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SDS-PAGE separation of dermatophyte antigens, and western immunoblotting in feline dermatophytosis.

Water-soluble antigens liberated from the disrupted mycelium of nine dermatophytes (seven isolates of Microsporum canis, one each of Microsporum gypseum and Trichophyton mentagrophytes) were compared by analytical slab SDS-PAGE. No substantial differences were observed between the protein bands of the M. Canis isolates, but certain distinctive bands were apparent in the other two species examined. Western immunoblotting using M. canis-derived antigens separated by SDS-PAGE was used to investigate the humoral immune response in 79 cats with naturally-occurring dermatophytosis (72 with M. canis, six with M. gypseum and one with T. mentagrophytes) and this information was compared to results of immunoblots from 46 control (non-dermatophyte exposed) cats. Seven dominant bands (bands which occurred frequently and stained heavily) were identified in immunoblots from the dermatophyte-infected cats with apparent molecular weights varying between 39 and 120 kD. None of these bands were totally specific markers for dermatophytosis as a variable proportion of the control cats showed reactivity to all these proteins. However, most (73%) of the dermatophyte-infected cats showed reactivity to six or seven of the identified bands whereas most (80%) of the control cats showed reactivity to between zero and three of these bands (p < 0.005). Western immunoblotting could be used to select individual immunodominant antigens for further evaluation of protective (cell-mediated) immunity.

Animals↗

PCR and PCR-RFLP techniques targeting the DNA topoisomerase II gene for rapid clinical diagnosis of the etiologic agent of dermatophytosis.

BACKGROUND: We have focused on the DNA topoisomerase II genes of several pathogenic fungi, and developed polymerase chain reaction (PCR) and PCR-restriction fragment length polymorphism (RFLP) methods targeting this gene for identification of dermatophytes. OBJECTIVE: To assess the availability of the PCR-based identification for an etiologic study of dermatophytosis, by testing these PCR and PCR-RFLP methods for stability and reproducibility. METHODS: Three hundred and fifty-six dermatophyte strains were isolated from 305 patients with tinea, and their genomic DNAs were used as templates for the PCR using primer mixes (PsT, PsME, dPsD1 or dPsD2) composed of gene-specific primers for identification of dermatophytes to the species level. The genomic DNAs of Trichophyton rubrum were further subjected to subrepeat element analysis of the nontranscribed spacer (NTS) of ribosomal DNA (rDNA). RESULTS: In this study, six dermatophyte species (T. rubrum, Trichophyton mentagrophytes, Trichophyton tonsurans, Microsporum canis, Microsporum gypseum, and Epidermophyton floccosum) were obtained. In all cases, the identifications obtained from the PCR and PCR-RFLP targeting the DNA topoisomerase II gene coincided with those from the conventional morphological features-based identification technique. The sensitivity of the PCR-based identification was found to be a colony of approximately 3mm in diameter. Furthermore, T. rubrum was divided into three groups (17 types) on the basis of the sizes and numbers of the products generated from the TRS-1 region, and three types from the TRS-2 region. CONCLUSION: The PCR and PCR-RFLP targeting the DNA topoisomerase II gene were rapid, stable, and reproducible for species identification of dermatophytes, and thus are convenient tools for an etiologic study of dermatophytosis.

Adolescent↗

Occurrence of keratinolytic fungi and related dermatophytes in soils in Cairo, Egypt.

120 soil samples collected from various sites of Cairo were processed for the isolation of keratinophilic fungi by "ToKaVa" hair baiting technique. 22 species belonging to 6 genera were isolated viz.: Chrysosporium tropicum, C. indicum, C. keratinophilum, C. queenslandicum, C. merdarium, C. anamorph of Arthroderma curreyi, C. pannicola, C. lobatum, C. anamorph of Renispora flavissima, C. pseudomerdarium, Microascus mangini, Malbranchea gypsea, Ml. State of Uncicarpus reesii, Ml. State of Coccidioides immitis, Microsporum gypseum, Mr. distortum Mr. audouinii, Mr. fulvum, Trichophyton mentagrophytes, T. terrestre, T. verrucosum and Epidermophyton floccosum. The frequency of occurrence of the isolated fungi was determined. Microsporum gypseum, Chrysosporium tropicum and Chrysosporium indicum were the most frequent species recovered from soil. Most species of keratinophilic fungi were isolated from university, public garden and zoo garden. The distribution of the isolates are discussed.

Arthrodermataceae↗

In vitro antimicrobial activity of ethanol and water extracts of Cassia alata.

Crude ethanol and water extract of leaves and barks from Cassia alata were tested in vitro against fungi, (Aspergillus fumigatus and Microsporum canis), yeast (Candida albicans) and bacteria (Staphylococcus aereus and Escherichia coli). C. albicans showed concentration-dependent susceptibility towards both the ethanol and water extracts from the barks, but resistant towards the extracts of leaves. The degree of susceptibility varied, the water extract from barks showed bigger inhibition zone than the ethanol extracts (12-16 and 10-14 mm, diameter respectively). The growth of Aspergillus fumigatus and Microsporum canis were not affected by all types of the plant extracts. Results were comparable to standard antifungal drug Tioconazole (18 mm diameter) at equivalent concentration. The anti-bacterial activity of C. alata extracts on S. aureus was detected with only the leaves extracts using water and ethanol. The water extract exhibited higher antibacterial activity than the ethanol extract from leaves (inhibition zones of 11-14 and 9-11 mm, respectively). E. coli showed resistance to all types of extracts. Based on the current findings, it can be concluded that this plant has antimicrobial activity, which is as potent as standard antimicrobial drugs against certain microorganisms.

Anti-Bacterial Agents↗

In vitro evaluation of antifungal properties of 8.O.4'-neolignans.

Eighteen racemic 8.O.4'-neolignans with six different substitution patterns in rings A and B, in their ketone and in their erythro and threo alcoholic forms, were evaluated for antifungal activity by the agar dilution method. Only the alcohols exhibited a broad spectrum of activities against Microsporum canis, Microsporum gypseum, Tricophyton mentagrophytes, Tricophyton rubrum, and Epidermophyton floccosum. (+/-)-erythro-3,4-(methylenedioxy) -7-hydroxy-1'-allyl-3',5'-dimethoxy-8.O.4'-neolignan (11) was the most active compound in the series, and E. floccosum was the most susceptible species.

Antifungal Agents↗

Efficacy of chlorhexidine against some strains of cultured and clinically isolated microorganisms.

The efficacy of chlorhexidine digluconate was determined against some strains of collected and clinically isolated bacteria and fungi. The efficacy was evaluated either by calculating a minimum inhibitory concentration (MIC) or by efficacy trials according to the guidelines of the European Committee for Standardization. The MIC values of chlorhexidine for Staphylococcus aureus, Microsporum gypseum, Microsporum canis and Trichophyton mentagrophytes were 0.625 microg/ml, 12.5 microg/ml, 50 microg/ml and 6.25 microg/ml, respectively. The in vitro efficacy of chlorhexidine was higher against ATCC strains of S. aureus and P. aeruginosa (0.5 mg/ml for 5 min and 0.5 mg/ml for 10 min, respectively) than against clinical isolates (0.5 mg/ml for 15 min and 1 mg/ml for 10 min, respectively). The antiseptic activity of aqueous solutions of chlorhexidine against spores of Bacillus subtilis, Bacillis sfericus and Clostridium perfringens required longer contact times than against the vegetative forms. Nevertheless, 5 mg/ml of chlorhexidine in water-ethanol 20:80 v/v was totally effective against the vegetative forms or spores of these microorganisms.

Anti-Infective Agents, Local↗

Occurrence of keratinophilic fungi and dermatophytes on domestic birds in Nigeria.

Feathers, nails and beaks of one hundred and twenty common birds in Nigeria, Chicken [50], Ducks [20], Turkeys [15] and Pigeons [35], were examined using the soil plate technique for their mycoflora. 15 species of fungi were recovered and they belong to the genera Chrysosporium, Trichophyton, Microsporum, Aspergillus, Fusarium, Mucor Rhizopus, Penicillium and Trichoderma. Microsporum gypseum was the species most frequently isolated (35% of the samples). The most common genus was Chrysosporium and C. keratinophilum was the species with the highest frequency in the genus (28.3%). The species isolated included potential pathogens and mycotoxin producing fungi (Aspergillus flavus, A. niger, Fusarium oxysporum).

Animals↗

Incidence of dermatophytes and cyclohexamide resistant fungi on healthy children hairs and nails in nurseries.

In order to estimate the prevalence of dermatophytes and other fungi on healthy children hairs and nails, 92 hair samples and 85 nail samples (groups of 10 finger nails from each child) were collected from 5 nurseries (children aged 9 months up to 4 years) in Assiut city. From hair samples 22 species were collected, Trichophyton (2 species) and Microsporum (2 species) were the only recovered dermatophytes in addition to well known keratinophilic genus Chrysosporium (4 species). From nail samples, 18 species were identified, Trichophyton was represented by 4 species, Microsporum, 2 species and Chrysosporium, 4 species. Also, several other saprophytes and cycloheximide resistant fungi were isolated.

Antifungal Agents↗

Molecular taxonomy of dermatophytes and related fungi by chitin synthase 1 (CHS1) gene sequences.

In the present study, the nucleotide sequences of the CHS1 gene from dermatophytes and related fungi in the genera Chrysosporium, Epidermophyton, Microsporum and Trichophyton were investigated using molecular methods. About 440-bp genomic DNA fragments of the CHS1 gene from 21 species were amplified by polymerase chain reaction (PCR) and sequenced. The CHS1 nucleotide sequences of these fungi showed more than 83% similarity. The molecular taxonomy of the CHS1 gene sequences revealed that Microsporum was genetically distinct from Chrysosporium and Trichophyton, as classified by morphological characteristics.

Arthrodermataceae↗

Tinea capitis in Basrah, Iraq.

Tinea capitis is the most common type of dermatophytosis constituting 35.2% (173/491) of all cases of dermatophytosis seen at two out-patients dermatological clinics of the Basrah General Hospital and at two private dermatological clinics between October 1994 and December 1995. Males were more commonly affected than females represented by 60 and 40%, respectively. Out of 173 mycologically positive cases, 143 specimens were culture-positive, represented by five dermatophyte species; Trichophyton violaceum was the most frequent causative agent (38.5%), followed by Trichophyton verrucosum (28.7%), Microsporum canis (26.5%), Trichophyton mentagrophytes var. mentagrophytes (5.6%) and Microsporum gypseum (0.7%). Six atypical cases appeared as cases of seborrheic dermatitis or dandruff and were proved to be atypical cases of tinea capitis caused by T. violaceum.

Adolescent↗

The aetiological agents of tinea capitis in Zaragoza (Spain).

Between 1977 and 1997, 190 cases of tinea capitis were observed. The age groups most commonly infected were 1-10 years old. The following dermatophytes were isolated: Microsporum canis (119), Trichophyton mentagrophytes (57), Trichophyton tonsurans (seven), Trichophyton verrucosum (three), Trichophyton violaceum (one), Trichophyton schoenleinii (one), Trichophyton soudanense (one) and Microsporum audouinii (one).

Adolescent↗

Dermatomycoses in the Rijeka area, Croatia, before, during and after the war 1990-1999.

The frequency of dermatomycoses as well as the spectrum of causative agents in the Rijeka area, northwestern part of Croatia, in the periods before and during the war (1990-1994) and in the postwar period (1995-1999) were analyzed. A total of 20 463 patients with clinically suspected dermatomycoses referred to the Department of Dermatovenerology, Clinical Hospital Center Rijeka, was examined. In the period from 1990 till 1994, 2462 samples were positive, compared with 2250 positive samples during the period from 1995 to 1999. The most frequently isolated dermatophytes were in both periods Trichophyton mentagrophytes and Microsporum canis, followed by Microsporum gypseum, Epidermophyton floccosum, Trichophyton verrucosum and in the first period T. rubrum. An intensive change in composition of the flora was observed in the postwar period, mainly connected to the migration of population from other regions to the Rijeka area. This was demonstrated by a disappearance of T. verrucosum and T. rubrum and by an eruptive occurrence of Trichophyton violaceum. This antropophilic species became third representative in the postwar period, followed by M. gypseum, and E. floccosum. Our data were compared with those obtained in the same area in previous studies, and with results obtained in other countries.

Croatia↗

Serological differences among the dermatophytes.

Antigens were prepared from young mycelial growth of 20 species of dermatophytes, and tested by double diffusion against homologous and heterologous antisera raised in rabbits. 48 distinct antigens were recognised by the procedures used. Although there were a considerable number of common reactions, there were significant differences between species and groups. Species of Microsporum, with the exception of M. gypseum and M. persicolor, form a coherent group distinct from Epidermophyton and Trichophyton. All Trichophyton species investigated with the exception of T. ajelloi, were serologically closely related. M. gypseum, T. ajelloi and M. persicolor showed affinities with both Microsporum and Trichophyton, and possibly form an intermediate group. The system used does not permit differentiation of species by serological means, but gives a new dimension to the inter-relationships of these fungi.

Antigen-Antibody Reactions↗

Etiology of ringworm of the scalp, beard and body in Rome, Italy.

From 1972 to 1977 the authors examined 1,311 patients in Rome affected by ringworm of the scalp, beard and body, comparing their findings with a study made in Milan between 1969 to 1973 and with earlier Italian studies from 1900 to date. It appears that at the beginning of the century scalp ringworm was usually caused by Trichophyton violaceum and T. tonsurans, whereas Microsporum canis was rarely found. Today the Trichophyton species are rarely isolated, whereas Microsporum species, particularly M. canis (88.9% of the cases in Rome, 74.6% in Milan), predominate in the etiology of scalp ringworm. Ringworm of the beard in Rome is mainly caused by T. mentagrophytes (50% of the cases) and T. verrucosum (25%). In Milan the frequency of T. verrucosum is 51.2% and of T. mentagrophytes 38.5%. Ringworm of the body today is mainly due to M. canis (47.8% of the cases in Rome and 65.7% of those in Milan). These ringworms in Rome and Milan are caused mostly by the zoophilic dermatophytes that have supplanted the anthropophilic dermatophytes, which were the main cause of dermatomycoses in the early years of this century. This change, particularly evident after World War II, appears to result from improved socio-economic and hygienic conditions, with resulting decrease of infections through interhuman contacts, and from increased numbers of pet animals and consequent increase of stray cats and dogs, which result in increase of infections of animal origin.

Epidermophyton↗

A study of tinea capitis in Sri Lanka.

Between January 1978 and December 1987, 106 cases of tinea capitis were detected in Sri Lanka. Diagnosis was confirmed mycologically by microscopic examination and by culture. Tinea capitis accounted for 3.4% of all superficial fungal infections seen during this period. This figure is low when compared with the incidence of this disease reported in other countries. Five main types of clinical manifestation were encountered. Inflammatory lesions were observed in 50% of the patients. Eighty-five percent of infections occurred in children below 15 years of age and 76% in children under 10 years of age. The 6-10 year age group was the most vulnerable. The zoophilic fungi Trichophyton mentagrophytes, Microsporum canis and the geophilic species Microsporum gypseum were together responsible for 81% of tinea capitis infections, and the zoophilic fungus Trichophyton simii accounted for a further 10% of infections. It is postulated that the Sri Lankan custom of frequent head baths with soap and water, especially among children, may account in part for the low prevalence of tinea capitis in the community.

Adolescent↗

Seasonal 4-year investigation into the role of the alpine marmot (Marmota marmota) as a carrier of zoophilic dermatophytes.

Two hundred and six samples of alpine marmot (Mamota marmota) hair (148 from adults and 58 from young subjects), 102 soil samples from the entrances to the burrows of the above individuals and 20 control specimens (obtained from adjoining areas away from the burrow systems where the rodents are not usually present) were examined from May 1994 to September 1997. Seventy-five isolates belonging to six species of dermatophytes were found in 69 of the 206 hair samples examined (33.5%). Two species were zoophilic, Microsporum canis (7.8%) and Trichophyton mentagrophytes (11.2.%), and four geophilic, Microsporum cookei (2%), M. gypseum (5.8%), Trichophyton ajelloi (3.9%) and T. terrestre (5.8%). The prevalence of each species in the hair samples did not change significantly according to year, season (chi-squared test [limit significance: P <0.05] gives no significant values [P>0.05] both in year and in season comparison) or age/sex (adult versus juvenile: P=0.1; male versus female: P=0.8) of the marmot. Twenty-three of the 102 soil samples (22.5%) were positive for dermatophytes found in the hair of marmots from the same burrow systems. Five of the 20 control soil samples (25%) were positive for dermatophytes. One isolate of M. gypseum, three of T. terrestre and one of T. mentagrophytes were obtained. Compared with other free-ranging rodent hosts studied in Europe, this mycoflora is characterized by the presence and relatively high prevalence of M. canis, frequently reported in symptomatic and asymptomatic cats, dogs and fur animals. M. canis has not been isolated in other rodents in the wild. However, it has recently been reported in asymptomatic foxes (Vulpes vulpes) from northern Italy. The close link between V vulpes and M. marmota, with the former representing the most important mammal predator of the latter in the Alps (only a fraction of the predator's attacks result in the death of the rodent) may have favoured the adaptation of M. canis to this rodent host. The stable character of the M. canis/M. marmota relationship (no seasonally or annually related difference in the prevalence of this dermatophyte has been found) suggests the inclusion of the alpine marmot in the reservoir of this zoophilic pathogenic agent. In this situation, hibernation in labyrinthine burrow systems, where temperature and moisture ranges are quite uniform the whole year round, may favour the viability of M. canis arthroconidia, whose survival in mountain habitat might otherwise be compromised. This seems to be confirmed by the fact that the fungus has never been found in the control samples collected at a distance of 300 m from the outer edge of the sampled burrow systems.

Animals↗

Utilization of standard laboratory methods in the laboratory diagnosis of problem dermatophytes.

The identification of certain dermatophytes may be simplified by using biochemical tests such as urease, nutritional requirements (with commercially available media), and the in-vitro hair penetration test. No study that combines these tests in a diagnostic scheme for identification of the common dermatophytes has been published. one to 20 isolates each of 29 species of dermatophytes (one Epidermophyton floccosum, 10 Microsporum species, and 18 Trichophyton species) were used. They were grown on Christensen's urea agar and the seven nutritional media for the differentiation of the Trichophyton species; the in-vitro hair penetration test was performed in duplicate. Patterns were developed that have been tested and proven to be useful for more than 22 months. In addition, the Microsporum species were all grown on polished rice, and color and sporulation were recorded. All dermatophytes in this study were grown on either Mycosel agar or 2% malt extract agar, and on modified potato dextrose agar and modified Sabouraud agar. Macroscopic and microscopic examinations were made to determine qualitatively the amounts of growth and sporulation on each medium.

Arthrodermataceae↗

Dermatophytosis in children and adolescents: epidemiological, clinical, and microbiological aspects changing with age.

During a four year period, a total of 84 cases of dermatophytosis were seen in patients from 0 to 17 years of age attending our out-patient department. The sex distribution was about equal in preadolescents. Males predominated among adolescents. A peak incidence at early school age was due to cases of tinea capitis, mainly caused by Microsporum canis. A peak among adolescents was due to cases of tinea pedis, mainly caused by Trichophyton rubrum. In general, patients with Trichophyton rubrum infections were older than those with other types of dermatophytosis (p less than 0.05). Dermatophytosis due to Microsporum canis was most frequently seen in January and in September (p less than 0.01). Although onychomycosis due to dermatophytes was a comparatively rare disease, it did occur within all age subgroups. Entities which were very frequent in former times and which have become rare during the last decades, such as tinea capitis due to Trichophyton schönleinii, still contribute to the spectrum of dermatophytosis in childhood.

Adolescent↗