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Effect of five pesticides on the mycelial growth of some soil and pathogenic fungi.

The effect of three doses (the lowest is the recommended dose in the field) of 5 pesticides namely: Ceresan, Orthocid (fungicides), VCS-438 (herbicide), and Dursban and Dipterex (insecticides) on the mycelial dry weight of Aspergillus fumigatus. Fusarium moniforme, Pencillium italicum, and Sclerotium ceprivorum was tested 2, 4, 6, 8 and 10 days after treatment. All the pesticides induced inhibition of mycelial growth of the test fungi. Only VCS-438 at the low dose was stimulatory to F. moniliforme after 4 and 6 days and to P. italicum after 4 days. The rate of inhibition was influenced by the type of fungus, the age of the mycelium, and the dose of the pesticide. At the end of the experiment the two fungicides were detoxified by the test fungi with a rate higher than that of the other pesticides.

Antifungal Agents↗

Cultural and nutritional studies of zoopathogenic fungi associated with livestock feeds in Nigeria.

Mycelial growth and nutritional physiology of zoopathogenic Aspergillus fumigatus FRES., Thermoascus aurantiacus MIEHE, and Thermomyces lanuginosus TSIK., obtained from livestock feeds and poultry droppings in Nigeria were studied. An optimal pH-temperature range of 5.5-6.5 and 37 degrees - 45 degrees C was recorded for the growth of these fungi. Various sources of carbon supplied in the growth medium, except sorbose and rhamnose, were utilized for growth. Only Aspergillus fumigatus caused considerable weight losses of the filter papers supplied and hydrolyzed the carboxy-methyl cellulose (CMC) in the medium. The culture filtrates of this fungus contained CM-cellulases which also hydrolyzed CMC to reducing sugar at 45 degrees C. D-glucose and L-asparagine concentrations in the range of 15-20 g/liter and 2.5-5 g/liter, respectively, were best for growth of the test fungi. Various forms of organic and inorganic nitrogen provided were also utilized. The inorganic sources of nitrogen could readily substitute the organic forms at the optimal growth conditions.

Amino Acids↗

Seasonal fluctuations of freshwater fungi in river Nile (Egypt).

The seasonal fluctuations of aquatic fungi were studied over 24 water sample collected monthly between May 1979 and April 1981 from the River Nile near Assiut (about 375 km, south from Cairo, Egypt). The richest periods in aquatic fungal genera and species were September--December 1979, January--March, October--December 1980, and January--April 1981. These periods represent low or moderate temperature months. The poorest periods were May--August 1979, April, May, July, August and September 1980 which are almost summer months. Twenty-nine species and one variety of Olpidiopsis saprolegniae which belong to ten genera of aquatic fungi were collected during this investigation. Three genera were of high seasonal occurrence (19-24 months) namely: Achlya, Saprolegnia and Dictyuchus. Three other genera were of moderate seasonal occurrence (6-11 months) namely: Isoachlya, Leptolegnia and Pythium. The rest of genera were of low or rare seasonal occurrence.

Egypt↗

Occurrence of mycotoxins (ochratoxin A, deoxynivalenol) and toxigenic fungi in Moroccan wheat grains: impact of ecological factors on the growth and ochratoxin A production.

The aim of the present work was to evaluate the contamination of some samples, taken from Moroccan wheat grains, by ochratoxin A (OTA), deoxynivalenol (DON) and the associated toxigenic fungi. Moreover, we focused on the influence of environmental factors on both the growth and OTA production by three strains of Aspergillus. The results showed that only few samples were contaminated by the two mycotoxins (2 samples for OTA and 7 for DON). The main isolated fungi belong to the Aspergillus, Penicillium and Fusarium genus; 74 Aspergillus and 28 Penicillium isolates were tested for their ability to produce OTA. Only 2 A. alliaceus and 14 A. niger were able to synthesize OTA. However, none of Penicillium isolates can produce this toxin under the conditions mentioned. In respect of the effects of the temperature and water activity (aw), the optimal conditions for the growth and OTA production were different. While the optimal conditions of growth for A. alliaceus and A. terreus are 30 degrees C and 0.98 aw, A. niger preferred 0.93-0.95 aw at 25 degrees C, whereas the optimal production of OTA was observed at 30 degrees C for both A. alliaceus and A. niger at 0.93 and 0.99 aw, respectively.

Aspergillus↗

Genotoxicity of fungi evaluated by SOS microplate assay.

By an introduction of sodium dodecylsulfate for cell lysis and immunomicroplate for mass assay, the modified SOS microplate assay method was established and applied for the evaluation of genotoxicity of mycotoxins and fungal cultures. Among 20 mycotoxins, the carcinogenic dihydrobisfuranoids such as aflatoxin B1, sterigmatocystin, and versicolorin A were positive in the presence of the activation system. While, the carcinogenic anthraquinones and lactones such as luteoskyrin, rugulosin, ochratoxin A, patulin, and citrinin were negative. The survey on genotoxic fungi revealed that, among 15 fungal isolates Aspergillus versicolor, Emericella acristata, and others were positive. Additional survey on 265 fungal isolates have revealed that various Aspergillus genera such as A. flavus, A. parasiticus, A. ustus, A. nidulans, and others were positive for SOS induction, along with several isolates of Fusarium moniliforme. The chemical analysis revealed that the dihydrobisfuranoids such as aflatoxin B1, and sterigmatocystin were the major genotoxic metabolites of several Aspergillus species. The SOS microplate assay system is a simple and rapid procedure for the mass screening of genotoxic fungi, particularly of the dihydrobisfuranoids-producing strains.

Animals↗

Ecology, metabolite production, and substrate utilization in endophytic fungi.

Endophytic fungi are a taxonomically and ecologically heterogenous group of organisms, mainly belonging to the Ascomycotina and Deuteromycotina. The isolation methods affect the species composition of the endophyte assemblage in a given host. The number of endophyte taxa isolated from a host species is usually large; however, only few, normally host specific species or strains are dominant. Endophyte assemblages are specific at the host species level, but species composition and frequencies are significantly affected by site-specific conditions. Moreover, the relative importance and number of endophytic species vary among individuals within sites. In some cases, each individual could be considered a separate ecosystem. In general, however, 40 individuals with 30 to 40 sampling units per organ and individual should be enough to detect 80% of taxa present in a given host at one site. Endophytes usually produce the enzymes necessary for the colonization of plant tissues. Substrate utilization studies and isozyme analysis have demonstrated that most endophytes are able to utilize most plant cell components. The production of growth promoting factors and of metabolites useful in the pharmaceutical and agricultural industry is widespread among endophytic fungi. The usefulness of endophytes in agricultural and pharmaceutical research is briefly discussed.

Ecology↗

Toxicity of cotton phytoalexins to zoopathogenic fungi.

The sesquiterpenoid phytoalexins desoxyhemigossypol, desoxymethoxyhemigossypol, and hemigossypolone formed in cotton (Gossypium hirsutum and G. barbadense) stem xylem infected with Verticillium dahliae were shown to be highly toxic to zoopathogenic fungi. This appears to be the first study of the toxicity of terpenoid phytoalexins to zoopathogenic fungi. The toxicities of the phytoalexins expressed as MIC (micrograms ml-1) values were 8 to 128 against four isolates of Candida albicans and one isolate of Cryptococcus neoformans. These highly toxic compounds or their derivatives may prove useful for the treatment of animal mycoses.

Antifungal Agents↗

Laboratory evaluation of temperature effects on the germination and growth of entomopathogenic fungi and on their pathogenicity to two aphid species.

As part of an approach to select potential mycoinsecticides for aphid biocontrol, we investigated the effects of temperature on the growth, germination and pathogenicity of some hyphomycete fungi. Commercially available mycoinsecticides (based on Beauveria bassiana (Balsamo) Vuillemin and Verticillium lecanii (Zimmermann) Viegas) and other isolates of B bassiana, V lecanii, Metarhizium anisopliae (Metschnikoff) Sorokin and Paecilomyces fumosoroseus (Wize) Brown & Smith were evaluated. The rate of in vitro conidial germination of all isolates was slower at 10 and 15 degrees C than at 20 and 25 degrees C. Similarly, in vitro growth of most isolates was adversely affected at 10 and 15 degrees C. The greatest reduction at 10 degrees C in rates of conidial germination and colony growth, compared with other temperatures, was for M anisopliae isolates. Germination of V lecanii (isolate HRI 1.72) was fastest at 10 degrees C compared with the other fungi. It was also the most pathogenic of three isolates tested against Aphis fabae Scopoli and Myzus persicae Sulzer at 10, 18 and 23 degrees C. Generally, A fabae was more susceptible than M persicae to infection by the fungal isolates tested. A significant interaction between aphid species and temperature indicated that the pathogenic nature of an isolate was dependent not only on the target aphid species but also the temperature conditions of the bioassay. The series of studies, detailed above, allowed a temperature profile to be formed for the different isolates. Verticillium lecanii isolate HRI 1.72 (commercialised as Vertalec) was the most promising isolate selected from results of the series of experiments. Temperature profiles in conjunction with infectivity assays can be useful in selecting appropriate isolates for a particular thermal environment.

Animals↗

United States Department of Agriculture-Agricultural Research Service research on pre-harvest prevention of mycotoxins and mycotoxigenic fungi in US crops.

Mycotoxins (ie toxins produced by molds) are fungal metabolites that can contaminate foods and feeds and cause toxic effects in higher organisms that consume the contaminated commodities. Therefore, mycotoxin contamination of foods and feeds results is a serious food safety issue and affects the competitiveness of US agriculture in both domestic and export markets. This article highlights research accomplished by Agricultural Research Service (ARS) laboratories on control of pre-harvest toxin contamination by using biocontrol, host-plant resistance enhancement and integrated management systems. Emphasis is placed on the most economically relevant mycotoxins, namely aflatoxins produced by Aspergillus flavus, Link, trichothecenes produced by various Fusarium spp and fumonisins produced by F verticillioides. Significant inroads have been made in establishing various control strategies such as development of atoxigenic biocontrol fungi that can outcompete their closely related, toxigenic cousins in field environments, thus reducing levels of mycotoxins in the crops. Potential biochemical and genetic resistance markers have been identified in crops, particularly in corn, which are being utilized as selectable markers in breeding for resistance to aflatoxin contamination. Prototypes of genetically engineered crops have been developed which: (1) contain genes for resistance to the phytotoxic effects of certain trichothecenes, thereby helping reduce fungal virulence, or (2) contain genes encoding fungal growth inhibitors for reducing fungal infection. Gene clusters housing the genes governing formation of trichothecenes, fumonisins and aflatoxins have been elucidated and are being targeted in strategies to interrupt the biosynthesis of these mycotoxins. Ultimately, a combination of strategies using biocompetitive fungi and enhancement of host-plant resistance may be needed to adequately prevent mycotoxin contamination in the field. To achieve this, plants may be developed that resist fungal infection and/or reduce the toxic effects of the mycotoxins themselves, or interrupt mycotoxin biosynthesis. This research effort could potentially save affected agricultural industries hundreds of millions of dollars during years of serious mycotoxin outbreaks.

Aspergillus↗

Field-emission scanning electron microscopy of the internal cellular organization of fungi.

Internal viewing of the cellular organization of hyphae by scanning electron microscopy is an alternative to observing sectioned fungal material with a transmission electron microscope. To study cytoplasmic organelles in the hyphal cells of fungi by SEM, colonies were chemically fixed with glutaraldehyde and osmium tetroxide and then immersed in dimethyl sulfoxide. Following this procedure, the colonies were frozen and fractured on a liquid nitrogen-precooled metal block. Next, the fractured samples were macerated in diluted osmium tetroxide to remove the cytoplasmic matrix and subsequently dehydrated by freeze substitution in methanol. After critical point drying, mounting, and sputter coating, fractured cells of several basidiomycetes were imaged with field-emission SEM. This procedure produced clear images of elongated and spherical mitochondria, the nucleus, intravacuolar structures, tubular- and plate-like endoplasmic reticulum, and different types of septal pore caps. This method is a powerful approach for studying the intracellular ultrastructure of fungi by SEM.

Fungi↗

Fungi use efficient algorithms for the exploration of microfluidic networks.

Fungi, in particular, basidiomycetous fungi, are very successful in colonizing microconfined mazelike networks (for example, soil, wood, leaf litter, plant and animal tissues), a fact suggesting that they may be efficient solving agents of geometrical problems. We therefore evaluated the growth behavior and optimality of fungal space-searching algorithms in microfluidic mazes and networks. First, we found that fungal growth behavior was indeed strongly modulated by the geometry of microconfinement. Second, the fungus used a complex growth and space-searching strategy comprising two algorithmic subsets: 1) long-range directional memory of individual hyphae and 2) inducement of branching by physical obstruction. Third, stochastic simulations using experimentally measured parameters showed that this strategy maximizes both survival and biomass homogeneity in microconfined networks and produces optimal results only when both algorithms are synergistically used. This study suggests that even simple microorganisms have developed adequate strategies to solve nontrivial geometrical problems.

Algorithms↗

Alkaline protease inhibitor: a novel class of antifungal proteins against phytopathogenic fungi.

A Streptomyces sp., which produces an alkaline protease inhibitor (API) exhibiting antifungal activity has been isolated from soil. The protein has been purified to homogeneity. The molecular characterization has revealed that it is a dimer (M(r) 28 kDa) with five disulphide linkages and has a pI of 3.8. API is a competitive type of inhibitor with a K(i) value of 2.5 x 10(-9) M. The inhibitor is stable over a pH range of 6 to 12 and a temperature range of 40 to 95 degrees C. API exhibits antifungal activity (in vitro) against phytopathogenic fungi such as Fusarium, Alternaria, and Rhizoctonia and also against Trichoderma, a saprophytic fungus. The antifungal activity of API appears to be associated with its ability to inhibit the fungal serine alkaline protease(s), which is indispensable for its growth. Retardation of the rate of fungal spore germination, as well as hyphal extention, was observed in the presence of API. Both the protease inhibitory and the antifungal activity were abolished on treatment of API with DTT (5 mM), suggestive of a common site for both the activities. This is the first report on API as a potential biocontrol agent against phytopathogenic fungi.

Alternaria↗

Possible side effects of airborne pesticides on fungi and vascular plants in The Netherlands.

A survey of the potential side effects of volatilized pesticides on fungi and vascular plants in The Netherlands has been made for two herbicides (atrazine and MCPA), a fungicide (captan) and a soil fumigant (metham sodium). By applying existing models, a prediction is presented of post-treatment dispersal and deposition due to volatilization, both at short and long ranges, and of the potential side effects on fungi and vascular plants. The general conclusion is that side effects (stunting and growth anomalies) are to be expected outside the target area, especially close (< 500 m) to treated plots with atrazine and metham sodium. In view of agricultural acreage in The Netherlands, these short-range effects may impinge on a relatively large scale. There may also be an impact at greater distances from treated plots, especially in the case of compounds that are slow to degrade such as MITC (methyl isothiocyanate), the active toxic moiety generated in soil from metham sodium.

2-Methyl-4-chlorophenoxyacetic Acid↗

Base composition of DNA from glomalean fungi: high amounts of methylated cytosine.

Glomales (Zygomycetes) are obligate fungal symbionts of roots of land plants and form arbuscular mycorrhiza. Sporal DNA of 10 isolates belonging to nine species was purified and the base composition was determined by RP-HPLC. Base composition fell in a narrow range between 30 and 35% G + C. A high amount of methylated cytosine (mC) accounting for 2-4% of the total nucleotides was found in all taxa. The DNA melting profile was defined for Scutellospora castanea. It corresponded to 32% G + C, and the shape of the denaturation curve suggested a heterogeneity in the GC content within the fungal genome. Knowledge of GC contents and variations between taxa are essential for evaluating nuclear DNA content using fluorimetric methods, and high proportions of mC/C + mC in the genomes of glomalean fungi could reflect the existence of repeated DNA families. Results are discussed in relation to data for other fungi and eukaryotes.

5-Methylcytosine↗

Cytochrome P450 enzyme systems in fungi.

The involvement of cytochrome P450 enzymes in many complex fungal bioconversion processes has been characterized in recent years. Accordingly, there is now considerable scientific interest in fungal cytochrome P450 enzyme systems. In contrast to S. cerevisiae, where surprisingly few P450 genes have been identified, biochemical data suggest that many fungi possess numerous P450 genes. This review summarizes the current information pertaining to these fungal cytochrome P450 systems, with emphasis on the molecular genetics. The use of molecular techniques to improve cytochrome P450 activities in fungi is also discussed.

Cytochrome P-450 Enzyme System↗

Serine/threonine protein kinases and phosphatases in filamentious fungi.

Protein phosphorylation and dephosphorylation are one of the central currencies by which living cells perceive and respond to environmental cues. A number of fundamental processes in fungi such as the cell cycle, transcription, and mating have been shown to require protein phosphorylation. The analysis of protein kinases and phosphatases in filamentous fungi is in its infancy; however, it has already become clear that kinases and phosphatases are likely to be important mediators of fungal proliferation and development as well as signal transduction and infection-related morphogenesis. In this review, we describe, summarize, and consider the rapidly expanding field of protein phosphorylation/dephosphorylation in various aspects of filamentous fungal growth and development.

Fungi↗

Mitosis in filamentous fungi: how we got where we are.

This review traces the principal advances in the study of mitosis in filamentous fungi from its beginnings near the end of the 19(th) century to the present day. Meiosis and mitosis had been accurately described and illustrated by the second decade of the present century and were known to closely resemble nuclear divisions in higher eukaryotes. This information was effectively lost in the mid-1950s, and the essential features of mitosis were then rediscovered from about the mid-1960s to the mid-1970s. Interest in the forces that separate chromatids and spindle poles during fungal mitosis followed closely on the heels of detailed descriptions of the mitotic apparatus in vivo and ultrastructurally during this and the following decade. About the same time, fundamental studies of the structure of fungal chromatin and biochemical characterization of fungal tubulin were being carried out. These cytological and biochemical studies set the stage for a surge of renewed interest in fungal mitosis that was issued in by the age of molecular biology. Filamentous fungi have provided model studies of the cytology and genetics of mitosis, including important advances in the study of mitotic forces, microtubule-associated motor proteins, and mitotic regulatory mechanisms.

Fungi↗

Development of a fungus-specific PCR assay for detecting low-level fungi in an indoor environment.

A fungus-specific PCR assay using only one primer set has been developed for detecting indoor fungi. Four fungal primer sets, NS3/NS4, NS5/NS6, FF1/FR1 and FF2/FR1, were tested with DNA from humans, rats, mice, bacteria, pollens and six commonly found fungal species (Alternaria chamydospora, Aspergillus flavus, Candida famata, Cladosporium fermentans, Penicillium chrycoIgenum and Stachybotrys chartarum). Results indicated that, although all four primer sets could amplify the fungal DNA, only FF2/FR1 demonstrated no cross-amplification with non-fungal DNA. In addition, these amplified fragments were sequenced to ensure that they indeed matched known fungal DNA sequences. Furthermore, besides the tested fungi, eighteen more genera of fungal sequences were examined and found to match the FF2/FR1. Here, the method of bead-beating was identified as the most effective way for spore breakage and fungal DNA release. The PCR amplification efficiency and potential inhibition were examined using different process solutions and preparation procedures. It was found that, when using 20% nutrient media and homogenization-first procedure, a higher amplification efficiency with less inhibition was achieved. Although positive bands were observed at 0.2 fungal spore/reaction using the homogenization-first procedure, the sensitivity of this assay would be two fungal spores/reaction for environmental samples.

Air Pollution, Indoor↗