simon; Scholte, Ernst-Jan; Butt, Tariq M

    2011-01-10

    The recent outbreak of bluetongue virus in northern Europe has led to an urgent need to identify control measures for the Culicoides (Diptera: Ceratopogonidae) biting midges that transmit it. Following successful use of the entomopathogenic fungus Metarhizium anisopliae against larval stages of biting midge Culicoides nubeculosus Meigen, we investigated the efficacy of this strain and other fungi (Beauveria bassiana, Isaria fumosorosea and Lecanicillium longisporum) as biocontrol agents against adult C. nubeculosus in laboratory and greenhouse studies. Exposure of midges to 'dry' conidia of all fungal isolates caused significant reductions in survival compared to untreated controls. Metarhizium anisopliae strain V275 was the most virulent, causing a significantly decrease in midge survival compared to all other fungal strains tested. The LT(50) value for strain V275 was 1.42 days compared to 2.21-3.22 days for the other isolates. The virulence of this strain was then further evaluated by exposing C. nubeculosus to varying doses (10(8)-10(11) conidia m(-2)) using different substrates (horse manure, damp peat, leaf litter) as a resting site. All exposed adults were found to be infected with the strain V275 four days after exposure. A further study exposed C. nubeculosus adults to 'dry' conidia and 'wet' conidia (conidia suspended in 0.03% aq. Tween 80) of strain V275 applied to damp peat and leaf litter in cages within a greenhouse. 'Dry' conidia were more effective than 'wet' conidia, causing 100% mortality after 5 days. This is the first study to demonstrate that entomopathogenic fungi are potential biocontrol agents against adult Culicoides, through the application of 'dry' conidia on surfaces (e.g., manure, leaf litter, livestock) where the midges tend to rest. Subsequent conidial transmission between males and females may cause an increased level of fungi-induced mortality in midges thus reducing the incidence of disease.

  10. MoTea4-Mediated Polarized Growth Is Essential for Proper Asexual Development and Pathogenesis in Magnaporthe oryzae▿†

    PubMed Central

    Patkar, Rajesh N.; Suresh, Angayarkanni; Naqvi, Naweed I.

    2010-01-01

    Polarized growth is essential for cellular development and function and requires coordinated organization of the cytoskeletal elements. Tea4, an important polarity determinant, regulates localized F-actin assembly and bipolar growth in fission yeast and directional mycelial growth in Aspergillus. Here, we characterize Tea4 in the rice blast fungus Magnaporthe oryzae (MoTea4). Similar to its orthologs, MoTea4-green fluorescent protein (MoTea4-GFP) showed punctate distribution confined to growth zones, particularly in the mycelial tips, aerial hyphae, conidiophores, conidia, and infection structures (appressoria) in Magnaporthe. MoTea4 was dispensable for vegetative growth in Magnaporthe. However, loss of MoTea4 led to a zigzag morphology in the aerial hyphae and a huge reduction in conidiation. The majority of the tea4Δ conidia were two celled, as opposed to the tricellular conidia in the wild type. Structure-function analysis indicated that the SH3 and coiled-coil domains of MoTea4 are necessary for proper conidiation in Magnaporthe. The tea4Δ conidia failed to produce proper appressoria and consequently failed to infect the host plants. The tea4Δ conidia and germ tubes showed disorganized F-actin structures with significantly reduced numbers of cortical actin patches. Compared to the wild-type conidia, the tea4Δ conidia showed aberrant germination, poor cytoplasmic streaming, and persistent accumulation of lipid droplets, likely due to the impaired F-actin cytoskeleton. Latrunculin A treatment of germinating wild-type conidia showed that an intact F-actin cytoskeleton is indeed essential for appressorial development in Magnaporthe. We show that MoTea4 plays an important role in organizing the F-actin cytoskeleton and is essentially required for polarized growth and morphogenesis during asexual and pathogenic development in Magnaporthe. PMID:20472691

  11. Neurospora crassa 1,3-α-glucan synthase, AGS-1, is required for cell wall biosynthesis during macroconidia development

    PubMed Central

    Fu, Ci; Tanaka, Asuma

    2014-01-01

    The Neurospora crassa genome encodes two 1,3-α-glucan synthases. One of these 1,3-α-glucan synthase genes, ags-1, was shown to be required for the synthesis of 1,3-α-glucan in the aerial hyphae and macroconidia cell walls. 1,3-α-Glucan was found in the conidia cell wall, but was absent from the vegetative hyphae cell wall. Deletion of ags-1 affected conidial development. Δags-1 produced only 5 % as many conidia as the WT and most of the conidia produced by Δags-1 were not viable. The ags-1 upstream regulatory elements were shown to direct cell-type-specific expression of red fluorescent protein in conidia and aerial hyphae. A haemagglutinin-tagged AGS-1 was found to be expressed in aerial hyphae and conidia. The research showed that 1,3-α-glucan is an aerial hyphae and conidia cell wall component, and is required for normal conidial differentiation. PMID:24847001

  12. Conidial Hydrophobins of Aspergillus fumigatus

    PubMed Central

    Paris, Sophie; Debeaupuis, Jean-Paul; Crameri, Reto; Carey, Marilyn; Charlès, Franck; Prévost, Marie Christine; Schmitt, Christine; Philippe, Bruno; Latgé, Jean Paul

    2003-01-01

    The surface of Aspergillus fumigatus conidia, the first structure recognized by the host immune system, is covered by rodlets. We report that this outer cell wall layer contains two hydrophobins, RodAp and RodBp, which are found as highly insoluble complexes. The RODA gene was previously characterized, and ΔrodA conidia do not display a rodlet layer (N. Thau, M. Monod, B. Crestani, C. Rolland, G. Tronchin, J. P. Latgé, and S. Paris, Infect. Immun. 62:4380-4388, 1994). The RODB gene was cloned and disrupted. RodBp was highly homologous to RodAp and different from DewAp of A. nidulans. ΔrodB conidia had a rodlet layer similar to that of the wild-type conidia. Therefore, unlike RodAp, RodBp is not required for rodlet formation. The surface of ΔrodA conidia is granular; in contrast, an amorphous layer is present at the surface of the conidia of the ΔrodA ΔrodB double mutant. These data show that RodBp plays a role in the structure of the conidial cell wall. Moreover, rodletless mutants are more sensitive to killing by alveolar macrophages, suggesting that RodAp or the rodlet structure is involved in the resistance to host cells. PMID:12620846

  13. Antifungal properties of wheat histones (H1-H4) and purified wheat histone H1.

    PubMed

    De Lucca, Anthony J; Heden, Lars-Olof; Ingber, Bruce; Bhatnagar, Deepak

    2011-07-13

    Wheat ( Triticum spp.) histones H1, H2, H3, and H4 were extracted, and H1 was further purified. The effect of these histones on specific fungi that may or may not be pathogenic to wheat was determined. These fungi included Aspergillus flavus , Aspergillus fumigatus , Aspergillus niger , Fusarium oxysporum , Fusarium verticillioides , Fusarium solani , Fusarium graminearum , Penicillium digitatum , Penicillium italicum , and Greeneria uvicola . Non-germinated and germinating conidia of these fungi were bioassayed separately. The non-germinated and germinating conidia of all Fusarium species were highly susceptible to the mixture (H1-H4) as well as pure H1, with viability losses of 99-100% found to be significant (p < 0.001) at ≤10 μM or less for the histone mixture and pure H1. F. graminearum was the most sensitive to histone activity. The histones were inactive against all of the non-germinated Penicillium spp. conidia. However, they significantly reduced the viability of the germinating conidia of the Penicillium spp. conidia, with 95% loss at 2.5 μM. Non-germinated and germinating conidia viability of the Aspergillus spp. and G. uvicola were unaffected when exposed to histones up to 10 μM. Results indicate that Fusarium spp. pathogenic to wheat are susceptible to wheat histones, indicating that these proteins may be a resistance mechanism in wheat against fungal infection.

  14. Morphological Characteristics of Conidiogenesis in Cordyceps militaris

    PubMed Central

    Shrestha, Bhushan; Han, Sang-Kuk; Yoon, Kwon-Sang

    2005-01-01

    Conidial development of Cordyceps militaris was observed from germinating ascospores and vegetative hyphae through light and scanning electron microscopy (SEM). Ascospores were discharged from fresh specimens of C. militaris in sterile water as well as Sabouraud Dextrose agar plus Yeast Extract (SDAY) plates. We observed ascospore germination and conidial formation periodically. Under submerged condition in sterile water, most part-spores germinated unidirectionally and conidia were developed directly from the tips of germinating hyphae of part-spores within 36 h after ascospore discharge, showing microcyclic conidiation. First-formed conidia were cylindrical or clavate followed by globose and ellipsoidal ones. Germination of ascospores and conidial development were observed on SDAY agar by SEM. Slimy heads of conidia on variously arranged phialides, from solitary to whorl, developed 5 days after ascospore discharge. Besides, two distinct types of conidia, elongated pyriform or cylindrical and globose, were observed in the same slimy heads by SEM. Conidia were shown to be uninucleate with 4,6-diamidino-2-phenylindole staining. Conidiogenous cells were more slender than vegetative hyphae, having attenuated tips. Microcyclic conidiation, undifferentiated conidiogenous hyphae (phialides), polymorphic conidia and solitary, opposite to whorled type of phialidic arrangement are reported here as the characteristic features of asexual stage of C. militaris, which can be distinguished from other Cordyceps species. PMID:24049477

  15. Effect of storage conditions (relative humidity, duration, and temperature) on the germination time of Aspergillus carbonarius and Penicillium chrysogenum.

    PubMed

    Lattab, Nadia; Kalai, Safaa; Bensoussan, Maurice; Dantigny, Philippe

    2012-11-01

    Fungal conidia are disseminated, often in the air, for a certain period of time, prior to contaminating food products. The objective of this study was to examine the effects of the relative humidity, RH (%), time (day), and temperature (°C) during this period of time, called "storage", on the germination time, τ (h), of Aspergillus carbonarius and Penicillium chrysogenum. A Doehlert design was used in the range, 20-100% RH, 2-28 days, and 5-25 °C. As compared to un-stored conidia, the germination time of conidia stored at 60% RH, 15 days, 5 °C was increased by 23 and 28%, for A. carbonarius and P. chrysogenum, respectively. Stored conidia exhibited a minimum τ value at 60% RH, and 100% RH for A. carbonarius and P. chrysogenum, respectively. For these species, τ was minimum for 2 days of storage. The effect of temperature was RH dependent for A. carbonarius. The germination time of stored conidia was clearly greater than that of fresh conidia obtained in the laboratory. This result should be taken into account to determine the mould free shelf-life of food products.

  16. Population genetics of the aquatic fungus Tetracladium marchalianum over space and time.

    PubMed

    Anderson, Jennifer L; Shearer, Carol A

    2011-01-14

    Aquatic hyphomycete fungi are fundamental mediators of energy flow and nutrient spiraling in rivers. These microscopic fungi are primarily dispersed in river currents, undergo substantial annual fluctuations in abundance, and reproduce either predominantly or exclusively asexually. These aspects of aquatic hyphomycete biology are expected to influence levels and distributions of genetic diversity over both spatial and temporal scales. In this study, we investigated the spatiotemporal distribution of genotypic diversity in the representative aquatic hyphomycete Tetracladium marchalianum. We sampled populations of this fungus from seven sites, three sites each in two rivers in Illinois, USA, and one site in a Wisconsin river, USA, and repeatedly sampled one population over two years to track population genetic parameters through two seasonal cycles. The resulting fungal isolates (N = 391) were genotyped at eight polymorphic microsatellite loci. In spite of seasonal reductions in the abundance of this species, genotypic diversity was consistently very high and allele frequencies remarkably stable over time. Likewise, genotypic diversity was very high at all sites. Genetic differentiation was only observed between the most distant rivers (∼450 km). Clear evidence that T. marchalianum reproduces sexually in nature was not observed. Additionally, we used phylogenetic analysis of partial β-tubulin gene sequences to confirm that the fungal isolates studied here represent a single species. These results suggest that populations of T. marchalianum may be very large and highly connected at local scales. We speculate that large population sizes and colonization of alternate substrates in both terrestrial and aquatic environments may effectively buffer the aquatic populations from in-stream population fluctuations and facilitate stability in allele frequencies over time. These data also suggest that overland dispersal is more important for structuring populations of T

  17. Population Genetics of the Aquatic Fungus Tetracladium marchalianum over Space and Time

    PubMed Central

    Anderson, Jennifer L.; Shearer, Carol A.

    2011-01-01

    Aquatic hyphomycete fungi are fundamental mediators of energy flow and nutrient spiraling in rivers. These microscopic fungi are primarily dispersed in river currents, undergo substantial annual fluctuations in abundance, and reproduce either predominantly or exclusively asexually. These aspects of aquatic hyphomycete biology are expected to influence levels and distributions of genetic diversity over both spatial and temporal scales. In this study, we investigated the spatiotemporal distribution of genotypic diversity in the representative aquatic hyphomycete Tetracladium marchalianum. We sampled populations of this fungus from seven sites, three sites each in two rivers in Illinois, USA, and one site in a Wisconsin river, USA, and repeatedly sampled one population over two years to track population genetic parameters through two seasonal cycles. The resulting fungal isolates (N = 391) were genotyped at eight polymorphic microsatellite loci. In spite of seasonal reductions in the abundance of this species, genotypic diversity was consistently very high and allele frequencies remarkably stable over time. Likewise, genotypic diversity was very high at all sites. Genetic differentiation was only observed between the most distant rivers (∼450 km). Clear evidence that T. marchalianum reproduces sexually in nature was not observed. Additionally, we used phylogenetic analysis of partial β-tubulin gene sequences to confirm that the fungal isolates studied here represent a single species. These results suggest that populations of T. marchalianum may be very large and highly connected at local scales. We speculate that large population sizes and colonization of alternate substrates in both terrestrial and aquatic environments may effectively buffer the aquatic populations from in-stream population fluctuations and facilitate stability in allele frequencies over time. These data also suggest that overland dispersal is more important for structuring populations of T

  18. Xylohypha emmonsii sp. nov., a new agent of phaeohyphomycosis.

    PubMed Central

    Padhye, A A; McGinnis, M R; Ajello, L; Chandler, F W

    1988-01-01

    Xylohypha emmonsii Padhye, McGinnis et Ajello is described as a new dematiaceous opportunistic pathogen of humans and lower animals. This hyphomycete differs from X. bantiana (Saccardo) McGinnis, Padhye, Borelli et Ajello by its inability to grow at 40 degrees C and by the development of short chains of asymmetrically bent to ellipsoid blastoconidia. In brain tissue of mice, experimentally infected with X. emmonsii, the host response to the invading fungus was similar to that observed in mice infected with X. bantiana. In tissues X. bantiana occurred predominantly as long, septate hyphae, in contrast to X. emmonsii, which was more polymorphic. Images PMID:3366865

  19. Tools for delivering entomopathogenic fungi to malaria mosquitoes: effects of delivery surfaces on fungal efficacy and persistence

    PubMed Central

    2010-01-01

    Background Entomopathogenic fungi infection on malaria vectors increases daily mortality rates and thus represents a control measure that could be used in integrated programmes alongside insecticide-treated bed nets (ITNs) and indoor residual spraying (IRS). Before entomopathogenic fungi can be integrated into control programmes, an effective delivery system must be developed. Methods The efficacy of Metarhizium anisopliae ICIPE-30 and Beauveria bassiana I93-825 (IMI 391510) (2 × 1010 conidia m-2) applied on mud panels (simulating walls of traditional Tanzanian houses), black cotton cloth and polyester netting was evaluated against adult Anopheles gambiae sensu stricto. Mosquitoes were exposed to the treated surfaces 2, 14 and 28 d after conidia were applied. Survival of mosquitoes was monitored daily. Results All fungal treatments caused a significantly increased mortality in the exposed mosquitoes, descending with time since fungal application. Mosquitoes exposed to M. anisopliae conidia on mud panels had a greater daily risk of dying compared to those exposed to conidia on either netting or cotton cloth (p < 0.001). Mosquitoes exposed to B. bassiana conidia on mud panels or cotton cloth had similar daily risk of death (p = 0.14), and a higher risk than those exposed to treated polyester netting (p < 0.001). Residual activity of fungi declined over time; however, conidia remained pathogenic at 28 d post application, and were able to infect and kill 73 - 82% of mosquitoes within 14 d. Conclusion Both fungal isolates reduced mosquito survival on immediate exposure and up to 28 d after application. Conidia were more effective when applied on mud panels and cotton cloth compared with polyester netting. Cotton cloth and mud, therefore, represent potential substrates for delivering fungi to mosquitoes in the field. PMID:20799967

  20. Inactivation of Aspergillus niger in mango nectar by high-pressure homogenization combined with heat shock.

    PubMed

    Tribst, Alline A L; Franchi, Mark A; Cristianini, Marcelo; de Massaguer, Pilar R

    2009-01-01

    This research evaluated the inactivation of a heat-resistant Aspergillus niger conidia in mango nectar by high-pressure homogenization (HPH) combined with heat shock. A. niger were inoculated in mango nectar (10(6) conidia mL(-1)) and subjected to HPH (300 to 100 MPa) and heat shock (80 degrees C for 5 to 20 min) before or after HPH. Processes were evaluated according to number of decimal reductions reached by each isolated or combined process. Scanning electron microscopy was performed to observe conidia wall after pressure treatment. Pressures below 150 MPa did not inactivate A. niger while pressures of 200 and 300 MPa resulted in 2 and more than 6 log reductions, respectively. D(80 degrees C) of A. niger was determined as 5.03 min. A heat shock of 80 degrees C/15 min, reaching 3 decimal conidia reductions, was applied before or after a 200 MPa pressure treatment to improve the decimal reduction to 5 log cycles. Results indicated that HPH inactivated A. niger in mango nectar at 300 MPa (>6.24 log cycles) and that, with pressure (200 MPa) combined with post heat shock, it was possible to obtain the same decimal reduction, showing a synergistic effect. On the other hand, pre heat shock associated with HPH resulted in an additive effect. The observation of A. niger conidia treated by HPH at 100 and 200 MPa by scanning electron microscopy indicated that HPH promoted intense cell wall damage, which can sensitize the conidia to post heat shock and possibly explain the synergistic effect observed. Practical Application: The results obtained in this paper are relevant to elucidate the mechanism of conidia inactivation in order to develop the application of HPH as an alternative pasteurization process for the fruit nectar industry.

  1. Bioassay and Scanning Electron Microscopic Observations Reveal High Virulence of Entomopathogenic Fungus, Beauveria bassiana, on the Onion Maggot (Diptera: Anthomyiidae) Adults.

    PubMed

    Zhang, Hui; Wu, Shengyong; Xing, Zhenlong; Wang, Xiaoqing; Lei, Zhongren

    2016-12-01

    When flies were dipped in 1 × 10(8) conidia/ml conidia suspensions and then kept in the incubator (22 ± 1 °C, 70 ± 5% RH), scanning electron microscope observations revealed that, at 2 h, the majority of adhering Beauveria bassiana conidia were attached to either the wing surface or the interstitial area between the macrochaetae on the thorax and abdomen of the onion maggot adults. Germ tubes were being produced and had oriented toward the cuticle by 18 h. Penetration of the insect cuticle had occurred by 36 h, and by 48 h, germ tubes had completely penetrated the cuticle. Fungal mycelia had emerged from the insect body and were proliferating after 72 h. The superficial area and structure of the wings and macrochaetae may facilitate the attachment of conidia and enable effective penetration. The susceptibility of adults to 12 isolates, at a concentration of 1 × 10(7) conidia/ml, was tested in laboratory experiments. Eight of the more potent strains caused in excess of 85% adult mortality 8 d post inoculation, while the median lethal time (LT50) of these strains was <6 d. The virulence of the more effective strains was further tested, and the median lethal concentrations (LC50) were calculated by exposing adults to doses ranging from 10(3)-10(7) conidia/ml. The lowest LC50 value, found in the isolate XJWLMQ-32, for the adults was 3.87 × 10(3) conidia/ml. These results demonstrate that some B. bassiana strains are highly virulent to onion maggot adults and should be considered as potential biocontrol agents against the adult flies.

  2. Synthesis of melanin-like pigments by Sporothrix schenckii in vitro and during mammalian infection.

    PubMed

    Morris-Jones, Rachael; Youngchim, Sirida; Gomez, Beatriz L; Aisen, Phil; Hay, Roderick J; Nosanchuk, Joshua D; Casadevall, Arturo; Hamilton, Andrew J

    2003-07-01

    Melanin has been implicated in the pathogenesis of several important human fungal pathogens. Existing data suggest that the conidia of the dimorphic fungal pathogen Sporothrix schenckii produce melanin or melanin-like compounds; in this study we aimed to confirm this suggestion and to demonstrate in vitro and in vivo production of melanin by yeast cells. S. schenckii grown on Mycosel agar produced visibly pigmented conidia, although yeast cells grown in brain heart infusion and minimal medium broth appeared to be nonpigmented macroscopically. However, treatment of both conidia and yeast cells with proteolytic enzymes, denaturant, and concentrated hot acid yielded dark particles similar in shape and size to the corresponding propagules, which were stable free radicals consistent with identification as melanins. Melanin particles extracted from S. schenckii yeast cells were used to produce a panel of murine monoclonal antibodies (MAbs) which labeled pigmented conidia, yeast cells, and the isolated particles. Tissue from hamster testicles infected with S. schenckii contained fungal cells that were labeled by melanin-binding MAbs, and digestion of infected hamster tissue yielded dark particles that were also reactive. Additionally, sera from humans with sporotrichosis contained antibodies that bound melanin particles. These findings indicate that S. schenckii conidia and yeast cells can produce melanin or melanin-like compounds in vitro and that yeast cells can synthesize pigment in vivo. Since melanin is an important virulence factor in other pathogenic fungi, this pigment may have a similar role in the pathogenesis of sporotrichosis.

  3. Virulence of Entomopathogenic Fungi Metarhizium anisopliae and Paecilomyces fumosoroseus for the Microbial Control of Spodoptera exigua

    PubMed Central

    Han, Ji Hee; Jin, Byung Rae; Lee, Sang Yeob

    2014-01-01

    The beet armyworm Spodoptera exigua (Lepidoptera: Noctuidae) is difficult to control using chemical insecticides because of the development of insecticide resistance. Several pest control agents are used to control the beet armyworm. Entomopathogenic fungi are one of the candidates for eco-friendly pest control instead of chemical control agents. In this study, among various entomopathogenic fungal strains isolated from soil two isolates were selected as high virulence pathogens against larva of beet armyworm. Control efficacy of fungal conidia was influenced by conidia concentration, temperature, and relative humidity (RH). The isolates Metarhizium anisopliae FT83 showed 100% cumulative mortality against second instar larvae of S. exigua 3 days after treatment at 1 × 107 conidia/mL and Paecilomyces fumosoroseus FG340 caused 100% mortality 6 days after treatment at 1 × 104 conidia/mL. Both M. anisopliae FT83 and P. fumosoroseus FG340 effectively controlled the moth at 20~30℃. M. anisopliae FT83 was significantly affected mortality by RH: mortality was 86.7% at 85% RH and 13.4% at 45% RH. P. fumosoroseus FG340 showed high mortality as 90% at 45% RH and 100% at 75% RH 6 days after conidia treatments. These results suggest that P. fumosoroseus FG340 and M. anisopliae FT83 have high potential to develop as a biocontrol agent against the beet armyworm. PMID:25606011

  4. Effect of different photoperiods on the growth, infectivity and colonization of Trinidadian strains of Paecilomyces fumosoroseus on the greenhouse whitefly, Trialeurodes vaporariorum, using a glass slide bioassay.

    PubMed

    Avery, Pasco B; Faull, Jane; Simmonds, Monique S J

    2004-01-01

    Growth, infectivity and colonization rates for blastospores and conidia of Trinidadian strains T, T10, and T11 of Paecilomyces fumosoroseus (Wize) Brown and Smith were assessed for activity against late fourth-instar nymphs of Trialeurodes vaporariorum (Westwood) (Homoptera:Aleyrodidae) under two different photoperiods (24 and 16 hour photophase). A glass-slide bioassay and a fungal development index, modified for both blastospores and conidia, were used to compare the development rates of the fungal strains on the insect hosts. Fewer adult whiteflies emerged from nymphs treated with blastospores and reared under a 16:8 hour light:dark photoperiod than a 24:0 hour photoperiod. Eclosion times of whitefly adults that emerged from nymphs treated with the different strains of conidia were similar over the 8 day experimental period at both light regimes. The percent eclosion of adult whiteflies seems to be directly correlated with the speed of infection of the blastospore or conidial treatment and the photoperiod regime. The longer photophase had a significant positive effect on development index for blastospores; however, a lesser effect was observed for the conidia at either light regime. Blastospore strain T11 offered the most potential of the three Trinidadian strains against T. vaporariorum fourth-instar nymphs, especially under constant light. The glass-slide bioassay was successfully used to compare both blastospores and conidia of P. fumosoroseus. It can be used to determine the pathogenicity and the efficacy of various fungal preparations against aleyrodid pests.

  5. Targeted destruction of fungal structures of Erysiphe trifoliorum on flat leaf surfaces of Marchantia polymorpha.

    PubMed

    Takikawa, Y; Senga, Y; Nonomura, T; Matsuda, Y; Kakutani, K; Toyoda, H

    2014-01-01

    In this study, we observed the germination behaviour of airborne conidia from powdery mildews that settle on thalloid surfaces. We inoculated thalli (flat, sheet-like leaf tissues) and gemmae (small, flat, sheet-like leaf tissues that propagate asexually via bud-like structures) of the common liverwort (Marchantia polymorpha) with conidia from tomato powdery mildew (Oidium neolycopersici; KTP-02) and red clover powdery mildew (Erysiphe trifoliorum; KRCP-4N) and examined their germination and subsequent appressorium formation under a high-fidelity digital microscope. Conidial bodies and germ tubes of the inoculated KRCP-4N conidia were destroyed on both the thalli and gemmae. The destruction of these fungal structures was observed only for KRCP-4N conidia inoculated onto M. polymorpha on both leaf surfaces. No differences in destruction of the KRCP-4N fungal structures between thalli and gemmae were observed. At 4 h post-inoculation, destruction of the germ tube tip was observed when it reached the gemmae leaf surface. At 6 h post-inoculation, the conidial bodies and germ tubes were destroyed. In contrast, KTP-02 conidia were not destroyed and formed normal, well-lobed appressoria on the surface of M. polymorpha gemmae. © 2013 German Botanical Society and The Royal Botanical Society of the Netherlands.

  6. A rapid knockdown effect of Penicillium citrinum for control of the mosquito Culex quinquefasciatus in Thailand.

    PubMed

    Maketon, Monchan; Amnuaykanjanasin, Alongkorn; Kaysorngup, Achirayar

    2014-02-01

    Twenty local isolates of entomopathogenic fungi were determined for control of the larvae and adults of Culex quinquefasciatus. In a laboratory experiment, a Penicillium sp. CM-010 caused 100% mortality of third-instar larvae within 2 h using a conidial suspension of 1 × 10⁶ conidia ml⁻¹. Its LC₅₀ was 3 × 10⁵ conidia ml⁻¹, and the lethal time (LT₅₀) was 1.06 h. Cloning and sequencing of its internal transcribed spacer region indicated that this Penicillium species is Penicillium citrinum (100% identity in 434 bp). Mortality of the adult was highest with Aspergillus flavus CM-011 followed with Metarhizium anisopliae CKM-048 from 1 × 10⁹ conidia ml⁻¹. P. citrinum CM-010 at 1 × 10⁶ conidia ml⁻¹ killed 100% larvae within 2 h while Bacillus thuringiensis var. israelensis at 5 ITU ml⁻¹ required 24 h. This P. citrinum CM-010 also greatly reduced survival of C. quinquefasciatus larvae in an unreplicated field test. Light and transmission electron micrographs showed that the fungal conidia were ingested by the larvae and deposited in the gut. The metabolite patulin was produced by P. citrinum CM-010 instead of citrinin.

  7. Virulence of a variant of Paracoccidioides brasiliensis that exists in the yeast form at room temperature.

    PubMed

    Villar, L A; Restrepo, A

    1989-01-01

    Paracoccidioides brasiliensis (ATCC 60885) produces conidia that, when plated in enriched media and incubated at 21-25 degrees C, give rise to a yeast-form variant (YRT) (ATCC 46678). The virulence of this variant for BALB/c mice has been compared with that of the yeast and conidia produced by the parent isolate. Adult female mice were inoculated intravenously with 10(7) viable cells of YRT or yeast, or 5 x 10(5) viable conidia. Mice were weighed weekly, and three mice were sacrificed at 1, 2, 3, 4 and 8 weeks post-inoculation and their organs weighed and prepared for determination of viable counts (colony forming units; c.f.u.). Animals infected with yeast cells or with conidia exhibited little weight loss in comparison with that seen in the YRT-infected mice which began to lose weight 1 week post-challenge. Organ weights increased in the YRT group, while the changes in the remaining two groups were minor. The c.f.u. from these organs also increased in the YRT-infected mice, whereas the infection in the yeast-and conidia-infected mice was self-limiting. A proportion (16.6%) of the YRT-infected animals died during the observation period. Conversely, no deaths occurred in the remaining groups. These results indicate that the YRT variant possesses increased virulence.

  8. A tritrophic effect of host plant on susceptibility of western flower thrips to the entomopathogenic fungus Beauveria bassiana.

    PubMed

    Ugine, Todd A; Wraight, Stephen P; Sanderson, John P

    2007-10-01

    Adult female western flower thrips (Frankliniella occidentalis) were exposed 12-24h to bean (Phaseolus vulgaris) and impatiens (Impatiens wallerana) leaf disks treated with Beauveria bassiana conidia and then transferred to clean bean or impatiens at various times post-treatment. Significantly greater levels of fungal infection were observed when thrips were treated on bean versus impatiens, but exposure to impatiens following treatment had no effect on fungal infection (percent mortality). This result, combined with observations of no inhibition of germination of conidia exposed to intact or macerated impatiens foliage, indicated that the negative effect of the impatiens host plant was not due to plant chemical compounds (antibiosis). Further observations revealed that insects acquired (picked-up) 75% more conidia from treated bean disks than from treated impatiens disks. This difference in dose acquisition was determined to account for the observed difference in percent mortality (15%) following treatment on the two host plants. Median lethal doses (LD(50)) of B. bassiana were not significantly different on the two host plants, but median lethal concentrations were nearly 7-fold greater on impatiens. This difference was explained by disproportionate rates of conidial acquisition at measured rates of conidial deposition (an inverse relationship was observed between application rate expressed as conidia/mm(2) and the number of conidia acquired). The mechanism underlying the differential rates of conidial acquisition from bean versus impatiens was not determined.

  9. Synthesis of Melanin-Like Pigments by Sporothrix schenckii In Vitro and during Mammalian Infection

    PubMed Central

    Morris-Jones, Rachael; Youngchim, Sirida; Gomez, Beatriz L.; Aisen, Phil; Hay, Roderick J.; Nosanchuk, Joshua D.; Casadevall, Arturo; Hamilton, Andrew J.

    2003-01-01

    Melanin has been implicated in the pathogenesis of several important human fungal pathogens. Existing data suggest that the conidia of the dimorphic fungal pathogen Sporothrix schenckii produce melanin or melanin-like compounds; in this study we aimed to confirm this suggestion and to demonstrate in vitro and in vivo production of melanin by yeast cells. S. schenckii grown on Mycosel agar produced visibly pigmented conidia, although yeast cells grown in brain heart infusion and minimal medium broth appeared to be nonpigmented macroscopically. However, treatment of both conidia and yeast cells with proteolytic enzymes, denaturant, and concentrated hot acid yielded dark particles similar in shape and size to the corresponding propagules, which were stable free radicals consistent with identification as melanins. Melanin particles extracted from S. schenckii yeast cells were used to produce a panel of murine monoclonal antibodies (MAbs) which labeled pigmented conidia, yeast cells, and the isolated particles. Tissue from hamster testicles infected with S. schenckii contained fungal cells that were labeled by melanin-binding MAbs, and digestion of infected hamster tissue yielded dark particles that were also reactive. Additionally, sera from humans with sporotrichosis contained antibodies that bound melanin particles. These findings indicate that S. schenckii conidia and yeast cells can produce melanin or melanin-like compounds in vitro and that yeast cells can synthesize pigment in vivo. Since melanin is an important virulence factor in other pathogenic fungi, this pigment may have a similar role in the pathogenesis of sporotrichosis. PMID:12819091

  10. Monitoring environmental Aspergillus spp. contamination and meteorological factors in a haematological unit.

    PubMed

    Cavallo, M; Andreoni, S; Martinotti, M G; Rinaldi, M; Fracchia, L

    2013-12-01

    The opportunistic pathogens belonging to the Aspergillus genus are present in almost all seasons of the year, and their concentration is related to meteorological conditions. The high density of Aspergillus spp. conidia in a haematological hospital ward may be a significant risk factor for developing invasive fungal diseases in immunocompromised patients. Aim of the present study was to evaluate the variability of airborne Aspergillus spp. conidia contamination in a Haematological Unit (HU) within a period of 16 months in relation with some meteorological parameters. An environmental Aspergillus surveillance was conducted in the HU in four rooms and their bathrooms, in the corridor and in three external sites using an agar impact sampler. During each sampling, temperature and relative humidity at each site were recorded and current wind speed and rainfall events were taken from the official weather service. Aspergillus spp. conidia concentration differed significantly across the sampling sites. Internal Aspergillus spp. loads were significantly dependent on temperature, internal relative humidity and rain. External conidia concentrations were significantly influenced by outdoor temperature and relative humidity. A suitable indicator was introduced to evaluate the seasonal distribution of Aspergillus spp. conidia in the sampling sites, and a significant dependence on this indicator was observed inside the HU. Seventeen different fungal species belonging to the Aspergillus genus were detected during the sampling period. Aspergillus fumigatus was the most frequently isolated species and its distribution depended significantly on the seasonal indicator both inside and outside the hospital ward.

  11. Agent-based modeling approach of immune defense against spores of opportunistic human pathogenic fungi.

    PubMed

    Tokarski, Christian; Hummert, Sabine; Mech, Franziska; Figge, Marc Thilo; Germerodt, Sebastian; Schroeter, Anja; Schuster, Stefan

    2012-01-01

    Opportunistic human pathogenic fungi like the ubiquitous fungus Aspergillus fumigatus are a major threat to immunocompromised patients. An impaired immune system renders the body vulnerable to invasive mycoses that often lead to the death of the patient. While the number of immunocompromised patients is rising with medical progress, the process, and dynamics of defense against invaded and ready to germinate fungal conidia are still insufficiently understood. Besides macrophages, neutrophil granulocytes form an important line of defense in that they clear conidia. Live imaging shows the interaction of those phagocytes and conidia as a dynamic process of touching, dragging, and phagocytosis. To unravel strategies of phagocytes on the hunt for conidia an agent-based modeling approach is used, implemented in NetLogo. Different modes of movement of phagocytes are tested regarding their clearing efficiency: random walk, short-term persistence in their recent direction, chemotaxis of chemokines excreted by conidia, and communication between phagocytes. While the short-term persistence hunting strategy turned out to be superior to the simple random walk, following a gradient of chemokines released by conidial agents is even better. The advantage of communication between neutrophilic agents showed a strong dependency on the spatial scale of the focused area and the distribution of the pathogens.

  12. Resting spore formation of aphid-pathogenic fungus Pandora nouryi depends on the concentration of infective inoculum.

    PubMed

    Huang, Zhi-Hong; Feng, Ming-Guang

    2008-07-01

    Resting spore formation of some aphid-pathogenic Entomophthorales is important for the seasonal pattern of their prevalence and survival but this process is poorly understood. To explore the possible mechanism involved in the process, Pandora nouryi (obligate aphid pathogen) interacted with green peach aphid Myzus persicae on cabbage leaves under favourable conditions. Host nymphs showered with primary conidia of an isolate (LC(50): 0.9-6.7 conidia mm(-2) 4-7 days post shower) from air captures in the low-latitude plateau of China produced resting spores (azygospores), primary conidia or both spore types. Surprisingly, the proportion of mycosed cadavers forming resting spores (P(CFRS)) increased sharply within the concentrations (C) of 28-240 conidia mm(-2), retained high levels at 240-1760, but was zero or extremely low at 0.3-16. The P(CFRS)-C relationship fit well the logistic equation P(CFRS) = 0.6774/[1 + exp(3.1229-0.0270C)] (r(2) = 0.975). This clarified for the first time the dependence of in vivo resting spore formation of P. nouryi upon the concentration of infective inoculum. A hypothesis is thus proposed that some sort of biochemical signals may exist in the host-pathogen interaction so that the fungal pathogen perceives the signals for prompt response to forthcoming host-density changes by either producing conidia for infecting available hosts or forming resting spores for surviving host absence in situ.

  13. Inactivation of plant-pathogenic fungus Colletotrichum acutatum with natural plant-produced photosensitizers under solar radiation.

    PubMed

    Fracarolli, Letícia; Rodrigues, Gabriela B; Pereira, Ana C; Massola Júnior, Nelson S; Silva-Junior, Geraldo José; Bachmann, Luciano; Wainwright, Mark; Bastos, Jairo Kenupp; Braga, Gilberto U L

    2016-09-01

    The increasing tolerance to currently used fungicides and the need for environmentally friendly antimicrobial approaches have stimulated the development of novel strategies to control plant-pathogenic fungi such as antimicrobial phototreatment (APT). We investigated the in vitro APT of the plant-pathogenic fungus Colletotrichum acutatum with furocoumarins and coumarins and solar radiation. The compounds used were: furocoumarins 8-methoxypsoralen (8-MOP) and 5,8-dimethoxypsoralen (isopimpinellin), coumarins 2H-chromen-2-one (coumarin), 7-hydroxycoumarin, 5,7-dimethoxycoumarin (citropten) and a mixture (3:1) of 7-methoxycoumarin and 5,7-dimethoxycoumarin. APT of conidia with crude extracts from 'Tahiti' acid lime, red and white grapefruit were also performed. Pure compounds were tested at 50μM concentration and mixtures and extracts at 12.5mgL(-1). The C. acutatum conidia suspension with or without the compounds was exposed to solar radiation for 1h. In addition, the effects of APT on the leaves of the plant host Citrus sinensis were determined. APT with 8-MOP was the most effective treatment, killing 100% of the conidia followed by the mixture of two coumarins and isopimpinellin that killed 99% and 64% of the conidia, respectively. APT with the extracts killed from 20% to 70% of the conidia, and the extract from 'Tahiti' lime was the most effective. No damage to sweet orange leaves was observed after APT with any of the compounds or extracts. Copyright © 2016 Elsevier B.V. All rights reserved.

  14. Alteration of protein patterns in black rock inhabiting fungi as a response to different temperatures

    PubMed Central

    Tesei, Donatella; Marzban, Gorji; Zakharova, Kristina; Isola, Daniela; Selbmann, Laura; Sterflinger, Katja

    2012-01-01

    Rock inhabiting fungi are among the most stress tolerant organisms on Earth. They are able to cope with different stressors determined by the typical conditions of bare rocks in hot and cold extreme environments. In this study first results of a system biological approach based on two-dimensional protein profiles are presented. Protein patterns of extremotolerant black fungi – Coniosporium perforans, Exophiala jeanselmei – and of the extremophilic fungus – Friedmanniomyces endolithicus – were compared with the cosmopolitan and mesophilic hyphomycete Penicillium chrysogenum in order to follow and determine changes in the expression pattern under different temperatures. The 2D protein gels indicated a temperature dependent qualitative change in all the tested strains. Whereas the reference strain P. chrysogenum expressed the highest number of proteins at 40 °C, thus exhibiting real signs of temperature induced reaction, black fungi, when exposed to temperatures far above their growth optimum, decreased the number of proteins indicating a down-regulation of their metabolism. Temperature of 1 °C led to an increased number of proteins in all of the analysed strains, with the exception of P. chrysogenum. These first results on temperature dependent reactions in rock inhabiting black fungi indicate a rather different strategy to cope with non-optimal temperature than in the mesophilic hyphomycete P. chrysogenum. PMID:22862921

  15. Phylogeny of mitosporic entomopathogenic fungi: is the genus Paecilomyces polyphyletic?

    PubMed

    Obornik, M; Jirku, M; Dolezel, D

    2001-09-01

    We analyzed sequences of the divergent domain at the 5' end of the large subunit rRNA gene from the mitosporic entomopathogenic fungi Paecilomyces sp., Paecilomyces fumosoroseus, Paecilomyces farinosus, Paecilomyces lilacinus, Verticillium lecanii, Verticillium psalliotae, Beauveria bassiana, Aschersonia sp., Aschersonia placenta, ascomycetous Cordyceps sp., and Cordyceps militaris. Phylogenetic analysis showed P. fumosorseus as the best characterized out of the analyzed species with the B. bassiana clade as its sister group. Two of the P. farinosus isolates were invariably placed within the Verticillium cluster, which also contained C. militaris. The only analyzed P. lilacinus isolate appeared on the root of the hyphomycetous fungi and was characterized as the most distinct from all the hyphomycetous fungi tested. Polyphyly of the genus Paecilomyces was well supported by the Kishino-Hasegawa test. In all trees based on the small subunit rRNA gene sequences obtained from the GenBank, V. lecanii, V. psalliotae, P. fumosoroseus, P. tenuipes and B. bassiana form, together with that of C. militaris, the best supported cluster in the tree. The rest of Cordyceps spp. constitute a distinct clade. Phylogenetic relationships derived from both tested DNA regions show polyphyly of the genus Paecilomyces and close relationships among entomopathogenic species of the genera Verticillium, Paecilomyces, and Beauveria.

  16. Fungi and bacteria involved in desert varnish formation

    NASA Technical Reports Server (NTRS)

    Taylor-George, S.; Palmer, F.; Staley, J. T.; Curtiss, B.; Adams, J. B.; Borns, D. J.

    1983-01-01

    Desert varnish is a coating of ferromanganese oxides and clays that develops on rock surfaces in arid to semi-arid regions. Active respiration but not photosynthesis was detected on varnished rock surfaces from the Sonoran Desert. Light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) observations, and cultivation experiments indicate that both fungi, primarily dematiaceous hyphomycetes, and bacteria are found on and within desert varnish coatings from the arid regions studied. Some fungi grow as microcolonial fungi (MCF) on rocks, and microscopic observations suggest MCF become incorporated in the varnish coating. SEM-EDAX (energy dispersive X-ray systems) analyses indicate the MCF contain 3 of the characteristic elements of varnish: iron, aluminum, and silicon. In some locations, MCF are also enriched in manganese relative to the rock substratum. Furthermore, some of the dematiaceous hyphomycetes that have been cultivated are able to oxidize manganese under laboratory conditions. It is possible that manganese-oxidizing bacteria, which are found in varnish, also play an important role in varnish formation.

  17. Ethanol and phenanthrene increase the biomass of fungal assemblages and decrease plant litter decomposition in streams.

    PubMed

    Barros, Diana; Oliveira, Patrícia; Pascoal, Cláudia; Cássio, Fernanda

    2016-09-15

    Fungi, particularly aquatic hyphomycetes, have been recognized as playing a dominant role in microbial decomposition of plant litter in streams. In this study, we used a microcosm experiment with different levels of fungal diversity (species number and identity) using monocultures and combinations with up to five aquatic hyphomycete species (Articulospora tetracladia, Tricladium splendens, Heliscus submersus, Tetrachaetum elegans and Flagellospora curta) to assess the effects of ethanol and phenanthrene on three functional measures: plant litter decomposition, fungal biomass accrual and reproduction. Alder leaves were conditioned by fungi for 7days and then were exposed to phenanthrene (1mgL(-1)) dissolved in ethanol (0.1% final concentration) or ethanol (at the concentration used to solubilise phenanthrene) for further 24days. Exposure to ethanol alone or in combination with phenanthrene decreased leaf decomposition and fungal reproduction, but increased fungal biomass produced. All aspects of fungal activity varied with species number. Fungal activity in polycultures was generally higher than that expected from the sum of the weighted performances of participating species in monoculture, suggesting complementarity between species. However, the activity of fungi in polycultures did not exceed the activity of the most productive species either in the absence or presence of ethanol alone or with phenanthrene.

  18. Fungi in freshwaters: ecology, physiology and biochemical potential.

    PubMed

    Krauss, Gerd-Joachim; Solé, Magali; Krauss, Gudrun; Schlosser, Dietmar; Wesenberg, Dirk; Bärlocher, Felix

    2011-07-01

    Research on freshwater fungi has concentrated on their role in plant litter decomposition in streams. Higher fungi dominate over bacteria in terms of biomass, production and enzymatic substrate degradation. Microscopy-based studies suggest the prevalence of aquatic hyphomycetes, characterized by tetraradiate or sigmoid spores. Molecular studies have consistently demonstrated the presence of other fungal groups, whose contributions to decomposition are largely unknown. Molecular methods will allow quantification of these and other microorganisms. The ability of aquatic hyphomycetes to withstand or mitigate anthropogenic stresses is becoming increasingly important. Metal avoidance and tolerance in freshwater fungi implicate a sophisticated network of mechanisms involving external and intracellular detoxification. Examining adaptive responses under metal stress will unravel the dynamics of biochemical processes and their ecological consequences. Freshwater fungi can metabolize organic xenobiotics. For many such compounds, terrestrial fungal activity is characterized by cometabolic biotransformations involving initial attack by intracellular and extracellular oxidative enzymes, further metabolization of the primary oxidation products via conjugate formation and a considerable versatility as to the range of metabolized pollutants. The same capabilities occur in freshwater fungi. This suggests a largely ignored role of these organisms in attenuating pollutant loads in freshwaters and their potential use in environmental biotechnology. © 2011 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

  19. A new species of Heliocephala from Mexico with an assessment of the systematic positions of the anamorph genera Heliocephala and Holubovaniella.

    PubMed

    Abarca, Gabriela Heredia; Ruiz, Rafael F Castañeda; Mota, Rosa María Arias; Hérnandez, Cinthya I Becerra; Gómez, Sergio; Bogale, Mesfin; Untereiner, Wendy A

    2011-01-01

    During a study of the microfungi on decaying leaves collected in México we encountered a fungus sharing features of the anamorph genera Heliocephala and Holubovaniella. This hyphomycete resembled Hol. elegans in possessing obclavate to fusiform, three-septate, rostrate phragmoconidia, but it can be distinguished by its much shorter, branched, determinate conidiophores. Unlike members of genus Heliocephala, which also possess determinant conidiophores, the phragmoconidia of this taxon are short-rostrate and have not been observed to germinate iteratively. Prompted by the discovery of this hyphomycete, we investigated the phylogenetic relationships among species of Heliocephala and Holubovaniella. Analysis of large subunit rDNA gene sequences positioned species of Heliocephala and Holubovaniella in the Dothideomycetes and identified Stomiopeltis betulae (Micropeltididaceae) as their closest relative. These results also indicated that Heliocephala and Holubovaniella are closely related taxa. Based on these findings and the similarity of the developmental and morphological characters of species of Heliocephala and Holubovaniella, we recognize these genera as synonymous and describe a new species, Heliocephala triseptata. A dichotomous key to the species of Heliocephala is provided.

  20. Alteration of protein patterns in black rock inhabiting fungi as a response to different temperatures.

    PubMed

    Tesei, Donatella; Marzban, Gorji; Zakharova, Kristina; Isola, Daniela; Selbmann, Laura; Sterflinger, Katja

    2012-08-01

    Rock inhabiting fungi are among the most stress tolerant organisms on Earth. They are able to cope with different stressors determined by the typical conditions of bare rocks in hot and cold extreme environments. In this study first results of a system biological approach based on two-dimensional protein profiles are presented. Protein patterns of extremotolerant black fungi -Coniosporium perforans, Exophiala jeanselmei - and of the extremophilic fungus -Friedmanniomyces endolithicus - were compared with the cosmopolitan and mesophilic hyphomycete Penicillium chrysogenum in order to follow and determine changes in the expression pattern under different temperatures. The 2D protein gels indicated a temperature dependent qualitative change in all the tested strains. Whereas the reference strain P. chrysogenum expressed the highest number of proteins at 40 °C, thus exhibiting real signs of temperature induced reaction, black fungi, when exposed to temperatures far above their growth optimum, decreased the number of proteins indicating a down-regulation of their metabolism. Temperature of 1 °C led to an increased number of proteins in all of the analysed strains, with the exception of P. chrysogenum. These first results on temperature dependent reactions in rock inhabiting black fungi indicate a rather different strategy to cope with non-optimal temperature than in the mesophilic hyphomycete P. chrysogenum.

  1. Fungi and bacteria involved in desert varnish formation

    NASA Technical Reports Server (NTRS)

    Taylor-George, S.; Palmer, F.; Staley, J. T.; Curtiss, B.; Adams, J. B.; Borns, D. J.

    1983-01-01

    Desert varnish is a coating of ferromanganese oxides and clays that develops on rock surfaces in arid to semi-arid regions. Active respiration but not photosynthesis was detected on varnished rock surfaces from the Sonoran Desert. Light microscopy, scanning electron microscopy (SEM) and transmission electron microscopy (TEM) observations, and cultivation experiments indicate that both fungi, primarily dematiaceous hyphomycetes, and bacteria are found on and within desert varnish coatings from the arid regions studied. Some fungi grow as microcolonial fungi (MCF) on rocks, and microscopic observations suggest MCF become incorporated in the varnish coating. SEM-EDAX (energy dispersive X-ray systems) analyses indicate the MCF contain 3 of the characteristic elements of varnish: iron, aluminum, and silicon. In some locations, MCF are also enriched in manganese relative to the rock substratum. Furthermore, some of the dematiaceous hyphomycetes that have been cultivated are able to oxidize manganese under laboratory conditions. It is possible that manganese-oxidizing bacteria, which are found in varnish, also play an important role in varnish formation.

  2. Pathogenicity and ultrastructural studies of the mode of penetration by Phoma strasseri in peppermint stems and rhizomes.

    PubMed

    Zimowska, Beata

    2012-01-01

    Pathogenicity and ultrastructural investigation of the inoculation of peppermint stems and rhizomes with Phoma strasseri conidia was undertaken using scanning and transmission electron microscopy to examine the host-parasite relationship. Pathogenicity experiments demonstrated that all tested P. strasseri isolates had infected the stems and rhizomes of peppermint. Of all inoculation methods, direct placement of colonized agar plugs on damaged epidermis and soaking stems and rhizomes in conidial suspension were the most effective. The behavior of the conidia deposited on the stems and rhizomes was investigated at different time intervals after inoculation: 6, 16, 24, 36 and 48 h. Conidia produced an appressorium directly at the end of a short germ tube. Appressoria were formed over the cuticle, but never over stomata. Direct penetration to host tissue through the cuticle was observed. The spore and hyphae were covered with a mucilaginous sheath.

  3. Identification and functional analysis of endogenous nitric oxide in a filamentous fungus

    PubMed Central

    Pengkit, Anchalee; Jeon, Seong Sil; Son, Soo Ji; Shin, Jae Ho; Baik, Ku Yeon; Choi, Eun Ha; Park, Gyungsoon

    2016-01-01

    In spite of its prevalence in animals and plants, endogenous nitric oxide (NO) has been rarely reported in fungi. We present here our observations on production of intracellular NO and its possible roles during development of Neurospora crassa, a model filamentous fungus. Intracellular NO was detected in hypha 8–16 hours after incubation in Vogel’s minimal liquid media and conidiophores during conidiation using a fluorescent indicator (DAF-FM diacetate). Treatment with cPTIO, an NO scavenger, significantly reduced fluorescence levels and hindered hyphal growth in liquid media and conidiation, whereas exogenous NO enhanced hyphal extension on VM agar media and conidia formation. NO scavenging also dramatically diminished transcription of con-10 and con-13, genes preferentially expressed during conidiation. Our results suggest that intracellular NO is generated in young hypha growing in submerged culture and during conidia development and regulate mycelial development and conidia formation. PMID:27425220

  4. Production of tremorgenic mycotoxins by isolates of Aspergillus fumigatus from sawmills in Sweden.

    PubMed

    Land, C J; Lundström, H; Werner, S

    1993-11-01

    One hundred and six strains of A. fumigatus were isolated from 21 sawmills in Sweden, and 73 of these strains were examined for production of fumitremorgen B and verruculogen (tremorgenic mycotoxins) on YES-medium using thin layer chromatography (TLC). Twenty-three strains (32%) were tremorgen producers and 50 strains (68%) were non-producers. Tremorgenic mycotoxins were detected in conidia of seven A. fumigatus strains. The amount of toxin varied between 0.6-8.0 microgram/10(8) conidia (mean value 2.3 micrograms/10(8) conidia, equivalent with 0.18%). No production of the mycotoxin gliotoxin was detected in 6 strains of A. fumigatus. No tremorgens were detected during mould growth on wood substrates, in spite of the use of different wood species (Scots pine, Pinus sylvestris: Norway spruce, Picea abies and birch, Betula spp.), dried versus non-dried wood, bark (pine), leached wood, and wood after various sterilization methods.

  5. Comment on: "Melanisation of Aspergillus terreus-Is Butyrolactone I Involved in the Regulation of Both DOPA and DHN Types of Pigments in Submerged Culture? Microorganisms 2017, 5, 22".

    PubMed

    Geib, Elena; Brock, Matthias

    2017-06-21

    A recent article by Palonen et al. describes the effect of butyrolactone I on the expression of a secondary metabolite biosynthesis gene cluster from Aspergillus terreus that shows similarities to fusarubin biosynthesis gene clusters from Fusarium species. The authors claim that two different types of pigments are formed in Aspergillus terreus conidia, whereby one pigment is termed a DOPA-type melanin and the second a DHN-type melanin. Unfortunately, the terminology of the classification of melanin-types requires revision as Asp-melanin present in A. terreus conidia is clearly distinct from DOPA-melanins. In addition, some hypotheses in this manuscript are based on questionable data published previously, resulting in incorrect conclusions. Finally, as biochemical data are lacking and metabolite production is only deduced from bioinformatics and transcriptomic data, the production of a second pigment type in A. terreus conidia appears highly speculative.

  6. Liberomyces gen. nov. with two new species of endophytic coelomycetes from broadleaf trees.

    PubMed

    Pazoutová, Sylvie; Srutka, Petr; Holusa, Jaroslav; Chudícková, Milada; Kubátová, Alena; Kolarík, Miroslav

    2012-01-01

    During a study of endophytic and saprotrophic fungi in the sapwood and phloem of broadleaf trees (Salix alba, Quercus robur, Ulmus laevis, Alnus glutinosa, Betula pendula) fungi belonging to an anamorphic coelomycetous genus not attributable to a described taxon were detected and isolated in pure culture. The new genus, Liberomyces, with two species, L. saliciphilus and L. macrosporus, is described. Both species have subglobose conidiomata containing holoblastic sympodial conidiogenous cells. The conidiomata dehisce irregularly or by ostiole and secrete a slimy suspension of conidia. The conidia are hyaline, narrowly allantoid with a typically curved distal end. In L. macrosporus simultaneous production of synanamorph with thin filamentous conidia was observed occasionally. The genus has no known teleomorph. Related sequences in the public databases belong to endophytes of angiosperms. Phylogenetic analysis revealed a position close to the Xylariales (Sordariomycetes), but family and order affiliation remained unclear.

  7. Ulocladium cantlous sp. nov. isolated from northwestern China: its morphology and molecular phylogenetic position.

    PubMed

    Wang, Yong; Pei, Yun-Fei; O'Neill, Nichole R; Zhang, Xiu-Guo

    2010-01-01

    A new species of Ulocladium was isolated from diseased leaves from two Cucumis sp. growing in Sinkiang and Gansu provinces of China. Conidia were isolated from necrotic leaves and used to establish single-spore pure cultures. Conidia were harvested from cultures 7 d after incubation for morphological comparisons. The morphology of this species resembles that of U. botrytis and U. consortiale. However it is distinguished from these two species by the sizes of obovoid to broadly ellipsoidal conidia and longer conidiophores. A taxonomic description of U. cantlous, comparison with related species in this genus, and a species phylogeny based on the partial nucleotide sequence of the glyceraldehyde-3-phosphate dehydrogenase (gpd) gene and the Alternaria alternata major allergen (Alt a 1) gene are provided.

  8. Lipid mobilization and acid phosphatase activity in lytic compartments during conidium dormancy and appressorium formation of Colletotrichum graminicola.

    PubMed

    Schadeck, R J; Leite, B; de Freitas Buchi, D

    1998-12-01

    Colletotrichum graminicola, a pathogen of sorghum and corn, was investigated prior and during germination as to certain aspects of acid phosphatase activity and lipid mobilization. Ungerminated conidia cytoplasm was filled with lipid deposits, which were mobilized during the germination process. Cytochemical ultrastructural examination showed that conidia vacuoles exhibit acid phosphatase activity, which is suggestive of lytic activity. Lipid bodies, stored in the ungerminated conidia cytoplasm, were internalized by vacuoles in a process analogous to microautophagy and were apparently digested inside them. The lipid bodies disappeared and vacuoles became enlarged in conidial cells during germination. Appressoria also showed acid phosphatase activity in multiple heterogeneous vesicles which were, in most cases, juxtaposed with lipid bodies. These results suggest that the vacuolar system plays an important role during C. graminicola germination and that the initial stages of lipid metabolization are taking place inside the vacuoles.

  9. In vitro activity of Melaleuca alternifolia (tea tree) oil against dermatophytes and other filamentous fungi.

    PubMed

    Hammer, K A; Carson, C F; Riley, T V

    2002-08-01

    The in vitro activity of Melaleuca alternifolia (tea tree) oil against dermatophytes (n = 106) and filamentous fungi (n = 78) was determined. Tea tree oil MICs for all fungi ranged from 0.004% to 0.25% and minimum fungicidal concentrations (MFCs) ranged from <0.03% to 8.0%. Time-kill experiments with 1-4 x MFC demonstrated that three of the four test organisms were still detected after 8 h of treatment, but not after 24 h. Comparison of the susceptibility to tea tree oil of germinated and non-germinated Aspergillus niger conidia showed germinated conidia to be more susceptible than non-germinated conidia. These data demonstrate that tea tree oil has both inhibitory and fungicidal activity.

  10. Rhinofacial conidiobolomycosis caused by Conidiobolus coronatus in a Chinese rice farmer.

    PubMed

    Yang, Xiumin; Li, Yadi; Zhou, Xiaoqian; Wang, Yuxin; Geng, Suying; Liu, Honggang; Yang, Qingwen; Lu, Xinxin; Hiruma, Masataro; Sugita, Takashi; Ikeda, Shigaku; Ogawa, Hideoki

    2010-07-01

    We provide the first report of rhinofacial conidiobolomycosis caused by Conidiobolus coronatus in China. The patient was a 36-year-old male rice farmer from Jiang Xi province who presented with diffuse swelling with classical centrofacial distribution and involvement of the right paranasal sinuses. Histopathology showed granulomas with hyphae surrounded by an eosinophilic sheath (Splendore-Hoeppli phenomenon). Culture of biopsy specimens on Sabouraud's dextrose agar led to the growth of fungi with microscopically visible conidiophores and terminal spherical conidia (primary conidium), with multiple secondary conidia and villose conidia. The patient was successfully treated with combination therapy, primarily itraconazole and terbinafine. We conclude with a brief literature review of the epidemiology of conidiobolomycosis.

  11. Pandora neoaphidis transmission and aphid foraging behaviour.

    PubMed

    Baverstock, J; Alderson, P G; Pell, J K

    2005-09-01

    Pandora neoaphidis is an aphid-specific entomopathogen that produces infective conidia. As aphid movement increases, so does the likelihood of contact with conidia. Volatile distress signals released in response to aphid infestation as an indirect defence against herbivory may affect aphid foraging and, therefore, the fungus-aphid interaction. In this study, two different methods were used to investigate the effect of plant volatiles and P. neoaphidis-sporulating cadavers on (1) the colonisation of Vicia faba plants by Acyrthosiphon pisum and (2) P. neoaphidis transmission. This study indicates that A. pisum does not avoid bean plants containing P. neoaphidis and that transmission of conidia occurs during plant colonisation and, to a lesser extent, during in situ feeding. Although significantly more aphids were recovered from damaged plants compared to undamaged plants, the likelihood of infection was not affected by previous infestation by aphids.

  12. Efficacy of Trichoderma asperellum oil formulations on the control of cocoa black pod disease (Phytophthora megakarya).

    PubMed

    Mbarga, J B; Ten Hoopen, G Martijn; Begoude, A D; Tondje, P R; Kuate, J; Ambang, Z; Amougou, A; Schiffers, B

    2012-01-01

    The objective of this study was therefore to develop a formulation of conidia of T. asperellum with the aim of improving its efficacy. The formulations developed were oily dispersions. It was a combination of solvents consisting of groundnut oil or palm oil with structural agents and emulsifying-dispersing agents. Emulsification tests were carried out and the stability of the emulsions evaluated. The evaluation of the effect of co-formulants on the growth of conidia of T. asperellum was done by reading the optical densities of the formulated samples on multi-plates using a plate reader. The test on detached cocoa pods was done by treating the cocoa pods with selected formulations at 1.10(7) conidia/ml and inoculation of the treated cocoa pods was done 24 hours later with zoospores of P. megakarya at 1.10(5) zoospores/ml. The growth of necrosis on the fruits was measured daily. The screening of co-formulants and emulsification tests ended up with the selection of two formulations. The first composed of conidia of T. asperellum, groundnut oil, Tensiofix NTM and Tensiofix 869. The second differed from the first by utilisation of palm oil as the solvent. These formulations proved stable when diluted in water with 1% and 0.5% of sedimentation respectively after 24 hours. The viability test of the conidia indicated that the different formulations selected did not have a fungitoxic effect. The test on detached cocoa pods showed an improved efficacy of T. asperellum to control the disease. The growth rates of necrosis were 6.29 mm/day, 7.25 mm/day and 31.6 mm/day for treatment with formulation 1, pure conidia and control treated with water respectively.

  13. N-Chlorotaurine Exhibits Fungicidal Activity against Therapy-Refractory Scedosporium Species and Lomentospora prolificans

    PubMed Central

    Lackner, Michaela; Binder, Ulrike; Reindl, Martin; Gönül, Beyhan; Fankhauser, Hannes; Mair, Christian

    2015-01-01

    N-Chlorotaurine (NCT), a well-tolerated endogenous long-lived oxidant that can be applied topically as an antiseptic, was tested on its fungicidal activity against Scedosporium and Lomentospora, opportunistic fungi that cause severe infections with limited treatment options, mainly in immunocompromised patients. In quantitative killing assays, both hyphae and conidia of Scedosporium apiospermum, Scedosporium boydii, and Lomentospora prolificans (formerly Scedosporium prolificans) were killed by 55 mM (1.0%) NCT at pH 7.1 and 37°C, with a 1- to 4-log10 reduction in CFU after 4 h and a 4- to >6-log10 reduction after 24 h. The addition of ammonium chloride to NCT markedly increased this activity. LIVE/DEAD staining of conidia treated with 1.0% NCT for 0.5 to 3 h increased the permeability of the cell wall and membrane. Preincubation of the test fungi in 1.0% NCT for 10 to 60 min delayed the time to germination of conidia by 2 h to >12 h and reduced their germination rate by 10.0 to 100.0%. Larvae of Galleria mellonella infected with 1.0 × 107 conidia of S. apiospermum and S. boydii died at a rate of 90.0 to 100% after 8 to 12 days. The mortality rate was reduced to 20 to 50.0% if conidia were preincubated in 1.0% NCT for 0.5 h or if heat-inactivated conidia were used. Our study demonstrates the fungicidal activity of NCT against different Scedosporium and Lomentospora species. A postantifungal effect connected with a loss of virulence occurs after sublethal incubation times. The augmenting effect of ammonium chloride can be explained by the formation of monochloramine. PMID:26239996

  14. Isolation of entomopathogenic fungi from Northern Thailand and their production in cereal grains.

    PubMed

    Mar, Thet Thet; Suwannarach, Nakarin; Lumyong, Saisamorn

    2012-12-01

    Spore productivity in six entomopathogenic fungal strains isolated from insect cadavers at four locations in Chiang Mai province was evaluated in five cereal grains: white-rice, wheat, rye, corn and sorghum. According to sequence analysis of the internal transcribed spacer regions of these isolates, they were closely related to Beauveria bassiana (2 isolates), Metarhizium flavoviride (1 isolate), Metarhizium anisopliae (1 isolate), Paecilomyces lilacinus (1 isolate) and Isaria tenuipes (1 isolate). Among all fungal isolates, the maximum amount of spores (530.0 × 10(9) conidia/g) was yielded P. lilacinus CMUCDMT02 on sorghum grain followed by white-rice (399.3 × 10(9) conidia/g). Moreover, the highest number of spore in M. flavoviride was 102.8 × 10(9) conidia/g sorghum whereas white-rice yielded the greatest amount of spore for B. bassiana CMUCDMF03 (141.0 × 10(9) conidia/g) after 60 days incubation. The fungal growth rate was found highest in corn for all strains and rye showed the lowest with the exception of P. lilacinus CMUCDMT02 among the tested grains. Spore viability was over 80 % for all isolates that had been inoculated for 60 days. Fungal conidia suspension of P. lilacinus obtained highest virulence against Bactrocera spp. at a concentration of 1 × 10(6) spore/ml. The strains isolated, exhibited good production of conidia suggesting a promising strategy for the mass production of inoculum as biocontrol agents with low production cost.

  15. Laboratory and field evaluation of the fungus Chrysosporium lobatum against the larvae of the mosquito Culex quinquefasciatus.

    PubMed

    Mohanty, Suman Sundar; Prakash, Soam

    2008-04-01

    Eleven fungal species in three genera were isolated from the soil at Agra, India by the feather-baiting technique. Out of the 11 species, Chrysosporium lobatum Scharapov, a deuteromycetous (Moniliales: Moniliaceae) fungus, caused high mortality of Culex quinquefasciatus Say (Diptera: Culicidae) larvae in the laboratory. Laboratory and field trials were carried out to study the efficacy of C. lobatum against various instars of C. quinquefasciatus. In the laboratory bioassays, first, second and third instar larvae were assessed separately. Conidia of C. lobatum that were 15 days old were used both in laboratory and field bioassays. Six different concentrations were used in the laboratory bioassays (10, 10(3), 10(4), 5x10(4), 10(5) and 10(6) conidia/mL), and one concentration was tested in the field trials (10(6) conidia/mL). The LC50 values with 95% fiducial limits and probit equations were calculated by the probit analysis. Significant (analysis o.f variance (ANOVA): P<0.0001) mortality difference between the instars of C. quinquefasciatus was observed after 72 h of exposure to conidia of C. lobatum. The third instar C. quinquefasciatus were 319- and 25-fold more susceptible to C. lobatum (0.47x10(3) conidia/mL) than the first and second instars, respectively. The cuticle of the abdominal region and anal papillae were densely covered by the C. lobatum conidia. In the field trials, populations had declined significantly (t test, P=0.003) 15 days after inoculation in the four test pools. Significantly (t test, P=0.03) higher mortality was observed in the pools with water quality that was lower in total dissolved solids, hardness, chemical oxygen demand and conductivity. Based on the performance and survival in the trial, C. lobatum may be considered as a bio-control agent of mosquitoes.

  16. Brown spot control on pear: infection models versus the inoculum pressure in Belgium.

    PubMed

    Van Laer, S; Vorstermans, B; Hauke, K; Creemers, P

    2006-01-01

    Stemphylium conidia and Pleospora ascospores were monitored in pear orchards in the region of Haspengouw in Belgium during the infectious periods of brown spot disease (end May - end August) in the years 2002, 2004 and 2005. The seasonal and daily dynamics of the captured fungal spores are discussed and a correlation analysis was performed to determine possible correlation with weather parameters. Furthermore the fungicide cover obtained by spraying upon climatological infection risk is compared with the inoculum pressure in the orchards. Pleospora ascospores were found until the end of May and discharge took place during rain events at the same time-points when Venturia inaequalis ascospores were detected. Although the fungi ejected during the same time period, the relative importance of the different spore peaks differed. The first conidia were detected at the end of May. After that date conidia were found almost every day. Seasonal dynamics of the conidia clearly differed between the years and also the number of spores retrieved differed. No spores were found below 7.5 degrees C and only at a temperature above 12.5 degrees C an increase in conidia was observed. When looking at the daily dynamics, a significant negative correlation was found between the aerial spore concentration and relative humidity and leaf wetness, and a significant positive correlation with wind, temperature and water vapor pressure deficit. When spraying upon a BSP-cast CR threshold value of 0.4, all days with spore discharges above 3 conidia/m3/day were covered in 2002 and 2004. In 2005, a year with very low infection risk and infestation, two periods with high spore discharge were not covered. The observations made show that incorporation of the inoculum pressure into the infection models will probably not lead to a big improvement of the infection models.

  17. Growth substrates and caleosin-mediated functions affect conidial virulence in the insect pathogenic fungus Beauveria bassiana.

    PubMed

    Ortiz-Urquiza, Almudena; Fan, Yanhua; Garrett, Timothy; Keyhani, Nemat O

    2016-11-01

    The entomopathogenic fungus, Beauveria bassiana, is a microbial biological control agent capable of infecting a wide range of insect hosts. Conidia (spores) initiate infection via adhesion, growth and penetration of the insect cuticle, whose outmost layer is rich in lipids. Conidial virulence was investigated in B. bassiana WT and caleosin mutants (ΔBbcal1), the latter a protein involved in lipid storage and turnover. Topical insect bioassays revealed that conidia of the WT strain showed up to 40-fold differences in LD50 values depending upon the growth substrate. The most virulent conidia were harvested from potato dextrose agar containing oleic acid, and the least potent were those derived from Sabouraud dextrose/yeast extract agar (SDAY). However, with the exception of conidia derived from SDAY and Czapek Dox agar, in which values were reduced, mean lethal times to kill (LT50) were essentially unaffected. In topical bioassays, the ΔBbcal1 mutant displayed LD50 values 5-40-fold higher than the WT depending upon the growth substrate, with ΔBbcal1 conidia derived from SDAY unable to effectively penetrate the host cuticle. The ΔBbcal1 mutant also showed concomitant dramatic increases in LT50 values from a mean of ~4.5 for WT to >8.5 days for the mutant. In contrast, intrahaemocoel injection bioassays that bypass cuticle penetration events revealed only minor effects on virulence for either WT or ΔBbcal1 conidia. These data highlight the importance of caleosin-dependent lipid mobilization and/or signalling in cuticle penetration events but suggest their dispensability for immune evasion and within-host growth.

  18. Stress tolerance and virulence of insect-pathogenic fungi are determined by environmental conditions during conidial formation.

    PubMed

    Rangel, Drauzio E N; Braga, Gilberto U L; Fernandes, Éverton K K; Keyser, Chad A; Hallsworth, John E; Roberts, Donald W

    2015-08-01

    The virulence to insects and tolerance to heat and UV-B radiation of conidia of entomopathogenic fungi are greatly influenced by physical, chemical, and nutritional conditions during mycelial growth. This is evidenced, for example, by the stress phenotypes of Metarhizium robertsii produced on various substrates. Conidia from minimal medium (Czapek's medium without sucrose), complex medium, and insect (Lepidoptera and Coleoptera) cadavers had high, moderate, and poor tolerance to UV-B radiation, respectively. Furthermore, conidia from minimal medium germinated faster and had increased heat tolerance and were more virulent to insects than those from complex medium. Low water-activity or alkaline culture conditions also resulted in production of conidia with high tolerance to heat or UV-B radiation. Conidia produced on complex media exhibited lower stress tolerance, whereas those from complex media supplemented with NaCl or KCl (to reduce water activity) were more tolerant to heat and UV-B than those from the unmodified complex medium. Osmotic and nutritive stresses resulted in production of conidia with a robust stress phenotype, but also were associated with low conidial yield. Physical conditions such as growth under illumination, hypoxic conditions, and heat shock before conidial production also induced both higher UV-B and heat tolerance; but conidial production was not decreased. In conclusion, physical and chemical parameters, as well as nutrition source, can induce great variability in conidial tolerance to stress for entomopathogenic fungi. Implications are discussed in relation to the ecology of entomopathogenic fungi in the field, and to their use for biological control. This review will cover recent technologies on improving stress tolerance of entomopathogenic fungi for biological control of insects.

  19. Membrane permeability and the loss of germination factor from Neurospora crassa at low water activities

    NASA Technical Reports Server (NTRS)

    Charlang, G.; Horowitz, N. H.

    1974-01-01

    Neurospora crassa conidia incubating in buffer at low water activities release a germination-essential component as well as 260-nm absorbing and ninhydrin-positive materials, regardless of whether an electrolyte or nonelectrolyte is used to reduce water activity. Chloroform and antibiotics known to increase cell-membrane permeability have a similar effect. This suggests that membrane damage occurs in media of low water activity and that an increase in permeability is responsible for the release of cellular components. The damage caused in media of low water activity is nonlethal in most cases, and the conidia recover when transferred to nutrient medium.

  20. Heat-induced changes in respiratory pathways and mitochondrial structure during microcycle conidiation of Neurospora crassa.

    PubMed

    Michéa-Hamzehpour, M; Grange, F; Ton That, T C; Turian, G

    1980-03-01

    Changes in both respiratory pathways and mitochondrial structure of Neurospora crassa occurred under conditions of microcycle conidiation. Upon heat-treatment at 46 degrees C, conidia developed a highly cyanide-insensitive, hydroxamate-sensitive respiration associated with morphological alterations in mitochondrial membranes; such changes were time-dependent. When heat-treated conidia were shifted down to 25 degrees C, the alternate, hydroxamate-sensitive respiration decreased significantly, paralleling the recovery of well-cristated mitochondria with an electron-dense matrix in the germ tubes. The decrease in hydroxamate-sensitivity was associated with two periods of increase in cyanide sensitivity corresponding to the events of germination and precocious proconidial budding.

  1. Membrane permeability and the loss of germination factor from Neurospora crassa at low water activities

    NASA Technical Reports Server (NTRS)

    Charlang, G.; Horowitz, N. H.

    1974-01-01

    Neurospora crassa conidia incubating in buffer at low water activities release a germination-essential component as well as 260-nm absorbing and ninhydrin-positive materials, regardless of whether an electrolyte or nonelectrolyte is used to reduce water activity. Chloroform and antibiotics known to increase cell-membrane permeability have a similar effect. This suggests that membrane damage occurs in media of low water activity and that an increase in permeability is responsible for the release of cellular components. The damage caused in media of low water activity is nonlethal in most cases, and the conidia recover when transferred to nutrient medium.

  2. Geomyces destructans sp. nov. associated with Bat white-nose syndrome

    USGS Publications Warehouse

    Gargas, A.; Trest, M.T.; Christensen, M.; Volk, T.J.; Blehert, D.S.

    2009-01-01

    We describe and illustrate the new species Geomyces destructans. Bats infected with this fungus present with powdery conidia and hyphae on their muzzles, wing membranes, and/or pinnae, leading to description of the accompanying disease as white-nose syndrome, a cause of widespread mortality among hibernating bats in the northeastern US. Based on rRNA gene sequence (ITS and SSU) characters the fungus is placed in the genus Geomyces, yet its distinctive asymmetrically curved conidia are unlike those of any described Geomyces species.

  3. Isolation and identification of the conidial germination factor of Neurospora crassa

    NASA Technical Reports Server (NTRS)

    Horowitz, N. H.; Charlang, G.; Horn, G.; Williams, N. P.

    1976-01-01

    The germination-essential substance (germination factor /GF/) that is lost from conidia of Neurospora crassa on exposure to solutions of low water activity has been isolated and identified as a group of iron-transport compounds, or siderochromes. The principal siderochrome of conidia is ferricrocin, a cyclic hexapeptide. A closely related substance, ferrichrome C, is tentatively identified as a minor constituent. The same substances are also present in extracts of mycelium along with small amounts of a third siderochrome, which has not been identified. The GF activity of culture filtrates is due to coprogen, the only siderochrome previously identified with N. crassa.

  4. A preliminary evaluation of the potential of Beauveria bassiana for bed bug control.

    PubMed

    Barbarin, Alexis M; Jenkins, Nina E; Rajotte, Edwin G; Thomas, Matthew B

    2012-09-15

    Residual biopesticide treatments of Beauveria bassiana were tested against the bed bug Cimex lectularius. An oil formulation of conidia was applied to different substrates. Bed bugs were exposed for 1 h, transferred to an unsprayed environment and monitored for mortality. Separate bioassays evaluated the effect of bed bug strain, sex, life stage, and exposure substrate on mortality. Rapid mortality was observed in all bioassays, with bed bugs exposed to treated jersey knit cotton dying most rapidly. A further assay demonstrated efficient autodissemination of conidia from exposed bed bugs to unexposed bed bugs within artificial harborages.

  5. Moisture Sources in Relation to Conidial Dissemination and Infection by Cladosporium carpophilum Within Peach Canopies.

    PubMed

    Lan, Z; Scherm, H

    2003-12-01

    ABSTRACT Cladosporium carpophilum, the causal agent of peach scab, overwinters in lesions on 1-year-old twigs, from which conidia infect the developing fruit during spring and early summer. Twig lesions constitute the sole source of initial inoculum; therefore, the mode of dissemination of conidia from such lesions to the fruit is of considerable interest. In a 4-year study, we determined the relative importance of air- versus water-borne conidia and their interaction with different fruit wetness sources (splash, twig runoff, and dew) in a peach orchard with areas that had been treated or not treated with fungicide the previous year. The rareness of scab twig lesions in the previously sprayed trees implied that fruit infection in these trees would occur primarily by airborne conidia from unsprayed trees nearby (located within the same tree row or the adjacent row). In the unsprayed areas, additional infections could occur by short-distance waterborne dissemination of conidia from locally abundant twig lesions via splashing or runoff. Beginning at calyx fall, individual fruit were protected from splash by rain shields, protected from runoff by cotton wicks placed proximal to the peduncle, or left untreated. Rain shields were adjustable, allowing rain or dew to be excluded selectively. Various combinations of the shield and wick treatments were implemented in the previously sprayed and unsprayed areas, and statistical comparison of fruit scab severity between individual treatments by linear contrasts allowed us to untangle the relative contributions of the various sources of inoculum and fruit wetness. Results showed that aerial dissemination of conidia contributed little to fruit scab development, even in the presence of fruit surface wetness caused by splashing, runoff, or dew. In contrast, waterborne conidia contributed considerably and significantly (P < 0.0001) to disease development. This was due primarily to the importance of splash in disseminating conidia

  6. Stachybotrys eucylindrospora, sp. nov. resulting from a re-examination of Stachybotrys cylindrospora.

    PubMed

    Li, De-Wei

    2007-01-01

    The holotype of Stachybotrys cylindrospora was examined and the morphological characters were found to fit the description of Stachybotrys chartarum. Thus Stachybotrys cylindrospora is a synonym of S. chartarum. However a number of isolates and specimens subsequently described and studied by several mycologists have typical cylindrical conidia with longitudinal striations. The conidia are much longer than those of S. chartarum. These conidial characters showed that those isolates and specimens are notably different from S. chartarum and of the holotype of S. cylindrospora. Therefore a new name, Stachybotrys eucylindrospora sp. nov., is proposed to accommodate these isolates and specimens.

  7. Isolation and identification of the conidial germination factor of Neurospora crassa

    NASA Technical Reports Server (NTRS)

    Horowitz, N. H.; Charlang, G.; Horn, G.; Williams, N. P.

    1976-01-01

    The germination-essential substance (germination factor /GF/) that is lost from conidia of Neurospora crassa on exposure to solutions of low water activity has been isolated and identified as a group of iron-transport compounds, or siderochromes. The principal siderochrome of conidia is ferricrocin, a cyclic hexapeptide. A closely related substance, ferrichrome C, is tentatively identified as a minor constituent. The same substances are also present in extracts of mycelium along with small amounts of a third siderochrome, which has not been identified. The GF activity of culture filtrates is due to coprogen, the only siderochrome previously identified with N. crassa.

  8. Antifungal effects of citronella oil against Aspergillus niger ATCC 16404.

    PubMed

    Li, Wen-Ru; Shi, Qing-Shan; Ouyang, You-Sheng; Chen, Yi-Ben; Duan, Shun-Shan

    2013-08-01

    Essential oils are aromatic oily liquids obtained from some aromatic plant materials. Certain essential oils such as citronella oil contain antifungal activity, but the antifungal effect is still unknown. In this study, we explored the antifungal effect of citronella oil with Aspergillus niger ATCC 16404. The antifungal activity of citronella oil on conidia of A. niger was determined by poisoned food technique, broth dilution method, and disc volatility method. Experimental results indicated that the citronella oil has strong antifungal activity: 0.125 (v/v) and 0.25 % (v/v) citronella oil inhibited the growth of 5 × 10⁵ spore/ml conidia separately for 7 and 28 days while 0.5 % (v/v) citronella oil could completely kill the conidia of 5 × 10⁵ spore/ml. Moreover, the fungicidal kinetic curves revealed that more than 90 % conidia (initial concentration is 5 × 10⁵ spore/ml) were killed in all the treatments with 0.125 to 2 % citronella oil after 24 h. Furthermore, with increase of citronella oil concentration and treatment time, the antifungal activity was increased correspondingly. The 0.5 % (v/v) concentration of citronella oil was a threshold to kill the conidia thoroughly. The surviving conidia treated with 0.5 to 2 % citronella oil decreased by an order of magnitude every day, and no fungus survived after 10 days. With light microscope, scanning electron microscope, and transmission electron microscope, we found that citronella oil could lead to irreversible alteration of the hyphae and conidia. Based on our observation, we hypothesized that the citronella oil destroyed the cell wall of the A. niger hyphae, passed through the cell membrane, penetrated into the cytoplasm, and acted on the main organelles. Subsequently, the hyphae was collapsed and squashed due to large cytoplasm loss, and the organelles were severely destroyed. Similarly, citronella oil could lead to the rupture of hard cell wall and then act on the sporoplasm to kill the

  9. Phosphorus availability modulates the toxic effect of silver on aquatic fungi and leaf litter decomposition.

    PubMed

    Funck, J Arce; Clivot, H; Felten, V; Rousselle, P; Guérold, F; Danger, M

    2013-11-15

    The functioning of forested headwater streams is intimately linked to the decomposition of leaf litter by decomposers, mainly aquatic hyphomycetes, which enables the transfer of allochthonous carbon to higher trophic levels. Evaluation of this process is being increasingly used as an indicator of ecosystem health and ecological integrity. Yet, even though the individual impacts of contaminants and nutrient availability on decomposition have been well studied, the understanding of their combined effects remains limited. In the current study, we investigated whether the toxic effects of a reemerging contaminant, silver (Ag), on leaf litter decomposition could be partly overcome in situations where microorganisms were benefitting from high phosphorus (P) availability, the latter being a key chemical element that often limits detritus decomposition. We also investigated whether these interactive effects were mediated by changes in the structure of the aquatic hyphomycete community. To verify these hypotheses, leaf litter decomposition by a consortium of ten aquatic hyphomycete species was followed in a microcosm experiment combining five Ag contamination levels and three P concentrations. Indirect effects of Ag and P on the consumption of leaf litter by the detritivorous crustacean, Gammarus fossarum, were also evaluated. Ag significantly reduced decomposition but only at the highest concentration tested, independently of P level. By contrast, P and Ag interactively affected fungal biomass. Both P level and Ag concentrations shaped microbial communities without significantly affecting the overall species richness. Finally, the levels of P and Ag interacted significantly on G. fossarum feeding rates, high [Ag] reducing litter consumption and low P availability tending to intensify the feeding rate. Given the high level of contaminant needed to impair the decomposition process, it is unlikely that a direct effect of Ag on leaf litter decomposition could be observed in

  10. Ecological study of the fungal populations of the acidic Tinto River in southwestern Spain.

    PubMed

    López-Archilla, A I; González, A E; Terrón, M C; Amils, R

    2004-11-01

    The characterization of the microbial ecology of the Tinto River, an extreme habitat with an extremely low pH and a high concentration of heavy metals, revealed an unexpected level of microbial richness. A variety of microbial eukaryotes was isolated, among them several fungal strains that were identified and their physiological characteristics studied. Ninety strains of yeast were isolated from the Tinto River. Fifty-two percent of them were capable of growth in vitro using medium amended with river water. They belong to 6 genera of basidiomycetes (Rhodotorula, Cryptococcus, Tremella, Holtermannia, Leucosporidium, and Mrakia) and 2 of ascomycetes (Candida and Williopsis). In addition, 349 strains of hyphomycetes belonging to 17 genera (most of them ascomycetes) were isolated and studied. Forty-four percent of the isolated filamentous fungi (154 strains) were capable of growing in vitro using medium amended with Tinto River water. Of this percentage, 19% (29 strains) belonged to the genus Penicillium (16 species) and 66% (102 strains) were included in the genera Scytalidium, Bahusakala, Phoma, and Heteroconium or showed dark sterile mycelia, which probably are of dematiaceous hyphomycetes. In addition, we characterized strains of the ascomycete genera Lecythophora and Acremonium and of the zygomycete genus Mortierella, all of them capable of growing in medium amended with river water. Statistical correlation of biological and physicochemical variables suggested a positive relationship between the dematiaceous hyphomycetes and the most extreme physicochemical conditions found in the Tinto River. Principal components analysis confirmed this relationship and also showed that the Acremonium and Lecythophora groups had environmental preferences similar to those of dematiaceous fungi. The spatial positions of the sampling sites were grouped in 2 main clusters: (i) sampling sites in the mine zone in which most of the dematiaceous, Acremonium, and Lecythophora strains were

  11. Host, Pathogen, and Environmental Characteristics Predict White-Nose Syndrome Mortality in Captive Little Brown Myotis (Myotis lucifugus)

    PubMed Central

    Johnson, Joseph S.; Reeder, DeeAnn M.; McMichael, James W.; Meierhofer, Melissa B.; Stern, Daniel W. F.; Lumadue, Shayne S.; Sigler, Lauren E.; Winters, Harrison D.; Vodzak, Megan E.; Kurta, Allen; Kath, Joseph A.; Field, Kenneth A.

    2014-01-01

    An estimated 5.7 million or more bats died in North America between 2006 and 2012 due to infection with the fungus Pseudogymnoascus destructans (Pd) that causes white-nose syndrome (WNS) during hibernation. The behavioral and physiological changes associated with hibernation leave bats vulnerable to WNS, but the persistence of bats within the contaminated regions of North America suggests that survival might vary predictably among individuals or in relation to environmental conditions. To investigate variables influencing WNS mortality, we conducted a captive study of 147 little brown myotis (Myotis lucifugus) inoculated with 0, 500, 5 000, 50 000, or 500 000 Pd conidia and hibernated for five months at either 4 or 10°C. We found that female bats were significantly more likely to survive hibernation, as were bats hibernated at 4°C, and bats with greater body condition at the start of hibernation. Although all bats inoculated with Pd exhibited shorter torpor bouts compared to controls, a characteristic of WNS, only bats inoculated with 500 conidia had significantly lower survival odds compared to controls. These data show that host and environmental characteristics are significant predictors of WNS mortality, and that exposure to up to 500 conidia is sufficient to cause a fatal infection. These results also illustrate a need to quantify dynamics of Pd exposure in free-ranging bats, as dynamics of WNS produced in captive studies inoculating bats with several hundred thousand conidia may differ from those in the wild. PMID:25409028

  12. O-Glycosylation in Cell Wall Proteins in Scedosporium prolificans Is Critical for Phagocytosis and Inflammatory Cytokines Production by Macrophages

    PubMed Central

    Xisto, Mariana I. D. S.; Bittencourt, Vera C. B.; Liporagi-Lopes, Livia Cristina; Haido, Rosa M. T.; Mendonça, Morena S. A.; Sassaki, Guilherme; Figueiredo, Rodrigo T.; Romanos, Maria Teresa V.; Barreto-Bergter, Eliana

    2015-01-01

    In this study, we analyze the importance of O-linked oligosaccharides present in peptidorhamnomannan (PRM) from the cell wall of the fungus Scedosporium prolificans for recognition and phagocytosis of conidia by macrophages. Adding PRM led to a dose-dependent inhibition of conidia phagocytosis, whereas de-O-glycosylated PRM did not show any effect. PRM induced the release of macrophage-derived antimicrobial compounds. However, O-linked oligosaccharides do not appear to be required for such induction. The effect of PRM on conidia-induced macrophage killing was examined using latex beads coated with PRM or de-O-glycosylated PRM. A decrease in macrophage viability similar to that caused by conidia was detected. However, macrophage killing was unaffected when beads coated with de-O-glycosylated PRM were used, indicating the toxic effect of O-linked oligosaccharides on macrophages. In addition, PRM triggered TNF-α release by macrophages. Chemical removal of O-linked oligosaccharides from PRM abolished cytokine induction, suggesting that the O-linked oligosaccharidic chains are important moieties involved in inflammatory responses through the induction of TNF-α secretion. In summary, we show that O-glycosylation plays a role in the recognition and uptake of S. prolificans by macrophages, killing of macrophages and production of pro- inflammatory cytokines. PMID:25875427

  13. Feasibility of detecting Aflatoxin B1 in single maize kernels using hyperspectral imaging

    USDA-ARS?s Scientific Manuscript database

    The feasibility of detecting Aflatoxin B1 (AFB1) in single maize kernel inoculated with Aspergillus flavus conidia in the field, as well as its spatial distribution in the kernels, was assessed using near-infrared hyperspectral imaging (HSI) technique. Firstly, an image mask was applied to a pixel-b...

  14. Agrobacterium tumefaciens-mediated transformation of Valsa mali: an efficient tool for random insertion mutagenesis.

    PubMed

    Wang, Caixia; Guan, Xiangnan; Wang, Hanyan; Li, Guifang; Dong, Xiangli; Wang, Guoping; Li, Baohua

    2013-01-01

    Valsa mali is a causal agent of apple and pear trees canker disease, which is a destructive disease that causes serious economic losses in eastern Asia, especially in China. The lack of an efficient transformation system for Valsa mali retards its investigation, which poses difficulties to control the disease. In this research, a transformation system for this pathogen was established for the first time using A. tumefaciens-mediated transformation (ATMT), with the optimal transformation conditions as follows: 10(6)/mL conidia suspension, cocultivation temperature 22°C, cocultivation time 72 hours, and 200  μ M acetosyringone (AS) in the inductive medium. The average transformation efficiency was 1015.00 ± 37.35 transformants per 10(6) recipient conidia. Thirty transformants were randomly selected for further confirmation and the results showed the presence of T-DNA in all hygromycin B resistant transformants and also revealed random and single gene integration with genetic stability. Compared with wild-type strain, those transformants exhibited various differences in morphology, conidia production, and conidia germination ability. In addition, pathogenicity assays revealed that 14 transformants had mitigated pathogenicity, while one had enhanced infection ability. The results suggest that ATMT of V. mali is a useful tool to gain novel insight into this economically important pathogen at molecular levels.

  15. Evidence for sexual reproduction in Fusicladium effusum

    USDA-ARS?s Scientific Manuscript database

    Fusicladium effusum is the causal agent of pecan scab, the most prevalent and often catastrophic disease of pecan in the southeastern USA. Despite earlier efforts to determine a sexual stage, reproduction in F. effusum has been observed only by asexually produced conidia. However, the degree and dis...

  16. Sporulation in Erysiphe necator: signals, differential gene expression and possible implications for disease management

    USDA-ARS?s Scientific Manuscript database

    Abundant production of conidia is a driving factor for epidemics of grape powdery mildew (Erysiphe necator (syn. Uncinula necator). Previous investigations revealed evidence for a signal that coordinates the onset of asexual reproduction. The genetic basis for this signal in powdery mildews had not ...

  17. Differential gene expression during conidiation in the grape powdery mildew fungus, Erysiphe necator

    USDA-ARS?s Scientific Manuscript database

    Asexual sporulation (conidiation) is coordinately regulated in the grape powdery mildew fungus Erysiphe necator, but nothing is known about its genetic regulation. We hypothesized that genes required for conidiation in other fungi would be up-regulated at conidiophore initiation and/or full conidia...

  18. In vitro colony interactions among species of Trichoderma with inference toward biological control.

    Treesearch

    Jimmy L. Reaves; Ralph H. Crawford

    1994-01-01

    Colony interactions among 15 isolates representing seven species of Trichoderma were evaluated in vitro. Interactions characterized by zones of inhibition, demarcation lines, ridges of conidia, overgrowth, intermingling, anastomosis, and hyphal coiling in self-pairings and intraspecific and interspecific pairings of the seven species were recorded...

  19. Performance of fogged disinfectants to inactivate conida of Penicillium digitatum within citrus degreening rooms

    USDA-ARS?s Scientific Manuscript database

    Fogging with formaldehyde of citrus packinghouses when the fruit are absent is a practice to control conidia of Penicillium digitatum (Pers.) Sacc., cause of citrus green mold. Replacements for formaldehyde in these facilities are needed because of worker and environmental health issues. To evaluate...

  20. Host deception: Predaceous fungus, esteya vermicola, entices pine wood nematode by mimicking the scent of its host pine for nutrient

    USDA-ARS?s Scientific Manuscript database

    A nematophagous fungus, Esteya vermicola, is recorded as the first endoparasitic fungus of pine wood nematode (PWN), Burasphelenchus xylophilus, in the last century. E. vermicola exhibited high infectivity toward PWN in the laboratory conditions and conidia spraying of this fungus on Japanese red pi...

  1. Population structure and genetic diversity of Fusicladium effusum in the USA

    USDA-ARS?s Scientific Manuscript database

    Scab (Fusicladium effusum) is the most destructive disease of pecan in the Southeast US. Infection is thought to occur solely through asexually produced conidia. To explore the population structure and genetic diversity of F. effusum, populations were hierarchically sampled from 11 orchards in Alaba...

  2. Potential of a strain of the entomopathogenic fungus Beauveria bassiana (Hypocreales: Cordycipitaceae) as a biological control agent against western flower thrips, Frankliniella occidentalis (Thysanoptera: Thripidae)

    USDA-ARS?s Scientific Manuscript database

    Five Beauveria bassiana strains were evaluated for control of western flower thrips. Strain RSB was the most virulent, causing 69-96% mortality at concentrations of 1×104 – 1×107 conidia mL-1, 10 days after inoculation of first instars. In greenhouse trials, RSB applied to broccoli foliage signifi...

  3. It’s a Jungle Out There! Abiotic and Biotic Factors That Affect Efficacy and Persistence of the Entomopathogenic Fungi

    USDA-ARS?s Scientific Manuscript database

    One might conclude the soil is a more congenial arena for using entomopathogenic fungi (EPF) than the phylloplane. No ultraviolet light, no rainfall washing conidia from foliage, no rapid attenuation of conidial deposits by rapid plant canopy expansion. The soil is cool, damp and dark – perfect fo...

  4. Near-infrared hyperspectral imaging for detecting Aflatoxin B1 of maize kernels

    USDA-ARS?s Scientific Manuscript database

    The feasibility of detecting the Aflatoxin B1 in maize kernels inoculated with Aspergillus flavus conidia in the field was assessed using near-infrared hyperspectral imaging technique. After pixel-level calibration, wavelength dependent offset, the masking method was adopted to reduce the noise and ...

  5. Ascospore dispersal of Ceratocystiopsis ranaculosus, a mycangial fungus of the southern pine beetle

    Treesearch

    John C. Moser; Thelma J. Perry; J. Robert Bridges; Hui-Fen Yin

    1995-01-01

    Ascospores of the heterothallic fungus Ceratocystiopsis ranaculosus were found in the sporothecae of three mite species of the genus Tarsonemus.These mites were phoretic on the coniferous bark beetles Dendroctonus frontalis, D. brevicomis, and Ips acuminatus.Ceratocystiopsis ranaculosus inhabits the mycangium of both Dendroctonus species as conidia in a budding yeast-...

  6. The Aspergillus flavus fluP-associated metabolite promotes sclerotial production

    USDA-ARS?s Scientific Manuscript database

    Aspergillus flavus is able to synthesize a variety of polyketide-derived secondary metabolites including the hepatocarcinogen, aflatoxin B1. The fungus reproduces and disseminates predominantly by production of conidia. It also produces hardened mycelial aggregates called sclerotia that are used to ...

  7. Use of acoustic technology to monitor the time course of Rhynchophorus ferrugineus larval mortality in date palms after treatments with Beauveria bassiana

    USDA-ARS?s Scientific Manuscript database

    Spectral and temporal patterns of insect sound impulses were monitored daily for 23-d periods in 8, 10, or 5 small date palm trees containing larvae dipped in 0 (control), 104 (low), or 108 (high) conidia/ml doses of entomopathogenic fungus, Beauveria bassiana (Bb 203), respectively. Each tree conta...

  8. DOSE-DEPENDENT ALLERGIC ASTHMA RESPONSES TO PENICILLIUM CHRYSOGENUM

    EPA Science Inventory

    ABSTRACT
    Indoor mold has been associated with development of allergic asthma. Penicillium chrysogenum, a common indoor mold, is known to have several allergens and its viable conidia can induce allergic responses in a mouse model of allergic penicilliosis. The hypothesis o...

  9. Exposure of bed bugs to metarhizium anisopliae, and the effect of defensive secretions on fungal growth in vitro

    USDA-ARS?s Scientific Manuscript database

    Bed bugs Cimex lectularius were treated with conidia of the entomopathogenic fungus Metarhizium anisopliae by topical, spray, and contact exposure. One week post-exposure, inconsistent mortalities were observed, averaging 30% across all treatment groups and replicates. Microscopic examination of top...

  10. Metacridamides A and B from the biocontrol fungus metarhizium acridum

    USDA-ARS?s Scientific Manuscript database

    Metarhizium acridum, an entomopathogenic fungus, has been commercialized and used successfully for biocontrol of grasshopper pests in Africa and Australia. As part of an effort to catalog the secondary metabolites of this fungus we discovered that its conidia produce two novel 17-membered macrocycl...

  11. Host, pathogen, and environmental characteristics predict white-nose syndrome mortality in captive little brown myotis (Myotis lucifugus).

    PubMed

    Johnson, Joseph S; Reeder, DeeAnn M; McMichael, James W; Meierhofer, Melissa B; Stern, Daniel W F; Lumadue, Shayne S; Sigler, Lauren E; Winters, Harrison D; Vodzak, Megan E; Kurta, Allen; Kath, Joseph A; Field, Kenneth A

    2014-01-01

    An estimated 5.7 million or more bats died in North America between 2006 and 2012 due to infection with the fungus Pseudogymnoascus destructans (Pd) that causes white-nose syndrome (WNS) during hibernation. The behavioral and physiological changes associated with hibernation leave bats vulnerable to WNS, but the persistence of bats within the contaminated regions of North America suggests that survival might vary predictably among individuals or in relation to environmental conditions. To investigate variables influencing WNS mortality, we conducted a captive study of 147 little brown myotis (Myotis lucifugus) inoculated with 0, 500, 5000, 50,000, or 500,000 Pd conidia and hibernated for five months at either 4 or 10°C. We found that female bats were significantly more likely to survive hibernation, as were bats hibernated at 4°C, and bats with greater body condition at the start of hibernation. Although all bats inoculated with Pd exhibited shorter torpor bouts compared to controls, a characteristic of WNS, only bats inoculated with 500 conidia had significantly lower survival odds compared to controls. These data show that host and environmental characteristics are significant predictors of WNS mortality, and that exposure to up to 500 conidia is sufficient to cause a fatal infection. These results also illustrate a need to quantify dynamics of Pd exposure in free-ranging bats, as dynamics of WNS produced in captive studies inoculating bats with several hundred thousand conidia may differ from those in the wild.

  12. Septoria Canker on Nursery Stock of Populus Deltoides

    Treesearch

    T. H. Filer; F. I. McCracken; C. A. Mohn; W. K. Randall

    1971-01-01

    Septoria musiva Peck is capable of establishing itself on unwounded first-year stems of eastern cottonwood. Natural infections have been observed since 1969 in three forest nurseries in Mississippi, and inoculations have confirmed that both conidia and ascospores are capable of causing stem infections.

  13. Successful Treatment of Osseous Blastomycosis without Pulmonary or Disseminated Disease and Review of the Literature

    PubMed Central

    Day, Shandra R.; Weiss, David B.; Hazen, Kevin C.; Moore, Christopher C.

    2014-01-01

    Blastomycosis commonly occurs following inhalation of Blastomyces dermatitidis conidia causing a pulmonary infection and can disseminate to extrapulmonary sites. Osseous involvement primarily results from hematogenous spread but in rare cases direct inoculation can occur. We describe a case of osseous blastomycosis without pulmonary or disseminated disease successfully treated with posaconazole. PMID:24703876

  14. Morphological and molecular variation among species of the Fusarium dimerum species group

    USDA-ARS?s Scientific Manuscript database

    The name Fusarium dimerum has been used in the past for saprotrophic fungi and opportunistic human pathogens with up to 3-septate but mostly 0- or 1-septate Fusarium-like conidia. On the basis of narrowly defined morphological characters, the varieties Pusillum, Nectrioides and Violaceum were disti...

  15. Inverse correlation of ability to produce aflatoxin and aspergillus colonization of maize seed

    USDA-ARS?s Scientific Manuscript database

    Seeds of aflatoxin-resistant and aflatoxin susceptible maize lines were inoculated with conidia of aflatoxin-producing Aspergillus flavus or A. parasiticus isolates or isogenic non-producing mutants. Conidial yields recovered from resistant maize seed after seven days were significantly lower for af...

  16. Impact of metarhizium brunneum petch clavicipitaceae (Hypocreales) on pre-imaginal Rhagoletis indifferens (Diptera: Tephritidae) within and on the surface of orchard soil

    USDA-ARS?s Scientific Manuscript database

    When last instar laboratory-reared Rhagoletis indifferens were allowed to pupate within non-sterile orchard soil containing Metarhizium brunneum isolate F52 conidia, a dose related proportion died from developmental abnormalities and mycosis. Similarly, when last instar larvae entered soil that was ...

  17. Impact of Metarhizium brunneum (Hypocreales: Clavicipitaceae) on pre-imaginal Rhagoletis indifferens (Diptera: Tephritidae) within and on the surface of orchard soil

    USDA-ARS?s Scientific Manuscript database

    When last instar laboratory-reared Rhagoletis indifferens (Cherry Fruit Fly) were allowed to pupate within non-sterile orchard soil containing incorporated Metarhizium brunneum isolate F52 conidia, a dose-related proportion died from developmental abnormalities and mycosis. Similarly, when prepupal ...

  18. Use of phosphite salts in laboratory and semi-commercial tests to control citrus postharvest decay

    USDA-ARS?s Scientific Manuscript database

    EC50 concentrations of potassium phosphite (KP) to inhibit germination of Penicillium digitatum conidia were 229, 334, 360, 469, 498, or 580 mg/liter at pH 3, 4, 5, 6, 7, or 8, respectively. Increasing phosphate content in media reduced phosphite toxicity. To control green or blue mold, fruit were i...

  19. Chitin synthases with a myosin motor-like domain control the resistance of Aspergillus fumigatus to echinocandins.

    PubMed

    Jiménez-Ortigosa, Cristina; Aimanianda, Vishukumar; Muszkieta, Laetitia; Mouyna, Isabelle; Alsteens, David; Pire, Stéphane; Beau, Remi; Krappmann, Sven; Beauvais, Anne; Dufrêne, Yves F; Roncero, César; Latgé, Jean-Paul

    2012-12-01

    Aspergillus fumigatus has two chitin synthases (CSMA and CSMB) with a myosin motor-like domain (MMD) arranged in a head-to-head configuration. To understand the function of these chitin synthases, single and double csm mutant strains were constructed and analyzed. Although there was a slight reduction in mycelial growth of the mutants, the total chitin synthase activity and the cell wall chitin content were similar in the mycelium of all of the mutants and the parental strain. In the conidia, chitin content in the ΔcsmA strain cell wall was less than half the amount found in the parental strain. In contrast, the ΔcsmB mutant strain and, unexpectedly, the ΔcsmA/ΔcsmB mutant strain did not show any modification of chitin content in their conidial cell walls. In contrast to the hydrophobic conidia of the parental strain, conidia of all of the csm mutants were hydrophilic due to the presence of an amorphous material covering the hydrophobic surface-rodlet layer. The deletion of CSM genes also resulted in an increased susceptibility of resting and germinating conidia to echinocandins. These results show that the deletion of the CSMA and CSMB genes induced a significant disorganization of the cell wall structure, even though they contribute only weakly to the overall cell wall chitin synthesis.

  20. Chitin Synthases with a Myosin Motor-Like Domain Control the Resistance of Aspergillus fumigatus to Echinocandins

    PubMed Central

    Jiménez-Ortigosa, Cristina; Aimanianda, Vishukumar; Muszkieta, Laetitia; Mouyna, Isabelle; Alsteens, David; Pire, Stéphane; Beau, Remi; Krappmann, Sven; Beauvais, Anne; Dufrêne, Yves F.

    2012-01-01

    Aspergillus fumigatus has two chitin synthases (CSMA and CSMB) with a myosin motor-like domain (MMD) arranged in a head-to-head configuration. To understand the function of these chitin synthases, single and double csm mutant strains were constructed and analyzed. Although there was a slight reduction in mycelial growth of the mutants, the total chitin synthase activity and the cell wall chitin content were similar in the mycelium of all of the mutants and the parental strain. In the conidia, chitin content in the ΔcsmA strain cell wall was less than half the amount found in the parental strain. In contrast, the ΔcsmB mutant strain and, unexpectedly, the ΔcsmA/ΔcsmB mutant strain did not show any modification of chitin content in their conidial cell walls. In contrast to the hydrophobic conidia of the parental strain, conidia of all of the csm mutants were hydrophilic due to the presence of an amorphous material covering the hydrophobic surface-rodlet layer. The deletion of CSM genes also resulted in an increased susceptibility of resting and germinating conidia to echinocandins. These results show that the deletion of the CSMA and CSMB genes induced a significant disorganization of the cell wall structure, even though they contribute only weakly to the overall cell wall chitin synthesis. PMID:22964252

  1. Biochemical analysis of plant protection afforded by a nonpathogenic endophytic mutant of Colletotrichum magna

    USGS Publications Warehouse

    Redman, R.S.; Freeman, S.; Clifton, D.R.; Morrel, J.; Brown, G.; Rodriguez, R.J.

    1999-01-01

    A nonpathogenic mutant of Colletotrichum magna (path-1) was previously shown to protect watermelon (Citrullus lanatus) and cucumber (Cucumis sativus) seedlings from anthracnose disease elicited by wild-type C. magna. Disease protection was observed in stems of path-1-colonized cucurbits but not in cotyledons, indicating that path-1 conferred tissue-specific and/or localized protection. Plant biochemical indicators of a localized and systemic (peroxidase, phenylalanine ammonia-lyase, lignin, and salicylic acid) 'plant-defense' response were investigated in anthracnose-resistant and -susceptible cultivars of cucurbit seedlings exposed to four treatments: (1) water (control), (2) path-1 conidia, (3) wild-type conidia, and (4) challenge conditions (inoculation into path-1 conidia for 48 h and then exposure to wild-type conidia). Collectively, these analyses indicated that disease protection in path-1 colonized plants was correlated with the ability of these plants to mount a defense response more rapidly and to equal or greater levels than plants exposed to wild-type C. magna alone. Watermelon plants colonized with path-1 were also protected against disease caused by Colletotrichum orbiculare and Fusarium oxysporum. A model based on the kinetics of plant-defense activation is presented to explain the mechanism of path-1-conferred disease protection.

  2. Aspergillus species in indoor environments and their possible occupational and public health hazards

    PubMed Central

    Mousavi, B; Hedayati, MT; Hedayati, N; Ilkit, M; Syedmousavi, S

    2016-01-01

    The genus Aspergillus, which consists of a few hundred opportunistic mold species found in various climatic conditions, causes diseases including localized infections, fatal diseases, allergic responses, and inhaled conidia in humans. Herein, we present an overview of the most common diseases and allergic infections caused by Aspergillus species and their associated health hazards in various indoor environments worldwide. PMID:28681011

  3. Antifungal activity of a synthetic cationic peptide against the plant pathogens Colletotrichum graminicola and three Fusarium species

    USDA-ARS?s Scientific Manuscript database

    A small cationic peptide (JH8944) was tested for activity against a number of pathogens of agricultural crops. JH8944 inhibited conidium growth in most of the tested plant pathogens with a dose of 50 µg ml 1, although one isolate of Fusarium oxysporum was inhibited at 5 µg ml 1. Most conidia of Fusa...

  4. Biotechnology of Aureobasidium pullulans: A phylogenetic perspective

    USDA-ARS?s Scientific Manuscript database

    Aureobasidium pullulans is a fungus historically included among the "black yeasts." Although many strains are predominantly yeast-like, the species is actually polymorphic, exhibiting complex forms ranging from blastic conidia and swollen cells to pseudophyphae, hyphae, and chlamydospores. A. pull...

  5. A role for the asexual spores in infection of Fraxinus excelsior by the ash-dieback fungus Hymenoscyphus fraxineus

    PubMed Central

    Fones, Helen Nicola; Mardon, Charlotte; Gurr, Sarah Jane

    2016-01-01

    The invasive pathogen, ash dieback fungus Hymenoscyphus fraxineus, is spreading rapidly across Europe. It shows high levels of outcrossing and limited population structure, even at the epidemic front. The anamorphic (asexual) form produces prolific conidia, thought to function solely as spermatia (male gametes), facilitating gene flow between sympatric strains. Here, we show that conidia are capable of germination on ash leaves and in vitro, and can infect seedlings via leaves or soil. In leaves, germlings form structures resembling fruiting bodies. Additionally, H. fraxineus colonises ash debris and grows in soil in the absence of ash tissues. We propose an amended life-cycle in which wind-dispersed, insect-vectored or water-spread conidia infect ash and may sporulate in planta, as well as in forest debris. This amplifies inoculum levels of different strains in ash stands. In combination with their function as spermatia, conidia thus act to maximise gene flow between sympatric strains, including those originally present at low inoculum. Such mixing increases evolutionary potential, as well as enhancing the likelihood of gene introgression from closely-related strains or assimilation of further genetic diversity from parental Asian populations. This scenario increases the adaptability of H. fraxineus to new climates and, indeed, onto new host species. PMID:27694963

  6. Successful treatment of osseous blastomycosis without pulmonary or disseminated disease and review of the literature.

    PubMed

    Day, Shandra R; Weiss, David B; Hazen, Kevin C; Moore, Christopher C

    2014-06-01

    Blastomycosis commonly occurs following inhalation of Blastomyces dermatitidis conidia causing a pulmonary infection and can disseminate to extrapulmonary sites. Osseous involvement primarily results from hematogenous spread, but in rare cases, direct inoculation can occur. We describe a case of osseous blastomycosis without pulmonary or disseminated disease successfully treated with posaconazole. Copyright © 2014 Elsevier Inc. All rights reserved.

  7. Effect of prior vegetative growth, inoculum density and light on conidiation in Erysiphe necator

    USDA-ARS?s Scientific Manuscript database

    A driving force in epidemics of grape powdery mildew is the abundant production of conidia. Our objective was to better define the three factors involved in the qualitative change that occurs when a mildew colony switches from vegetative growth to sporulation –inoculum density, light, and a sporulat...

  8. Biochemical analysis of plant protection afforded by a nonpathogenic endophytic mutant of Colletotrichum magna

    SciTech Connect

    Redman, R.S.; Rodriguez, R.J. Univ. of Washington, Seattle, WA . Dept. of Botany); Clifton, D.R.; Morrel, J.; Brown, G. ); Freeman, S. . Dept. of Plant Pathology)

    1999-02-01

    A nonpathogenic mutant of Colletotrichum magna (path-1) was previously shown to protect watermelon (Citrullus lanatus) and cucumber (Cucumis sativus) seedlings from anthracnose disease elicited by wild-type C. magna. Disease protection was observed in stems of path-1-colonized cucurbits but not in cotyledons, indicating that path-1 conferred tissue-specific and/or localized protection. Plant biochemical indicators of a localized and systemic (peroxidase, phenylalanine ammonia-lyase, lignin, and salicylic acid) plant-defense response were investigated in anthracnose-resistant and-susceptible cultivars of cucurbit seedlings exposed to four treatments: (1) water (control), (2) path-1 conidia, (3) wild-type conidia, and (4) challenge conditions (inoculation into path-1 conidia for 48 h and then exposure to wild-type conidia). Collectively, these analyses indicated that disease protection in path-1-colonized plants was correlated with the ability of these plants to mount a defense response more rapidly and to equal or greater levels than plants exposed to wild-type C. magna alone. Watermelon plants colonized with path-1 were also protected against disease caused by Colletotrichum orbiculare and Fusarium oxysporum. A model based on the kinetics of plant-defense activation is presented to explain the mechanism of path-1-conferred disease protection.

  9. Aspergillus fumigatus evades immune recognition during germination through loss of toll-like receptor-4-mediated signal transduction.

    PubMed

    Netea, Mihai G; Warris, Adilia; Van der Meer, Jos W M; Fenton, Matthew J; Verver-Janssen, Trees J G; Jacobs, Liesbeth E H; Andresen, Tonje; Verweij, Paul E; Kullberg, Bart Jan

    2003-07-15

    Peritoneal macrophages from Toll-like receptor (TLR) 4-deficient ScCr mice produced less tumor necrosis factor, interleukin (IL)-1alpha, and IL-1beta than did macrophages of control mice, when stimulated with conidia, but not with hyphae, of Aspergillus fumigatus, a finding suggesting that TLR4-mediated signals are lost during germination. This hypothesis was confirmed by use of a TLR4-specific fibroblast reporter cell line (3E10) that responded to the conidia, but not to the hyphae, of A. fumigatus. In contrast, macrophages from TLR2-knockout mice had a decreased production of proinflammatory cytokines in response to both Aspergillus conidia and Aspergillus hyphae, and these results were confirmed in 3E10 cells transfected with human TLR2. In addition, Aspergillus hyphae, but not Aspergillus conidia, stimulated production of IL-10 through TLR2-dependent mechanisms. In conclusion, TLR4-mediated proinflammatory signals, but not TLR2-induced anti-inflammatory signals, are lost on Aspergillus germination to hyphae. Therefore, phenotypic switching during germination may be an important escape mechanism of A. fumigatus that results in counteracting the host defense.

  10. Proteome profiling of the dimorphic fungus Penicillium marneffei extracellular proteins and identification of glyceraldehyde-3-phosphate dehydrogenase as an important adhesion factor for conidial attachment.

    PubMed

    Lau, Susanna K P; Tse, Herman; Chan, Joanna S Y; Zhou, Anna C; Curreem, Shirly O T; Lau, Candy C Y; Yuen, Kwok-Yung; Woo, Patrick C Y

    2013-12-01

    Despite being the most important thermal dimorphic fungus causing systemic mycosis in Southeast Asia, the pathogenic mechanisms of Penicillium marneffei remain largely unknown. By comparing the extracellular proteomes of P. marneffei in mycelial and yeast phases, we identified 12 differentially expressed proteins among which glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and heat shock protein 60 (HSP60) were found to be upregulated in mycelial and yeast phases respectively. Based on previous findings in other pathogens, we hypothesized that these two extracellular proteins may be involved in adherence during P. marneffei-host interaction. Using inhibition assays with recombinant GAPDH (rGAPDH) proteins and anti-rGAPDH sera, we demonstrated that adhesion of P. marneffei conidia to fibronectin and laminin was inhibited by rGAPDH or rabbit anti-rGAPDH serum in a dose-dependent manner. Similarly, a dose-dependent inhibition of conidial adherence to A549 pneumocytes by rGAPDH or rabbit anti-rGAPDH serum was observed, suggesting that P. marneffei GAPDH can mediate binding of conidia to human extracellular matrix proteins and pneumocytes. However, HSP60 did not exhibit similar inhibition on conidia adherence, and neither GAPDH norHSP60 exhibited inhibition on adherence to J774 or THP-1 macrophage cell lines. This report demonstrates GAPDH as an adherence factor in P. marneffei by mediating conidia adherence to host bronchoalveolar epithelium during the early establishment phase of infection.

  11. Genetic diversity and population structure of Fusicladium effusum on pecan in the U.S.A.

    USDA-ARS?s Scientific Manuscript database

    Fusicladium effusum causes pecan scab, which is the most destructive disease of pecan in the U.S.A. The disease is known to be spread through asexually produced conidia in rain splash and wind. The fungus has demonstrated pathogenic diversity, yet there is no information on its genetic diversity or ...

  12. Primary in vitro culture of porcine tracheal epithelial cells in an air-liquid interface as a model to study airway epithelium and Aspergillus fumigatus interactions.

    PubMed

    Khoufache, Khaled; Cabaret, Odile; Farrugia, Cécile; Rivollet, Danièle; Alliot, Annie; Allaire, Eric; Cordonnier, Catherine; Bretagne, Stéphane; Botterel, Françoise

    2010-12-01

    Since the airway epithelium is the first tissue encountered by airborne fungal spores, specific models are needed to study this interaction. We developed such a model using primary porcine tracheal epithelial cells (PTEC) as a possible alternative to the use of primary human cells. PTEC were obtained from pigs and were cultivated in an air-liquid interface. Fluorescent brightener was employed to quantify the internalization of Aspergillus fumigatus conidia. Potential differences (Vt) and transepithelial resistances (Rt) after challenge with the mycotoxin, verruculogen, were studied. Primers for porcine inflammatory mediator genes IL-8, TNF-alpha, and GM-CSF were designed for a quantitative real-time PCR procedure to study cellular responses to challenges with A. fumigatus conidia. TEM showed the differentiation of ciliated cells and the PTEC ability to internalize conidia. The internalization rate was 21.9 ± 1.4% after 8 h of incubation. Verruculogen (10(-6) M) significantly increased Vt without having an effect on the Rt. Exposure of PTEC to live A. fumigatus conidia for 24 h induced a 10- to 40-fold increase in the mRNA levels of inflammatory mediator genes. PTEC behave similarly to human cells and are therefore a suitable alternative to human cells for studying interaction between airway epithelium and A. fumigatus.

  13. Presence of Fusarium graminearum in air associated with sorghum fields

    USDA-ARS?s Scientific Manuscript database

    Sorghum can be included in crop rotations with wheat. However, there are no known reports on the effects of sorghum grown in rotation with wheat on the epidemiology of head scab caused by Fusarium graminearum. Conidia in air samples within two sorghum fields were collected by passive spore trapping ...

  14. Horizontal transmission of Beauveria bassiana in Anastrepha ludens (Diptera: Tephritidae) under laboratory and field cage conditions.

    PubMed

    Toledo, Jorge; Campos, Sergio E; Flores, Salvador; Liedo, Pablo; Barrera, Juan F; Villaseñior, Antonio; Montoya, Pablo

    2007-04-01

    The virulence of two products of the fungus Beauveria bassiana (LCPP and Bassianil) on adult Anastrepha ludens (Loew) (Diptera: Tephritidae) and their effect on the mating performance of infected males was evaluated in laboratory and field cage tests. The horizontal transmission capacity of the fungus during copulation or attempted copulation also was quantified using inoculated males as well as the impact of infection on female fecundity and longevity. Both fungal products were found to be highly virulent (LCPP, mortality = 98.7% at 1 x 10(8) conidia per ml, LT50 = 4.20 d, LC50 = 9.35 x 10(5) conidia per ml; Bassianil, mortality = 99.3% at 1 X 10(8) conidia per ml, LT50 = 4.04 d, LC50 = 2.69 x 10(7) conidia per ml). Mating success of inoculated males was not affected compared with the control group during the 3 d postinoculation. Horizontal transmission to females during the first day was 80.6 and 84.3% through mating and 15.4 and 21.6% through attempts to mate and contact during courtship for the LCPP and Bassianil products, respectively. The fertility of infected females was notably reduced, and longevity did not extend beyond 15 d. Our results suggest the possibility of using sterile flies as fungus vectors in sterile insect technique programs, but the potential benefits and shortcomings of this approach require further investigation.

  15. Metacridamide B methanol-d4 monosolvate

    USDA-ARS?s Scientific Manuscript database

    The title compound was extracted from conidia of the fungus Metarhizium acridum. Crystals were obtained as a methanol-d4 solvate. The tail part of the 4-methylhexan-2-yl group exhibits disorder over two positions, with an occupancy ratio of 0.682 (9):0.318 (9). The crystal structure confirms the abs...

  16. Lack of acute pathogenicity and toxicity in mice of an isolate of Metarhizium anisopliae var. anisopliae from spittlebugs.

    PubMed

    Toriello, C; Pérez-Torres, A; Burciaga-Díaz, A; Navarro-Barranco, H; Pérez-Mejía, A; Lorenzana-Jiménez, M; Mier, T

    2006-10-01

    A monospore strain of Metarhizium anisopliae var. anisopliae (EH-479/2), isolated in Mexico from Aeneolamia sp., was tested for oral acute intragastric pathogenicity and toxicity in CD-1 mice, including a thorough histological study. Animals were inoculated by gavage with one dose (10(8) conidia/animal) of viable (72 mice) and nonviable (24 mice) conidia and compared to 18 control mice. Clinical observations were done daily; mycological and histological tests were performed during necropsies at days 3, 10, 17, and 21 after the inoculation. At the end of the study, no mice showed clinical symptoms of illness, and the animals' mean weight corresponded to that of healthy adults. No inflammatory reactions were identified in analyzed organs, suggesting the nonimmunogenic status of this fungal strain. Evidence of fungal germination was noted in two lymph nodes and in liver and lung of one dead mouse, out of 72 viable-conidia treated mice. There was no evidence of toxicity symptoms in mice inoculated with nonviable conidia. The results obtained support the nonpathogenic and nontoxic status of this fungal strain when administered in a sole intragastric dose in mice.

  17. Ranking cultivated blueberry for Mummy Berry Blight and Fruit Infection Incidence

    USDA-ARS?s Scientific Manuscript database

    Mummy berry is an important disease of cultivated blueberry. The disease has two distinct phases; a blighting phase initiated by ascospores and a fruit infection stage initiated by conidia. In this study we investigated the resistance of more than 100 blueberry cultivar to both phases of the disease...

  18. Mummy Berry Fruit Rot and Shoot Blight Incidence in Blueberry: Prediction, Ranking, and Stability in a Long-term Study

    USDA-ARS?s Scientific Manuscript database

    Mummy berry is an important disease of cultivated blueberry. The disease has two distinct phases; a blighting phase initiated by ascospores and a fruit infection stage initiated by conidia. In this study we investigated blueberry cultivar resistance to both phases of the disease and, utilizing ‘stan...

  19. Metarhizium microsclerotia and hydrogel versus hydromulch: testing fungal formulations against Asian longhorned beetles

    USDA-ARS?s Scientific Manuscript database

    The efficacy of microsclerotia of Metarhizium brunneum (Petch) (Hypocreales: Clavicipitaceae) strain F52 (ARSEF 7711) was tested using samples that had been exposed on forest trees, allowing time for conidia to be produced. An aqueous mixture of microsclerotial granules (61.3% of dry mass), a straw ...

  20. A monograph of the entomopathogenic genera Hypocrella, Moelleriella, and Samuelsia gen. nov. (Ascomycota, Hypocreales, Clavicipitaceae), and their aschersonia-like anamorphs in the Neotropics.

    PubMed

    Chaverri, P; Liu, M; Hodge, K T

    2008-01-01

    The present taxonomic revision deals with Neotropical species of three entomopathogenic genera that were once included in Hypocrella s. l.: Hypocrella s. str. (anamorph Aschersonia), Moelleriella (anamorph aschersonia-like), and Samuelsia gen. nov (anamorph aschersonia-like). Species of Hypocrella, Moelleriella, and Samuelsia are pathogens of scale insects (Coccidae and Lecaniidae, Homoptera) and whiteflies (Aleyrodidae, Homoptera) and are common in tropical regions. Phylogenetic analyses of DNA sequences from nuclear ribosomal large subunit (28S), translation elongation factor 1-alpha (TEF 1-alpha), and RNA polymerase II subunit 1 (RPB1) and analyses of multiple morphological characters demonstrate that the three segregated genera can be distinguished by the disarticulation of the ascospores and shape and size of conidia. Moelleriella has filiform multi-septate ascospores that disarticulate at the septa within the ascus and aschersonia-like anamorphs with fusoid conidia. Hypocrella s. str. has filiform to long-fusiform ascospores that do not disarticulate and Aschersonia s. str. anamorphs with fusoid conidia. The new genus proposed here, Samuelsia, has filiform to long-fusiform ascospores that do not disarticulate and aschersonia-like anamorphs with small allantoid conidia. In addition, the present study presents and discusses the evolution of species, morphology, and ecology in Hypocrella, Moelleriella, and Samuelsia based on multigene phylogenetic analyses.

  1. Entomopathogenic Fungi (Hypocreales) for Control of Potato Psyllid

    USDA-ARS?s Scientific Manuscript database

    Laboratory and field trials of the four isolates of entomopathogenic fungi, Isaria fumosorosea and Metarhizium anisopliae were conducted to evaluate their potential for control of the potato psyllid. In the laboratory, 2 ml aqueous suspension of 107conidia/ml in a of each isolate applied in a Potte...

  2. Heat-stressed Metarhizium anisopliae: Viability (in vitro) and virulence (in vivo) assessments against the tick Rhipicephalus sanguineus

    USDA-ARS?s Scientific Manuscript database

    The current study investigated the thermotolerance of Metarhizium anisopliae s.l. conidia from the commercial products Metarril® SP Organic and Metarril® WP. The efficacy of these M. anisopliae formulations against the tick Rhipicephalus sanguineus s.l. was studied in laboratory under optimum or hea...

  3. Three new hecogenin glycosides from fermented leaves of Agave americana.

    PubMed

    Jin, Jian-Ming; Liu, Xi-Kui; Yang, Chong-Ren

    2003-06-01

    Eight steroidal compounds, including three new hecogenin glycosides, agamenosides D-F, were isolated from the fermented leaves of Agave americana. The structures of the new steroidal saponins were elucidated by spectroscopic data and chemical methods. The activity of the isolated compounds on deformations of mycelia germinated from conidia of Pyricularia oaryzae P-2b was evaluated.

  4. Efficacy of spray applications of entomopathogenic fungi against western flower thrips infesting greenhouse impatiens under variable moisture conditions

    USDA-ARS?s Scientific Manuscript database

    Efficacy tests of three entomopathogenic fungi (Beauveria bassiana strain GHA, Metarhizium brunneum strain F52, and Metarhizium anisopliae s.l. strain ESC-1) were conducted against thrips infesting greenhouse crops of single impatiens under variable moisture conditions. Fungal conidia suspended in 0...

  5. A monograph of the entomopathogenic genera Hypocrella, Moelleriella, and Samuelsia gen. nov. (Ascomycota, Hypocreales, Clavicipitaceae), and their aschersonia-like anamorphs in the Neotropics

    PubMed Central

    Chaverri, P.; Liu, M.; Hodge, K.T.

    2008-01-01

    The present taxonomic revision deals with Neotropical species of three entomopathogenic genera that were once included in Hypocrella s. l.: Hypocrella s. str. (anamorph Aschersonia), Moelleriella (anamorph aschersonia-like), and Samuelsia gen. nov (anamorph aschersonia-like). Species of Hypocrella, Moelleriella, and Samuelsia are pathogens of scale insects (Coccidae and Lecaniidae, Homoptera) and whiteflies (Aleyrodidae, Homoptera) and are common in tropical regions. Phylogenetic analyses of DNA sequences from nuclear ribosomal large subunit (28S), translation elongation factor 1-α (TEF 1-α), and RNA polymerase II subunit 1 (RPB1) and analyses of multiple morphological characters demonstrate that the three segregated genera can be distinguished by the disarticulation of the ascospores and shape and size of conidia. Moelleriella has filiform multi-septate ascospores that disarticulate at the septa within the ascus and aschersonia-like anamorphs with fusoid conidia. Hypocrella s. str. has filiform to long-fusiform ascospores that do not disarticulate and Aschersonia s. str. anamorphs with fusoid conidia. The new genus proposed here, Samuelsia, has filiform to long-fusiform ascospores that do not disarticulate and aschersonia-like anamorphs with small allantoid conidia. In addition, the present study presents and discusses the evolution of species, morphology, and ecology in Hypocrella, Moelleriella, and Samuelsia based on multigene phylogenetic analyses. PMID:18490956

  6. DOSE-DEPENDENT ALLERGIC ASTHMA RESPONSES TO PENICILLIUM CHRYSOGENUM

    EPA Science Inventory

    ABSTRACT
    Indoor mold has been associated with development of allergic asthma. Penicillium chrysogenum, a common indoor mold, is known to have several allergens and its viable conidia can induce allergic responses in a mouse model of allergic penicilliosis. The hypothesis o...

  7. Conidiogenic effects of mannose-binding lectins isolated from cotyledons of red kidney bean (Phaseolus vulgaris) on Alternaria alternata.

    PubMed

    Alizadeh, Hossein; Leung, David W M; Cole, Anthony L J

    2011-01-01

    Effect of proteinaceous extracts from red kidney bean cotyledons on mycelium of Alternaria alternata growing on potato dextrose agar (PDA) plates was investigated. Unexpectedly, conidia formation was induced in response to applied crude extracts. A PDA disc method was developed to quantify conidia formed. A purified fraction retaining conidiation inducing effect (CIE) was obtained following several protein purification procedures including the last step of eluting bound proteins from an Affi-gel blue gel column. Based on MALDI (matrix assisted laser desorption/ionization) mass spectrometric analysis, a previously identified mannose-binding lectin (MBL) called PvFRIL (Phaseolus vulgaris fetal liver tyrosine kinase 3-receptor interacting lectin) was present in this conidiation inducing fraction. The PvFRIL was subsequently purified using a single step mannose-agarose affinity column chromatography. When the lectin was applied exogenously to A. alternata, increased conidiation resulted. The conidia produced in response to the MBL were similar to those induced by other methods and their germ tubes were longer after 12 h growth than those induced under white light. To our knowledge this is the first report of exogenous application of a PvFRIL or another purified protein from a plant inducing conidia formation in a fungus.

  8. Physiological and antioxidant response by Beauveria bassiana Bals (Vuill.) to different oxygen concentrations.

    PubMed

    Garza-López, Paul Misael; Konigsberg, Mina; Gómez-Quiroz, Luis Enrique; Loera, Octavio

    2012-01-01

    The effect of three levels of oxygen (normal atmosphere (21% O(2)), low oxygen (16% O(2)) and enriched oxygen (26% O(2))) on the production and germination of conidia by Beauveria bassiana was evaluated using rice as a substrate. The maximum yield of conidia was achieved under hypoxia (16% O(2)) after 8 days of culture (1.51 × 10(9) conidia per gram of initial dry substrate), representing an increase of 32% compared to the normal atmosphere. However, germination was reduced by at least 27% due to atmospheric modifications. Comparison of antioxidant enzyme activity (superoxide dismutases and catalases) with the oxidation profiles of biomolecules (proteins and lipids) showed that a decrease in catalase activity in the final days of culture coincided with an increase in the amount of oxidized lipids, showing that oxidative stress was a consequence of pulses of different concentrations of O(2). This is the first study describing oxidative stress induction by atmospheric modification, with practical implications for conidia production.

  9. Effects of catalase on the accumulation of H(2)O(2) in rice cells inoculated with rice blast fungus, Magnaporthe oryzae.

    PubMed

    Tanabe, Shigeru; Nishizawa, Yoko; Minami, Eiichi

    2009-10-01

    Roles of H(2)O(2) in the infection process of Magnaporthe oryzae on rice were investigated. In a leaf sheath assay for up to 48 h post-inoculation, the absence or presence of catalase in the conidia suspension was correlated with the level of accumulated H(2)O(2) in infected leaf cells, as observed by staining with 3',3-diaminobenzidine tetrahydrochloride. In the incompatible interaction, the appearance of autofluorescence or frequency of cell death characterized by granulation (symptoms characteristic of hypersensitive responses) was not significantly affected by the presence of catalase in the conidia suspension. In the leaf blade assay, inoculation of compatible conidia in the presence of catalase produced more severe symptoms than that of conidia in the absence of catalase at 6 days post-inoculation. These results suggest that, in this host-parasite interaction, the primary role of host-produced H(2)O(2) is in limiting hyphal growth after penetration through toxic action. Furthermore, in incompatible interactions, H(2)O(2) is implied not to be a major mediator of hypersensitive cell death.

  10. Direct Polymerase Chain Reaction-Based Detection of Cercospora Beticola in Field Soils

    USDA-ARS?s Scientific Manuscript database

    Cercospora beticola, the causal agent of Cercospora leaf spot of sugar beet, survives as pseudostromata in infected beet leaf residues in the soil. Under optimal conditions, overwintering propagules germinate and produce conidia that are dispersed as primary inoculum to initiate infection in sugar b...

  11. Transcriptome of Aspergillus flavus aswA (AFLA_085170) deletion strain related to sclerotial development and production of secondary metabolites

    USDA-ARS?s Scientific Manuscript database

    Aspergillus flavus produces many secondary metabolites including aflatoxins. Besides conidia, the fungus uses sclerotia as another type of propagule. We obtained transcriptomes from four growth conditions of the aswA mutant, a strain impaired in sclerotial development and production of sclerotium-sp...

  12. Phylogeny of Hirsutella species (Ophiocordycipitaceae) from the USA: remedying the paucity of Hirsutella sequence data

    USDA-ARS?s Scientific Manuscript database

    Hirsutella (Ophiocordycipitaceae: Hypocreales) is a genus of insect, mite, and nematode pathogens with an asexual morph, which generally produce a mucilaginous cluster of one or several conidia on phialides that are basally subulate and taper to a fine neck. The generic name Hirsutella has been prop...

  13. Silver enhances the in vitro antifungal activity of the saponin, CAY-1

    USDA-ARS?s Scientific Manuscript database

    The separate fungicidal properties of purified CAY-1, dissolved silver, and EDTA were studied, as were the abilities of silver or EDTA to enhance CAY-1 fungicidal properties. Nongerminated and germinating conidia of A. flavus, A. fumigatus, A. niger, F. moniliforme, F. oxysporum and F. solani were ...

  14. Effects of surfactants on conidial germination of Myrothecium verrucaria

    USDA-ARS?s Scientific Manuscript database

    Myrothecium verrucaria has been employed as a unique biological control agent because it is highly effective against several annual and perennial weeds, including red vine, trumpet creeper, redroot pigweed, kudzu, hempsesbania and sicklepod. Although aerial conidia of M. verrucaria are hydrophilic, ...

  15. Spicellum ovalisporum

    USDA-ARS?s Scientific Manuscript database

    A new species of fungi was discovered that is associated with the leaf-cutter bee Megachile rotundata. It is described as Spicellum ovalisporum and is similar to the type and only species in Spicellum, namely S. roseum except that the conidia are ellipsoid to ovate, 3.5–5.5 x 2–2.5 µm. As the neig...

  16. Comparative transcriptomic analysis of the heat stress response in the filamentous fungus Metarhizium anisopliae using RNA-Seq.

    PubMed

    Wang, Zhang-Xun; Zhou, Xia-Zhi; Meng, Hui-Min; Liu, Yu-Jun; Zhou, Quan; Huang, Bo

    2014-06-01

    The entomopathogenic fungus Metarhizium anisopliae is widely used for biological control of a variety of insect pests. The effectiveness of the microbial pest control agent, however, is limited by poor thermotolerance. The molecular mechanism underlying the response to heat stress in the conidia of entomopathogenic fungi remains unclear. Here, we conducted high-throughput RNA-Seq to analyze the differential gene expression between control and heat treated conidia of M. anisopliae at the transcriptome level. RNA-Seq analysis generated 6,284,262 and 5,826,934 clean reads in the control and heat treated groups, respectively. A total of 2,722 up-regulated and 788 down-regulated genes, with a cutoff of twofold change, were identified by expression analysis. Among these differentially expressed genes, many were related to metabolic processes, biological regulation, cellular processes and response to stimuli. The majority of genes involved in endocytic pathways, proteosome pathways and regulation of autophagy were up-regulated, while most genes involved in the ribosome pathway were down-regulated. These results suggest that these differentially expressed genes may be involved in the heat stress response in conidia. As expected, significant changes in expression levels of genes encoding heat shock proteins and proteins involved in trehalose accumulation were observed in conditions of heat stress. These results expand our understanding of the molecular mechanisms of the heat stress response of conidia and provide a foundation for future investigations.

  17. Selection of entomopathogenic fungi for aphid control.

    PubMed

    Vu, Van Hanh; Hong, Suk Il; Kim, Keun

    2007-12-01

    Twelve strains of entomopathogenic fungi such as Lecanicillium lecanii, Paecilomyces farinosus, Beauveria bassiana, Metarhizium anisopliae, Cordyceps scarabaeicola, and Nomuraea rileyi were screened for aphid control. At 25 degrees C and 75% relative humidity (RH), among tested entomopathogenic fungi, L. lecanii 41185 showed the highest virulent pathogenicity for both Myzus persicae and Aphis gossypii, and their control values were both nearly 100% 5 and 2 d after treatment, respectively. Moreover, at an RH of 45% and in a wide temperature range (20-30 degrees C), L. lecanii 41185 also exhibited the highest virulence to M. persicae. The control value of M. persicae and the 50% lethal time (LT50) decreased significantly as the applied conidial concentration increased. The 50% lethal concentration (LC50) of the conidial suspension of this fungus was determined to be 6.55x10(5) conidia/ml. The control values of M. persicae resulting from the application of 1x10(7) and 1x10(8) conidia/ml were nearly the same and were significantly higher than that of 1x10(6) conidia/ml. The tested entomopathogenic fungi grew in a broad temperature range (15-30 degrees C). Lecanicillium strains showed optimum growth at 25 degrees C. The aerial conidia of Lecanicillium strains also could germinate in a broad temperature range (15-30 degrees C) and L. lecanii 41185 was the only strain with conidial germination at 35 degrees C.

  18. Efficacy of Tolypocladium cylindrosporum against Aedes aegypti eggs, larvae and adults.

    PubMed

    Rocha, L F N; Sousa, N A; Rodrigues, J; Catão, A M L; Marques, C S; Fernandes, É K K; Luz, C

    2015-11-01

    The aim of this study was to test the activity of seven Tolypocladium cylindrosporum isolates against different developmental stages of Aedes aegypti. Ovicidal activity (≤30% of eclosion; at a 84% eclosion of control eggs) and lowest cumulative emergence of adults (≤30%; at a 66·3% control emergence) originating from eclosing larvae was found 15 and 30 days, respectively, after applying conidia of ARSEF 962 or 1580 onto the eggs. All isolates induced larval mortality when third-instar larvae were treated with conidia (≤5% control mortality); the lowest lethal concentration (LC50) at 5 days after treatment was found with ARSEF 1580 (9·2 × 10(5) conidia ml(-1)), and the shortest lethal time (LT50 ) with ARSEF 2912 (2·8 days at 3·3 × 10(6) conidia ml(-1)). The pathogenicity of T. cylindrosporum to A. aegypti adults is corroborated. ARSEF 1027, 1580 and 2912 induced the highest cumulative mortality in adults (≥90%) 15 days after application (≤5% control mortality). Findings emphasized an isolate- and developmental stage-related virulence of T. cylindrosporum. ARSEF 1580, in the group of tested isolates, is the most promising isolate for use against terrestrial eggs, aquatic larvae and adults of this important vector. © 2015 The Society for Applied Microbiology.

  19. Laboratory and field evaluation of an entomopathogenic fungus, Isaria cateniannulata strain 08XS‐1, against Tetranychus urticae (Koch)

    PubMed Central

    Zhang, Xiaona; Zou, Xiao; Guo, Jianjun

    2016-01-01

    Abstract BACKGROUND The two‐spotted mite, Tetranychus urticae Koch, is one of the most serious mite pests of crops throughout the world. Biocontrol of the mite with fungal agents has long been paid much attention because of the development of insecticide resistance and the severe restriction of chemical pesticides. In this study, the efficacy of submerged conidia of the entomopathogenic fungus Isaria cateniannulata strain 08XS‐1 against T. urticae eggs, larvae and female adults was evaluated at different temperatures and humidity in the laboratory and under field conditions. RESULTS The results showed that a suspension of 2 × 107 submerged conidia mL −1 caused the highest mortalities of mite eggs, larvae and females (100, 100 and 70% respectively) at 100% relative humidity and 25 °C in the laboratory. In the field experiments against the mites, a suspension of 2 × 108 submerged conidia mL −1 achieved significant efficiency – the relative control effects were 88.6, 83.8 and 83%, respectively, in cucumber, eggplant and bean fields after 10 days of treatment. CONCLUSION The results suggest that the I. cateniannulata strain 08XS‐1 is a potential fungal agent, with acceptable production cost of conidia, against T. urticae in the field in an area such as southwestern China with higher air humidity. © 2016 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry. PMID:26775712

  20. Phylogeny and ecology of the ubiquitous saprobe Cladosporium sphaerospermum, with descriptions of seven new species from hypersaline environments

    PubMed Central

    Zalar, P.; de Hoog, G.S.; Schroers, H.-J.; Crous, P.W.; Groenewald, J.Z.; Gunde-Cimerman, N.

    2007-01-01

    Saprobic Cladosporium isolates morphologically similar to C. sphaerospermum are phylogenetically analysed on the basis of DNA sequences of the ribosomal RNA gene cluster, including the internal transcribed spacer regions ITS1 and ITS2, the 5.8S rDNA (ITS) and the small subunit (SSU) rDNA as well as β-tubulin and actin gene introns and exons. Most of the C. sphaerospermum-like species show halotolerance as a recurrent feature. Cladosporium sphaerospermum, which is characterised by almost globose conidia, is redefined on the basis of its ex-neotype culture. Cladosporium dominicanum, C. psychrotolerans, C. velox, C. spinulosum and C. halotolerans, all with globoid conidia, are newly described on the basis of phylogenetic analyses and cryptic morphological and physiological characters. Cladosporium halotolerans was isolated from hypersaline water and bathrooms and detected once on dolphin skin. Cladosporium dominicanum and C. velox were isolated from plant material and hypersaline water. Cladosporium psychrotolerans, which grows well at 4 °C but not at 30 °C, and C. spinulosum, having conspicuously ornamented conidia with long digitate projections, are currently only known from hypersaline water. We also newly describe C. salinae from hypersaline water and C. fusiforme from hypersaline water and animal feed. Both species have ovoid to ellipsoid conidia and are therefore reminiscent of C. herbarum. Cladosporium langeronii (= Hormodendrum langeronii) previously described as a pathogen on human skin, is halotolerant but has not yet been recorded from hypersaline environments. PMID:18490999

  1. Ascochyta blight of chickpea reduced 38% by application of Aureobasidium pullulans (anamorphic Dothioraceae, Dothideales) to post harvest debris.

    USDA-ARS?s Scientific Manuscript database

    In 2004-2005, early winter application of suspensions of Aureobasidium pullulans (AuP) conidia to post-harvest chickpea debris resulted in significantly fewer spring-time Ascochyta blight lesions on chickpea test plants relative to controls. Survival of plants adjacent to treated debris was higher t...

  2. Pathogenicity of Metarhizium anisopliae for Ceratitis capitata (Wied.) (Diptera: Tephritidae) in soil with different pesticides.

    PubMed

    Mochi, Dinalva A; Monteiro, Antonio C; De Bortoli, Sergio A; Dória, Háyda O S; Barbosa, José C

    2006-01-01

    This research intended to investigate if the presence of pesticides in the soil could affect the pathogenicity of Metarhizium anisopliae Metsch. (Sorokin) for Ceratitis capitata (Wied.) and assess the effect of conidia application as suspension or dry conidia. The fungicides chlorothalonyl and tebuconazol, the acaricide abamectin, the insecticide trichlorfon, and the herbicide ametrin were applied at the manufacturer-recommended doses. Soil samples were placed in glass flasks and were given the fungus as conidial suspension or dry. After pesticide application, 20 3rd-instar larvae were placed in the soil. The flasks were sealed with voile fabric and incubated at 27 +/- 0.5 masculineC for nine days, until adult emergence; incubation continued for four more days at room temperature. The total insect survival was significantly affected and pathogenic activity was detected from the pupa stage on. Pupa survival was reduced (P<0.05); the same occurred during the adult phase. No effect was observed at the larval stage. The pesticides applied to the soil affected the activity of M. anisopliae slightly: only in the dry conidia assay the fungicides chlorothalonyl and tebuconazole reduced (86.2% and 82.5%, respectively) the survival period of C. capitata compared to the control (95.0%). The techniques used for conidia application did not influence the total insect survival rate, but conidial suspension applied on soil surface reduced survival during the pupae and adult phases.

  3. Pseudomonas contamination of a fungal-based biopesticide: implications for honey bee (Hymenoptera: Apidae) health and varroa mite (Acari: Varroidae) control

    USDA-ARS?s Scientific Manuscript database

    A biopesticide, formulated with commercially-prepared conidia of a strain of Beauveria bassiana isolated from Varroa mites, was tested in a field experiment in southern France to compare published results from the same geographic region with the same biopesticide but made with laboratory-prepared co...

  4. A Trichophyton Rubrum Infection Model Based on the Reconstructed Human Epidermis - Episkin®.

    PubMed

    Liang, Pan-Pan; Huang, Xin-Zhu; Yi, Jin-Ling; Chen, Zhi-Rui; Ma, Han; Ye, Cong-Xiu; Chen, Xian-Yan; Lai, Wei; Chen, Jian

    2016-01-05

    Trichophyton rubrum represents the most common infectious fungus responsible for dermatophytosis in human, but the mechanism involved is still not completely understood. An appropriate model constructed to simulate host infection is the prerequisite to study the pathogenesis of dermatophytosis caused by T. rubrum. In this study, we intended to develop a new T. rubrum infection model in vitro, using the three-dimensional reconstructed epidermis - EpiSkin ®, and to pave the way for further investigation of the mechanisms involved in T. rubrum infection. The reconstructed human epidermis (RHE) was infected by inoculating low-dose (400 conidia) and high-dose (4000 conidia) T. rubrum conidia to optimize the infection dose. During the various periods after infection, the samples were processed for pathological examination and scanning electron microscopy (SEM) observation. The histological analysis of RHE revealed a fully differentiated epidermis with a functional stratum corneum, which was analogous to the normal human epidermis. The results of hematoxylin and eosin staining and the periodic acid-Schiff staining showed that the infection dose of 400 conidia was in accord with the pathological characteristics of host dermatophytosis caused by T. rubrum. SEM observations further exhibited the process of T. rubrum infection in an intuitionistic way. We established the T. rubrum infection model on RHE in vitro successfully. It is a promising model for further investigation of the mechanisms involved in T. rubrum infection.

  5. A Trichophyton Rubrum Infection Model Based on the Reconstructed Human Epidermis - Episkin®

    PubMed Central

    Liang, Pan-Pan; Huang, Xin-Zhu; Yi, Jin-Ling; Chen, Zhi-Rui; Ma, Han; Ye, Cong-Xiu; Chen, Xian-Yan; Lai, Wei; Chen, Jian

    2016-01-01

    Background: Trichophyton rubrum represents the most common infectious fungus responsible for dermatophytosis in human, but the mechanism involved is still not completely understood. An appropriate model constructed to simulate host infection is the prerequisite to study the pathogenesis of dermatophytosis caused by T. rubrum. In this study, we intended to develop a new T. rubrum infection model in vitro, using the three-dimensional reconstructed epidermis - EpiSkin®, and to pave the way for further investigation of the mechanisms involved in T. rubrum infection. Methods: The reconstructed human epidermis (RHE) was infected by inoculating low-dose (400 conidia) and high-dose (4000 conidia) T. rubrum conidia to optimize the infection dose. During the various periods after infection, the samples were processed for pathological examination and scanning electron microscopy (SEM) observation. Results: The histological analysis of RHE revealed a fully differentiated epidermis with a functional stratum corneum, which was analogous to the normal human epidermis. The results of hematoxylin and eosin staining and the periodic acid-Schiff staining showed that the infection dose of 400 conidia was in accord with the pathological characteristics of host dermatophytosis caused by T. rubrum. SEM observations further exhibited the process of T. rubrum infection in an intuitionistic way. Conclusions: We established the T. rubrum infection model on RHE in vitro successfully. It is a promising model for further investigation of the mechanisms involved in T. rubrum infection. PMID:26712433

  6. Population balance modeling of the conidial aggregation of Aspergillus niger.

    PubMed

    Lin, P-J; Grimm, L H; Wulkow, M; Hempel, D C; Krull, R

    2008-02-01

    Numerous biotechnological production processes are based on the submerse cultivation of filamentous fungi. Process design, however, is often hampered by the complex growth pattern of these organisms. In the morphologic development of coagulating filamentous fungi, like Aspergillus niger, conidial aggregation is the first step of filamentous morphogenesis. For a proper description of this phenomenon it is necessary to characterize conidial populations. Kinetic studies performed with an in-line particle size analyzer suggested that two distinct aggregation steps have to be considered. The first step of conidial aggregation starts immediately after inoculation. Both the rate constants of formation and disintegration of aggregates have been determined by measuring the concentration of conidia at the beginning of the cultivation and the concentration of particles at steady state during the first hours of cultivation. In contrast to the first aggregation step, where the collision of conidia is presumed to be responsible for the process, the second aggregation step is thought to be initiated by germination of conidia. Growing hyphae provide additional surface for the attachment of non- germinated conidia, which leads to a strong decrease in particle concentration. The specific hyphal length growth rate and the ratio of particle concentration to the growing adhesion hyphal surface are decisive matters of the second aggregation step. Both aggregation steps can be described by population dynamics and simulated using the program package PARSIVAL (PARticle SIze eVALution) for the treatment of general particle population balances.

  7. Photoprotective and Antimutagenic Activity of Agaricus subrufescens Basidiocarp Extracts.

    PubMed

    da Costa, M C D; Regina, M; Cilião Filho, M; Linde, G A; do Valle, J S; Paccola-Meirelles, L D; Colauto, N B

    2015-10-01

    The photoprotective and antimutagenic activity of opened and closed basidiocarps of Agaricus subrufescens (=A. blazei; =A. brasiliensis) obtained by different extraction methods were evaluated on Aspergillus nidulans conidia submitted to ultraviolet (UV) light. The aqueous extracts were obtained by three extraction methods: maceration, infusion, and decoction, at two different extraction times. The extracts of A. subrufescens did not present toxicity for A. nidulans conidia. A suspension of A. nidulans conidia was submitted to extracts before and after the exposure to UV light. All basidiocarp extracts, regardless of the extraction method or development stage, protected A. nidulans conidia against the damaging effects of the mutagenic agent. The antimutagenic and photoprotective activity was strengthened with extracts obtained by 168-h maceration, followed by 24-h maceration and 60-min infusion and, at last, by 30-min infusion. Although the extracts presented protector effect as well as recoverer effect to the action of UV light, the preventive effect was more evident. Differences in the biological activity in function of the different development stages were detected with greater antimutagenic and photoprotective activity for the opened basidiocarps. However, the extraction method is the most important factor to be considered when compared to the basidiocarp development stage to obtain better antimutagenic and photoprotective activity of A. subrufescens basidiocarps.

  8. WetA Is Required for Conidiogenesis and Conidium Maturation in the Ascomycete Fungus Fusarium graminearum

    PubMed Central

    Son, Hokyoung; Kim, Myung-Gu; Min, Kyunghun; Lim, Jae Yun; Choi, Gyung Ja; Kim, Jin-Cheol; Chae, Suhn-Kee

    2014-01-01

    Fusarium graminearum, a prominent fungal pathogen that infects major cereal crops, primarily utilizes asexual spores to spread disease. To understand the molecular mechanisms underlying conidiogenesis in F. graminearum, we functionally characterized the F. graminearum ortholog of Aspergillus nidulans wetA, which has been shown to be involved in conidiogenesis and conidium maturation. Deletion of F. graminearum wetA did not alter mycelial growth, sexual development, or virulence, but the wetA deletion mutants produced longer conidia with fewer septa, and the conidia were sensitive to acute stresses, such as oxidative stress and heat stress. Furthermore, the survival rate of aged conidia from the F. graminearum wetA deletion mutants was reduced. The wetA deletion resulted in vigorous generation of single-celled conidia through autophagy-dependent microcycle conidiation, indicating that WetA functions to maintain conidial dormancy by suppressing microcycle conidiation in F. graminearum. Transcriptome analyses demonstrated that most of the putative conidiation-related genes are expressed constitutively and that only a few genes are specifically involved in F. graminearum conidiogenesis. The conserved and distinct roles identified for WetA in F. graminearum provide new insights into the genetics of conidiation in filamentous fungi. PMID:24186953

  9. Hemolysis, Toxicity, and Randomly Amplified Polymorphic DNA Analysis of Stachybotrys chartarum Strains

    PubMed Central

    Vesper, Stephen J.; Dearborn, Dorr G.; Yike, Iwona; Sorenson, W. G.; Haugland, Richard A.

    1999-01-01

    Stachybotrys chartarum is an indoor air, toxigenic fungus that has been associated with a number of human and veterinary health problems. Most notable among these has been a cluster of idiopathic pulmonary hemorrhage cases that were observed in the Cleveland, Ohio, area. In this study, 16 strains of S. chartarum isolated from case (n = 8) or control (n = 8) homes in Cleveland and 12 non-Cleveland strains from diverse geographic locations were analyzed for hemolytic activity, conidial toxicity, and randomly amplified polymorphic DNA banding patterns. In tests for hemolytic activity, strains were grown at 23°C on wet wallboard pieces for an 8-week test period. Conidia from these wallboard pieces were subcultured on sheep’s blood agar once a week over this period and examined for growth and clearing of the medium at 37 or 23°C. Five of the Cleveland strains (all from case homes) showed hemolytic activity at 37°C throughout the 8-week test compared to 3 of the non-Cleveland strains. Five of the Cleveland strains, compared to two of the non-Cleveland strains, produced highly toxic conidia (>90 μg of T2 toxin equivalents per g [wet weight] of conidia) after 10 and 30 days of growth on wet wallboard. Only 3 of the 28 strains examined both were consistently hemolytic and produced highly toxic conidia. Each of these strains was isolated from a house in Cleveland where an infant had idiopathic pulmonary hemorrhage. PMID:10388719

  10. Multigene analysis suggests ecological speciation in the fungal pathogen Claviceps purpurea

    USDA-ARS?s Scientific Manuscript database

    Claviceps purpurea is an important pathogen of grasses and source of novel chemical compounds. Three groups within this species (G1, G2, and G3) have been recognized based on habitat association, sclerotia and conidia morphology, and alkaloid production. These groups have further been supported by g...

  11. CCR4-Not Complex Subunit Not2 Plays Critical Roles in Vegetative Growth, Conidiation and Virulence in Watermelon Fusarium Wilt Pathogen Fusarium oxysporum f. sp. niveum

    PubMed Central

    Dai, Yi; Cao, Zhongye; Huang, Lihong; Liu, Shixia; Shen, Zhihui; Wang, Yuyan; Wang, Hui; Zhang, Huijuan; Li, Dayong; Song, Fengming

    2016-01-01

    CCR4-Not complex is a multifunctional regulator that plays important roles in multiple cellular processes in eukaryotes. In the present study, the biological function of FonNot2, a core subunit of the CCR4-Not complex, was explored in Fusarium oxysporum f. sp. niveum (Fon), the causal agent of watermelon wilt disease. FonNot2 was expressed at higher levels in conidia and germinating conidia and during infection in Fon-inoculated watermelon roots than in mycelia. Targeted disruption of FonNot2 resulted in retarded vegetative growth, reduced conidia production, abnormal conidial morphology, and reduced virulence on watermelon. Scanning electron microscopy observation of infection behaviors and qRT-PCR analysis of in planta fungal growth revealed that the ΔFonNot2 mutant was defective in the ability to penetrate watermelon roots and showed reduced fungal biomass in root and stem of the inoculated plants. Phenotypic and biochemical analyses indicated that the ΔFonNot2 mutant displayed hypersensitivity to cell wall perturbing agents (e.g., Congo Red and Calcofluor White) and oxidative stress (e.g., H2O2 and paraquat), decreased fusaric acid content, and reduced reactive oxygen species (ROS) production during spore germination. Our data demonstrate that FonNot2 plays critical roles in regulating vegetable growth, conidiogenesis and conidia morphology, and virulence on watermelon via modulating cell wall integrity, oxidative stress response, ROS production and FA biosynthesis through the regulation of transcription of genes involved in multiple pathways. PMID:27695445

  12. GERMINATION, VIABILITY AND CLEARANCE OF STACHYBOTRYS CHARTARUM IN THE LUNGS OF INFANT RATS

    EPA Science Inventory

    The fungus Stachybotrys chartarum has been associated with many adverse health effects including the condition known as idiopathic pulmonary hemorrhage in infants. In order to gain some insight into possible mechanisms, viable conidia of S. chartarum were instilled into the lung...

  13. Occurrence of toxicity among protease, amylase, and color mutants of a nontoxic soy sauce koji mold

    SciTech Connect

    Kalayanamitr, A.; Bhumiratana, A.; Flegel, T.W.; Glinsukon, T.; Shinmyo, A.

    1987-08-01

    A soy sauce koji mold, Aspergillus flavus var. columnaris Raper and Fennel (ATCC 44310), was treated with UV irradiation to obtain mutant strains possessing high protease activities, high amylase activities, and light-colored conidia. Selected mutant strains were tested for toxicity, and some were found acutely toxic to weanling rats, although all were negative for aflatoxin production.

  14. Transcriptome analysis of the blueberry-mummy berry pathosystem

    USDA-ARS?s Scientific Manuscript database

    Mummy berry disease of blueberry, casual agent Monilinia vaccinii-corymbosi, has two distinct phases- a blight stage of young foliage and flowers and a flower infection stage that leads to mummified fruit (pseudoscelrotia). The flower infections stage requires conidia to germinate on the style and g...

  15. Predatory activity of Butlerius nematodes and nematophagous fungi against Haemonchus contortus infective larvae.

    PubMed

    Silva, Manoel Eduardo da; Uriostegui, Miguel Angel Mercado; Millán-Orozco, Jair; Gives, Pedro Mendoza de; Hernández, Enrique Liébano; Braga, Fabio Ribeiro; Araújo, Jackson Victor de

    2017-01-26

    The purpose of this study was to evaluate the predatory activity of the nematode Butlerius spp. and fungal isolates of Duddingtonia flagrans, Clonostachys rosea, Arthrobotrys musiformis and Trichoderma esau against H. contortus infective larvae (L3) in grass pots. Forty-eight plastic gardening pots containing 140 g of sterile soil were used. Panicum spp. grass seeds (200 mg) were sown into each pot and individually watered with 10 mL of tap water. Twelve days after seeding, the pots were randomly divided into 6 groups (n=8). Two thousand H. contortus infective larvae (L3) were added to each group. Additionally, the following treatments were established: Group 1 - 2000 Butlerius spp. larvae; group 2 - A. musiformis (1x107 conidia); group 3 - T. esau (1x107 conidia); group 4 - C. rosea (1x107 conidia), group 5 - D. flagrans (1x107conidia) and Group 6 - no biological controller (control group). The larval population of H. contortus exposed to Butlerius spp. was reduced by 61.9%. Population reductions of 90.4, 66.7, 61.9 and 85.7% were recorded in the pots containing A. musiformis, T. esau, C. rosea and D. flagrans, respectively. The results of this study indicate that the predatory nematode Butlerius spp. and the assessed fungi display an important predatory activity can be considered suitable potential biological control agents.

  16. Fungicidal and bactericidal properties of bisabolol and dragosantol

    USDA-ARS?s Scientific Manuscript database

    a-bisabolol, a natural plant sesquiterpene alcohol, and dragosantol, a racemic mixture of synthetic bisabolol, were studied for their fungicidal properties separately against the nongerminated and germinating conidia of several species of Aspergillus and Fusarium which are problems in agriculture an...

  17. GERMINATION, VIABILITY AND CLEARANCE OF STACHYBOTRYS CHARTARUM IN THE LUNGS OF INFANT RATS

    EPA Science Inventory

    The fungus Stachybotrys chartarum has been associated with many adverse health effects including the condition known as idiopathic pulmonary hemorrhage in infants. In order to gain some insight into possible mechanisms, viable conidia of S. chartarum were instilled into the lung...

  18. Serinocyclins A and B, Cyclic Heptapeptides from Metarhizium anisopliae

    USDA-ARS?s Scientific Manuscript database

    Two new cyclic heptapeptides, serinocyclins A (1) and B (2), were isolated from conidia of the entomopathogenic fungus Metarhizium anisopliae. Structures were elucidated by a combination of mass spectrometric, NMR, and X-ray diffraction techniques. Serinocyclin A (1) contains three serine units, a...

  19. Does methyl salicylate, a component of herbivore-induced plant odour, promote sporulation of the mite-pathogenic fungus Neozygites tanajoae?

    PubMed

    Hountondji, Fabien C C; Hanna, Rachid; Sabelis, Maurice W

    2006-01-01

    Blends of volatile chemicals emanating from cassava leaves infested by the cassava green mite were found to promote conidiation of Neozygites tanajoae, an entomopathogenic fungus specific to this mite. Methyl salicylate (MeSA) is one compound frequently present in blends of herbivore-induced plant volatiles (HIPV) as well as that of mite-infested cassava. Here, we investigated the effect of methyl salicylate in its pure form on the production of pre-infective spores (conidia), and the germination of these spores into infective spores (capilliconidia), by a Brazilian isolate and a Beninese isolate of N. tanajoae. Mummified mites previously infected by the fungal isolates were screened under optimal abiotic conditions for sporulation inside tightly closed boxes with or without methyl salicylate diffusing from a capillary tube. Production of conidia was consistently higher (37%) when the Beninese isolate was exposed to MeSA than when not exposed to it (305.5 +/- 52.62 and 223.2 +/- 38.13 conidia per mummy with and without MeSA, respectively). MeSA, however, did not promote conidia production by the Brazilian isolate (387.4 +/- 44.74 and 415.8 +/- 57.95 conidia per mummy with and without MeSA, respectively). Germination of the conidia into capilliconidia was not affected by MeSA for either isolate (0.2%, 252.6 +/- 31.80 vs. 253.0 +/- 36.65 for the Beninese isolate and 4.2%, 268.5 +/- 37.90 vs. 280.2 +/- 29.43 for the Brazilian isolate). The effects of MeSA on the production of conidia were similar to those obtained under exposure to the complete blends of HIPV for the case of the Beninese isolate, but dissimilar (no promoting effect of MeSA) for the case of the Brazilian isolate. This shows that MeSA, being one compound out of many HIPV, can be a factor promoting sporulation of N. tanajoae, but it may not be the only factor as its effect varies with the fungal isolate under study.

  20. First report of the infection of insecticide-resistant malaria vector mosquitoes with an entomopathogenic fungus under field conditions

    PubMed Central

    2011-01-01

    Background Insecticide-resistant mosquitoes are compromising the ability of current mosquito control tools to control malaria vectors. A proposed new approach for mosquito control is to use entomopathogenic fungi. These fungi have been shown to be lethal to both insecticide-susceptible and insecticide-resistant mosquitoes under laboratory conditions. The goal of this study was to see whether entomopathogenic fungi could be used to infect insecticide-resistant malaria vectors under field conditions, and to see whether the virulence and viability of the fungal conidia decreased after exposure to ambient African field conditions. Methods This study used the fungus Beauveria bassiana to infect the insecticide-resistant malaria vector Anopheles gambiae s.s (Diptera: Culicidae) VKPER laboratory colony strain. Fungal conidia were applied to polyester netting and kept under West African field conditions for varying periods of time. The virulence of the fungal-treated netting was tested 1, 3 and 5 days after net application by exposing An. gambiae s.s. VKPER mosquitoes in WHO cone bioassays carried out under field conditions. In addition, the viability of B. bassiana conidia was measured after up to 20 days exposure to field conditions. Results The results show that B. bassiana infection caused significantly increased mortality with the daily risk of dying being increased by 2.5× for the fungus-exposed mosquitoes compared to the control mosquitoes. However, the virulence of the B. bassiana conidia decreased with increasing time spent exposed to the field conditions, the older the treatment on the net, the lower the fungus-induced mortality rate. This is likely to be due to the climate because laboratory trials found no such decline within the same trial time period. Conidial viability also decreased with increasing exposure to the net and natural abiotic environmental conditions. After 20 days field exposure the conidial viability was 30%, but the viability of control

  1. Neem oil increases the efficiency of the entomopathogenic fungus Metarhizium anisopliae for the control of Aedes aegypti (Diptera: Culicidae) larvae.

    PubMed

    Gomes, Simone A; Paula, Adriano R; Ribeiro, Anderson; Moraes, Catia O P; Santos, Jonathan W A B; Silva, Carlos P; Samuels, Richard I

    2015-12-30

    Entomopathogenic fungi are potential candidates for use in integrated vector management and many isolates are compatible with synthetic and natural insecticides. Neem oil was tested separately and in combination with the entomopathogenic fungus Metarhizium anisopliae against larvae of the dengue vector Aedes aegypti. Our aim was to increase the effectiveness of the fungus for the control of larval mosquito populations. Commercially available neem oil was used at concentrations ranging from 0.0001 to 1%. Larval survival rates were monitored over a 7 day period following exposure to neem. The virulence of the fungus M. anisopliae was confirmed using five conidial concentrations (1 × 10(5) to 1 × 10(9) conidia mL(-1)) and survival monitored over 7 days. Two concentrations of fungal conidia were then tested together with neem (0.001%). Survival curve comparisons were carried out using the Log-rank test and end-point survival rates were compared using one-way ANOVA. 1% neem was toxic to A. aegypti larvae reducing survival to 18% with S50 of 2 days. Neem had no effect on conidial germination or fungal vegetative growth in vitro. Larval survival rates were reduced to 24% (S50 = 3 days) when using 1 × 10(9) conidia mL(-1). Using 1 × 10(8) conidia mL(-1), 30% survival (S50 = 3 days) was observed. We tested a "sub-lethal" neem concentration (0.001%) together with these concentrations of conidia. For combinations of neem + fungus, the survival rates were significantly lower than the survival rates seen for fungus alone or for neem alone. Using a combination of 1 × 10(7) conidia mL(-1) + neem (0.001%), the survival rates were 36%, whereas exposure to the fungus alone resulted in 74% survival and exposure to neem alone resulted in 78% survival. When using 1 × 10(8) conidia mL(-1), the survival curves were modified, with a combination of the fungus + neem resulting in 12% survival, whilst the fungus alone at this concentration also

  2. The Neurospora crassa mutant NcΔEgt-1 identifies an ergothioneine biosynthetic gene and demonstrates that ergothioneine enhances conidial survival and protects against peroxide toxicity during conidial germination.

    PubMed

    Bello, Marco H; Barrera-Perez, Viviana; Morin, Dexter; Epstein, Lynn

    2012-02-01

    Ergothioneine (EGT) is a histidine derivative with sulfur on the imidazole ring and a trimethylated amine; it is postulated to have an antioxidant function. Although EGT apparently is only produced by fungi and some prokaryotes, it is acquired by animals and plants from the environment, and is concentrated in animal tissues in cells with an EGT transporter. Monobromobimane derivatives of EGT allowed conclusive identification of EGT by LC/MS and the quantification of EGT in Colletotrichum graminicola and Neurospora crassa conidia and mycelia. EGT concentrations were significantly (α=0.05) higher in conidia than in mycelia, with approximately 17X and 5X more in C. graminicola and N. crassa, respectively. The first EGT biosynthetic gene in a fungus was identified by quantifying EGT in N. crassa wild type and knockouts in putative homologs of actinomycete EGT biosynthetic genes. NcΔEgt-1, a strain with a knockout in gene NCU04343, does not produce EGT, in contrast to the wild type. To determine the effects of EGT in vivo, we compared NcΔEgt-1 to the wild type. NcΔEgt-1 is not pleiotropically affected in rate of hyphal elongation in Vogel's medium either with or without ammonium nitrate and in the rate of germination of macroconidia on Vogel's medium. The superoxide-producer menadione had indistinguishable effects on conidial germination between the two strains. Cupric sulfate also had indistinguishable effects on conidial germination and on hyphal growth between the two strains. In contrast, germination of NcΔEgt-1 conidia was significantly more sensitive to tert-butyl hydroperoxide than the wild type; germination of 50% (GI(50)) of the NcΔEgt-1 conidia was prevented at 2.7 mM tert-butyl hydroperoxide whereas the GI(50) for the wild type was 4.7 mM tert-butyl hydroperoxide, or at a 1.7X greater concentration. In the presence of tert-butyl hydroperoxide and the fluorescent reactive oxygen species indicator 5-(and-6)-carboxy-2',7'-dichlorodihydrofluorescein

  3. The revenge of time: fungal deterioration of cultural heritage with particular reference to books, paper and parchment.

    PubMed

    Sterflinger, Katja; Pinzari, Flavia

    2012-03-01

    Hyphomycetous fungi - so called 'mould'- are the most important agents of biodeterioration in museums, museums' storage rooms, in libraries, collections and restoration studios. Fungi are able to live at low water activities, they are perfectly adapted to indoor environments and thrive in microclimatic niches caused by condensation, lack of ventilation or water retention by hygroscopic materials. Fungi spoil valuable pieces of art aesthetically, mechanically, chemically and by degradation of organic components. Historical material made of paper and oil paintings with high amounts of organic binders are especially susceptible to fungal deterioration. In order to prevent fungal contamination or to treat already contaminated objects an integrated approach including climate control, material-specific cleaning and application of carefully selected biocides is necessary. © 2011 Society for Applied Microbiology and Blackwell Publishing Ltd.

  4. Nothing Special in the Specialist? Draft Genome Sequence of Cryomyces antarcticus, the Most Extremophilic Fungus from Antarctica

    PubMed Central

    Sterflinger, Katja; Lopandic, Ksenija; Pandey, Ram Vinay; Blasi, Barbara; Kriegner, Albert

    2014-01-01

    The draft genome of the Antarctic endemic fungus Cryomyces antarcticus is presented. This rock inhabiting, microcolonial fungus is extremely stress tolerant and it is a model organism for exobiology and studies on stress resistance in Eukaryots. Since this fungus is a specialist in the most extreme environment of the Earth, the analysis of its genome is of important value for the understanding of fungal genome evolution and stress adaptation. A comparison with Neurospora crassa as well as with other microcolonial fungi shows that the fungus has a genome size of 24 Mbp, which is the average in the fungal kingdom. Although sexual reproduction was never observed in this fungus, 34 mating genes are present with protein homologs in the classes Eurotiomycetes, Sordariomycetes and Dothideomycetes. The first analysis of the draft genome did not reveal any significant deviations of this genome from comparative species and mesophilic hyphomycetes. PMID:25296285

  5. Evaluation of the fungus Beauveria bassiana as a potential biological control agent against phlebotomine sand flies in Colombian coffee plantations.

    PubMed

    Reithinger, R; Davies, C R; Cadena, H; Alexander, B

    1997-09-01

    In Colombia, the entomopathogenic fungus Beauveria bassiana (Deuteromycotina: Hyphomycetes) is widely used to control the coffee berry borer Hypothenemus hampei (Coleoptera: Scolytidae) in coffee plantations. Recent studies suggested that this fungus is also pathogenic to several important vectors of disease, including Phlebotomus papatasi and Lutzomyia longipalpis (Diptera: Psychodidae). The present study evaluated the use of B. bassiana as a potential biological control agent against phlebotomine sand flies in Colombian coffee plantations. Histopathologic examination indicates that B. bassiana is unable to infect sand flies under natural conditions, although dead sand flies were shown to be readily infected. In addition, laboratory bioassays where flies were exposed to the fungus applied onto coffee plants (though not filter paper) showed lower mean survival times than the control.

  6. Cadmium induces a novel metallothionein and phytochelatin 2 in an aquatic fungus

    SciTech Connect

    Jaeckel, Petra; Krauss, Gudrun; Menge, Sieglinde; Krauss, Gerd-Joachim . E-mail: krauss@biochemtech.uni-halle.de

    2005-07-22

    Cadmium stress response was measured at the thiol peptide level in an aquatic hyphomycete (Heliscus lugdunensis). In liquid culture, 0.1mM cadmium increased the glutathione (GSH) content and induced the synthesis of additional thiol peptides. HPLC, electrospray ionization mass spectrometry, and Edman degradation confirmed that a novel small metallothionein as well as phytochelatin (PC2) were synthesized. The metallothionein has a high homology to family 8 metallothioneins (http://www.expasy.ch/cgi-bin/lists?metallo.txt). The bonding of at least two cadmium ions to the metallothionein was demonstrated by mass spectrometry (MALDI MS). This is the first time that simultaneous induction of metallothionein and phytochelatin accompanied by an increase in GSH level has been shown in a fungus under cadmium stress, indicating a potential function of these complexing agents for in vivo heavy metal detoxification. The method presented here should be applicable as biomarker tool. ol.

  7. Nothing special in the specialist? Draft genome sequence of Cryomyces antarcticus, the most extremophilic fungus from Antarctica.

    PubMed

    Sterflinger, Katja; Lopandic, Ksenija; Pandey, Ram Vinay; Blasi, Barbara; Kriegner, Albert

    2014-01-01

    The draft genome of the Antarctic endemic fungus Cryomyces antarcticus is presented. This rock inhabiting, microcolonial fungus is extremely stress tolerant and it is a model organism for exobiology and studies on stress resistance in Eukaryots. Since this fungus is a specialist in the most extreme environment of the Earth, the analysis of its genome is of important value for the understanding of fungal genome evolution and stress adaptation. A comparison with Neurospora crassa as well as with other microcolonial fungi shows that the fungus has a genome size of 24 Mbp, which is the average in the fungal kingdom. Although sexual reproduction was never observed in this fungus, 34 mating genes are present with protein homologs in the classes Eurotiomycetes, Sordariomycetes and Dothideomycetes. The first analysis of the draft genome did not reveal any significant deviations of this genome from comparative species and mesophilic hyphomycetes.

  8. Microbial specialists in below-grade foundation walls in Scandinavia.

    PubMed

    Nunez, M; Hammer, H

    2014-10-01

    Below-grade foundation walls are often exposed to excessive moisture by water infiltration, condensation, leakage, or lack of ventilation. Microbial growth in these structures depends largely on environmental factors, elapsed time, and the type of building materials and construction setup. The ecological preferences of Actinomycetes (Actinobacteria) and the molds Ascotricha chartarum, Myxotrichum chartarum (Ascomycota), Geomyces pannorum, and Monocillium sp. (Hyphomycetes) have been addressed based on analyses of 1764 samples collected in below-grade spaces during the period of 2001-2012. Our results show a significant correlation between these taxa and moist foundation walls as ecological niches. Substrate preference was the strongest predictor of taxa distribution within the wall, but the taxa's physiological needs, together with gradients of abiotic factors within the wall structure, also played a role. Our study describes for the first time how the wall environment affects microbial growth.

  9. Subcutaneous phaeohyphomycosis caused by Geniculosporium species; a new fungal pathogen.

    PubMed

    Suzuki, Y; Udagawa, S; Wakita, H; Yamada, N; Ichikawa, H; Furukawa, F; Takigawa, M

    1998-02-01

    A 70-year-old Japanese timberworker dealing with imported timber from the U.S.A. and Russia had an asymptomatic subcutaneous nodule with a small fistula on his left knee. Histopathological examination of the nodule revealed brownish hyphal elements in encapsulated pyogranuloma. The fungus isolated from a discharge of the fistula and an excised specimen of the lesion was identified as Geniculosporium sp., which represents a conidial state (anamorph) of several genera such as Anthostomella, Biscogniauxia, Euepixylon, Leprieuria, Nemania, Phylacia and Rosellina in the Xylariaceae. Whereas this dematiaceous hyphomycete is commonly found on decaying wood and bark of various trees, to our knowledge, this is the first case of a phaeomycotic cyst caused by fungi belonging to the genus Geniculosporium.

  10. Persistence versus Escape: Aspergillus terreus and Aspergillus fumigatus Employ Different Strategies during Interactions with Macrophages

    PubMed Central

    Slesiona, Silvia; Gressler, Markus; Mihlan, Michael; Zaehle, Christoph; Schaller, Martin; Barz, Dagmar; Hube, Bernhard; Jacobsen, Ilse D.; Brock, Matthias

    2012-01-01

    Invasive bronchopulmonary aspergillosis (IBPA) is a life-threatening disease in immunocompromised patients. Although Aspergillus terreus is frequently found in the environment, A. fumigatus is by far the main cause of IBPA. However, once A. terreus establishes infection in the host, disease is as fatal as A. fumigatus infections. Thus, we hypothesized that the initial steps of disease establishment might be fundamentally different between these two species. Since alveolar macrophages represent one of the first phagocytes facing inhaled conidia, we compared the interaction of A. terreus and A. fumigatus conidia with alveolar macrophages. A. terreus conidia were phagocytosed more rapidly than A. fumigatus conidia, possibly due to higher exposure of β-1,3-glucan and galactomannan on the surface. In agreement, blocking of dectin-1 and mannose receptors significantly reduced phagocytosis of A. terreus, but had only a moderate effect on phagocytosis of A. fumigatus. Once phagocytosed, and in contrast to A. fumigatus, A. terreus did not inhibit acidification of phagolysosomes, but remained viable without signs of germination both in vitro and in immunocompetent mice. The inability of A. terreus to germinate and pierce macrophages resulted in significantly lower cytotoxicity compared to A. fumigatus. Blocking phagolysosome acidification by the v-ATPase inhibitor bafilomycin increased A. terreus germination rates and cytotoxicity. Recombinant expression of the A. nidulans wA naphthopyrone synthase, a homologue of A. fumigatus PksP, inhibited phagolysosome acidification and resulted in increased germination, macrophage damage and virulence in corticosteroid-treated mice. In summary, we show that A. terreus and A. fumigatus have evolved significantly different strategies to survive the attack of host immune cells. While A. fumigatus prevents phagocytosis and phagolysosome acidification and escapes from macrophages by germination, A. terreus is rapidly phagocytosed, but

  11. Recognition of a core fragment of Beauveria bassiana hydrophobin gene promoter (P hyd1) and its special use in improving fungal biocontrol potential

    PubMed Central

    Wang, Zheng-Liang; Ying, Sheng-Hua; Feng, Ming-Guang

    2013-01-01

    To identify a suitable promoter for use in engineering fungal entomopathogens to improve heterologous gene expression and fungal biocontrol potential, a 1798 bp promoter (Phyd1) upstream of Beauveria bassiana class I hydrophobin gene (hyd1) was optimized by upstream truncation and site-directed mutation. A truncated 1290 bp fragment (Phyd1-t1) drove eGFP expression in B. bassiana much more efficiently than full-length Phyd1. Further truncating Phyd1-t1 to 1179, 991 and 791 bp or mutating one of the binding domains of three transcription factors in Phyd1-t1 reduced significantly the expression of eGFP (enhanced green fluorescence protein). Under Phyd1-t1 control, eGFP was expressed more abundantly in conidiogenic cells and conidia than in mycelia. Therefore, Phyd1-t1 was used to integrate a bacterium-derived, insect midgut-specific toxin (vip3Aa1) gene into B. bassiana, yielding a transgenic strain (BbHV8) expressing 9.8-fold more toxin molecules in conidia than a counterpart strain (BbV28) expressing the toxin under the control of PgpdA, a promoter widely used for gene expression in fungi. Consequently, BbHV8 showed much higher per os virulence to Spodoptera litura larvae than BbV28 in standardized bioassays with normal conidia for both cuticle penetration and ingestion or heat-killed conidia for ingestion only. Conclusively, Phyd1-t1 is a useful tool for enhancing beneficial protein expression, such as vip3Aa1, in fungal conidia, which are the active ingredients of mycoinsecticides. PMID:22639846

  12. Recognition of a core fragment ofBeauveria bassiana hydrophobin gene promoter (P hyd1) and its special use in improving fungal biocontrol potential.

    PubMed

    Wang, Zheng-Liang; Ying, Sheng-Hua; Feng, Ming-Guang

    2013-01-01

    To identify a suitable promoter for use in engineering fungal entomopathogens to improve heterologous gene expression and fungal biocontrol potential, a 1798 bp promoter (P hyd1) upstream of Beauveria bassiana class I hydrophobin gene (hyd1) was optimized by upstream truncation and site-directed mutation. A truncated 1290 bp fragment (P hyd1-t1) drove eGFP expression in B. bassiana much more efficiently than full-length P hyd1. Further truncating P hyd1-t1 to 1179, 991 and 791 bp or mutating one of the binding domains of three transcription factors in P hyd1-t1 reduced significantly the expression of eGFP (enhanced green fluorescence protein). Under P hyd1-t1 control, eGFP was expressed more abundantly in conidiogenic cells and conidia than in mycelia. Therefore, P hyd1-t1 was used to integrate a bacterium-derived, insect midgut-specific toxin (vip3Aa1) gene into B. bassiana, yielding a transgenic strain (BbHV8) expressing 9.8-fold more toxin molecules in conidia than a counterpart strain (BbV28) expressing the toxin under the control of P gpdA, a promoter widely used for gene expression in fungi. Consequently, BbHV8 showed much higher per os virulence to Spodoptera litura larvae than BbV28 in standardized bioassays with normal conidia for both cuticle penetration and ingestion or heat-killed conidia for ingestion only. Conclusively, P hyd1-t1 is a useful tool for enhancing beneficial protein expression, such as vip3Aa1, in fungal conidia, which are the active ingredients of mycoinsecticides.

  13. Spatial separation of semiochemical Lurem-TR and entomopathogenic fungi to enhance their compatibility and infectivity in an autoinoculation system for thrips management.

    PubMed

    Mfuti, David K; Subramanian, Sevgan; van Tol, Rob W H M; Wiegers, Gerrie L; de Kogel, Willem J; Niassy, Saliou; du Plessis, Hannalene; Ekesi, Sunday; Maniania, Nguya K

    2016-01-01

    The effect of spatial separation of the semiochemical Lurem-TR, which has been found to inhibit conidia of entomopathogenic fungi when put together, on the persistence of conidia of Metarhizium brunneum and M. anisopliae was evaluated in the greenhouse and field in order to develop an autodissemination strategy for the management of Megalurothrips sjostedti on cowpea crop. Influence of spatial separation of the semiochemical on thrips attraction and conidial acquisition by thrips from the autoinoculation device was also investigated in the field. Persistence of conidia of M. brunneum and M. anisopliae increased with distance of separation of Lurem-TR. Direct exposure of fungus without separation from Lurem-TR recorded the lowest conidial germination as compared with the other treatments. Attraction of thrips to the device also varied significantly according to distance between device and semiochemical, with a higher number of thrips attracted when Lurem-TR was placed in a container below the device and at 10 cm distance. There was no significant difference in conidial acquisition between spatial separation treatments of conidia and Lurem-TR. Attraction of other insect pests to the device did not significantly vary between treatments. Positive correlations were found between conidial acquisition and thrips attraction. This study suggests that spatial separation of fungal conidia from Lurem-TR in an autoinoculation device could provide a low-cost strategy for effective management of thrips in grain legume cropping systems. © 2015 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

  14. Infection of malaria (Anopheles gambiae s.s.) and filariasis (Culex quinquefasciatus) vectors with the entomopathogenic fungus Metarhizium anisopliae

    PubMed Central

    Scholte, Ernst-Jan; Njiru, Basilio N; Smallegange, Renate C; Takken, Willem; Knols, Bart GJ

    2003-01-01

    Background Current intra-domiciliary vector control depends on the application of residual insecticides and/or repellents. Although biological control agents have been developed against aquatic mosquito stages, none are available for adults. Following successful use of an entomopathogenic fungus against tsetse flies (Diptera: Glossinidae) we investigated the potency of this fungus as a biological control agent for adult malaria and filariasis vector mosquitoes. Methods In the laboratory, both sexes of Anopheles gambiae sensu stricto and Culex quinquefasciatus were passively contaminated with dry conidia of Metarhizium anisopliae. Pathogenicity of this fungus for An. gambiae was further tested for varying exposure times and different doses of oil-formulated conidia. Results Comparison of Gompertz survival curves and LT50 values for treated and untreated specimens showed that, for both species, infected mosquitoes died significantly earlier (p < 0.0001) than uninfected control groups. No differences in LT50 values were found for different exposure times (24, 48 hrs or continuous exposure) of An. gambiae to dry conidia. Exposure to oil-formulated conidia (doses ranging from 1.6 × 107 to 1.6 × 1010 conidia/m2) gave LT50 values of 9.69 ± 1.24 (lowest dose) to 5.89 ± 0.35 days (highest dose), with infection percentages ranging from 4.4–83.7%. Conclusion Our study marks the first to use an entomopathogenic fungus against adult Afrotropical disease vectors. Given its high pathogenicity for both adult Anopheles and Culex mosquitoes we recommend development of novel targeted indoor application methods for the control of endophagic host-seeking females. PMID:14565851

  15. Unraveling the Nanoscale Surface Properties of Chitin Synthase Mutants of Aspergillus fumigatus and Their Biological Implications

    PubMed Central

    Alsteens, David; Aimanianda, Vishukumar; Hegde, Pushpa; Pire, Stéphane; Beau, Rémi; Bayry, Jagadeesh; Latgé, Jean-Paul; Dufrêne, Yves F.

    2013-01-01

    Understanding the surface properties of the human opportunistic pathogen Aspergillus fumigatus conidia is essential given the important role they play during the fungal interactions with the human host. Although chitin synthases with myosin motor-like domain (CSM) play a major role in cell wall biosynthesis, the extent to which deletion of the CSM genes alter the surface structural and biophysical-biological properties of conidia is not fully characterized. We used three complementary atomic force microscopy techniques—i.e., structural imaging, chemical force microscopy with hydrophobic tips, and single-molecule force spectroscopy with lectin tips—to gain detailed insights into the nanoscale surface properties (ultrastructure, hydrophobicity) and polysaccharide composition of the wild-type and the chitin synthase mutant (ΔcsmA, ΔcsmB, and ΔcsmA/csmB) conidia of A. fumigatus. Wild-type conidia were covered with a highly hydrophobic layer of rodlet nanostructures. By contrast, the surface of the ΔcsmA mutant was almost completely devoid of rodlets, leading to loss of hydrophobicity and exposure of mannan and chitin polysaccharides. The ΔcsmB and ΔcsmA/csmB mutants showed a different behavior, i.e., the surfaces featured poorly organized rodlet layers, yet with a low hydrophobicity and substantial amounts of exposed mannan and chitin at the surface. As the rodlet layer is important for masking recognition of immunogenic fungal cell wall components by innate immune cells, disappearance of rodlet layers in all three chitin synthase mutant conidia was associated with an activation of human dendritic cells. These nanoscale analyses emphasize the important and distinct roles that the CSMA and CSMB genes play in modulating the surface properties and immune interactions of A. fumigatus and demonstrate the power of atomic force microscopy in fungal genetic studies for assessing the phenotypic characteristics of mutants altered in cell surface organization. PMID

  16. Unraveling the nanoscale surface properties of chitin synthase mutants of Aspergillus fumigatus and their biological implications.

    PubMed

    Alsteens, David; Aimanianda, Vishukumar; Hegde, Pushpa; Pire, Stéphane; Beau, Rémi; Bayry, Jagadeesh; Latgé, Jean-Paul; Dufrêne, Yves F

    2013-07-16

    Understanding the surface properties of the human opportunistic pathogen Aspergillus fumigatus conidia is essential given the important role they play during the fungal interactions with the human host. Although chitin synthases with myosin motor-like domain (CSM) play a major role in cell wall biosynthesis, the extent to which deletion of the CSM genes alter the surface structural and biophysical-biological properties of conidia is not fully characterized. We used three complementary atomic force microscopy techniques-i.e., structural imaging, chemical force microscopy with hydrophobic tips, and single-molecule force spectroscopy with lectin tips-to gain detailed insights into the nanoscale surface properties (ultrastructure, hydrophobicity) and polysaccharide composition of the wild-type and the chitin synthase mutant (ΔcsmA, ΔcsmB, and ΔcsmA/csmB) conidia of A. fumigatus. Wild-type conidia were covered with a highly hydrophobic layer of rodlet nanostructures. By contrast, the surface of the ΔcsmA mutant was almost completely devoid of rodlets, leading to loss of hydrophobicity and exposure of mannan and chitin polysaccharides. The ΔcsmB and ΔcsmA/csmB mutants showed a different behavior, i.e., the surfaces featured poorly organized rodlet layers, yet with a low hydrophobicity and substantial amounts of exposed mannan and chitin at the surface. As the rodlet layer is important for masking recognition of immunogenic fungal cell wall components by innate immune cells, disappearance of rodlet layers in all three chitin synthase mutant conidia was associated with an activation of human dendritic cells. These nanoscale analyses emphasize the important and distinct roles that the CSMA and CSMB genes play in modulating the surface properties and immune interactions of A. fumigatus and demonstrate the power of atomic force microscopy in fungal genetic studies for assessing the phenotypic characteristics of mutants altered in cell surface organization. Copyright

  17. Oxygen and an Extracellular Phase Transition Independently Control Central Regulatory Genes and Conidiogenesis in Aspergillus fumigatus

    PubMed Central

    Chi, Myoung-Hwan; Craven, Kelly D.

    2013-01-01

    Conidiogenesis is the primary process for asexual reproduction in filamentous fungi. As the conidia resulting from the conidiogenesis process are primarily disseminated via air currents and/or water, an outstanding question has been how fungi recognize aerial environments suitable for conidial development. In this study, we documented the somewhat complex development of the conidia-bearing structures, termed conidiophores, from several Aspergillus species in a subsurface (gel-phase) layer of solid media. A subset of the isolates studied was able to develop conidiophores in a gel-phase environment, but exposure to the aeriform environment was required for the terminal developmental transition from phialide cells to conidia. The remaining Aspergilli could not initiate the conidiogenesis process until they were exposed to the aeriform environment. Our observations of conidiophore development in high or low oxygen conditions in both aeriform and gel-phase environments revealed that oxygen and the aeriform state are positive environmental factors for inducing conidiogenesis in most of the aspergilli tested in this study. Transcriptional analysis using A. fumigatus strain AF293 confined to either the aeriform or gel-phase environments revealed that expression of a key regulatory gene for conidiophore development (AfubrlA) is facilitated by oxygen while expression of another regulatory gene controlling conidia formation from phialides (AfuabaA) was repressed regardless of oxygen levels in the gel-embedded environment. Furthermore, by comparing the developmental behavior of conidiation-defective mutants lacking genes controlling various regulatory checkpoints throughout the conidiogenesis pathway, we propose that this aerial response by the fungus requires both oxygen and the phase transition (solid to aeriform), with these environmental signals integrating into the upstream regulatory pathway and central regulatory pathway of conidiogenesis, respectively. Our findings

  18. Density and Molecular Epidemiology of Aspergillus in Air and Relationship to Outbreaks of Aspergillus Infection

    PubMed Central

    Leenders, Alexander C. A. P.; van Belkum, Alex; Behrendt, Myra; Luijendijk, Ad; Verbrugh, Henri A.

    1999-01-01

    After five patients were diagnosed with nosocomial invasive aspergillosis caused by Aspergillus fumigatus and A. flavus, a 14-month surveillance program for pathogenic and nonpathogenic fungal conidia in the air within and outside the University Hospital in Rotterdam (The Netherlands) was begun. A. fumigatus isolates obtained from the Department of Hematology were studied for genetic relatedness by randomly amplified polymorphic DNA (RAPD) analysis. This was repeated with A. fumigatus isolates contaminating culture media in the microbiology laboratory. The density of the conidia of nonpathogenic fungi in the outside air showed a seasonal variation: higher densities were measured during the summer, while lower densities were determined during the fall and winter. Hardly any variation was found in the numbers of Aspergillus conidia. We found decreasing numbers of conidia when comparing air from outside the hospital to that inside the hospital and when comparing open areas within the hospital to the closed department of hematology. The increase in the number of patients with invasive aspergillosis could not be explained by an increase in the number of Aspergillus conidia in the outside air. The short-term presence of A. flavus can only be explained by the presence of a point source, which was probably patient related. Genotyping A. fumigatus isolates from the department of hematology showed that clonally related isolates were persistently present for more than 1 year. Clinical isolates of A. fumigatus obtained during the outbreak period were different from these persistent clones. A. fumigatus isolates contaminating culture media were all genotypically identical, indicating a causative point source. Knowledge of the epidemiology of Aspergillus species is necessary for the development of strategies to prevent invasive aspergillosis. RAPD fingerprinting of Aspergillus isolates can help to determine the cause of an outbreak of invasive aspergillosis. PMID:10325319

  19. Cell Wall Modifications during Conidial Maturation of the Human Pathogenic Fungus Pseudallescheria boydii

    PubMed Central

    Ghamrawi, Sarah; Rénier, Gilles; Saulnier, Patrick; Cuenot, Stéphane; Zykwinska, Agata; Dutilh, Bas E.; Thornton, Christopher; Faure, Sébastien; Bouchara, Jean-Philippe

    2014-01-01

    Progress in extending the life expectancy of cystic fibrosis (CF) patients remains jeopardized by the increasing incidence of fungal respiratory infections. Pseudallescheria boydii (P. boydii), an emerging pathogen of humans, is a filamentous fungus frequently isolated from the respiratory secretions of CF patients. It is commonly believed that infection by this fungus occurs through inhalation of airborne conidia, but the mechanisms allowing the adherence of Pseudallescheria to the host epithelial cells and its escape from the host immune defenses remain largely unknown. Given that the cell wall orchestrates all these processes, we were interested in studying its dynamic changes in conidia as function of the age of cultures. We found that the surface hydrophobicity and electronegative charge of conidia increased with the age of culture. Melanin that can influence the cell surface properties, was extracted from conidia and estimated using UV-visible spectrophotometry. Cells were also directly examined and compared using electron paramagnetic resonance (EPR) that determines the production of free radicals. Consistent with the increased amount of melanin, the EPR signal intensity decreased suggesting polymerization of melanin. These results were confirmed by flow cytometry after studying the effect of melanin polymerization on the surface accessibility of mannose-containing glycoconjugates to fluorescent concanavalin A. In the absence of melanin, conidia showed a marked increase in fluorescence intensity as the age of culture increased. Using atomic force microscopy, we were unable to find rodlet-forming hydrophobins, molecules that can also affect conidial surface properties. In conclusion, the changes in surface properties and biochemical composition of the conidial wall with the age of culture highlight the process of conidial maturation. Mannose-containing glycoconjugates that are involved in immune recognition, are progressively masked by polymerization of

  20. Biocontrol of the Brown-Banded Cockroach, Supella longipalpa F. (Blattaria: Blattellidae), with Entomopathogenic Fungus, Metharhizium anisopliae

    PubMed Central

    Sharififard, Mona; Mossadegh, Mohammad Saeed; Vazirianzadeh, Babak; Latifi, Seyed Mahmood

    2016-01-01

    Background: Considering to the high distribution of cockroaches as urban pests, the efficacy of different formulations of Metarhizium anisopliae strain Iran 437C were assessed against the brown-banded cockroach, Supella longipalpa F. under laboratory and field conditions. Methods: Metarhizium anisopliae isolates were screened with immersing adults of the brown-banded cockroachs in aqueous suspension of 108 conidia ml−1 followed by surface or bait treated with different doses of the most virulent isolate against the nymphs. Then formulations of conidia oil-in-water were examined versus cockroach nymphs using different plant oils and paraffin. Then they were evaluated and compared with aqueous suspension and control group. On a large-scale, the sunflower oil-in-water formulation of conidia was sprayed at houses using a hand sprayer. Results: Metarhizium anisopliae IRAN 437C was the most virulent isolate against the brown-banded cockroach, causing 100% mortality in adults at seven days post-exposure. Inoculated bait with this isolate was not enough pathogenic against the cockroach even at two weeks after treatment. Treated surface with conidia as aqueous suspension or oil-in-water formulation was more effective than the bait formulation against the cockroach caused 39.4–97.2% mortality compared with 2.5% mortality in control group after two days. Spraying the conidia formulated with sunflower oil was an effective formulation causing 76.1% reduction in the cockroach density on the third day post treatment in the houses. Conclusion: The oil-in-water formulation of M. anisopliae IRAN 437C could be recommended as a promising alternative for cockroach control. PMID:27308292