Molecular confirmation of a Trichophyton violaceum isolate from human black-dot ringworm

A clinical isolate from a black-dot ringworm lesion of a 28-year-old female Japanese was investigated by morphological and biochemical analyses as well as molecular analyses. The isolate grew well onthiamine enriched agar and did not produce violetpigment, macroconidia or microconidia on Sabouraud's dextrose agar. Approximately 620-bp genomic DNA fragments of the CHS1 gene were amplified from Trichophyton mentagrophytes, T. rubrum, T. tonsurans and T. violaceum by polymerase chain reaction (PCR) and sequenced. The chitin synthase 1 (CHS1) nucleotide sequences of the clinical isolate showed more than 97% similarity to that of T. violaceum and less than 96% similarity to that of T. mentagrophytes, T. rubrum and T. tonsurans. The phylogenetic analysis of their sequences revealed that the clinical isolate was genetically close to T. violaceum and distinct from T. mentagrophytes, T. rubrum and T. tonsurans. Therefore, the isolate was confirmed as T. violaceum by mycological examination and molecular analyses. This revised version was published online in June 2006 with corrections to the Cover Date.     

Antifungal potential of essential oil and various organic extracts of <Emphasis Type="Italic">Nandina domestica</Emphasis> Thunb. against skin infectious fungal pathogens

This study was undertaken to assess the in vitro antifungal potential of the essential oil and n-hexane, chloroform, ethyl acetate, and methanol extracts of Nandina domestica Thunb. against dermatophytes, the casual agents of superficial infections in animals and human beings. The oil (1,000 μg/disc) and extracts (1,500 μg/disc) revealed 31.1–68.6% and 19.2–55.1% antidermatophytic effect against Trichophyton rubrum KCTC 6345, T. rubrum KCTC 6375, T. rubrum KCTC 6352, Trichophyton mentagrophytes KCTC 6085, T. mentagrophytes KCTC 6077, T. mentagrophytes KCTC 6316, Microsporum canis KCTC 6591, M. canis KCTC 6348, and M. canis KCTC 6349, respectively, along with their respective minimum inhibitory concentration values ranging from 62.5 to 500 and 125 to 2,000 μg/ml. Also, the oil had strong detrimental effect on spore germination of all the tested dermatophytic fungi as well as concentration and time-dependent kinetic inhibition of T. rubrum KCTC 6375. The present results demonstrated that N. domestica mediated oil and extracts could be potential sources of natural fungicides to control certain important dermatophytic fungi.     

Sur l'état actuel des dermatophytes en Roumanie

Summary The actual state of dermatophytes in Romania is briefly reviewed. From the 20 species discovered, up to now, 14 belonged to the group of human and animal origin, the other 6 to geophilic dermatophytes.The most frequently isolated wereT. mentagrophytes, (var.interdigitale),M. audouinii, T. violaceum andT. rubrum, which altogether caused more than 50 % of the human ringworm found in the last 15 years.WhileT. mentagrophytes still remains the agent no. 1 of tinea pedis in Romania andT. rubrum keeps the first place in the nail involvements by dermatophytes, the first organism of tinea capitis areM. audouinii andT. violaceum, the former causing mainly the epidemic foci. Such epidemics may also be due toT. ferrugineum. The number of cases byT. schönleinii is more and more reduced. The infections byT. verruccosum are increasing. The rarity of cases due toT. gallinae, toT. equinum and toT. mengninii are pointed out.From the so called geophilic dermatophytes,M. gypseum, K. ajelloi andChrysosporium keratinophilum are mostly isolated from the Romanian soil.T. terrestre, T. evolceanui, T. indicum andM. cookei are infrequently found. A few isolates ofT. mentagrophytes andT. quinckeanum were also discovered in the soil.     

Incidence of dermatophytosis in Jordan with special reference to tinea capitis

Clinical data on 1068 cases of dermatophytosis as well as mycological data on 382 of these cases seen from March 1983 to September 1984 are reported. Tinea cruris was present in 34.1%, with Trichophyton rubrum and Epidermophyton floccosum isolated from 31% and 25% respectively. Tinea corporis was found in 17.9% of all patients of dermatophytosis with T. tonsurans responsible in 30.8%. Tinea pedis and Tinea unguium were present in 7.2% and 2% respectively, T. rubrum being the main causative agent; nail involvement was present in 27.3% of these cases. Tinea capitis was observed in 38.7% of 1068 dermatophytosis cases with T. violaceum and M. canis causing 48% and 32% of all T.C. cases respectively.The dermatophytic flora of the West Bank of Jordan apparently comprises about 14 dermatophytes, T. violaceum being the predominant agent of infection (in more than 34%). Other most common agents found in this survey were M. canis, T. rubrum, E. floccosum, T. schoenleini, T. tonsurans, and T. mentagrophytes.Formerly Assistant Professor at the Department of Internal Medicine, Faculty of Medical Sciences, University of Jordan, Amman     

Review of dermatophytoses in Galicia from 1951 to 1987, and comparison with other areas of Spain

We have reviewed all the dermatophytoses diagnosed in Galicia during four consecutive 9-year periods 1951–86 and 1987. From 4571 patients, we isolated 3351 fungal strains belonging, in decreasing order of frequency, to the following dermatophyte species: Microsporum canis (25.5%), Trichophyton rubrum (24.6%), T. mentagrophytes (21.4%), Epidermophyton floccosum (11.8%), M. gypseum (5.2%), T. tonsurans (3.9%), T. verrucosum (3.1%), T. schoenleinii (2.5%), T. violaceum (1.2%), T. mengninii (0.3%), M. audouinii (0.2%), T. equinum (0.1%) and T. soudanense (0.1%). Tinea capitis has diminished in frequency since 1951, though there was been a slight increase since 1978; M. canis has always been the most common agent, and between 1951 and 1959 T. schoenleinii was also very frequent but is no longer found. The frequency of tinea corporis, on the other hand, has experienced a considerable increase. Its most common causal agents in the last few years have been T. mentagrophytes, M. canis and T. rubrum. Until 1977 the most common tinea cruris dermatophyte was E. floccosum, but since then it has been T. rubrum. The commonest tinea pedis dermatophytes have been T. rubrum and T. mentagrophytes. Tinea unguium and tinea barbae have been the most frequent dermatophytoses, and their commonest causal agents T. rubrum and T. mentagrophytes respectively. We have documented the distribution of the various causal agents by location of the lesions, age and source of the patients (private or National Health Service patients), and we have compared the results with those obtained in other regions of Spain.     

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.

     

Dermatophytes isolated from patients in University Hospital,Kuala Lumpur,Malaysia

A total of 576 dermatophytes were isolated from patients with a variety of skin infections from January 1993 to May 2000. Ten species of dermatophytes were identified: Epidermophyton floccosum (0.7%), Microsporum audouinii (1.1%), M. canis (3.1%), M. gypseum (0.3%), Trichophyton concentricum(3.5%), T. equinum (0.2%), T. mentagrophytes (36.%), T . rubrum (53.8%), T. verrucosum (0.2) and T. violaceum (1.0%). The body sites most frequently affected by dermatophytes were the buttocks, nails and trunk. Anthropophilic dermatophytes made up 60.1% of the isolates; the most common species was T. rubrum, T. mentagrophytes and M. canis were the two main zoophilic dermatophytes. T. mentagrophytes was isolated from all body sites except the scalp. M. canis was found to be associated with domestic dogs and wasnot isolated from ethnic Malays. The only geophilic dermatophyte was M. gypseum, an uncommon dermatophyte associated with tinea pedis.This revised version was published online in October 2005 with corrections to the Cover Date.     

Parvalbumin isoforms in chicken muscle and thymus. Amino acid sequence analysis of muscle parvalbumin by tandem mass spectrometry

Parvalbumins are high-affinity Ca(2+)-binding proteins characterized by an EF-hand structure. Muscles of lower vertebrates contain up to five isoparvalbumins whereas higher vertebrates were believed to contain only one isoform per species. Recently Brewer et al. [Brewer, J.M., Wunderlich, J.K., & Ragland, W. (1990) Biochimie 72, 653-660] purified and sequenced a protein that they named avian thymic hormone, from chicken thymus. This protein, promoting immunological maturation of bone marrow cells in culture, was identified as a parvalbumin. The amino acid composition of this thymic parvalbumin was, however, considerably different from those of chicken muscle parvalbumin [Strehler, E.E., Eppenberger, H.M., & Heizman, C.W. (1977) FEBS Lett. 75, 127-133], suggesting the existence of two tissue-specific parvalbumins in chicken. We purified parvalbumin from chicken muscle, determined its complete amino acid sequence by tandem mass spectrometry, and showed that this protein is rather homologous to muscle parvalbumins from other species but different in 45 positions from the thymic parvalbumin. We discuss the possibility that a parvalbumin gene family might exist in higher vertebrates, expressed in a tissue-specific and developmentally regulated manner.     

A short review on the onychomycoses by dermatophytes in Romania

In 525 patients with onychomycoses by dermatophytes, collected in Bucharest (45% of the cases being females), the toe-nails were found to be affected in 65% of the cases, while the finger-nails only in 20%. In order of frequency, the following agents were isolated:T. rubrum (55%),T. mentagrophytes (interdigitale — 26%),T. violaceum (12%),E. floccosum (3%),T. schonleinii (1.3%). Only sporadically were found other species, viz.:M. canis (4),T. tonsurans (3),T. verruccosum (2),T. quinckeanum (1) andM. audouinii (1). Tinea pedis was the main manifestation associated with the respective cases.     

Effective photodynamic treatment of Trichophyton species with Rose Bengal

Photodynamic therapy (PDT) with Rose Bengal has previously achieved eradication of Trichophyton rubrum infections causing toenail onychomycosis; however, its antifungal activity against other clinically relevant dermatophytes has yet to be studied. Here, we test the efficacy of PDT using Rose Bengal (140 μM) and 532 nm irradiation (101 J/cm²) against Trichophyton mentagrophytes and Trichophyton interdigitale spores, in comparison to T. rubrum. A significant reduction (>99%) of T. mentagrophytes and T. interdigitale was observed, while actual eradication of viable T. rubrum was achieved (99.99%). Laser irradiation alone inhibited growth of T. rubrum (55.2%) and T. mentagrophytes (45.2%) significantly more than T. interdigitale (25.5%) (P = .0086), which may indicate an increased presence of fungal pigments, xanthomegnin and melanin. The findings suggest that Rose Bengal-PDT can act against a broader spectrum of fungal pathogens, and with continued development may be employed in a wider range of clinical antifungal applications.     

Dermatomycose experimental, étude clinico-pathologique

Using rabbits, we have studied the effect of reinfection of T. mentagrophytes var granulosum and T. rubrum into lesions that were previously infected and resolved. Clinical mycological and histopathological studies were done for 16 weeks. Timentagrophytes produced a more severe infection than T. rubrum. The clinical lesions produced by reinoculation were less intense and long lasting.     

Prevalence of tinea pedis,tinea unguium of toenails and tinea capitis in school children from Barcelona

ObjectiveTo evaluate the prevalence of tinea capitis, tinea pedis, and tinea unguium in children from several schools of Barcelona city.MethodsDuring the period of 2003–2004, a prospective cross-sectional study was carried out in 1,305 children (9% immigrant population) between the ages 3 and 15 in 17 schools in Barcelona. A systematic examination of the feet, (including nails and scalp), was performed to identify lesions compatible with tinea. Cultures of scalp and feet samples were done and analysis of environmental samples was performed for dermatophyte isolation.ResultsDermatophytes were isolated in 2.9% of the samples with a prevalence of 2.5% in feet, 0.23% in scalp, and 0.15% in nails of the feet. The predominant etiologic agents in feet were Trichophyton mentagrophytes in 45.7% of the cases and Trichophyton rubrum in 31.4%. In the nails, T. rubrum and Trichophyton tonsurans were isolated, while T. mentagrophytes (2 cases) and Trichophyton violaceum (1 case) were identified in scalp samples. Forty-five per cent of dermatophytes were isolated from healthy feet, the majority of cases in children 13–15 years old (p<0.05). Microsporum gypseum was the only agent identified in the environmental samples, and was also found in one of the cases of tinea pedis.ConclusionThe results of this study demonstrate a low prevalence of tinea capitis and tinea unguium in school children of Barcelona. On the contrary, high prevalence of dermatophytes in feet was found. It highlights the high prevalence of healthy carriers of dermatophytes in feet.     

Fish allergy in patients with parvalbumin-specific immunoglobulin E depends on parvalbumin content rather than molecular differences in the protein among fish species

Allergenic characteristics of purified parvalbumins from different fish species have not been thoroughly investigated. We revealed that purified parvalbumins from nine different fish species have identical IgE-reactivities and high cross-reactivities. We also showed that fish allergenicity is associated with the parvalbumin content of the fish species, rather than species-specific differences in the molecular characteristics of the individual parvalbumin proteins.     

(−)-5,6-Dehydrocamphor from the antifungal essential oil of Zuccagnia punctata

The essential oils from aerial parts of nine populations of Zuccagnia punctata Cav. (Fabaceae) were obtained by hydrodistillation and analyzed by GC–FID, GC–MS and ¹³C NMR. A total of 80 constituents, mainly oxygenated monoterpenes, were identified representing from 79.0 to 95.2% of the total oils which showed different composition patterns. The unusual compound (?)-5,6-dehydrocamphor, the major constituent in six of the samples investigated with percentages ranging from 12.3% to 56.5%, was isolated for the first time from nature and its structure was completely defined from NMR spectrometric data. Microscope studies of leaves indicated that not only capitate glands but also non-glandular trichomes, not previously described in this species, were present on the leaf surfaces.The essential oil showed antifungal activity against the dermatophytes Microsporum gypseum, Tricophyton rubrum and T. mentagrophytes with MIC values between 15.6 and 125 μg ml^?1, being T. rubrum the most susceptible species.     

A Cross-Reactive Human Single-Chain Antibody for Detection of Major Fish Allergens,Parvalbumins, and Identification of a Major IgE-Binding Epitope

Fish allergy is associated with moderate to severe IgE-mediated reactions to the calcium binding parvalbumins present in fish muscle. Allergy to multiple fish species is caused by parvalbumin-specific cross-reactive IgE recognizing conserved epitopes. In this study, we aimed to produce cross-reactive single chain variable fragment (scFv) antibodies for the detection of parvalbumins in fish extracts and the identification of IgE epitopes. Parvalbumin-specific phage clones were isolated from the human ETH-2 phage display library by three rounds of biopanning either against cod parvalbumin or by sequential biopanning against cod (Gad m 1), carp (Cyp c 1) and rainbow trout (Onc m 1) parvalbumins. While biopanning against Gad m 1 resulted in the selection of clones specific exclusively for Gad m 1, the second approach resulted in the selection of clones cross-reacting with all three parvalbumins. Two clones, scFv-gco9 recognizing all three parvalbumins, and scFv-goo8 recognizing only Gad m 1 were expressed in the E. coli non-suppressor strain HB2151 and purified from the periplasm. scFv-gco9 showed highly selective binding to parvalbumins in processed fish products such as breaded cod sticks, fried carp and smoked trout in Western blots. In addition, the scFv-gco9-AP produced as alkaline phosphatase fusion protein, allowed a single-step detection of the parvalbumins. In competitive ELISA, scFv-gco9 was able to inhibit binding of IgE from fish allergic patients’ sera to all three β-parvalbumins by up to 80%, whereas inhibition by scFv-goo8 was up to 20%. ¹H/¹⁵N HSQC NMR analysis of the rGad m 1:scFv-gco9 complex showed participation of amino acid residues conserved among these three parvalbumins explaining their cross-reactivity on a molecular level. In this study, we have demonstrated an approach for the selection of cross-reactive parvalbumin-specific antibodies that can be used for allergen detection and for mapping of conserved epitopes.     

A survey of dermatophytes isolated from human patients in the United States from 1982 to 1984

A survey of dermatophytes isolated from patients seeking medical advice was made from 1982 to 1984 in the United States. The survey included 59 locations with data from 49 cities and one state. Listing of the isolated dermatophytes and the frequency given by percentage of total follows: Trichophyton rubrum 46.8%, T. tonsurans 33.3%, T. mentagrophytes 10.1%, Microsporum canis 4.5%, Epidermophyton floccosum 3,5%, M. gypseum and T. verrucosum both 0.7%, M. audouinii and T. terrestre both 0.1%, and T. violaceum 0.06%. No isolations of M. ferrugineum or T. schoenleinii were reported.Temporal increases were observed for frequencies of T. mentagrophytes, T. tonsurans and M. canis, and decreases occurred for frequencies of E. floccosum and T. rubrum when the data from this survey were compared by the goodness of fit test to data of the 1979 to 1981 survey. The percent of dermatophyte isolations identified as T. tonsurans correlated significantly with the percentage of blacks in cities of 100000 or more people.     

In vitro hair cultures for differentiating between atypical isolates ofTrichophyton mentagrophytes andTrichophyton rubrum

Summary Isolates ofT. mentagrophytes andT. rubrum are encountered that cannot be distinguished from each other solely on the basis of morphological criteria.Since these two species fundamentally differ in the manner in which they attack hairin vitro, this property can be used as a diagnostic aid, when correlated with their morphologic characteristics as developed on a variety of media. T. mentagrophytes radially penetrates hair segments immersed in water forming wedge-shaped perforations.T. rubrum does not perforate hair.The method of determining the ability or inability ofT. mentagrophytes orT. rubrum to perforate hair and the use of this test in identifying those species are described.     

Enzyme make-up of trichophyton rubrum and t. mentagrophytes

Summary Aldolase activity in the cell-free extracts of two dermatophytes,T. mentagrophytes andT. rubrum, was investigated. The kinetics of the enzyme and the effects of metal ions and metal-binders are also reported. The enzyme was more active inT. mentagrophytes than inT. rubrum. The optimum pH for the enzyme action was 7.2 and it was completely inactivated at 60° C. Cobalt and magnesium ions and cysteine activated the enzyme. Inhibition caused by EDTA and o-phenanthroline was partially reversed by cobalt ions. The dermatophyte aldolase resembles bacterial aldolase in its properties.     

The Use of Albino Adult Hair and Blond Prepubertal Hair Yields Equivalent Results in an In Vitro Hair Perforation Test to Differentiate Between Different Dermatophytic Fungi

An in vitro hair perforation test is used to differentiate isolates of Trichophyton mentagrophytes and Trichophyton rubrum complexes because morphological criteria are insufficient. Here, we performed in vitro hair perforation tests using blond prepubertal hair and albino adult hair to determine whether they differentiate between fungal species. We tested 43 well-characterized dermatophyte strains, Arthroderma spp. [n = 4], Epidermophyton floccosum [n = 1], Microsporum spp. [n = 8], and Trichophyton spp. [n = 30], and examined hair perforation at 3–30 days postinoculation (p.i.). The perforation times were not significantly different between the two hair types (P > 0.05). The T. mentagrophytes complex strains perforated hair 4–5 days p.i., whereas T. rubrum complex strains perforated hair 13–30 days p.i., except for Trichophyton violaceum, which perforated hair after 6–7 days. Thus, the hair perforation test is highly sensitive (100 %) and specific (100 %) for differentiating T. mentagrophytes from T. rubrum complexes 5 days p.i. At 14 and 30 days, the sensitivity and negative predictive value of the test remained unchanged (100 %), but the specificity was reduced (64.3 and 14.3 %, respectively). Consistent with previous reports, we observed “perforating organs” of zoophilic Microsporum canis and geophilic Microsporum gypseum at 4 and 3 days, respectively. This paper offers a “low-cost” and “low-tech” alternative to differentiating dermatophyte species where standard morphological techniques fail and/or where molecular techniques are not a viable option.     

Solanum melongena: A potential source of antifungal agent

The antifungal activity of Solanum melongena leaf, extracted with petroleum ether, chloroform, methanol and water was evaluated against three human pathogenic dermatophytes namely Trichophyton mentagrophytes, T. rubrum and T. tonsurans and two opportunistic fungi Candida albicans and Trichosporon beigelii. Maximum yield of plant components was 4.32 g, extracted in water and minimum 1.07 g in petroleum ether from 150 g of dry plant material. Except water extract, all the extracts possessed significant antifungal property. All the test pathogens showed highest sensitivity towards chloroform extract, exhibiting maximum inhibition zone diameter of 50.0 mm in T. mentagrophytes and minimum 30.0 mm in C. albicans at 2 × 10⁵ μg/ml concentration. Chloroform extract at lower concentration 2.5 × 10⁴ μg/ml was inhibitory for all the test pathogens, exhibiting inhibition zone diameter 21.0 mm against T. tonsurans and 15.0 mm against C. albicans and T. beigelii. The activity of the different solvent extracts against the test pathogens in terms of inhibition zone diameter in decreasing order was as followsChloroform extract > Petroleum ether extract > Methanol extract for T. mentagrophytes, T. rubrum and T. tonsurans.Chloroform extract > Methanol extract > Petroleum ether extract for C. albicans and T. beigelii.