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Background: Malassezia (M.) pachydermatis as a frequent reason for dermatological consultation in dogs and cats was recently shown to be lipid-dependent, too. Lipolytic activity is a prerequisite for activating antimicrobial effectivity of fatty acid esters., Objectives: It was therefore of interest whether it is possible to induce this mechanism in M. pachydermatis and to identify possible differences between minimal and strong lipid-dependent strains., Methods: In an agar dilution test, the minimal inhibitory concentrations of six fatty acid esters were determined for seventeen M. pachydermatis strains. GC analysis of parent compounds and liberated fatty acids was used to quantify ester cleavage., Results: Hydrolysis was observed in all test strains in a homogenous manner but was dependent on the chemical structure. Lowest MICs (500 ppm after 14 days of incubation) were obtained applying glyceryl monocaprylate and 3-hydroxylpropyl caprylate, while the corresponding esters of undecylenic acid showed nearly twice the value. As shown by GC analysis with the reference strains CBS 1879 and CBS 1892 and 3-hydroxypropyl caprylate, hydrolysis and caprylic acid formation starts immediately and was dependent on yeast density. Furthermore, nine azole-resistant strains isolated from dogs with treatment failures showed MIC values comparable to the other strains and no resistance to monohydric fatty acid esters., Conclusions: Medium-chain fatty acid esters may represent a new therapeutic option for veterinary use even in azole-resistant strains. The in vivo verification in M. pachydermatis-associated dermatitis in dogs and cats will be the next step for the successful development of new therapeutics., (© 2022 Wiley-VCH GmbH.)

Approximately 70-75% of women will have vulvovaginal candidosis (VVC) at least once in their lifetime. In premenopausal, pregnant, asymptomatic and healthy women and women with acute VVC, Candida albicans is the predominant species. The diagnosis of VVC should be based on clinical symptoms and microscopic detection of pseudohyphae. Symptoms alone do not allow reliable differentiation of the causes of vaginitis. In recurrent or complicated cases, diagnostics should involve fungal culture with species identification. Serological determination of antibody titres has no role in VVC. Before the induction of therapy, VVC should always be medically confirmed. Acute VVC can be treated with local imidazoles, polyenes or ciclopirox olamine, using vaginal tablets, ovules or creams. Triazoles can also be prescribed orally, together with antifungal creams, for the treatment of the vulva. Commonly available antimycotics are generally well tolerated, and the different regimens show similarly good results. Antiseptics are potentially effective but act against the physiological vaginal flora. Neither a woman with asymptomatic colonisation nor an asymptomatic sexual partner should be treated. Women with chronic recurrent Candida albicans vulvovaginitis should undergo dose-reducing maintenance therapy with oral triazoles. Unnecessary antimycotic therapies should always be avoided, and non-albicans vaginitis should be treated with alternative antifungal agents. In the last 6 weeks of pregnancy, women should receive antifungal treatment to reduce the risk of vertical transmission, oral thrush and diaper dermatitis of the newborn. Local treatment is preferred during pregnancy., (© 2021 The Authors.Mycoses published by Wiley-VCH GmbH.)

Background: For fatty acid esters of monohydric alcohols, cleavage by exo-enzymes of Malassezia (M.) spp. and release of fatty acids with antimicrobial activity have been shown recently. On skin surface, this selective activation of antimicrobial activity might result in a 'self-kill' targeted locally at the site with the highest M. density., Objectives: As for the disadvantage of strong odour, use of these esters for topical therapy is limited to low concentrations. Therefore, cleavage was also tested for monoesters of octanoic and undec-10-enoic acid with the bihydric alcohol propane-1,3-diol or the trihydric glycerol., Methods: In an agar dilution test, the minimal inhibitory concentrations of these compounds were determined for M. furfur, M. globosa, M. sympodialis and M. restricta, respectively. GC analysis of parent compounds and liberated fatty acids was used to reveal ester cleavage., Results: Ester cleavage started immediately. MICs for the test compounds ranged between ~1000-8000 ppm after 14 days of incubation. 1,3-propanediol esters, especially 3-hydroxypropyl octanoate and 3-hydroxypropyl undecylenate were most effective, binary combinations exerted synergistic effects., Conclusions: The new substances are advantageous in terms of odour and substantivity and have also beneficial skin caring properties if not hydrolysed by M. spp. As a different panel of hydrolases of each single M. species is responsible for variation in efficacy among the test substances, tailored products to treat preferentially single species or blends with a broader effectivity can be designed. In vivo verification will be the next step for the successful development of this new therapeutical concept for M.-associated diseases., (© 2020 Blackwell Verlag GmbH.)

Miconazole is a broad-spectrum antifungal used in topical preparations. In the present investigation the minimal inhibitory concentration (MIC) of miconazole for eighty wild type strains of gram-positive and gram-negative bacteria isolated from infected skin lesions was assessed using a modified agar dilution test (adapted to CLSI, Clinical Laboratory Standards Institute). 14 ATCC reference strains served as controls. Miconazole was found efficacious against gram-positive aerobic bacteria (n=62 species), the MICs against Staphylococcus (S.) aureus, S. spp., Streptococcus spp. und Enterococcus spp. ranged between 0.78 and 6.25 μg/mL. Interestingly, there were no differences in susceptibility between methicillin-susceptible (MSSA, 3) methicillin-resistant (MRSA, 6) and fusidic acid-resistant (FRSA, 2) S. aureus isolates. Strains of Streptococcus pyogenes (A-streptococci) (8) were found to be slightly more sensitive (0.78-1.563 μg/mL), while for gram-negative bacteria, no efficacy was found within the concentrations tested (MIC >200 μg/mL). In conclusion, for the gram-positive aerobic bacteria the MICs of miconazole were found within a range which is much lower than the concentration of miconazole used in topical preparations (2%). Thus topically applied miconazole might be a therapeutic option in skin infections especially caused by gram-positive bacteria even by those strains which are resistant to antibiotics., (© 2017 The Authors. Mycoses Published by Blackwell Verlag GmbH.)

In the late 1980's, the incidence of invasive fungal diseases (IFDs) in Germany was estimated with 36.000 IFDs per year. The current number of fungal infections (FI) occurring each year in Germany is still not known. In the actual analysis, data on incidence of fungal infections in various patients groups at risk for FI were calculated and mostly estimated from various (mostly national) resources. According to the very heterogenous data resources robust data or statistics could not be obtained but preliminary estimations could be made and compared with data from other areas in the world using a deterministic model that has consistently been applied in many countries by the LIFE program ( www.LIFE-worldwide.org). In 2012, of the 80.52 million population (adults 64.47 million; 41.14 million female, 39.38 million male), 20% are children (0-14 years) and 16% of population are ≥65 years old. Using local data and literature estimates of the incidence or prevalence of fungal infections, about 9.6 million (12%) people in Germany suffer from a fungal infection each year. These figures are dominated (95%) by fungal skin disease and recurrent vulvo-vaginal candidosis. In general, considerable uncertainty surrounds the total numbers because IFDs do not belong to the list of reportable infectious diseases in Germany and most patients were not hospitalised because of the IFD but a distinct underlying disease., (© 2015 Blackwell Verlag GmbH.)

Free medium and short chain fatty acids are known to have broad antimicrobial activity. However, their practical use in topical therapy is limited by their intensive smell and acidity. Surprisingly, a nearly identical antimicrobial effect was found with the ethyl ester derivatives of these fatty acids, but only against Malassezia (M.) yeast, not against Candida spp. Obviously, these esters are hydrolysed by M. enzymes, thus generating a selective activation of antimicrobial activity especially in areas well populated with these yeast ('targeting'). Octanoic acid ethyl ester (CAS 106-32-1) was found to be most suitable. In an agar dilution test, the minimal inhibitory concentrations against M. globosa, M. pachydermatis and M. sympodialis, respectively, ranged between ~5 and 10 mmol l(-1) after 10 days of incubation. The effect started immediately and was not delayed by other lipid sources applied simultaneously. Based on these data, fatty acid monoesters may represent a new therapeutic concept in M.-associated diseases., (© 2015 Blackwell Verlag GmbH.)

The oestrogenised vagina is colonised by Candida species in at least 20% of women; in late pregnancy and in immunosuppressed patients, this increases to at least 30%. In most cases, Candida albicans is involved. Host factors, particularly local defence mechanisms, gene polymorphisms, allergies, serum glucose levels, antibiotics, psycho-social stress and oestrogens influence the risk of candidal vulvovaginitis. Non-albicans species, particularly Candida glabrata, and in rare cases also Saccharomyces cerevisiae, cause less than 10% of all cases of vulvovaginitis with some regional variation; these are generally associated with milder signs and symptoms than normally seen with a C. albicans-associated vaginitis. Typical symptoms include premenstrual itching, burning, redness and odourless discharge. Although itching and redness of the introitus and vagina are typical symptoms, only 35-40% of women reporting genital itching in fact suffer from vulvovaginal candidosis. Medical history, clinical examination and microscopic examination of vaginal content using 400× optical magnification, or preferably phase contrast microscopy, are essential for diagnosis. In clinically and microscopically unclear cases and in chronically recurring cases, a fungal culture for pathogen determination should be performed. In the event of non-C. albicans species, the minimum inhibitory concentration (MIC) should also be determined. Chronic mucocutaneous candidosis, a rarer disorder which can occur in both sexes, has other causes and requires different diagnostic and treatment measures. Treatment with all antimycotic agents on the market (polyenes such as nystatin; imidazoles such as clotrimazole; and many others including ciclopirox olamine) is easy to administer in acute cases and is successful in more than 80% of cases. All vaginal preparations of polyenes, imidazoles and ciclopirox olamine and oral triazoles (fluconazole, itraconazole) are equally effective (Table ); however, oral triazoles should not be administered during pregnancy according to the manufacturers. C. glabrata is not sufficiently sensitive to the usual dosages of antimycotic agents approved for gynaecological use. In other countries, vaginal suppositories of boric acid (600 mg, 1-2 times daily for 14 days) or flucytosine are recommended. Boric acid treatment is not allowed in Germany and flucytosine is not available. Eight hundred-milligram oral fluconazole per day for 2-3 weeks is therefore recommended in Germany. Due to the clinical persistence of C. glabrata despite treatment with high-dose fluconazole, oral posaconazole and, more recently, echinocandins such as micafungin are under discussion; echinocandins are very expensive, are not approved for this indication and are not supported by clinical evidence of their efficacy. In cases of vulvovaginal candidosis, resistance to C. albicans does not play a significant role in the use of polyenes or azoles. Candida krusei is resistant to the triazoles, fluconazole and itraconazole. For this reason, local imidazole, ciclopirox olamine or nystatin should be used. There are no studies to support this recommendation, however. Side effects, toxicity, embryotoxicity and allergies are not clinically significant. Vaginal treatment with clotrimazole in the first trimester of a pregnancy reduces the rate of premature births. Although it is not necessary to treat a vaginal colonisation of Candida in healthy women, vaginal administration of antimycotics is often recommended in the third trimester of pregnancy in Germany to reduce the rate of oral thrush and napkin dermatitis in healthy full-term newborns. Chronic recurrent vulvovaginal candidosis continues to be treated in intervals using suppressive therapy as long as immunological treatments are not available. The relapse rate associated with weekly or monthly oral fluconazole treatment over 6 months is approximately 50% after the conclusion of suppressive therapy according to current studies. Good results have been achieved with a fluconazole regimen using an initial 200 mg fluconazole per day on 3 days in the first week and a dosage-reduced maintenance therapy with 200 mg once a month for 1 year when the patient is free of symptoms and fungal infection (Table ). Future studies should include Candida autovaccination, antibodies to Candida virulence factors and other immunological experiments. Probiotics with appropriate lactobacillus strains should also be examined in future studies on the basis of encouraging initial results. Because of the high rate of false indications, OTC treatment (self-treatment by the patient) should be discouraged., (© 2015 Blackwell Verlag GmbH.)

Dermatomycosis is one of the most common dermatological infectious diseases. In recent years, the incidence of tinea pedum, a fungal infection of the feet, was increasing due to changing lifestyles. The risk of tinea pedum infections is associated with the use of sport shoes as well as contact with public sports facilities. Transmission of dermatomycosis occurs almost exclusively through indirect contacts, meaning that contagious material initially contaminates the patients' environment and subsequently facilitates the spread of infection to others. A suitable disinfection procedure for 'fungal reservoirs' is very important in order to reduce the risk of reinfection of tinea pedum. This study investigates the effect of microwave radiation on various dermatophytes- (Trichophyton rubrum, T. rubrum var. nigricans, T. interdigitale and Microsporum canis infected cork and polyethylene sponge shoe insoles. The contaminated insoles were irradiated with various intensities and durations of microwaves. In each case, 10 colonies on cork and polyethylene sponge insoles were irradiated with the same intensity and duration, and subsequently compared with those of corresponding non-irradiated control groups. Results of three independent experiments were statistically verified using Chi-squared test for significance. We found significant differences between the various dermatophytes on polyethylene sponge insoles and also partly on cork insoles for the same irradiation intensity and duration. We were also able to show that a complete growth inhibition of all four dermatophytes occurs on both types of insoles after a 30 s exposure at 560 W, including a maximum temperature of 60 °C., (© 2013 Blackwell Verlag GmbH.)

Malassezia (M.) furfur, a commensal organism found on the human skin, produces a wide range of pigments and fluorochromes when cultured with tryptophan as a sole nitrogen source. Some compounds of this pigment metabolism may provide an explanation for clinical characteristics of pityriasis versicolor (PV), a frequent skin disease in humans characterised by long-lasting pigmentary changes. Malassezia globosa is currently regarded as the causative agent of PV, but tryptophan-dependent pigment production has not yet been demonstrated in this species. In a previous study, we identified M. furfur genes that were differentially expressed 3 and 5 h, respectively, after induction of tryptophan-dependent pigment production. The recent publication of the genome of M. globosa prompted us to check the M. furfur sequences for homologues in M. globosa. The 3-h pool contained 79 sequences and the 5-h pool contained 91 sequences. A translated vs. translated BLAST search resulted in 62 sequences (78%) of the 3-h pool and 61 sequences (67%) of the 5-h pool showing similarity to a sequence from M. globosa. It appears that M. globosa possesses homologues to most of the genes that are differentially expressed during pigment production in M. furfur., (© 2010 Blackwell Verlag GmbH.)

The effects of the addition of different amino nitrogens on growth, morphology and secondary metabolism of Malassezia furfur were investigated. After primary culture on Dixon agar, M. furfur CBS 1878 was transferred into a fluid medium together with the nitrogen sources, glycine (Gly) or tryptophan (Trp), or a combination of both. Growth was measured by means of a direct cell counting method and pigment synthesis was photometrically assessed. Addition of glycine resulted in an exponential increase in biomass, but not in pigment production. Tryptophan as the sole nitrogen source caused distinct brown staining of the medium, without increasing biomass. Simultaneous equimolar addition of both amino acids resulted in an initial increase in biomass as a sign of preferential metabolism of glycine, followed by a growth plateau and pigment production which, caused by higher biomass, occurred more rapidly than after addition of tryptophan alone. The yeast-cell morphology changed from round to oval. Addition of glycine to the tryptophan-containing liquid culture stopped pigment formation with simultaneous growth induction. These in vitro on-off phenomena depending on the nitrogen source might be significant in the pathogenesis of pityriasis versicolor: hyperhidrosis followed by preferential consumption of individual nitrogen sources such as glycine with exponential growth and thereafter transamination of tryptophan and TRP-dependent pigment synthesis., (© 2009 Blackwell Verlag GmbH.)

The guideline on onychomycosis, as passed by the responsible German medical societies, is presented in the present study.

The guideline tinea capitis, as passed by three German medical societies, is presented in the present study.

In diabetic patients, mycotic infections may increase the risk of developing diabetic foot syndrome. However, little data are available on the prevalence of fungal foot infections in patients with diabetes. In a first study published using data obtained during a conference attended by patients with long-term diabetes mellitus type 1 (DM1), 78/95 patients (82.1%) showed probable pedal fungal infections, of which 84.6% (66/78) were mycologically confirmed by direct microscopy and/or culture. The dermatophyte Trichophyton rubrum was the most common (69.2% of isolates). Significant correlation was found between infection and the gender (men more frequently affected) and the age of the patients. Marked mycoses on the soles of the feet were often considered to be dry skin by the patients. In a second study, 174 [31 DM1, 112 DM2 and 29 healthy accompanying persons (HAP), family members without DM] participants at a regional patients' symposium on diabetes took part in an examination for fungal infections and neuropathy of the feet. In addition to the items of the first study, we gathered data on the quality of blood glucose control (HbA1c), peripheral neuropathy (neuropathy symptome and deficit score) and measurement of sudomotoric activity by Neuropad. Mean duration of disease was 23.6 (DM1) and 11.2 (DM2) years, mean HbA1c 7.56% (DM1) and 6.89% (DM2) and fungal foot infections were confirmed at 35.5% (DM1), 53.1% (DM2) and 37.9% (HAP) respectively. In DM2, the prevalence of positive fungal samples is significantly higher for participants with less controlled blood glucose (higher HbA1c) (P = 0.04). Mycotic foot infection is also correlated with age, gender and duration of diabetes disease. Of special interest is the finding of relatively high numbers of black fungi ('Dematiaceae') (n = 10), Phialophora europea (n = 3) being the most common one. The sudomotoric activity was impaired in a very high number of participants [107/171 (61.5%)], and was found positively correlated with the prevalence of fungal foot infection in DM2 but not in DM1 and HAP. The high prevalence of fungal infections detected in DM1 as well as in DM2 diabetics is remarkable, especially considering this highly motivated collective. Therefore, it appears that the feet of diabetics require more diagnostic, therapeutic and preventive care in terms of mycotic infections and sudomotoric dysfunction than previously thought.

In Malassezia furfur, tryptophan as the main nitrogen source induces production of the potent ultraviolet-absorbing indole compound pityriacitrin. An in vitro study about the effects of pityriacitrin on other human skin microorganisms is presented, with special focus on Candida albicans and staphylococci in which its toxicity and UV-protective capacity were investigated. Candida albicans was irradiated with UVB light either in the presence or in the absence of pityriacitrin (11 mmol) and the growth rate was determined. A UVB dose of 1 J cm(-2) caused death of the fungi without pityriacitrin, whereas those in the presence of pityriacitrin showed almost unaffected growth. A diffusion test in staphylococci revealed no antibiotic effects of pityriacitrin. For testing of an ultraviolet-protective effect, staphylococci were either inoculated and irradiated in a plate model for visual assessment of growth or inoculated and irradiated in square quartz cylinders for quantitative measurement of cell density, each time in the absence or presence of pityriacitrin. Cell density of the bacterial suspensions exhibited nearly no influence of pityriacitrin on growth rates, while again a UV-protective effect was observed. In summary, pityriacitrin has an ultraviolet-protective effect on Candida albicans and staphylococci with no toxicity in the range tested.

A total of 500 combs of adult chickens from two different locations in Germany (Hessen and Schleswig-Holstein) were clinically and mycologically examined. The chickens came from three battery cages (n = 79), one voliere system (n=32), six flocks maintained on deep litter (n = 69) and 12 flocks kept on free outdoor range (n=320). Twenty-two of the 500 chicken combs (4.4%) were found to have clinical signs: only non-specific lesions neither typical of mycosis nor of avian pox such as desquamation with crust formation, yellow to brown or black dyschromic changes, alopecia in the surrounding area and moist inflammation. Only seven of the 22 clinically altered combs showed a positive mycological result; the non-pathogenic and geophilic Trichophyton terrestre in one case and non-pathogenic yeast in six cases. The following fungi were seen in the different housing systems: 13 dermatophytes (2.6% of 500 samples): 12 x T. terrestre, 1 x Trichophyton mentagrophytes, 11 isolates of Chrysosporium georgiae (2.2% of 500 samples) and 149 isolates of yeasts (29.8%): Malassezia sympodialis: n = 52, Kloeckera apiculata: n = 33, Trichosporon capitatum (syn. Geotrichum capitatum): n = 23, Trichosporon cutaneum/Trichosporon mucoides: n = 12, Trichosporon inkin (syn. Sarcinosporon inkin): n = 8 and Candida spp.: n = 21, including pathogenic or possibly pathogenic species: Candida albicans: n = 3, Candida famata: n = 4, Candida guilliermondii: n = 3, Candida lipolytica: n = 3, Candida dattila: n = 2 and one isolate each of Candida glabrata, Candida parapsilosis, Candida aaseri, Candida catenulata sive brumpti, Candida fructus and Candida kefyr sive pseudotropicalis. There is no stringent correlation between the clinical symptoms diagnosed on the chicken combs and the species of yeasts isolated. The causative agent of favus in chickens, Trichophyton gallinae, and the saprophytic yeast in pigeons, Cr. neoformans were not isolated. The most frequently isolated yeasts M. sympodialis and Kloeckera apiculata are suggested to be classified as members of the resident flora of the chicken comb.

The yeast Malassezia furfur is a part of the resident flora of human skin. It causes various diseases such as pityriasis versicolor, which hardly shows signs of inflammation despite marked clinical symptoms (e.g. hypopigmentation). The pathophysiology related morphological picture might give a clue to this phenomenon. As a part of the literature data are controversial, the present study compared the inflammatory infiltrate of pityriasis versicolor with that of tinea corporis in 40 human skin preparations each from diagnostic specimens. All preparations were stained with HE and PAS. Neutrophilic granulocytes were counted in the HE stain, and hyphae and spores in the PAS stain. The number of counted cells was related to the size of the respective area and the values were compared between pityriasis and tinea corporis. Significantly, more neutrophilic granulocytes were found with tinea corporis (P > 0.01), while they were virtually not demonstrable with pityriasis versicolor. It is surprising that fungal load in the stratum corneum is significantly higher with pityriasis versicolor (P > 0.01). Obviously the immune response involving neutrophilic granulocytes does not occur despite high bacterial load. This might be explained by reduced immunogenicity because of high content of lipids in the cell membrane. Furthermore, pityriarubins that are produced during tryptophan metabolism might be involved, which, in a stimulus-dependent manner, can suppress the ROS production of neutrophilic granulocytes in vivo.

Reference as well as field strains of Malassezia furfur (30), M. sympodialis (49), M. globosa (52), M. obtusa (one), M. restricta (one), M. slooffiae (seven), and M. pachydermatis (373) were investigated for their ability to produce pigment and fluorochromes when tryptophan (Trp) is offered as the main nitrogen source. Only the M. furfur strains produced pigment on a pigment-inducing medium (p-medium). Remarkably, the optical activity of Trp was not significant for pigment synthesis. Other nitrogen sources that are structurally similar to Trp (gramine, tryptamine, serotonin) did not induce pigment formation. All lipophilic non-furfur species failed to grow and to form pigment on this agar. However, growth of all lipid-dependent species was achieved on a modified Dixon agar in which peptone had been substituted by an equal amount of l-Trp. Here, too, the M. furfur colonies were characterized by rapidly developing dark brown halos. Furthermore, about 11% of the M. pachydermatis strains tested produced pigment formation on the p-medium, which was enhanced by addition of d-glucose. In contrast to M. furfur, pigment formation occurred after a markedly longer incubation time (4 weeks unlike 3-5 days) with a lower yield and limited color spectrum (thin layer chromatography, TLC). The UV filter pityriacitrine recently described for M. furfur was also demonstrated for M. pachydermatis by extraction, high-performance liquid chromatography (HPLC) analysis with co-elution and mass spectroscopy. The phenotypic feature of pigment formation in some strains of M. pachydermatis may confirm recent molecular-genetic findings suggesting a relationship between some strains of M. pachydermatis and M. furfur.

A new species, Phialophora europaea, member of the P. verrucosa complex, is introduced. It is distinguished from existing species by reduced, flaring phialidic collarettes and inability to assimilate melibiose as sole source of carbon. Analysis of ITS1 and 2 rDNA of six strains attributed to the species show it to be clearly individualized. All strains originated from cutaneous and nail infections of humans in North-western Europe. A key to morphologically similar taxa is provided.

The historical development of the taxonomic classification of Malassezia yeasts until today yielded the description of seven different species based upon molecularbiological, morphological and biochemical parameters (M. furfur, M. pachydermatis, M. sympodialis, M. globosa, M. obtusa, M. restricta, M. slooffiae). Tests like Cremophor EL assimilation, esculin degradation, catalase test, pigment production and determination of polidocanol sensitivity appear to be suitable procedures for routine identification of the different species. Their colonization of clinically healthy humans (in particular M. sympodialis, M. globosa, M. obtusa) and animals (M. pachydermatis) skin renders interpretation difficulties, when isolating Malassezia spp. from clinical specimens. Out of the seven species, in man the clinical significance of M. furfur for pityriasis versicolor and systemic infections appears to be accepted largely. In dogs and cats M. pachydermatis has been regarded as cause of otitis externa and seborrhoeic dermatitis. But, due to geno- and phenotypic variabilities described for M. pachydermatis field isolates further investigations concerning the taxonomic uniformity of the species are necessary.

In this double-blind clinical trial 429 patients (217 terbinafine and 212 clotrimazole) were randomized to receive twice daily terbinafine 1% topical solution for 1 week followed by a vehicle application for 3 weeks, or 1% clotrimazole solution for 4 weeks. Patients were evaluated clinically and mycologically at baseline and then at weeks one, two, four (end of treatment), and eight (end of follow-up). To be evaluable the patient needed to have a positive culture for a dermatophyte and positive KOH microscopy and a clinical diagnosis of tinea pedis (interdigital type) at baseline. Effective treatment of tinea pedis was recorded in 181 of 217 (83%) of patients treated for 1 week with terbinafine 1% solution and 174 of 212 (82%) of patients treated for 4 weeks with clotrimazole 1% solution. Mycological cure and disappearance of signs and symptoms were similar at each assessment visit in the two groups. In the subgroup of patients without any protocol violation the mycological cure rate was 95% (164 of 173) with terbinafine solution and 91% (159 of 174) with clotrimazole solution (P = 0.05). Adverse events believed to be drug-related occurred in 13 patients in the terbinafine group and 11 in the clotrimazole group (4 to 5% in each group). The events were primarily local skin reactions of mild to moderate intensity. It can be concluded that terbinafine 1% solution used for 1 week to treat tinea pedis is well tolerated and at least as effective as clotrimazole 1% solution used for 4 weeks.

In the present study, 47 wild-type isolates of the genus Malassezia were isolated from dog and cat specimens by means of a simple differentiating system recently published. The purpose was to determine whether any of the other seven Malassezia spp. apart from M. pachydermatis occur in carnivores. Of the 47 isolates, three had been obtained from cats (ear 2, skin 1) and 44 from dogs (ear 37, skin 3, faeces 2, claw and paw 2). After primary isolation, they were subcultured on mDixon agar and then purified and differentiated by means of assimilation of Cremophor EL, splitting of esculin, growth on lipid-free medium and formation of tryptophan-dependent pigments and fluorochromes. Thus, a total of 100 strains could be obtained from the 47 primary isolates. Referring to the source material, M. pachydermatis was found in 83%, M. furfur in 45% and M. sympodialis in 75%. More than 80% of cultures were mixed, comprising two or all three species; a single species was isolated in only nine cases. This shows that animals are not colonized by M. pachydermatis alone, as has been thought until now, but in nearly all cases by mixed cultures. Thus, (domestic) animals could well be a reservoir for other Malassezia species such as M. furfur and M. sympodialis. Surprisingly, Malassezia yeasts were also isolated from dog faeces, indicating that they apparently pass through the gastrointestinal tract in unchanged form after having been taken up by licking colonized areas. The survivability of Malassezia yeasts in highly acid milieu was also demonstrated in vitro. The study confirms the usefulness of the new test procedures and allows new statements concerning the epidemiology of Malassezia yeasts.

A new minimal medium consisting only of L-tryptophan (L-Trp) and a lipid source induced formation of brown pigmentation only in the species Malassezia furfur, which diffuses into the agar. Strains of the species M. sympodialis and M. pachydermatis failed to grow on this medium. On mDixon medium, however, after replacement of peptone by L-Trp, growth of all three Malassezia species was achieved. Under these conditions pigment production was observed with all M. furfur strains tested, although the results for M. pachydermatis strains were inconsistent. M. sympodialis strains showed no pigment production. On the minimal medium pigmentogenesis was induced in M. furfur by only 0.01 g% tryptophan; the pH optimum was pH 5. In all M. furfur strains, alternative amino nitrogen sources given concurrently with Trp suppressed pigmentogenesis. Furthermore, there were differences in the optimal temperature among the individual M. furfur strains. CBS 7019, CBS 6000 and CBS 6001 failed to produce pigment at 37 degrees C. The extract of the culture exhibited remarkable fluorescence, and several indole derivatives with a broad spectrum of colours were detected. This finding may have an impact on the clinical appearance of pityriasis versicolor, a very common skin disease caused by lipophilic yeasts of the genus Malassezia. We hypothesize that in pityriasis versicolor metabolic adaptation of Malassezia yeasts to altered nitrogen conditions on superficial skin might be of patho-physiological importance. Tryptophan as an inducer of pigmentogenesis probably accumulates during excessive sweating, a well-known manifestation of pityriasis versicolor.

A new minimal medium consisting only of L-tryptophan (L-trp) and a lipid source induced formation of brown pigmentation only in the species Malassezia furfur. Strains of the species M. sympodialis and M. pachydermatis failed to grow on this medium. Pigmentogenesis was already induced in M. furfur by 0.01 g% tryptophan, the pH optimum was pH = 5. Alternative amino nitrogen sources given concurrently with trp suppressed pigmentogenesis. The extract of the culture exhibited remarkable fluorescence, and several indole derivatives with a broad spectrum of colors were detected by means of mass spectroscopy and NMR. This finding may have an impact on the clinical appearance of pityriasis versicolor, a very common skin disease caused by lipophilic yeasts of the genus Malassezia. We hypothesize that in pityriasis versicolor metabolic adaptation of Malassezia yeasts to altered nitrogen conditions on superficial skin might be of pathophysiological importance. Tryptophan as inductor of pigmentogenesis probably cumulates during excessive sweating, a well known manifestation factor of pityriasis versicolor.

In recent years, the genus Malassezia was reclassified based on molecular data; in addition to M. furfur, M. pachydermatis and M. sympodialis, four new species (M. globosa, M. obtusa, M. restricta, M. slooffiae) were described. Primary keys for routine identification have recently been presented. Polidocanol was shown to have specific inhibitory effects against Malassezia spp. In an agar diffusion test, type strains of all Malassezia species were incubated with polidocanol concentrations between 0.01% and 10%. M. furfur strains were most resistant, with minimal inhibitory concentrations (MIC) ranging from 7.5% to 10%. Inhibitory concentrations of the other strains were lower by at least one factor of ten. Most sensitive were strains of M. pachydermatis (0.05%). In a further test, polidocanol-containing olive oil was used to determine the sensitivity of Malassezia furfur and M. sympodialis. Again, the inhibitory concentrations for strains of M. sympodialis were one tenth of those found for M. furfur. In addition to its antifungal effects, polidocanol might therefore be a useful tool in differentiating Malassezia species.

Apart from pityriasis versicolor, Malassezia furfur is thought to play a significant role in the pathogenesis of seborrhoic eczema and Malassezia folliculitis. However, it has not been clarified whether in addition to host factors (e.g. immune status, greasy skin), yeast-dependent activities are responsible for manifestation of the disease. In this context interstrain variability of hydrolase activity of Malassezia isolates might be significant. For determination of hydrolase activity, washed yeast suspension was applied to selective agar for pathogenic fungi containing 8% (v/v) Tween 80 or Tween 60 and was incubated at 37 degrees C for 10 days. Growth was accompanied by formation of a dense white zone around the colony, in which free fatty acids corresponding to Tween 80 (C18:1) or Tween 60 (16:0, 18:0) were demonstrated by means of thin layer and gas chromatography. Thus, this phenomenon is a parameter for yeast-dependent hydrolysis of Tween 80 and Tween 60. Considering different growth behaviour, a "hydrolase zone' (H2) was determined using the quotient colony diameter/(ring+colony) diameter in each of the 150 strains tested. Although no significant H2 variations were observed in strains of different clinical origin, the present study revealed that in addition to a known enzyme, which is located within the cell wall and/or membrane systems, these Malassezia isolates produce a very active hydrolase diffusing into the medium.

The tea fungus 'Kombucha' is a symbiosis of Acetobacter, including Acetobacter xylinum as a characteristic species, and various yeasts. A characteristic yeast species or genus has not yet been identified. Kombucha is mainly cultivated in sugared black tea to produce a slightly acidulous effervescent beverage that is said to have several curative effects. In addition to sugar, the beverage contains small amounts of alcohol and various acids, including acetic acid, gluconic acid and lactic acid, as well as some antibiotic substances. To characterize the yeast spectrum with special consideration given to facultatively pathogenic yeasts, two commercially available specimens of tea fungus and 32 from private households in Germany were analysed by micromorphological and biochemical methods. Yeasts of the genera Brettanomyces, Zygosaccharomyces and Saccharomyces were identified in 56%, 29% and 26% respectively. The species Saccharomycodes ludwigii and Candida kefyr were only demonstrated in isolated cases. Furthermore, the tests revealed pellicle-forming yeasts such as Candida krusei or Issatchenkia orientalis/occidentalis as well as species of the apiculatus yeasts (Kloeckera, Hanseniaspora). Thus, the genus Brettanomyces may be a typical group of yeasts that are especially adapted to the environment of the tea fungus. However, to investigate further the beneficial effects of tea fungus, a spectrum of the other typical genera must be defined. Only three specimens showed definite contaminations. In one case, no yeasts could be isolated because of massive contamination with Penicillium spp. In the remaining two samples (from one household), Candida albicans was demonstrated. The low rate of contamination might be explained by protective mechanisms, such as formation of organic acids and antibiotic substances. Thus, subjects with a healthy metabolism do not need to be advised against cultivating Kombucha. However, those suffering from immunosuppression should preferably consume controlled commercial Kombucha beverages.

In vitro antifungal properties of polidocanol (Thesi), a hydroxypolyethoxydecan, were investigated against various yeasts at concentrations of 0.05-10% (w/w) by means of the agar diffusion test; in the case of lipophilic Malassezia furfur, polidocanol-containing olive oil was used additionally for sensitivity study. Six strains of M. furfur of different clinical origin were tested as well as strains of C. albicans (3), C. krusei (3), C. parapsilosis (2), C. robusta (2), Tr. cutaneum (2) and one strain each of C. guilliermondii, C. glabrata, C. tropicalis and M. pachydermatis. Both test systems revealed predominantly fungistatic activity against M. furfur, with a minimal inhibitory concentration (MIC) of 1% polidocanol (w/w), while yeasts of other genera (Candida species, Trichosporum species) showed no or only transitory (C. krusei) inhibition of growth. M. pachydermatis was also found to be sensitive with a MIC of 0.1%; this suggests a specific inhibitory effect against the genus Malassezia. Polidocanol, which has been used for decades as an antipruritic and analgesic in various topical preparations at concentrations between 3% and 5%, might therefore be suitable for prophylaxis of recurrent pityriasis versicolor or in veterinary medicine.