Richard C. Summerbell

Fungus-growing ants use antibiotic-producing bacteria to control garden parasites

The well-studied, ancient and highly evolved mutualism between fungus-growing ants and their fungi has become a model system in the study of symbiosis. Although it is thought at present to involve only two symbionts, associated with each other in near isolation from other organisms, the fungal gardens of attine ants are in fact host to a specialized and virulent parasitic fungus of the genus Escovopsis (Ascomycotina). Because the ants and their fungi are mutually dependent, the maintenance of stable fungal monocultures in the presence of weeds or parasites is critical to the survival of both organisms. Here we describe a new, third mutualist in this symbiosis, a filamentous bacterium (actinomycete) of the genus Streptomyces that produces antibiotics specifically targeted to suppress the growth of the specialized garden-parasite Escovopsis. This third mutualist is associated with all species of fungus-growing ants studied, is carried upon regions of the ants cuticle that are genus specific, is transmitted vertically (from parent to offspring colonies), and has the capacity to promote the growth of the fungal mutualist, indicating that the association of Streptomyces with attine ants is both highly evolved and of ancient origin.

Comparative Genome Analysis of Trichophyton rubrum and Related Dermatophytes Reveals Candidate Genes Involved in Infection

ABSTRACT The major cause of athlete’s foot is Trichophyton rubrum, a dermatophyte or fungal pathogen of human skin. To facilitate molecular analyses of the dermatophytes, we sequenced T. rubrum and four related species, Trichophyton tonsurans, Trichophyton equinum, Microsporum canis, and Microsporum gypseum. These species differ in host range, mating, and disease progression. The dermatophyte genomes are highly colinear yet contain gene family expansions not found in other human-associated fungi. Dermatophyte genomes are enriched for gene families containing the LysM domain, which binds chitin and potentially related carbohydrates. These LysM domains differ in sequence from those in other species in regions of the peptide that could affect substrate binding. The dermatophytes also encode novel sets of fungus-specific kinases with unknown specificity, including nonfunctional pseudokinases, which may inhibit phosphorylation by competing for kinase sites within substrates, acting as allosteric effectors, or acting as scaffolds for signaling. The dermatophytes are also enriched for a large number of enzymes that synthesize secondary metabolites, including dermatophyte-specific genes that could synthesize novel compounds. Finally, dermatophytes are enriched in several classes of proteases that are necessary for fungal growth and nutrient acquisition on keratinized tissues. Despite differences in mating ability, genes involved in mating and meiosis are conserved across species, suggesting the possibility of cryptic mating in species where it has not been previously detected. These genome analyses identify gene families that are important to our understanding of how dermatophytes cause chronic infections, how they interact with epithelial cells, and how they respond to the host immune response. IMPORTANCE Athlete’s foot, jock itch, ringworm, and nail infections are common fungal infections, all caused by fungi known as dermatophytes (fungi that infect skin). This report presents the genome sequences of Trichophyton rubrum, the most frequent cause of athlete’s foot, as well as four other common dermatophytes. Dermatophyte genomes are enriched for four gene classes that may contribute to the ability of these fungi to cause disease. These include (i) proteases secreted to degrade skin; (ii) kinases, including pseudokinases, that are involved in signaling necessary for adapting to skin; (iii) secondary metabolites, compounds that act as toxins or signals in the interactions between fungus and host; and (iv) a class of proteins (LysM) that appear to bind and mask cell wall components and carbohydrates, thus avoiding the host’s immune response to the fungi. These genome sequences provide a strong foundation for future work in understanding how dermatophytes cause disease. Athlete’s foot, jock itch, ringworm, and nail infections are common fungal infections, all caused by fungi known as dermatophytes (fungi that infect skin). This report presents the genome sequences of Trichophyton rubrum, the most frequent cause of athlete’s foot, as well as four other common dermatophytes. Dermatophyte genomes are enriched for four gene classes that may contribute to the ability of these fungi to cause disease. These include (i) proteases secreted to degrade skin; (ii) kinases, including pseudokinases, that are involved in signaling necessary for adapting to skin; (iii) secondary metabolites, compounds that act as toxins or signals in the interactions between fungus and host; and (iv) a class of proteins (LysM) that appear to bind and mask cell wall components and carbohydrates, thus avoiding the host’s immune response to the fungi. These genome sequences provide a strong foundation for future work in understanding how dermatophytes cause disease.

An overview of topical antifungal therapy in dermatomycoses. A North American perspective.

SummaryDermatophytes cause fungal infections of keratinised tissues, e.g. skin, hair and nails. The organisms belong to 3 genera, Trichophyton, Epidermophyton and Microsporum. Dermatophytes may be grouped into 3 categories based on host preference and natural habitat. Anthropophilic species predominantly infect humans, geophilic species are soil based and may infect both humans and animals, zoophilic species generally infect non-human mammals.It is important to confirm mycologically the clinical diagnosis of onychomycosis and other tinea infections prior to commencing therapy. The identity of the fungal organism may provide guidance about the appropriateness of a given topical antifungal agent. Special techniques may be required to obtain the best yield of fungal organisms from a given site, especially the scalp and nails.It is also important to realise the limitations of certain diagnostic aids e.g., Wood’s light examination is positive in tinea capitis due to M. canis and M. audouinii (ectothrix organisms); however, Wood’s light examination is negative in T. tonsurans (endothrix organism). Similarly, it is important to be aware that cicloheximide in culture medium will inhibit growth of non-dermatophytes. Appropriate media are therefore required to evaluate the growth of some significant non-dermatophyte moulds.For tinea infections other than tinea capitis and tinea unguium, topical antifungals may be considered. For effective therapy of tinea capitis an oral antifungal is generally necessary. Similarly, oral antifungals are the therapy of choice, especially if onychomycosis is moderate to severe. Furthermore, where the tinea infection involves a large area, in an immunocompromised host or if infection is recurrent with poor response to topical agents, then oral antifungal therapy may be necessary.Topical antifungal agents may be broadly divided into specific and nonspecific agents. The former group includes the polyenes, azoles, allylamines, amorolfine, ciclopirox and butenafine. Generally the topical agent is available as a cream, sometimes for use intravaginally. Less commonly, the formulation may be in the form of a powder, lacquer, spray, gel or solution. Many of these agents have a broad spectrum of activity, being effective against dermatophytes, yeasts and Malassezia furfur. For the treatment of tinea corporis, tinea cruris tinea versicolor and cutaneous candidosis, once or twice daily application may be required, the most common duration of therapy being 2 to 4 weeks. For tinea pedis the most common treatment duration is 4 to 6 weeks.

Arthroconidial formation in Trichophyton raubitschekii

Arthroconidia produced by dermatophytic fungi are considered to be the primary cause of skin and nail infections in humans and animals. Trichophyton rubrum is currently the most common cause of tinea pedis all over the world. The common form of T. rubrum produces a cottony colony in cultures that is characteristically low in conidia formation. The attempts to produce arthroconidia in T. rubrum have shown little success so far. Recently, Trichophyton raubitschekii, an anthropophilic dermatophyte prevalent in Asia, Africa, and the Mediterranean, has been recognized as a variant of T. rubrum. In cultures, T. raubitschekii is characterized by a granular colony form, and an abundance of both micro‐ and macroconidia. The present study reveals a predominance of arthroconidia in two T. raubitschekii cultures isolated from clinical materials. These isolates were able to maintain arthroconidiation in bimonthly subcultures throughout the entire course of this study. The growth parameters for in vitro cultivation of arthroconidia are described here. Arthroconidia prepared from T. raubitschekii cultures showed greater than 95% germination within 21u2003h of suspension in phosphate‐buffered saline. The availability of arthroconidia in T. raubitschekii cultures appears to offer a practical means of characterizing infective cells in T. rubrum.

THE STAINING OF FILAMENTOUS FUNGI WITH DIAZONIUM BLUE B

The diazonium blue B staining reaction, commonly used to determine the subdivisional affinities of anamorphic yeasts, is adapted for staining cultures of filamentous fungi. When unprocessed mycelium from shake culture is treated with the reagent, Zygomycetes and some Basidiomycetes stain positively; Ascomycetes and most Basidiomycetes do not. After alkaline hydrolysis of the mycelium, Zygomycetes, Basidiomycetes, and some AsGomycetes react positively. Strains reacting negatively are exclusively ascomycetous. Apart from the standard positive and negative reactions, two anomalous reactions occur. The first is a bright yellow reaction found in certain Basidiomycetes; the second is a red reaction of the conidial slime of Hainesia lythri, a species with ascomycetous affinities.

Comparison of efficacy criteria across onychomycosis trials: need for standardization

Background The last 10 years have seen a substantial increase in the number of studies reporting the efficacy of the various antifungal agents used to treat onychomycosis.

Ethanol physiology in the warehouse-staining fungus, Baudoinia compniacensis

The fungus Baudoinia compniacensis colonizes the exterior surfaces of a range of materials, such as buildings, outdoor furnishings, fences, signs, and vegetation, in regions subject to periodic exposure to low levels of ethanol vapour, such as those in the vicinity of distillery aging warehouses and commercial bakeries. Here we investigated the basis of ethanol metabolism in Baudoinia and investigate the role of ethanol in cell germination and growth. Germination of mycelia of Baudoinia was enhanced by up to roughly 1d exposure to low ethanol concentrations, optimally 10ppm when delivered in vapour form and 5mm in liquid form. However, growth was strongly inhibited following exposure to higher ethanol concentrations for shorter durations (e.g., 1.7m for 6h). We found that ethanol was catabolized into central metabolism via alcohol dehydrogenase (ADH) and acetaldehyde dehydrogenase (ACDH). Isocitrate dehydrogenases (IDHs) were active in cells grown on glucose, but these enzymes were not expressed when ethanol was provided as a sole or companion carbon source. The glyoxylate cycle enzymes isocitrate lyase (ICL) and malate synthase (MS) activities observed in cells grown on acetate were comparable to those reported for other microorganisms. By replenishing tricarboxylic acid (TCA) cycle intermediates, it is likely that the functionality of the glyoxylate cycle is important in the establishment of luxuriant growth of Baudoinia compniacensis on ethanol-exposed, nutrient-deprived, exposed surfaces. In other fungi, such as Saccharomyces cerevisiae, ADH II catalyses the conversion of ethanol to acetaldehyde, which then can be metabolized via the TCA cycle. ADH II is known to be strongly repressed in the presence of glucose.

Studies on the systematics of ectomycorrhizal fungi in axenic culture. Reactions of mycelia to diazonium blue B staining.

Diazonium blue B reagent was used to test staining reactions of mycelia of 96 species of ectomy? corrhizal and ectomycorrhizal-like fungi grown for five weeks in liquid culture. Red to violet staining was seen in harvested mycelia of basidiomycetes treated in cold KOH while negative reactions indicated fungi with ascomycetous affinities. Non-treated mycelia gave a variety of different color responses which were generally species-specific. Mycelia of Cortinarius species and most boletes did not stain while most species of Hebeloma, Hygrophorus, Laccaria and Tricholoma stained yellow. A purely red staining reaction was restricted mainly to a few species within the Boletaceae. Results reveal potential value of diazonium blue B staining as a taxonomic character for identification of mycelia isolated from fieldcollected ectomycorrhizae.

A new inoperculate discomycete associated with roots of Picea

A new inoperculate discomycete, Neocudoniella radicella, producing ascomata on rootlets, probably ofPicea spp., is described and its taxonomic relationships discussed. In vitro resynthesis studies between isolates of the new fungus and seedlings of Picea mariana show that the fungus produces irregular mantle and Hartig net-like structures on rootlets. It is, however, unclear whether the association is mycorrhizal or parasitic.

Taxonomic annotation of public fungal ITS sequences from the built environment – a report from an April 10–11, 2017 workshop (Aberdeen, UK)

Abstract Recent DNA-based studies have shown that the built environment is surprisingly rich in fungi. These indoor fungi – whether transient visitors or more persistent residents – may hold clues to the rising levels of human allergies and other medical and building-related health problems observed globally. The taxonomic identity of these fungi is crucial in such pursuits. Molecular identification of the built mycobiome is no trivial undertaking, however, given the large number of unidentified, misidentified, and technically compromised fungal sequences in public sequence databases. In addition, the sequence metadata required to make informed taxonomic decisions – such as country and host/substrate of collection – are often lacking even from reference and ex-type sequences. Here we report on a taxonomic annotation workshop (April 10–11, 2017) organized at the James Hutton Institute/University of Aberdeen (UK) to facilitate reproducible studies of the built mycobiome. The 32 participants went through public fungal ITS barcode sequences related to the built mycobiome for taxonomic and nomenclatural correctness, technical quality, and metadata availability. A total of 19,508 changes – including 4,783 name changes, 14,121 metadata annotations, and the removal of 99 technically compromised sequences – were implemented in the UNITE database for molecular identification of fungi (https://unite.ut.ee/) and shared with a range of other databases and downstream resources. Among the genera that saw the largest number of changes were Penicillium, Talaromyces, Cladosporium, Acremonium, and Alternaria, all of them of significant importance in both culture-based and culture-independent surveys of the built environment.