In vitro susceptibility of dermatomycoses agents to six antifungal drugs and evaluation by fractional inhibitory concentration index of combined effects of amorolfine and itraconazole in dermatophytes

To investigate the antifungal drug susceptibility of fungi responsible for dermatomycoses, minimum inhibition concentration (MIC) tests were performed in 44 strains of dermatophytes, including Trichophyton rubrum, Trichophyton mentagrophytes, Trichophyton verrucosum, Trichophyton tonsurans, Microsporum canis, Microsporum gypseum and Epidermophyton floccosum, with six antifungal drugs (amorolfine, terbinafine, butenafine, ketoconazole, itraconazole and bifonazole) by broth microdilution assay according to Clinical Laboratory Standard Institute protocols. Six possible dermatomycosis‐causing non‐dermatophytic fungi were also tested. The two major causes of tinea, T. rubrum and T. mentagrophytes, showed significantly different sensitivities to ketoconazole and bifonazole. Clinically derived dermatophytes were sensitive to the six antifungal drugs tested. However, non‐dermatophytes, especially Fusarium spp., tended to be resistant to these antifungal drugs. In Trichophyton spp., the MICs of non‐azole drugs had narrower distributions than those of azoles. To evaluate the effects of antifungal drug combinations, the fractional inhibitory concentration index was calculated for the combination of amorolfine and itraconazole as representative external and internal drugs for dermatophytes. It was found that this combination had synergistic or additive effects on most dermatophytes, and had no antagonistic effects. The variation in susceptibility of clinically derived fungal isolates indicates that identification of causative fungi is indispensable for appropriately choosing effective antifungal drugs in the early stages of infection. The results of combination assay suggest that multiple drugs with different antifungal mechanisms against growth of dermatophytes should be used to treat refractory dermatomycoses, especially onychomycosis.     

Antimicrobial agents

In a study of the effect of progesterone, its hydroxy derivatives and various sterols on the growth of dermatophytes, it was demonstrated that all the steroid compounds employed inhibited growth of the 51 strains of dermatophytes tested. No significant differences were found in sensitivity to the given steroids, either in dermatophyte strains of the same species, but of different origin, ior in different species of the four genera used (Trichophyton, Microsporum, Epidermophyton andKeratinomyces). Hydroxylation of the progesterone molecule in any except the C-21 position lowered the inhibitory effect. The course of the transformation reaction on the progesterone or cholesterol molecule was likewise of a uniform character from the aspect of species specificity. Progesterone was simultaneously hydroxylated in positions 15α and 15β by all the strains, giving rise to monohydroxy-derivatives as the main metabolites, and cholesterol was oxidized to cholestenone.     

Sequenced dermatophyte strains: growth rate, conidiation, drug susceptibilities, and virulence in an invertebrate model

Although dermatophytes are the most common cause of fungal infections in the world, their basic biology is not well understood. The recent sequencing and annotation of the genomes of five representative dermatophyte species allows for the creation of hypotheses as to how they cause disease and have adapted to their distinct environments. An understanding of the microbiology of these strains will be essential for testing these hypotheses. This study is the first to generally characterize these five sequenced strains of dermatophytes for their microbiological aspects. We measured the growth rate on solid medium and found differences between species, with Microsporum gypseum CBS118893 having the fastest growth and Trichophyton rubrum CBS118892 the slowest. We also compared different media for conidia production and found that the highest numbers of conidia were produced when dermatophytes were grown on MAT agar. We determined the Minimum Inhibitory Concentration (MIC) of nine antifungal agents and confirmed susceptibility to antifungals commonly used as selectable markers. Finally, we tested virulence in the Galleria mellonella (wax moth) larvae model but found the results variable. These results increase our understanding of the microbiology and molecular biology of these dermatophyte strains and will be of use in advancing hypothesis-driven research about dermatophytes.     

Antigenic characterization of Microsporum canis,M. distortum,M. equinum,M. ferrugineum and Trichophyton soudanense cultures

The authors investigated the antigenic properties of a group of closely related dermatophytes whose components may be morphologically confused with one another: Microsporum canis, M. distortum, M. equinum, M. ferrugineum and Trichophyton soudanense. By using reference antigens and adsorbed monospecific antisera, it was possible to serologically distinguish the reference strains by the exoantigen technique. Their common and specific antigenic determinants were visualized by cross-immunoelectrophoresis tests with intermediate gel.     

Epidemiology of Dermatophytes in Belgium: A 5 Years’ Survey

Dermatophytes are among the most common fungal agents causing superficial skin infections worldwide. Epidemiology of these infections is evolving and variable in every country. This report presents the Belgian epidemiological data regarding the distribution of dermatophytes species isolated by the two national reference centers for mycosis during a period of 5 years (2012–2016). Trichophyton rubrum was the most frequently isolated species, considering all sampling sites (60.3% on average between 2012 and 2016). More precisely, this dermatophyte was the major agent of Tinea unguium and Tinea corporis during this period, followed by species of the Trichophyton mentagrophytes complex. Moreover, Microsporum audouinii was the main etiological agent of Tinea capitis (TC) with a frequency of 52.5% on average between 2012 and 2016. Other African dermatophytes species such as Trichophyton soudanense and Trichophyton violaceum were also agents of TC with a respective prevalence of 11.6% and 11.5% on average. This study highlights a different dermatophyte distribution in Belgium in comparison with other European countries.

     

Lipolytic activity of dermatophytes

Lipolytic activity of dermatophytes was tested by the method generally used forCandida lipolytica. Most of the freshly isolated strains ofMicrosporum canis, M. gypseum, Epidermophyton floccosum andTrichophyton mentagrophytes gave positive reactions, whereas, only few strains ofT. schoenleini, T. violaceum, T. megnini, T. rubrum andT. tonsurans yielded such reactions. Among subcultured strains, it was found a decreased lipolytic activity inMicrosporum species and an increased lipolytic activity inEpidermophyton and someTrichophyton species.Part of this work was performed at Serviço de Doenças Infecto-Contagiosas (Secção de Dermatologia), Hospital de Curry Cabral - Lisboa.     

Action of griseofulvin on geophilic dermatophytes and related keratinophilic fungi

Summary Growth response to griseofulvin has been studied in 24 strains of 16 species of geophilic dermatophytes and related keratinophilic fungi. Based on their sensitivity to griseofulvin these fungi fall into three groups: (1) Resistant group includesKeratinomyces ajelloi, Trichophyton terrestre andTrichophyton evolceanui which can grow upto 40µg per ml concentration of griseofulvin. (2) Fairly resistant group comprisingMicrosporon cookei, Microsporon vanbreuseghemii, a strain ofMicrosporon gypseum andTrichophyton sp., with growth occurring upto 8–16µg per ml concentration. (3) Sensitive group includes species ofTrichophyton, Keratinophyton, Microsporon, Nannizzia, Chrysosporium andCtenomyces which are completely inhibited at 4–8µg or lower griseofulvin concentrations. Griseofulvin inhibited formation of cleistothecia inK. terreum, delayed pleomorphic growth inT. indicum andN. incurvata, and influenced pigment production in several strains.     

Exocellular enzyme activity of dermatophytes and other fungi isolated from ruminants in Southern Iraq

Sixteen fungal species were isolated from 182 specimens collected from four ruminants (buffalo, camel, cattle and sheep) in Southern Iraq. Fungi represented by five species of dermatophytes and eleven species of other fungi were screened for the activity of four enzymes; keratinase, proteinase, lipase and amylase. Keratinase was found to be produced by all of the dermatophytes and non-dermatophytes, except for Paecillomyces variottii and Scytalidium lignicola. However, high keratinase activity was expressed by the dermatophytic species particularly by Trichophyton mentagrophytes var. erinacei and Microsporum gypseum. Three dermatophytes viz. M. gypseum, T. verrucosum and T. mentagrophytes var. nodulare were capable of producing protease, lipase and amylase. Although, T. mentagrophytes var. erinacei showed high protease activity, it did not produce lipase and amylase. On the contrary most of the non-dermatophytic species revealed protease and lipase activities higher than the dermatophytes. The Curvularia spp. isolates showed the highest protease and amylase activity, while Aspergillus parasiticus revealed the highest activity of lipase and amylase. No correlation was observed between enzyme activity and the growth rate of the examined fungi. This revised version was published online in June 2006 with corrections to the Cover Date.     

Species diversity of keratinophilic fungi in various soil types

Keratinophilic fungi are a highly specialized, keratin-degrading ecological group. They live in natural environments, mostly in the keratinrich remains of dead animal in the soil. We investigated species diversity in four types of soils with different physico-chemical properties. The strain material was identified based on morphological characters. Different representatives of Chrysosporium and geophilic dermatophytes dominated depending on soil pH. Geophilic dermatophytes were represented by one species, Trichophyton ajelloi, and the Chrysosporium group was represented by Chrysosporium keratinophilum. The frequency of Trichophyton ajelloi increased with an increase in pH, and it reached the maximum in strongly acidic soil (podzol), unlike the Chrysosporium group. The frequency of Chrysosporium keratinophilum was positively correlated with the content of humus, nitrogen, CaCO₃ and phosphorus in the soils.     

Degradation of keratin substrates by fungi isolated from sewage sludge

Four fungal species including two dermatophytes and two saprophytes were isolated from sewage sludge samples at Basrah (Iraq) they were tested for their degradative ability towards three types of keratin substrates (human hair, chicken feathers and wool). The rate of keratin degradation was expressed as weight loss over three weeks of incubation using a liquid culture medium. Human hair had the highest degradation rate by colonization of Chrysosporium pannicola and Microsporum gypseum at a rate of 62% and 4% respectively. Chicken feathers were highly degraded by Aspergillus flavus (32%) while wool degradation was highest by C. pannicola (45.5%) and Trichophyton mentagrophytes var. erinacei (38%). There was a significant difference (p < 0.00l) in keratin substrate degradation rates by the examined fungi. Keratinase activity was highest for C. pannicola and M. gypseum in the culture medium baited with human hair. Aspergillus flavus revealed the highest activity of this enzyme in cultures amended with chicken feathers while T. mentagrophytes var. erinacei showed highest keratinase activity in cultures with wool substrate. The amount of protein released into the culture medium varied among the tested fungi. The medium's alkalinity increased over incubation time from 6.5 to 7.8. Microscopic examination showed maceration of the keratin substrates by the fungi. This revised version was published online in June 2006 with corrections to the Cover Date.     

Onychomycosis in Malaysia

The common etiological agents of onychomycosis are dermatophytes, molds and yeasts. A mycological nail investigation of onychomycosis using direct microscopy and culture was conducted by the Mycology Unit, Department of Medical Microbiology, University of Malaya from March 1996 to November 1998. The study involved 878 nail clippings or subungal scrapings from subjects with onychomycosis. On direct microcopy examination, 50% of the specimens were negative for fungal elements. On culture, 373 specimens had no growth; bacteria were isolated from 15 nail specimens. Among the 490 specimens with positive fungal cultures, 177 (36.1%) were dermatophytes, 173 (35.5%) were molds and 130 (26.5%) were Candida. There were 2% (10/490) mixed infections of molds, yeasts and dermatophytes. Trichophyton rubrum (115/177) and Trichophyton mentagrophytes (59/177) were the main dermatophytes isolated. The molds isolated were predominantly Aspergillus niger (61/173), Aspergillus nidulans (30/173), Hendersonula toruloidea (26/173) and Fusarium species (16/173). 96.9% of the Candida species identified were Candida albicans.This revised version was published online in October 2005 with corrections to the Cover Date.     

Molecular differentiation of Keratinomyces (Trichophyton) species

The taxonomy of the form-genus Keratinomyces (Trichophyton) within the group of the dermatophytes is based on morphological features which remain insufficient for the distinction of these anamorphic species. The three species included in the genus Keratinomyces, namely K. ajelloi, K. ceretanicus and K. longifusus were examined by means of their mitochondrial-like DNA diversity and compared to few other dermatophytes. The analysis of the mtDNA restriction fragments confirmed that the three species are different and well separate from the other dermatophytes.     

Taxonomic study of keratinophilic fungi isolated from soil and some mammals in France

In France, most of the researches have showed the distribution of dermatophytes and other keratinophilic fungi in the soil and in the coat of wild mammals. During these last years, we have also practiced numerous investigations about the distribution of such fungi. The isolated species are: Trichophyton mentagrophytes, Trichophyton terrestre, Trichophyton ajelloï, Trichophyton simii, Microsporum gypseum, Microsporum cookei, Microsporum nanum, Microsporum persicolor, Anixiopsis stercoraria, Chrysosporium keratinophilum, Chrysosporium tropicum, Chrysosporium state of Arthroderma curreyi, Chrysosporium state of Arthroderma cuniculi, Chrysosporium state of Arthroderma tuberculatum, Chrysosporium state of Arthroderma multifidum, Ctenomycès serratus, Chrysosporium parvum, Chrysosporium indicum, Chrysosporium georgii, Chrysosporium merdarium, Chrysosporium asperatum, Chrysosporium pannorum.A key to these species is proposed: it attempts to provide an aid in identifying geophilic dermatophytes and related fungi (belonging to the Chrysosporium genus).     

The fungistatic activities of vitamin K on dermatophytes

Summary and conclusions Four commercial preparations of vitamin K were evaluated for their fungistatic activities on the following dermatophytes:Microsporum canis, Epidermophyton floccosum, Trichophyton mentagrophytes, andTrichophyton Schönleinii.Table 1 lists the maximum nonfungistatic and the minimum fungistatic concentrations of each vitamin K.The fungistatic activities of the vitamins K are higher in the liquid than in the solid medium.VitaminIV was the most active on all the dermatophytes in both, the solid and the liquid medium. The fungistatic activities of the other vitamins K vary, in each medium, according to the dermatophyte. Sodium bisulfite decreases the fungistatic activities of vitamin K. Epidermophyton floccosum was the most sensitive to all the vitamins K used.     

Antifungal activity of 5 new synthetic compounds vs. Trichophyton rubrum and Epidermophyton floccosum

The antifungal activity of five new synthetic compounds was evaluated on two dermatophytes: Epidermophyton floccosum and Trichophyton rubrum. The data showed that the imidazo-pyrazole and pyrazolo-thiazoles were not particularly effective, while the two pyrazole-thiocyanates proved highly active on both fungi. The most active 5-amino-3-methyl-1-phenylpyrazolo-4-thiocyanate was chosen to perform SEM and TEM morphological studies on both fungi. Both SEM and TEM observations revealed interesting alterations on the two dermatophytes, particularly involving the endomembrane system. This revised version was published online in June 2006 with corrections to the Cover Date.     

Les mycoses superficielles en zone forestière gabonaise: bilan des champignons isolés

Résumé Les auteurs, effectuant une rétrospective de 2 années d'activité en mycologie à Libreville (Gabon), exposent et analysent l'ensemble des résultats positifs obtenus à partir des teignes et épidermomycoses qu'ils ont eu l'occasion d'observer. Signalant simplement l'existence du Pityriasis versicolor, ils replacent les agents des différentes mycoses selon leur importance respective.Ce premier inventaire des dermatophytes au Gabon, (13 espèces au total) en zone forestière humide et chaude, montre la nette prédominance de Microsporum langeroni, Trichophyton soudanense, Trichophyton rubrum, Epidermophyton floccosum. Trichophyton violaceum est isolé dans une proportion non négligeable de cas. D'autres espèces, plus rares, sont mentionnées.

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The authors draw up an inventory of the dermatophytes in Gabon (13 species isolated from 593 people in the wet and hot forest). They show the clear prevalence of Microsporum langeroni, Trichophyton soudanense, Trichophyton rubrum, and Epidermophyton floccosum, Trichophyton violaceum also is fairly often isolated.More infrequently, they find T. mentagrophytes, T. mentagrophytes var. interdigitale, T. gourvili, M. canis, M. audouini, T. yaoundei, M. gypseum.

     

A survey of dermatophytes isolated from human patients in the United States from 1979 to 1981 with chronological listings of worldwide incidence of five dermatophytes often isolated in the United States

A survey of dermatophytes isolated from patients seeking medical advice was made from 1979 to 1981 in the United States. The survey included 54 locations with data from 40 cities and 2 states. Correlations of these data with that of the other localities of the world were made to illustrate the dynamic epidemiology of several common dermatophytes. The most often isolated dermatophyte in this survey was Trichophyton rubrum having 53.66% of the total for these three calendar years. In a chronological listing of ringworm infections caused by this organism, many areas of the world have reported similar increased incidence of this pathogen. Trichophyton tonsurans was isolated 27.85% of the total. A dramatic increase of this pathogen as a cause of tinea capitis has been observed in most cities of the United States. It has been isolated in 25 different countries of the world. The percentage of isolation of Trichophyton mentagrophytes was 8.56%. This percentage may not be near the true incidence of infection by this dermatophyte because the infections are mild and respond to treatment without the individual seeking medical advice. Since the 1950s the percentage of isolations of the total has dropped for T. mentagrophytes in the United States. Epidermophyton floccosum accounted for 4.36% of the total. In a few areas of the world it causes over 30% of the total of dermatophytoses. Microsporum canis was isolated 3.72% of the total in the United States. It has recently been reported to be the dominant agent of tinea capitis in several South American countries, Tucson, Arizona and Kuwait. Once the dominant pathogen of tinea capitis in children in the United States, it was replaced by Microsporum audouinii before 1960. Today in the United States, M. audouinii only accounts for 0.30% of the total. It is considered eliminated as a pathogen in England. In this survey, isolated less than 1.0% of the total were Microsporum gypseum, Microsporum ferrugineum, Microsporum nanum, Microsporum fulvum and Trichophyton schoenleinii. Trichophyton meginii and Trichophyton terrestre were reported isolated but no numerical data were available.     

Investigation of keratinophilic fungi from soils in Western Australia a preliminary survey

In order to determine which species of geophilic dermatophytes were present in Western Australian soils 299 samples were investigated. These samples were collected from a range of locations, 208 from towns throughout the state and 91 samples from the Perth Metropolitan area.Most samples were collected from areas frequented by people and animals, such as home gardens, parks and animal yards.Of the total 299 soils, 271 (90.6 %) yielded keratinophilic fungi. A total of 181 dermatophytes were isolated, and there were 205 isolations of other keratinophilic fungi. Microsporum gypseum (30.7 %) was the most prevalent dermatophyte recovered from soil followed byMicrosporum cookei (21.7 %) and thenTrichophyton ajelloi (8.0 %). No other dermatophytes were recovered.Chrysosporium indicum was the most common of all the keratinophilic fungi and was isolated from 50.1 % of the samples. Mixed growth was obtained from 33.5 % of the soil samples.     

The routine use of modified Borelli's lactritmel agar (MBLA)

The original formula of Borelli's lactritmel agar (BLA)(3) which contains wheat flour, milk and honey, has been modified by replacing the wheat flour with dehydrated Bacto Corn Meal Agar (Difco) and by slightly altering the concentrations of the milk and honey. The modified medium (MBLA) is less turbid, less particulate, and easier to prepare than BLA. Although Trichophyton rubrum usually produces a wine-red pigment with BLA, most strains initially produce a yellow pigment, with the red pigment developing later. The corn meal in MBLA reduces this tendency and stimulates the early formation of deep wine red pigment. MBLA enhances sporulation of dermatophytes and various fungi which fail to sporulate on other media, and maintains characteristic growth without developing pleomorhic degeneration. It has been used routinely since 1972 as a reliable aid to the differentiation of T. rubrum and T. mentagrophytes. Since 1975 selective MBLA has been used as a routine primary isolation medium for dermatophytes, and has proved to be most useful.     

Human mycoses in Portugal [1960–1973]

A critical survey is made of human mycoses diagnosed in European Portugal and in the Portuguese Overseas Provinces. Dermatophyte infections and pityriasis versicolor are commom in the entire territory. The more frequently isolated dermatophytes in Continental Portugal wereTrichophyton violaceum, Microsporum canis, Trichophyton tonsurans andTrichophyton schoenleinii, in the scalp andTrichophyton rubrum, Trichophyton mentagrophytes, Trichophyton megninii andEpidermophyton floccosum in the other body sites. In the Overseas Provinces the species found in white people were very much the same, whereas in negroes mainlyMicrosporum audouinii andT. violaceum in Mozambique,M. audouinii in Angola, andTrichophyton soudanense, M. audouinii andT. rubrum in Guinea were identified.In Continental Portugal cases of candidiasis, mycetoma, aspergillosis, sporotrichosis and cryptococcosis were described in patients who had never been outside Europe; and tinea nigra, African histoplasmosis and South-American blastomycosis in individuals who live or lived in India, Africa and Brazil.The first case of North-American blastomycosis was reported in Mozambique.