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Saprophytic and cycloheximide resistant fungi isolated from golden hamster.

Healthy hair samples from golden hamsters were examined for the presence of dermatophytes and non-dermatophytes using baiting technique and direct inoculation. Thirty-four species and 2 varieties attributed to 17 genera were recovered. Paecilomyces variotii (isolated from 84.4% of the examined hair) and Aspergillus niger (81.3%) were the more frequent isolates on Sabouraud's dextrose agar (SDA) without cycloheximide. Our results have clearly demonstrated that the hair of hamster was free from true dermatophytes. Using the dilution plate method many fungal species were isolated from cage material (7 genera and 10 species + 1 variety); from faeces (10 genera and 17 species); from standard chow (3 genera and 6 species) of hamster. P. variotii which was the most frequent fungus in the preceding 3 substrates was completely absent in the presence of cycloheximide in SDA. The present study has demonstrated for the first time the isolation of Trichophyton rubrum from hamster faeces. Also, several saprophytic and cycloheximide resistant fungi were isolated. In the air of hamster cage Cladosporium cladosporioides, Penicillium chrysogenum, Alternaria alternata and Scopulariopsis brevicaulis were the most dominant species on SDA with or without cycloheximide. Using the agar diffusion method, Aloe sap, onion oil, garlic bulb extract and aqueous leaf extracts of Andropogon citratus, Euphorbia sp. and Ruta graveolens were tested for their antifungal activity on 10 fungal species. It was observed that onion oil exhibited a high inhibitory effect against most of the tested fungi.

Animal Feed↗

Acute pulmonary hemorrhage in infants associated with exposure to Stachybotrys atra and other fungi.

BACKGROUND: A geographic cluster of 10 cases of pulmonary hemorrhage and hemosiderosis in infants occurred in Cleveland, Ohio, between January 1993 and December 1994. STUDY DESIGN: This community-based case-control study tested the hypothesis that the 10 infants with pulmonary hemorrhage and hemosiderosis were more likely to live in homes where Stachybotrys atra was present than were 30 age- and ZIP code-matched control infants. We investigated the infants' home environments using bioaerosol sampling methods, with specific attention to S atra. Air and surface samples were collected from the room where the infant was reported to have spent the most time. RESULTS: Mean colony counts for all fungi averaged 29 227 colony-forming units (CFU)/m3 in homes of patients and 707 CFU/m3 in homes of controls. The mean concentration of S atra in the air was 43 CFU/m3 in homes of patients and 4 CFU/m3 in homes of controls. Viable S atra was detected in filter cassette samples of the air in the homes of 5 of 9 patients and 4 of 27 controls. The matched odds ratio for a change of 10 units in the mean concentration of S atra in the air was 9.83 (95% confidence interval, 1.08-3 X 10(6)). The mean concentration of S atra on surfaces was 20 X 10(6) CFU/g and 0.007 x 10(6) CFU/g in homes of patients and controls, respectively. CONCLUSION: Infants with pulmonary hemorrhage and hemosiderosis were more likely than controls to live in homes with toxigenic S atra and other fungi in the indoor air.

Acute Disease↗

Lignocellulolysis by ascomycetes (fungi) of a saltmarsh grass (smooth cordgrass).

Lignocellulose (LC) makes up greater than 70% of the mature shoots of the prodigiously photosynthetically productive saltmarsh grass Spartina alterniflora. Naturally decaying shoots of this cordgrass were examined by transmission electron microscopy (after high-pressure freezing and freeze-substitution) as a means of directly detecting lysis of the LC-rich tissues. Portions of the cordgrass were selected that contained ascomata (sexual reproductive structures) of only one of each of four species of fungi (Kingdom Fungi; Subdivision Ascomycotina): Phaeosphaeria spartinicola and Buergenerula spartinae from leaf blades, Phaeosphaeria spartinae from leaf sheaths, and Passeriniella obiones from naked stems. All four of the ascomycetes were LC-lytic. Phaeosphaeria spartinicola caused both thinning of LC-rich secondary walls of fiber cells from cell lumina outwards (type 2 soft rot, akin to white rot) and digestion extending from hyphae within longitudinal cavities in the secondary walls (type 1 soft rot). The other three species caused either one or the other type of soft rot. Bacterial erosion of cordgrass cells was found only in the samples of naked stems. Ascomycetous decomposers of standing-dead grasses may have potential for biotechnological applications involving alterations of lignocellulose or toxic polyphenolic substances.

Ascomycota↗

Toxigenic fungi in a water-damaged building: an intervention study.

In an investigation of health complaints among employees of a water-damaged office building, the environment showed evidence of fungal contamination with the isolation of Stachybotrys chartarum in one of five bulk samples tested for fungal growth. In response, a public health official recommended that employees be relocated from the building. Employees were subsequently moved to a different environment. A focused environmental investigation of microbial growth within the building followed, revealing moderate to high levels of fungi (Penicillium, Aspergillus versicolor) and bacteria in bulk and surface samples. S. Chartarum was identified in one of 19 (5%) environmental samples using Czapek agar. A health survey of building occupants revealed a high prevalence of multiple symptoms, with the predominance of neurobehavioral and upper respiratory tract complaints. The majority of symptoms were significantly less prevalent after relocation from the water-damaged environment. The initial hypothesis that exposure to toxigenic fungi was responsible for the high prevalence of reported symptoms is difficult to investigate and confirm given the current limits of epidemiological knowledge regarding exposure to these organisms and building-related illness. Future interventions where mycotoxin exposure is suspected should emphasize the importance of risk assessment and risk communication.

Adult↗

Identification of hallucinogenic fungi from the genera Psilocybe and Panaeolus by amplified fragment length polymorphism.

Unambiguous identification of the hallucinogenic fungi of the genera Psilocybe and Panaeolus is required by national and international drug control legislation. We report on a DNA-based test using the technique of amplified fragment length polymorphism (AFLP). AFLP can differentiate species of the two genera Psilocybe and Panaeolus by using different primer sets. The identification of hallucinogenic fungi using a DNA-based test, which can be used in conjunction with morphological features, will assist in forensic investigations.

Agaricales↗

Effects of carbohydrates, ethanol and selected cell wall phenolics on in vitro growth of necrotrophic fungi--Heterobasidion annosum and Fusarium avenaceum.

In this study, a comparative assessment was undertaken of nutritional factors that are likely to influence in vivo growth of the S and P-types of necrotrophic root rot fungus Heterobasidion annosum and of the damping off pathogen Fusarium avenaceum respectively. The results indicate that glucose and fructose but not cellobiose or sucrose supported growth of these fungi when used as sole carbon source. Particularly for P-types and F. avenaceum, however, the utilization of cellobiose and sucrose was considerably enhanced by several fold in the presence of ethanol. In addition, key plant cell wall phenolics (ferulic acid, caffeic acid, p-coumaric acid) at 0.025% inhibited growth of the test fungi. But this toxicity was reversed in the presence of exogenously added cell wall sugars (glucose, fructose, sucrose, cellobiose) except for the P-types where sucrose addition was apparently slightly effective. Similarly, ethanol whether alone or in combination with the phenolics was poorly utilized except for F. avenaceum. The importance of these results in understanding factors influencing growth and development during pathogenesis of the necrotrophs on their conifer hosts is discussed.

Basidiomycota↗

Competition strategies for the decolorization of a textile-reactive dye with the white-rot fungi Trametes versicolor under non-sterile conditions.

A variety of white-rot fungi can oxidize textile dyes under sterile conditions; however, an important consideration for their use in treating wastewater containing textile dyes is whether similar degrees of treatment can be achieved under non-sterile conditions. Four strategies were investigated for their potential in optimizing the use of the fungus Trametes versicolor in non-sterile culture for treating wastewater containing the diazo textile dye C.I. Reactive Black 5 (RB5). Three strategies with suspended culture were designed to increase the decolorization activity in suspended culture from a given amount of T. versicolor inoculum based on its tolerance of low pH (pH reduction in medium), production of extracellular enzymes (use of suspended enzymes alone), and its ability to produce enzymes independent of growth (nitrogen limitation in medium). The results showed that reduction of the medium pH to 3 did not suppress bacterial growth, while enzyme production by T. versicolor ceased. The use of the extracellular enzymes alone would allow the decoupling of the process of fungal growth from wastewater treatment; however, the enzyme activity of an enzyme suspension decreased rapidly under non-sterile conditions. The strategy of limiting nitrogen in the medium to suppress bacterial growth has potential together with the fourth strategy, the cultivation of fungi on organic solids to produce inocula for a decolorization process under non-sterile conditions. A high degree of decolorization of RB5 under non-sterile conditions was achieved with T. versicolor grown on grains as sole substrate. The rate of decolorization was dependent on the amount of fungal inoculum used.

Basidiomycota↗

Degradation of aromatic hydrocarbons by white-rot fungi in a historically contaminated soil.

Phanerochaete chrysosporium NRRL 6361 and Pleurotus pulmonarius CBS 664.97 were tested for their ability to grow under nonsterile conditions and to degrade various aromatic hydrocarbons in an aged contaminated soil that also contained high concentrations of heavy metals. After 24 days fungal incubation, carbon-CO2 liberated, an indicator of microbial activity, reached a plateau. At the end of the incubation time (30 days), fungal colonization was clearly visible and was confirmed by ergosterol and cell organic carbon determinations. In spite of unfavorable pH (around 7.4) and the presence of heavy metals, both fungi produced Mn-peroxidase activity. In contrast, laccase and aryl-alcohol oxidase were detected only in the soil treated with P. pulmonarius CBS 664.97 and lignin-peroxidase in that with P. chrysosporium NRRL 6361. No lignin-modifying enzyme activities were present in non-inoculated soil incubated for 30 days (control microcosm). Regardless of the fungus employed, a total removal of naphtalene, tetrachlorobenzene, and dichloroaniline isomers, diphenylether and N-phenyl-1-naphtalenamine, was observed. Significant release of chloride ions was also observed in fungal-treated soil, in comparison with that recorded in the control microcosm. Both fungi led to a significant decrease in soil toxicity, as assessed using two different soil contact assays, including the Lepidium sativum L. germination test and the Collembola mortality test.

Biodegradation, Environmental↗

Growth and production of laccases by the ligninolytic fungi, Pleurotus ostreatus and Botryosphaeria rhodina , cultured on basal medium containing the herbicide, Scepter (imazaquin).

The herbicide, Scepter, whose active principle is imazaquin, is commonly used in soybean farming to combat wide-leaf weeds. The basidiomycete, Pleurotus ostreatus , and the ascomycete, Botryosphaeria rhodina , were evaluated for their growth and laccase production when cultured on basal media containing Scepter. Both fungi could grow on the herbicide when cultivated in solid and submerged liquid culture in the presence of Scepter at concentrations of 0-6% (v/v) for P. ostreatus , and up to 0-50% (v/v) for B. rhodina , and in each case produced laccases when assayed against ABTS [2,2(1)-azino-bis(3-ethylbenzthiazoline-6-sulfonic acid)] and 2,6-dimethoxyphenol. P . ostreatus could tolerate up to 6% of Scepter before it became toxic to the fungus, while in the case of B. rhodina , 50% (v/v) Scepter was the highest amount that supported grow, and laccase activity was detectable up to 25% (v/v). An inverse relationship existed between the level of Scepter in the culture medium that supported fungal growth and laccase production. Analysis of the results showed that the fungi studied presented different behaviour towards Scepter in the culture environment.

Ascomycota↗

Effect of phenol on the mycelial growth and fructification in some of basidiomycetous fungi.

Cometabolic growth studies with phenol were undertaken to screen 32 strains of white and brown rot fungi. All the cultures studied grew well up to 4 mM of phenol on Czapekdox agar except Agaricus bisporus (white button mushroom) and Pleurotus cystidiosus. Most of them could grow even up to 6 mM of phenol. Phenol induced a brown pigmentation of the culture medium. P. flabellatus and P. pulmonarius metabolized 67 and 64 mg/l phenol in 10 days. Studies have indicated that phenol (0.1 to 1.0 mM) incorporated in malt-extract agar has no inhibitory effect on fruitbody formation. Preliminary studies indicate that soaking of wheat straw with phenol solution up to 1600 mg/l give better mycelial growth and fructification of P. cornucopiae, P. ostreatus Z-15 and Calocybe indica than water soaked. Soaking of wheat straw in phenol inhibited the growth of common competitor weed fungi like Stachybotrys sp. and Coprinus sp.

Basidiomycota↗

Discrimination against 13C during degradation of simple and complex substrates by two white rot fungi.

Changes in isotopic 13C signatures of CO2-C evolved during decomposition of a sugar (glucose), a fatty acid (palmitic acid), a protein (albumin), a structural biopolymer (lignin) and bulk plant tissue (aerial shoots from Lolium perenne) were monitored over a period of 76 days. All materials were sterilized and inoculated with either of two different species of white rot fungi, Phanerochaete chrysosporium or Coriolus versicolor, and incubated in sealed bottles at 28 degrees C. The CO2 concentration in the jars was periodically determined using an infrared gas analyzer and its isotopic (13C) signature was assessed using a trace gas (ANCA TGII) module coupled to an isotope ratio mass spectrometer (IRMS, Europa 20-20). L. perenne material inoculated with C. versicolor showed the highest C mineralization activity with approximately 70% of total C evolved as CO2 after 76 days of incubation, followed by glucose. Substrates inoculated with C. versicolor generally decomposed faster than when degraded by P. chrysosporium, except for lignin, where no significant differences between the two fungi types were found and CO2-C released was less than 2% of the initial C. Considerable 13C isotopic fractionation during the degradation of plant tissue and of pure biochemical compounds was revealed as well as progressive shifts in cumulative CO2-13C isotopic signatures over time. During the first stages of decomposition, the CO2-C released was usually depleted in 13C as compared with the initial solid substrate, but with ongoing decomposition the CO2-C evolved became progressively more enriched in 13C. P. chrysosporium usually showed a slightly higher 13C fractionation than C. versicolor during the first decomposition phase. At posterior decomposition stages isotopic discrimination was often stronger by C. versicolor. These findings on isotopic 13C discrimination during microbial degradation both of simple biochemical compounds and of complex vegetal tissue confirmed not only the existence of significant 13C isotopic fractionation during plant residue decomposition, but also the existence of non-random isotopic distribution within substrates. They also demonstrated the ability of microorganisms to selectively discriminate against 13C even when degrading an isolated simple substrate.

Albumins↗

Extracellular biosynthesis of magnetite using fungi.

The development of synthetic processes for oxide nanomaterials is an issue of considerable topical interest. While a number of chemical methods are available and are extensively used, the collaborations are often energy intensive and employ toxic chemicals. On the other hand, the synthesis of inorganic materials by biological systems is characterized by processes that occur at close to ambient temperatures and pressures, and at neutral pH (examples include magnetotactic bacteria, diatoms, and S-layer bacteria). Here we show that nanoparticulate magnetite may be produced at room temperature extracellularly by challenging the fungi, Fusarium oxysporum and Verticillium sp., with mixtures of ferric and ferrous salts. Extracellular hydrolysis of the anionic iron complexes by cationic proteins secreted by the fungi results in the room-temperature synthesis of crystalline magnetite particles that exhibit a signature of a ferrimagnetic transition with a negligible amount of spontaneous magnetization at low temperature.

Crystallization↗

Organelle transport and molecular motors in fungi

Polarized growth, secretion of exoenzymes, organelle inheritance, and organelle positioning require vectorial transport along cytoskeletal elements. The discovery of molecular motors and intensive studies on their biological function during the past 3 years confirmed a central role of these mechanoenzymes in morphogenesis and development of yeasts and filamentous fungi. Saccharomyces cerevisiae proved to be an excellent model system, in which the complete set of molecular motors is presumed to be known. Genetic studies combined with cell biological methods revealed unexpected functional relationships between these motors and has greatly improved our understanding of nuclear migration, exocytosis, and endocytosis in yeasts. Tip growth of elongated hyphae, compared to budding, however, does require vectorial transport over long distances. The identification of ubiquitous motors that are not present in yeast indicates that studies on filamentous fungi might be helpful to elucidate the role of motors in long-distance organelle transport within higher eukaryotic cells. Copyright 1998 Academic Press.

Journal Article↗

Biodegradation of lignin by white rot fungi.

A review is presented related to the biochemistry of lignocellulose transformation. The biodegradation of wood constituents is currently understood as a multienzymatic process with the mediation of small molecules; therefore, this review will focus on the roles of these small molecular compounds and radicals working in concert with enzymes. Wood rotting basidiomycetous fungi penetrate wood and lead to more easily metabolized, carbohydrate constituents of the complex. Having a versatile machinery of enzymes, the white rot fungi are able to attack directly the "lignin barrier." They also use a multienzyme system including so-called "feed back" type enzymes, allowing for simultaneous transformation of both lignin and cellulose. These enzymes may function separately or cooperatively.

Basidiomycota↗

Purification and characterization of chitinases from the nematophagous fungi Verticillium chlamydosporium and V. suchlasporium.

Culture filtrates of the nematophagous fungi Verticillium chlamydosporium and V. suchlasporium growing on colloidal chitin showed increasing chitinolytic activity and production of two (32- and 43-kDa) main proteins. Maximum activity was found 18-20 days after inoculation, but V. suchlasporium always displayed higher activity. Zymography of such filtrates on carboxymethyl-chitin-Remazol brilliant violet 5R/acrylamide gels showed five bands of substrate degradation for V. suchlasporium and three for V. chlamydosporium. Filtrates with maximum activity were chromatographed on macroporous cross-linked chitin affinity matrix, showing a peak of main (50-60%) activity, which only contained a 43-kDa protein for both fungi. Zymography and colloidal chitin degradation showed that it was a single endochitinase (CHI43) with optimum pH range of 5.2-5.7. The main isoforms had pIs of 7.6 for V. suchlasporium and 7.9 for V. chlamydosporium. Eggs of the nematode Globodera pallida treated with CHI43 and the serine protease P32 from V. suchlasporium alone or in combination showed surface damage in comparison with controls when examined by scanning electron microscopy.

Animals↗

Water and temperature relations of growth of the entomogenous fungi beauveria bassiana, metarhizium anisopliae, and paecilomyces farinosus

The effects of temperature (5-50 degrees C), water availability (0. 998-0.88 water activity, a(w)), and a(w) x temperature interactions (15-45 degrees C) on growth of three entomogenous fungi, Beauveria bassiana, Metarhizium anisopliae, and Paecilomyces farinosus, were evaluated on a Sabouraud dextrose-based medium modified with the ionic solute KCl, the non-ionic solute glycerol, and an inert solute, polyethylene glycol (PEG) 600. The temperature ranges for growth of B. bassiana, M. anisopliae, and P. farinosus were 5-30, 5-40, and 5-30 degrees C, and optimum growth temperatures were 25, 30, and 20 degrees C, respectively. All three species grew over a similar a(w) range (0.90-0.998) at optimum temperatures for growth. However, there were significant interspecies variations in growth rates on media modified with each of the three a(w)-modifying solutes. Growth a(w) optima ranged between 0.99 and 0.97 on KCl-, glycerol-, and PEG 600-modified media for M. anisopliae and P. farinosus. B. bassiana grew optimally at 0.998 a(w), regardless of a(w). Comprehensive two-dimensional profiles of a(w) x temperature relations for growth of these three species were constructed for the first time. The results are discussed in relation to the environmental limits that determine efficacy of entomogenous fungi as biocontrol agents in nature. Copyright 1999 Academic Press.

Journal Article↗

Functional consequences of nutrient translocation in mycelial fungi.

Fungi are of fundamental importance for plant and microbial nutrition with primary roles in decomposition and nutrient recycling. They also have great potential for use in areas of biotechnology such as bioremediation of organic and inorganic pollutants and biocontrol of plant pathogens. In all these contexts, environmental heterogeneity has a strong influence on growth and function. A large class of fungi overcome the difficulties encountered in such environments by the mechanism of translocation which results in the internal redistribution of nutrients within the fungal mycelium. In this paper, we use a combination of experimental techniques and mathematical modelling to examine fungal growth in general, and in particular, translocation in the common soil saprophytic fungus Rhizoctonia solani. A detailed mathematical model is presented where translocation is considered to have both diffusive and metabolically-driven components. A calibration experiment provided the necessary parameter values. Growth experiments were compared with model solutions and thus we provide strong evidence that diffusion is the dominant mechanism for translocation in homogeneous environments. In heterogeneous environments, we conclude that diffusion is still vital for exploration, i.e. the expansion of the fungal network into the surrounding area. However, we also conclude that localized resources may be utilized faster if energy is invested, i.e. when exploitation of the fungal microenvironment is enhanced by metabolically driven translocation.

Food Chain↗

Microwave-stimulated Jones-Marres method for staining fungi in brain tissue of immunocompromised patients.

Life-threatening fungal infections have increased significantly in the past decade due to the rising number of immunocompromised patients. Serological diagnosis of most fungal infections is unreliable and blood cultures are positive in only 50% of premortem cases; therefore, tissue sampling together with fast, reliable staining of fungi should be carried out to reach the correct, timely diagnosis. We developed, partly serendipitously, a microwave silver staining method for fungi in histological sections. During a differentiation step in periodic acid, background staining is removed. This rapid staining method can be combined with immunostaining, for example, the alpha smooth muscle actin method, to visualize blood vessels. Silver staining results were optimized using the recently developed MicroMED BASIC microwave labstation for histology (Milestone srl, Italy), featuring no-touch temperature measurements and PC control.

Aspergillosis↗