Study of onychomycosis in Córdoba,Spain: Prevailing fungi and pattern of infection

From a total of 20 004 patients seen during two years, we carried out a mycologic nail investigation (direct microscopy and repeated cultures). Ninety-three (43.2%) of the nails were judged to be infected by their clinical appearance. They fulfilled the laboratory criteria required to start antifungal treatment (isolation of the same fungus in culture on two consecutive occasions), but only in 64 cases (29.7%) was there a clinical and mycological recovery once antifungal treatment and follow up were completed.Yeasts were isolated in two thirds of the cases of onychomycosis, mainly from fingernails. Candida albicans, C. parapsilosis or both were the most prevalent species. Dermatophytes were found in 18.8% of the samples, especially from toenails. Trichophyton rubrum was the predominant species. Non-dermatophytic filamentous fungi were cultured in 17.2%, Scopulariopsis brevicaulis being the most prevalent species.The highest prevalence of onychomycosis was found in patients between 50 and 70 years of age. Females were affected more frequently than males. Fingernails were affected more frequently than toenails. Proximal subungual onychomycosis, secondary to paronychia (PSOp), was the most prevalent clinical type, although primary distal and lateral subungual onychomycosis (DLSO) and total dystrophic onychomycosis (TDO) were also frequent. PSOp was only observed in fingernails, while DLSO was almost only seen in toenails and TDO in both fingernails and toenails. All the clinical types were more frequent in women except TDO, which showed a similar prevalence in both sexes.This revised version was published online in October 2005 with corrections to the Cover Date.     

Dermatomycosis in lower limbs of diabetic patients followed by podiatry consultation

BackgroundDiabetic patients are particularly susceptible to fungal infections due to modifications that occur in their immunological system. These modifications compromise natural defences, such as skin and nails, especially from lower limbs.AimsAssessing the presence of dermatomycosis in lower limbs of Portuguese diabetic patients followed on Podiatry consultation. Determination of possible predisposing factors and the most frequent fungal species associated with the cases are included in the study.MethodsA six-month prospective study was carried out in 163 diabetic patients with signs and symptoms of dermatomycosis followed by Podiatry at the Portuguese Diabetes Association in Lisbon. Samples from the skin and/or nails of the lower limbs were collected and demographic and clinical data of those patients were recorded.ResultsTrichophyton rubrum was the most frequently isolated dermatophyte (12.1%), followed by Trichophyton mentagrophytes (7.7%) and Trichophyton tonsurans (4.4%). Our study showed positive associations between type 2 diabetes and the presence of dermatomycosis in the studied population (p = 0.013); this association was also shown between the occurrence of dermatomycosis and the localization of the body lesion (p = 0.000). No other predisposing factor tested was positively associated with infection (p > 0.05).ConclusionsData on superficial fungal infections in diabetic patients are scarce in Portugal. This study provides information on the characterization of dermatomycosis in lower limbs of diabetic patients.     

Onychomycosis: Multicentre epidemiological, clinical and mycological study

BackgroundOnychomycosis accounts for up to 50% of all nail disorders. They can be caused by: yeasts, dermatophytes and non-dermatophyte moulds.Objectives and methodsA multicentre study designed to determine the prevalence, mycological test results, aetiological agents, and clinical presentation of onychomycosis was carried out. All fingernail and toenail samples taken during a one year period at 9 diagnostic centres were included.ResultsA total of 5,961 samples were analysed, of which 82.3% were from toenails and 17.7% from fingernails. The mean age of the patients was 49.7 years, and 66% were females. Direct microscopic examination was positive in 61% of the samples. In adults, 61.2% of toenails were positive using potassium hydroxide (KOH), and 43.7% were positive in cultures. The prevailing aetiological agents belong to the dermatophyte group (82.8%), and distal subungual was the most common clinical form. In fingernails, direct examination showed 59.8% positive samples, and cultures were positive in 52.9%. The prevailing agents were yeasts belonging to Candida species, and onycholysis was the most common lesion.ConclusionsDirect mycological examinations were positive in 61%, a higher value than that found in other series. Dermatophytes were prevalent in toenails of both sexes, and in finger nails yeast were prevalent in females, and dermatophytes in males. Non-dermatophyte moulds corresponded to 4.8% of toenail and 2.05% of fingernails isolates.     

Onychomycosis in Ceará (Northeast Brazil): epidemiological and laboratory aspects

Knowledge of epidemiological and mycological characteristics of onychomycosis has been noted by many authors as being an important tool for control of these fungal infections. This study seeks to improve knowledge of onychomycosis epidemiology and mycological features. Samples were taken from infected fingernails and toenails of 976 patients undergoing treatment at a respected Dermatology Center in Ceará, Fortaleza, CE, Brazil. Specimens from 512 patients (52%) were positive for onychomycosis. From the culture-positive samples, yeasts of the genus Candida (C. albicans, C. tropicalis, C. krusei, C. parapsilosis) were dominant. The dermatophytes isolated (Trichophyton rubrum, T. tonsurans, T. mentagrophytes var. mentagrophytes) were dominant in 46 patients (12.99%). The mould Fusarium spp. was isolated from 29 patients (8.19%). Yeast of the genus Candida is the main causal factor in onychomycosis in our region. Also, the study showed the importance of performing direct examination and culture in diagnosis of onychomycosis.     

<Emphasis Type="Italic">Trichophyton rubrum</Emphasis> Infection Characterized by Majocchi’s Granuloma and Deeper Dermatophytosis: Case Report and Review of Published Literature

Infections caused by Trichophyton rubrum are very common in dermatological disease. It most often appears as superficial cutaneous mycosis, such as tinea manuum, tinea pedis, and tinea corporis. However, deep infection caused by T. rubrum was rarely reported. We describe a case of mixed type of deep infection caused by T. rubrum in a 45-year-old man with no significant immunodeficiency. This patient had a history of onychomycosis on the toenails without regular treatment for nearly 6 years. And, he had erythema, papule, and nodules on the submandibular area, neck, and chest for almost 1 year. After treated with intravenous infusion of cefotiam for 2 weeks, the lesion aggravated. The fungal direct microscopic examination of pyogenic fluid was positive, and the fungal cultures that produced reddish-brown and yellow pigment showed cottony, wooly, and white colony. After the DNA sequencing, it was identified as T. rubrum. We gave the patient oral terbinafine 250 mg per day and bifonazole cream for external use. Six months later, the patient’s skin lesion was disappeared, and healthy nail growth was seen in two-thirds of nail bed. The terbinafine is effective against deep infection caused by T. rubrum.     

ONYCHOMYCOSIS OF THE FEET—Treatment with Griseofulvin

Griseofulvin, a new orally administered antifungal antibiotic which has proved to be effective for the treatment of a wide variety of superficial fungus infections of man, was used in the treatment of 51 patients with infections of the toenails due to T. rubrum. Thirty-four of the patients were treated with griseofulvin alone and seven were treated with griseofulvin combined with surgical avulsion of all involved toenails. The remaining ten had bilateral infections, and avulsion was done on one foot but not the other before griseofulvin therapy was begun.Of 34 patients who were treated with griseofulvin alone, few had complete cure even after prolonged treatment. Some nails showed improvement for a time, then no further gain; some showed no improvement; some showed resistant wedges of infection which penetrated proximally toward the posterior nail fold.In the instances of surgical avulsion, clinically normal nails regrew during griseofulvin therapy. This simple procedure, with thorough removal of all underlying keratinous debris, apparently did away with foci of possible reinfection.The results of the study indicated that surgical avulsion of the toenails in combination with griseofulvin therapy is an effective and practical method of treating onychomycosis of the toenails due to T. rubrum.     

An in vitro study of the photodynamic effect of rose bengal on trichophyton rubrum

Onychomycosis, a fungal infection of the finger or toenails, is predominantly caused by Trichophyton rubrum. Treatment is difficult due to high recurrence rates and problems with treatment compliance. For these reasons, alternative therapies are needed. Here we describe the photoactivation of Rose Bengal (RB) using a green laser (λ = 532 nm) at fluences of 68, 133 and 228 J/cm², and assess its fungicidal activity on T. rubrum spore suspensions. A 140 µM RB solution was able to induce a fungicidal effect on T. rubrum when photosensitized with the fluence of 228 J/cm². RB photosensitization using a green laser provides a potential novel treatment for T. rubrum infections. (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Erratum to: The Interplay Between NSAIDs and Candida albicans on the Gastrointestinal Tract of Guinea Pigs

Trichophyton rubrum is an anthropophilic species that is the most frequent etiologic agent of human dermatophytosis throughout the world. No teleomorph has been identified for T. rubrum strains. This study used PCR analysis to confirm the presence of a mating type locus in the genome of Japanese isolates of T. rubrum. To clarify the epidemiological and ecological characteristics of this fungus, mating type sequences were tested for correlation of MAT genotype to mating type. This study examined clinical isolates of T. rubrum that had been obtained from 206 human cases of tinea pedis and tinea unguium in Japan, including those from Fukuoka (29 strains), Gifu (23 strains), Kanazawa (63 strains), and Tokyo (91 strains), along with 10 isolates derived from 10 cases of canine dermatophytosis. PCR detected the presence of MAT1-1 in all of the human and animal isolates. Therefore, all isolates examined were expected to react as (?) type on the mating test and not as (+) type.     

Clinico-mycological Profile of Dermatophytosis in a Reference Centre for Leprosy and Dermatological Diseases in Addis Ababa

To assess the clinical and fungal species spectrum of dermatophyte infection in a reference centre in Addis Ababa, 539 dermatological patients with signs of dermatophytosis were investigated. Seventy-one percent were female and 29% male, aged 2–66 years (median 9). Four hundred-fifteen (77%) had at least one skin lesion. Tinea capitis was diagnosed in 138/155 males (89%) as compared to 214/384 females (40%) (p<0.05). T. capitis was diagnosed in 69% of the 374 children. Fingernails were affected in 132/145 (91%) of onychomycosis, 118 (90%) of these patients were females and 14 males (p<0.05). Tinea corporis was observed in 45, and other types of tinea in 12 patients. Thirty-six percent of all patients had also other skin lesions, mostly impetigo. Of 490 cultured samples 364 (74%) grew dermatophytes: Trichophyton violaceum in 84%, Trichophyton verrucosum in 9.6%, Trichophyton tonsurans in 1.4% and T. rubrum in 0.5%. Additionally, 15 isolates were identified as white variants of T. violaceum, in 3 cases confirmed by sequencing of the rDNA ITS 2 region. T. capitis in young males and T. unguium of fingernails in females were the most common manifestations of dermatophytosis in Addis Ababa, usually caused by T.violaceum.     

The frequency of Candida species in onychomycosis

Mycological investigation of 108 nail specimens taken from a total of 41 patients examined over three years included direct microscopy and repeated cultures. A higher incidence of onychomycosis of the fingernails (75%) was observed in women while afflictions of the toenails (71%) prevailed in men. The highest prevalence of onychomycosis was found in patients between 50 and 70 years of age.Candida albicans was the dominant organism causing onychomycosis (prevalence rate 60.9%), followed byC. parapsilosis (19.6%),C. tropicalis (9.8),C. krusei (4.9),C. guilliermondii andC. zeylanoides (2.4% each).     

Dermatophytosis in Military in the Central-West Region of Brazil: Literature Review

Background

Dermatophytes represent a group of keratinophilic fungi capable of invading the superficial layer of the skin, hair, and nails of humans and animals. There is a high prevalence of dermatomycosis in tropical regions, and military personnel are susceptible to this kind of infection due to the type of occupational activities.

Objective

This study was to investigate dermatophytosis in military, in addition to predisposing factors to such infections.

Methods

The direct examination of the 221 samples obtained was conducted by preparing fresh slides, clarified with KOH. The clinical materials were seeded in duplicate in SDA and in Mycosel agar medium. The identification of the etiologic agents was performed according to the Riddell technique.

Results

99/221 (44.8 %) of the dermatophyte infection agents distributed in the three taxonomic genera that cause Tinea were recovered. The Trichophyton genus was the most representative and T. rubrum species 33 (33.3 %), the most prevalent. The other species found were as follows: T. tonsurans 13 (13.1 %), T. verrucosum 11 (11.1 %), T. interdigitale 9 (9.1 %), and T. mentagrophytes 6 (6.1 %). Among the most affected anatomical sites were skin 83 (83.8 %) and nails 17 (17.2 %).

Conclusions

Dermatophyte infections are common disorders in tropical countries. These infections lead to a variety of clinical manifestations. This study reports the incidence of dermatophytosis in the military personnel in the Central-West Region of Brazil. The occupational activities of the military individuals, in addition to the hot and humid climate of the region, can predispose them to infection by these fungal entities.     

Incidence of dermatophytosis in a public hospital of São Bernardo do Campo,São Paulo State,Brazil

The incidence of cases of dermatophytosis in dermatology outpatient departments has increased in recent years. Infection control is essential to prevent transmission, and accurate diagnosis of this type of infection is important to avoid confusion with other dermatological processes caused by non-fungal agents. The objective of this study was to determine the incidence of dermatophytosis and its etiological agents in a public hospital of São Bernardo do Campo, Brazil. From February 2005 to May 2006, 273 samples were collected from 191 patients in a public dermatology outpatient department. The samples were collected according to conventional methods used in clinical mycology and direct examinations and cultures were performed. Of the samples suspected of fungal infection, 19% produced positive cultures, and the largest number of samples was from female patients between 31 and 40 years old. The most commonly infected area was the toenails, and the most common infective agents according to microscopic analysis and biochemical tests were the Candida species, which were responsible for 61.6% of the infections. These data agree with findings reported in the literature indicating a lower prevalence of filamentous fungi, such as Epidermophyton, Trichophyton, and Microsporum, in dermatomycosis.     

Genotyping of Trichophyton rubrum by analysis of ribosomal-DNA intergenic spacer regions

An attempt was made to explore the genotyping of Trichophyton rubrum (T. rubrum) and the relationship between genotype and geographical origin using ribosomal restriction endonuclease polymorphic analysis. The total DNA was extracted by cetyltrimethyl ammonium bromide (CTAB). The probe was amplified from part of the 18S, ITSI, 5.8S, and ITSII region of T. rubrum standard strain with the universal fungal primers NS5 [5′-AACTT AAAGG AATTG ACGGA AG-3′] and ITS4 [5′-TCCTC CGCTT ATTGA TATGC-3′]. The genomic DNA of 49 clinical T. rubrum isolates digested by EcoR1 were hybridized with this probe, and the hybridization patterns were used as the basis of genotyping. Of the data from 49 strains of T. rubrum studied (21 from Nanjing, 26 from Dalian, and two from Beijing), 20 individual patterns (DNA Type A–T) were identified, among which Type A–C accounted for 48.98% of all the strains. The DNA patterns of Nanjing strains were represented by three bands, those of Dalian strains were represented by four bands. The DNA typing of T. rubrum by Southern blotting was highly sensitive and highly distinguishable. The DNA patterns of Nanjing strains were obviously different from those of Dalian strains.     

Onychomycosis in Cali,Colombia

This study presents the epidemiological and mycological aspects of 299 patients with nail lesions who were referred to three diagnostic laboratories in the city of Cali. The diagnosis of mycoses was established through visualization of mycotic structures in a direct microscopic examination of skin scrapings and by isolation. Onychomycosis was found in 183 cases (61.2%), of which 141 were in toenails (44 in males and 97 in females), 38 in fingernails (9 males and 29 females), and 4 cases in toenails and fingernails simultaneously (all females). No statistically significant relation was found between sex and onychomycosis. Yeasts accounted for 40.7% of the mycoses, dermatophytes for 38%, nondermatophyte molds for 14% and the etiology was mixed in the remaining cases (7.3%). Candida albicans was the most commonly isolated yeast species; the most common dermatophyte was Trichophyton rubrum and Fusarium spp. and Scytalidium dimidiatum were the most common nondermatophytic molds. Them common fungi found in fingernails were yeasts; in toenails dermatophytes were more prevalent (chi2 with Yates' correction = 19.75, P= 0.000088). Yeasts were observed more frequently in females while dermatophytes were more common in males. The difference between these two etiologic groups was statistically significant (chi2 with Yates' correction = 7.43, P = 0.0064); no relation was observed according to age.     

Dermatophytes in Portugal (1972–1981)

For the years 1972–1981, 7 333 isolates of dermatophytes belonging to 14 species were obtained from glabrous skin (32%), feet (28%), groin (19%), scalp (8%), toenails (7%), fingernails (3%) and beard (1%)., T. rubrum represented 50% of all the isolates and was the most frequent species on glabrous skin, groin and nails. T. mentagrophytes (24%) was mainly obtained from the feet, E. floccosum (9%) from the groin and T. megninii (4%) from uncovered areas of the skin, fingernail and beard. These 4 species predominated in men. M. canis was the commonest agent on the scalp and in children up to 11 years. T. violaceum, previously the main cause of tinea capitis, and T. tonsurans have been decreasing for the period of this study, just as T. schoenleinii for the years 1962–71.The rising prevalence of T. rubrum was observed since 1962. In the whole it seems stable after 1969, but the analysis of the main sites involved shows that in the glabrous skin this species increased from 1962 to 1974; in the groin it was gone up from 30% during 1962–1965, to 64% in the years 1969–1971; in the feet the evolution was slower and only in 1980 T. rubrum became more frequent than T. mentagrophytes.The increase in certain species, whereas others become rare, lacks a satisfactory explanation.

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Résumé Depuis 1972 jusqu'à 1981, 7 333 isolements de dermatophytes concernant 14 espèces ont été obtenus de la peau glabre (32%), des pieds (28%), des plis inguinaux (19%), des ongles des orteils (7%), des ongles des mains (3%) et de la barde (1%). T. rubrum a réprésenté 50% de tous les isolements, étant l'espèce la plus fréquente dans la peau glabre, aux plis inguinaux et dans les ongles. T. mentagrophytes (24%) a été isolé surtout des pieds; E.floccosum (9%) des plis inguinaux; et T. megninii (4%) des régions découvertes de la peau, des ongles de la main et de la barbe. Ces 4 espèces ont prédominé chez des adultes du sexe masculin. M. canis a été l'agent le plus commun dans le cuir chevelu et chez des enfants jusqu'à 11 ans. T. violaceum (auparavant la cause principale des teignes du cuir chevelu) et T. tonsurans sont devenus beaucoup moins fréquents pendant cette étude, comme d'ailleurs T. schoenleinii était devenu rare au cours d'une enquête anterieur (1962–1971).La prévalence de T. rubrum s'est beaucoup elevée dès 1962. Dans l'ensemble, elle parait stabilisée depuis 1969, mais l'analyse des principales localisations de la maladie démontre que dans la peau glabre cette espèce a augmenté depuis 1962 jusqu'à 1974; aux plis inguinaux elle a changé de 30% en 1962–1965 pour 64% en 1969–1971; aux pieds l'évolution a été moins rapide et seulement dès 1980 T. rubrum est devenu plus fréquent que T. mentagrophytes.L'augmentation de certaines espèces tandis que d'autres deviennent rares n'a pas trouvée une interpretation satisfaisante.

     

Sensibilidad a fluconazol y voriconazol de especies de Candida aisladas de pacientes provenientes de unidades de cuidados intensivos en Medellín,Colombia (2001–2007)

BackgroundDisseminated candidiasis is caused by different Candida species and mainly affects immunocompromised patients and those hospitalized in intensive care units (ICU).ObjectiveOur aim was to determine the frequency and susceptibility of Candida spp. isolates to fluconazole and voriconazole, obtained from patients hospitalized in ICU in the city of Medellin during the years 2001–2007.MethodsThe agar diffusion technique based on the protocols recommended by the CLSI from the United States (M44A) was used. The Chi² test and the Kruskal Wallis statistical methods were used to compare changes in the frequency of Candida spp. isolates and their susceptibility to azoles by year of isolation.ResultsA total of 337 isolates were analyzed, 147 (43.6%) of which corresponded to Candida albicans, followed by 79 (23.4%) Candida tropicalis, 47 (13.9%) Candida parapsilosis, 32 (9.5%) Candida glabrata, 12 (3.6%) Candida guilliermondii and 11 (3.3%) Candida krusei. The remaining isolates (2.7%) were distributed among other species (Candida famata, Candida lusitaniae, Candida lipolytica, Candida pelliculosa and Candida spp.) Most of these isolates (78.3%) were susceptible; 11.9% were dose-dependent susceptible (DDS) and 9.8% resistant to fluconazole. For voriconazole, we observed that 94.1% of the isolates were susceptible, 2.4% DDS and 3.6% resistant.ConclusionsThese data indicate a notable change in the species frequency, as well as a new susceptibility patterns that requires the precise identification of the causative organism and susceptibility testing in order to determine the characteristics of the isolates circulating in ICUs and then to treat them appropriately.     

Efecto inhibitorio in vitro de ajoeno sobre aislamientos de Candida recuperados de secreciones vaginales

AimsThe main purpose of this work was to evaluate the in vitro activity of ajoene of the Candida, obtained from vaginal discharges.MethodsFor this, 136 samples were analyzed. The yeasts were recovered and identified by conventional mycological methods. The susceptibility to ajoene (at 20, 15, 12.5, 10, 6.25 and 3.125 μg/ml) was performed according to the CLSI M27-A2 document with the EUCAST modifications. The ATCC reference strains 90028 (Candida albicans), 22019 (Candida parapsilosis), and 6258 (Candida krusei) were included in this study. The minimal inhibitory concentration (MIC) was considered as the minimal concentration of ajoena able to inhibit 80% of the fungal growth.ResultsFifty five yeasts were recovered, 36 (65.4%) of them were causing candidosis and 19 (34.5%) were colonizing. C. albicans was the most frequent (81.8%) of the six isolated species, prevailing on the patients with candidosis (54.5%). The non-albicans species were less frequently isolated (18.2%), and Candida glabrata was the prevailing agent (7.3%) followed by Candida tropicalis (3.6%), C. krusei, C. parapsilosis, Candida guilliermondii and Candida sp. (1.8% each of them). The susceptibility tests to ajoeno showed inhibition of fungal growth in 98.2% of the isolates, showing MIC values ?15 μg/ml, and in (one isolate of C. glabrata) (1.8%) this value was >20 μg/ml. The reference strains showed MIC values of 3.125 and 10 μg/ml.ConclusionsThe results here presented, obtained from a significant number of isolates, mainly C. albicans, demonstrate, once more, the potential of ajoeno as an antifungal agent.     

Onychomycosis in Tehran,Iran: Prevailing fungi and treatment with itraconazole

A total of 187 Patients with suspected onychomycosis were examined for causative fungal agents between 1996 and 1997. Laboratory examination confirmed onychomycosis in 115 patients, of which 97 cases were presented with positive microscopic and cultural examinations, and they were selected for itraconazole pulse therapy. From an etiological point of view, 48.4% of the nail infections, mainly toenail infections, were caused by dermatophytes, 43.3% were infected with Candida spp, specially infected fingernails, and 8.2% by non-dermatophytic molds. Trichophyton mentagrophytes var. interdigital and T. violaceum were the most prevalent species. Candida albicans and C. parapsilosis were the predominant species of the Genus Candida. Scopolariopsis brevicaulis was the most common non-dermatophyte molds observed. Female affected more frequently than male and in both sexes, those who were 30–49 years old, more infected. Toenails were affected more frequently than fingernails. In this study, itraconazole pulse therapy (400 mg daily) gave during the first week of per month for 3 months. The study included 51 patients with toenail onychomychosis (group 1) and 46 patients with fingernail infections (group 2). Patients were followed up for 9 months after the last treatment. Clinical response rates were 83% in the group 1, 95% in the group 2 at month 12; the corresponding mycological cure rates were 71 and 87%, respectively. This revised version was published online in June 2006 with corrections to the Cover Date.     

Prevalence of Candida nivariensis and Candida bracarensis in Vulvovaginal Candidiasis

Candida nivariensis and Candida bracarensis were isolated from patients with vulvovaginal candidiasis (VVC). Candida nivariensis and Candida bracarensis were found in presumptive Candida glabrata isolates, which were identified using the API Candida system. We retrospectively re-examined vaginal presumptive Candida glabrata isolates for Candida nivariensis and Candida bracarensis from January 1, 2003, through December 31, 2012, via detection of the ITS1 region and the 5.8S ribosomal RNA gene. Among 301 presumptive Candida glabrata isolates, 293 isolates were confirmed as C. glabrata (97.34 %), 7 isolates were identified as C. nivariensis (2.33 %) and 1 isolate was identified as C. bracarensis (0.33 %). The C. nivariensis and C. bracarensis isolates were confirmed by sequencing. All C. nivariensis isolates were susceptible to nystatin and susceptible or susceptible dose-dependent to fluconazole, itraconazole, miconazole, and clotrimazole. The C. bracarensis isolate was susceptible to nystatin and the tested azoles. Among the seven patients with VVC caused by C. nivariensis and who were treated with various antifungal agents, only one patient achieved mycological eradication at both the day 7–14 and day 30–35 follow-ups. The C. bracarensis isolate was isolated from a symptomatic pregnant woman; additional data for this patient were unavailable. We conclude that C. nivariensis and C. bracarensis existed in the vaginal samples of patients with VVC. Therapeutic efficacy in the patients with C. nivariensis was poor and inconsistent with the observed in vitro antifungal susceptibility, which requires further study.     

A Study on Stability of Phenotype and Genotype of Trichophyton rubrum

Objective: To explore the stability of phenotype and genotype in Trichophyton rubrum. Methods: All the strains were cultured on Sabouraud’s dextrose agar slopes, and identified to species level. Strains isolated recently were subcultured on Sabouraud agar slopes four times at an interval of 4 weeks. DNA was extracted with CTAB method. A probe consisting of 3′ end of 18S rDNA, adjacent ITS1, 5.8S rDNA and ITS2 regions was amplified from template DNA of the T. rubrum standard strain using fungal universal primers NS5 and ITS4, labelled by P³² and hybridized with EcoR I-digested T. rubrum genomic DNA. Results: (1) Four phenotypes were isolated from 207 T. rubrum strains, with downy type (45.4%) in the first place, and granular type not found. After 1 year of conservation, 54 strains showed morphological variations with the total variation rate of 26.1%. (2) Eleven strains showed variations in colony morphology or pigment upon subculture. (3) AP-PCR analysis of 10 T. rubrum isolates and one T. rubrum standard strain showed similar DNA patterns with main bands at 2.2, 1.7, 1.3, 0.9 and 0.7 kb. No changes in DNA pattern were found upon subculture. (4) Hybridization analysis revealed that all the 11 T. rubrum strains presented three bands and were identified into two types (2.4, 3.9, 5.9 kb and 2.4, 4.4, 6.5 kb). No changes in band pattern were found upon subculture. Conclusions: Phenotype of T. rubrum was instable and the colonial morphology and pigment easily changed during conservation or subculture, while its genotype was relatively stable.