Methods for isolation and cultivation of filamentous fungi

Filamentous fungi are important organisms for basic discovery, industry, and human health. Their natural growth environments are extremely variable, a fact reflected by the numerous methods developed for their isolation and cultivation. Fungal culture in the laboratory is usually carried out on agar plates, shake flasks, and bench top fermenters starting with an inoculum that typically features fungal spores. Here we discuss the most popular methods for the isolation and cultivation of filamentous fungi for various purposes with the emphasis on enzyme production and molecular microbiology.

Highly effective fungal inactivation in He+O2 atmospheric-pressure nonequilibrium plasmas

Highly effective (more than 99.9%) inactivation of a pathogenic fungus Candida albicans commonly found in oral, respiratory, digestive, and reproduction systems of a human body using atmospheric-pressure plasma jets sustained in He+ O2 gas mixtures is reported. The inactivation is demonstrated in two fungal culture configurations with open (Petri dish without a cover) and restricted access to the atmosphere (Petri dish with a cover) under specific experimental conditions. It is shown that the fungal inactivation is remarkably more effective in the second configuration. This observation is supported by the scanning and transmission electron microscopy of the fungi before and after the plasma treatment. The inactivation mechanism explains the experimental observations under different experimental conditions and is consistent with the reports by other authors. The results are promising for the development of advanced health care applications.

Molecular characterization of the EhaG and UpaG trimeric autotransporter proteins from pathogenic Escherichia coli

Trimeric autotransporter proteins (TAAs) are important virulence factors of many Gram-negative bacterial pathogens. A common feature of most TAAs is the ability to mediate adherence to eukaryotic cells or extracellular matrix (ECM) proteins via a cell surface-exposed passenger domain. Here we describe the characterization of EhaG, a TAA identified from enterohemorrhagic Escherichia coli (EHEC) O157:H7. EhaG is a positional orthologue of the recently characterized UpaG TAA from uropathogenic E. coli (UPEC). Similarly to UpaG, EhaG localized at the bacterial cell surface and promoted cell aggregation, biofilm formation, and adherence to a range of ECM proteins. However, the two orthologues display differential cellular binding: EhaG mediates specific adhesion to colorectal epithelial cells while UpaG promotes specific binding to bladder epithelial cells. The EhaG and UpaG TAAs contain extensive sequence divergence in their respective passenger domains that could account for these differences. Indeed, sequence analyses of UpaG and EhaG homologues from several E. coli genomes revealed grouping of the proteins in clades almost exclusively represented by distinct E. coli pathotypes. The expression of EhaG (in EHEC) and UpaG (in UPEC) was also investigated and shown to be significantly enhanced in an hns isogenic mutant, suggesting that H-NS acts as a negative regulator of both TAAs. Thus, while the EhaG and UpaG TAAs contain some conserved binding and regulatory features, they also possess important differences that correlate with the distinct pathogenic lifestyles of EHEC and UPEC.

Biology Aotearoa : Unique Flora, Fauna and Fungi

As a large, isolated and relatively ancient landmass, New Zealand occupies a unique place in the biological world, with distinctive terrestrial biota and a high proportion of primitive endemic forms. Biology Aotearoa covers the origins, evolution and conservation of the New Zealand flora, fauna and fungi. Each chapter is written by specialists in the field, often working from different perspectives to build up a comprehensive picture. Topics include: the geological history of our land origins, and evolution of our plants, animals and fungi current status of rare and threatened species past, present and future management of native species the effect of human immigration on the native biota. Colour diagrams and photographs are used throughout the text. This book is suitable for all students of biology or ecology who wish to know about the unique nature of Aotearoa New Zealand and its context in the biological world.

Airborne viable fungi in school environments in different climatic regions – A review

Elevated levels of fungi in indoor environments have been linked with mould/moisture damage in building structures. However, there is a lack of information about “normal” concentrations and flora as well as guidelines of viable fungi in the school environment in different climatic conditions. We have reviewed existing guidelines for indoor fungi and the current knowledge of the concentrations and flora of viable fungi in different climatic areas, the impact of the local factors on concentrations and flora of viable fungi in school environments. Meta-regression was performed to estimate the average behaviour for each analysis of interest, showing wide variation in the mean concentrations in outdoor and indoor school environments (range: 101-103 cfu/m3). These concentrations were significantly higher for both outdoors and indoors in the moderate than in the continental climatic area, showing that the climatic condition was a determinant for the concentrations of airborne viable fungi. The most common fungal species both in the moderate and continental area were Cladosporium spp. and Penicillium spp. The suggested few quantitative guidelines for indoor air viable fungi for school buildings are much lower than for residential areas. This review provides a synthesis, which can be used to guide the interpretation of the fungi measurements results and help to find indications of mould/moisture in school building structures.

Airborne culturable fungi in naturally ventilated primary school environments in a subtropical climate

There is currently a lack of reference values for indoor air fungal concentrations to allow for the interpretation of measurement results in subtropical school settings. Analysis of the results of this work established that, in the majority of properly maintained subtropical school buildings, without any major affecting events such as floods or visible mould or moisture contamination, indoor culturable fungi levels were driven by outdoor concentration. The results also allowed us to benchmark the “baseline range” concentrations for total culturable fungi, Penicillium spp., Cladosporium spp. and Aspergillus spp. in such school settings. The measured concentration of total culturable fungi and three individual fungal genera were estimated using Bayesian hierarchical modelling. Pooling of these estimates provided a predictive distribution for concentrations at an unobserved school. The results indicated that “baseline” indoor concentration levels for indoor total fungi, Penicillium spp., Cladosporium spp. and Aspergillus spp. in such school settings were generally ≤ 1450, ≤ 680, ≤ 480 and ≤ 90 cfu/m3, respectively, and elevated levels would indicate mould damage in building structures. The indoor/outdoor ratio for most classrooms had 95% credible intervals containing 1, indicating that fungi concentrations are generally the same indoors and outdoors at each school. Bayesian fixed effects regression modeling showed that increasing both temperature and humidity resulted in higher levels of fungi concentration.

Pathogens penetrating the central nervous system: Infection pathways and the cellular and molecular mechanisms of invasion

The brain is well protected against microbial invasion by cellular barriers, such as the blood-brain barrier (BBB) and the blood-cerebrospinal fluid barrier (BCSFB). In addition, cells within the central nervous system (CNS) are capable of producing an immune response against invading pathogens. Nonetheless, a range of pathogenic microbes make their way to the CNS, and the resulting infections can cause significant morbidity and mortality. Bacteria, amoebae, fungi, and viruses are capable of CNS invasion, with the latter using axonal transport as a common route of infection. In this review, we compare the mechanisms by which bacterial pathogens reach the CNS and infect the brain. In particular, we focus on recent data regarding mechanisms of bacterial translocation from the nasal mucosa to the brain, which represents a little explored pathway of bacterial invasion but has been proposed as being particularly important in explaining how infection with Burkholderia pseudomallei can result in melioidosis encephalomyelitis.

Further new smut fungi (Ustilaginomycetes) from Australia

Nine new species of smut fungi, belonging to eight genera, are described from Australia: Dermatosorus schoenoplecti Vánky & R.G. Shivas, on Schoenoplectus mucronatus, Entyloma grampiansis Vánky & R.G. Shivas, on Hydrocotyle laxiflora, Macalpinomyces brachiariae Vánky, C. Vánky & R.G. Shivas, on Brachiaria holosericea, M. digitariae Vánky & R.G. Shivas, on Digitaria gibbosa, Restiosporium baloskionis Vánky & R.G. Shivas, on Baloskion tetraphyllum, Thecaphora maireanae R.G. Shivas & Vánky, on Maireana pentagona, Tilletia cape yorkensis Vánky & R.G. Shivas, on Whiteochloa airoides, Urocystis chorizandrae J. Cunnington, R.G. Shivas & Vánky, on Chorizandra enodis, and Ustanciosporium tenellum R.G . Shivas & Vánky, on Cyperus tenellus. New combinations are: Macalpinomyces ordensis(R.G. Shivas & Vánky) Vánky & R.G. Shivas (based on Sporisorium ordense, type on Brachiaria pubigera, Australia), and Sporisorium setariae (McAlpine) Vánky & R.G. Shivas (based on Sorosporium setariae, type on Setaria glauca, Australia).

Biodiversity of Australian smut fungi.

There are about 250 species of smut fungi known from Australia of which 95 are endemic. Fourteen of these endemic species were first collected in the period culminating with the publication of Daniel McAlpine's revision of Australian smut fungi in 1910. Of the 68 species treated by McAlpine, 10 were considered to be endemic to Australia at that time. Only 23 of the species treated by McAlpine have names that are currently accepted . During the following eighty years until 1990, a further 31 endemic species were collected and just 11 of these were named and described in that period. Since 1990, 50 further species of endemic smut fungi have been collected and named in Australia . There are 115 species that are restricted to either Australia or to Australia and the neighbouring countries of Indonesia, New Zealand, Papua New Guinea and the Philippines . These 115 endemic species occur in 24 genera, namely Anthracoidea (1 species), Bauerago (1), Cintractia (3), Dermatosorus (1), Entyloma (3), Farysporium (1), Fulvisporium (1), Heterotolyposporium (1), Lundquistia (1), Macalpinomyces (4), Microbotryum (2), Moreaua (20), Pseudotracya (1), Restiosporium (5), Sporisorium (26), Thecaphora (2), Tilletia (12), Tolyposporella (1), Tranzscheliella (1), Urocystis (2), Ustanciosporium (1), Ustilago (22), Websdanea (1) and Yelsemia (2). About a half of these local and regional endemic species occur on grasses and a quarter on sedges . The northern tropical savannah region of Australia offers most promise for the discovery of new endemic species . The agricultural, quarantine and environmental significance to Australia of some introduced species is discussed.

Identification and characterization of non-pathogenic Fusarium oxysporum capable of increasing and decreasing Fusarium wilt severity

Fusarium wilt of banana is a potentially devastating disease throughout the world. Options for control of the causal organism, Fusarium oxysporum f.sp. cubense (Foc) are limited. Suppressive soil sites have previously been identified where, despite the presence of Foc, Fusarium wilt does not develop. In order to understand some aspects of this disease suppression, endophytic Fusarium oxysporum isolates were obtained from banana roots. These isolates were genetically characterized and compared with an isolate of Fusarium oxysporum previously identified as being capable of suppressing Fusarium wilt of banana in glasshouse trials. Three additional isolates were selected for glasshouse trials to assess suppression of Fusarium wilt in two different cultivars of banana, Cavendish and Lady Finger. One isolate (BRIP 29089) was identified as a potential biocontrol organism, reducing the disease severity of Fusarium wilt in Lady Finger and Cavendish cultivars. Interestingly, one isolate (BRIP 45952) increased Fusarium wilt disease severity on Cavendish. The implications of an isolate of Fusarium oxysporum, non-pathogenic on banana, increasing disease severity and the potential role of non-pathogenic isolates of Fusarium oxysporum in disease complexes are discussed.

An annotated checklist of smut fungi (Ustilaginomycetes) from Thailand

An annotated checklist of the smut fungi (Ustilaginomycetes) from Thailand has been compiled after an examination of the scientific literature, previously deposited herbarium specimens and specimens collected by the authors during a survey in December 2005. Fifty-two species of smut fungi are listed, including 11 species which were newly discovered during our survey. Most of these smut fungi are reported for the first time from Thailand. Several species are very rare, being known only from the type material or a limited number of collections.

Six new smut fungi (Ustilaginomycotina) from Australia

Six new smut fungi, Sporisorium rarum (type on Eulalia aurea), S. vermiculum (type on Sarga plumosa), S. xerofasciculatum (type on Xerochloa laniflora), Tilletia xerochloae (type on Xerochloa laniflora), T. yakirrae (type on Yakirra majuscula) and Ustilago lunata (type on Triodia longiceps), are described and illustrated from central and western Australia. Keys are provided for the smut fungi on Sarga and Xerochloa.

Is organic acid required for nutrition of thermophilic fungi?

In contrast to a published report [Wali et al. Arch Microbiol 118:49–53 (1978)], an organic acid is not essential for the growth of thermophilic fungi. The thermophilic fungus, Thermomyces lanuginosus, grows satisfactorily in a synthetic medium containing glucose as carbon source if the pH of the medium is controlled. The control of pH is essential for the concentration of carbon dioxide in the growth medium and the activity of anaplerotic enzyme, pyruvate carboxylase.

Mycotoxins and toxigenic fungi in sago starch from Papua New Guinea

Aims: To assay sago starch from Papua New Guinea (PNG) for important mycotoxins and to test fungal isolates from sago for mycotoxin production in culture. Methods and Results: Sago starch collected from Western and East Sepik Provinces was assayed for aflatoxins, ochratoxin A, cyclopiazonic acid, sterigmatocystin, citrinin and zearalenone and all 51 samples were negative. Frequently isolated species of Penicillium (13), Aspergillus (five) and Fusarium (one) were cultured on wheat grain, and tested for the production of ochratoxin A, cyclopiazonic acid, sterigmatocystin, citrinin, patulin and penicillic acid. All 12 isolates of P. citrinin and one of two A. flavipes isolates produced citrinin. A single isolate of A. versicolor produced sterigmatocystin. No other mycotoxins were detected in these cultures. Conclusions: No evidence was found of systemic mycotoxin contamination of sago starch. However, the isolation of several mycotoxigenic fungi shows the potential for citrinin and other mycotoxins to be produced in sago stored under special conditions. Significance and Impact of the study: Sago starch is the staple carbohydrate in lowland PNG and the absence of mycotoxins in freshly prepared sago starch is a positive finding. However, the frequent isolation of citrinin-producing fungi indicates a potential health risk for sago consumers, and food safety is dependant on promoting good storage practices.