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1.
Mycopathologia ; 189(4): 59, 2024 Jun 18.
Article in English | MEDLINE | ID: mdl-38890181

ABSTRACT

Dermatophyte biofilms frequently count for inadequate responses and resistance to standard antifungal treatments, resulting in refractory chronic onychomycosis infection. Although antimicrobial photodynamic therapy (aPDT) has clinically proven to exert significant antifungal effects or even capable of eradicating dermatophyte biofilms, considerably less is known about the molecular mechanisms underlying aPDT and the potential dysregulation of signaling networks that could antagonize its action. The aim of this study is to elucidate the molecular mechanisms underlining aPDT combat against dermatophyte biofilm in recalcitrant onychomycosis and to decipher the potential detoxification processes elicited by aPDT, facilitating the development of more effective photodynamic interventions. We applied genome-wide comparative transcriptome analysis to investigate how aPDT disrupting onychomycosis biofilm formed by three distinct dermatophytes, including Trichophyton rubrum, Trichophyton mentagrophytes, and Microsporum gypseum, the most frequently occurring pathogenic species. In total, 352.13 Gb of clean data were obtained for the transcriptomes of dermatophyte biofilms with or without aPDT treatment, resulting in 2,422.42 million reads with GC content of 51.84%, covering 99.9%, 98.5% and 99.4% of annotated genes of T. rubrum, T. mentagrophytes, and M. gypseum, respectively. The genome-wide orthologous analysis identified 6624 transcribed single-copy orthologous genes in all three species, and 36.5%, 6.8% and 17.9% of which were differentially expressed following aPDT treatment. Integrative orthology analysis demonstrated the upregulation of oxidoreductase activities is a highly conserved detoxification signaling alteration in response to aPDT across all investigated dermatophyte biofilms. This study provided new insights into the molecular mechanisms underneath anti-dermatophyte biofilm effects of aPDT and successfully identified a conserved detoxification regulation upon the aPDT application.


Subject(s)
Arthrodermataceae , Biofilms , Gene Expression Profiling , Photochemotherapy , Biofilms/drug effects , Arthrodermataceae/drug effects , Arthrodermataceae/genetics , Microsporum/drug effects , Microsporum/genetics , Humans , Antifungal Agents/pharmacology , Onychomycosis/microbiology , Onychomycosis/drug therapy , Transcriptome
2.
Article in English | MEDLINE | ID: mdl-38747851

ABSTRACT

Microsporum canis, one of the most widespread dermatophytes worldwide, is a zoonotic microorganism that transmits infection from reservoirs such as cats and dogs to humans. This microorganism is associated with Tinea corporis and other clinical manifestations; however, few studies have used genetic surveillance to determine and characterize the process of zoonotic transmission. In this study, we show a clear example of zoonotic transmission from a cat to an intrafamilial environment, where it caused Tinea corporis by infection with M. canis. Molecular characterization using the b-tubulin gene and Random Amplified Polymorphic DNA analysis made it possible to determine that the six isolates of M. canis obtained in this study belonged to the same genetic variant or clone responsible for reservoir-reservoir or reservoir-human transmission.


Subject(s)
Cat Diseases , Microsporum , Tinea , Zoonoses , Microsporum/isolation & purification , Microsporum/genetics , Microsporum/classification , Cats/microbiology , Animals , Tinea/microbiology , Tinea/transmission , Tinea/veterinary , Cat Diseases/microbiology , Cat Diseases/transmission , Zoonoses/microbiology , Zoonoses/transmission , Pets/microbiology , Humans , Dogs , Random Amplified Polymorphic DNA Technique , Male , Female , Dog Diseases/microbiology , Dog Diseases/transmission , DNA, Fungal/genetics
3.
Acta Trop ; 255: 107237, 2024 Jul.
Article in English | MEDLINE | ID: mdl-38723739

ABSTRACT

The surge in domestic cat adoption across India, particularly the rising preference for high-pedigree cats, coupled with environmental factors, has resulted in increased incidence of dermatophytosis among feline companions. Despite this growing concern, there is a noticeable scarcity of studies in India delving into the etiological factors contributing to dermatophytosis in cats. This disease is a threat to animal health and carries public health significance, given that cats are recognized reservoir hosts for Microsporum canis, a common dermatophyte affecting humans and animals. This study endeavours to identify the dermatophytes affecting cats and establish a standardized therapeutic regimen while accounting for the local stigma surrounding the regular bathing of cats. The study involved the examination of 82 cats presenting dermatological lesions, when subjected to cultural examination in dermatophyte test medium revealed 36 afflicted with dermatophytes. Isolates were presumptively identified by staining using lactophenol cotton blue, Chicago sky blue 6B, and Calcofluor white stains. Molecular-level identification of the isolates was confirmed through PCR-RFLP, amplifying the Internal Transcribed Spacer Sequence of 16 s rDNA, followed by restriction digestion using the Mva1 enzyme. Among the thirty-six isolates, 29 were identified as M. canis, while the remaining 7 were M. gypseum. The cases were categorized into five groups and treated with Lime Sulphur dip, 4 % chlorhexidine shampoo, a shampoo containing 2 % miconazole and 4 % chlorhexidine, oral itraconazole alone, and a combination of oral itraconazole with lime-Sulphur dip. Statistical analysis revealed that the response was notably swifter with lime Sulphur dip when considering only topical therapy. Moreover, the mycological cure was most expeditious when combining Lime Sulphur dip with oral itraconazole. These findings underscore the pivotal role of topical biocides in feline dermatophytosis treatment, potentially reducing the reliance on specific antifungals and thereby contributing to the mitigation of antimicrobial resistance emergence.


Subject(s)
Antifungal Agents , Cat Diseases , Microsporum , Tinea , Cats/microbiology , Animals , Cat Diseases/microbiology , Cat Diseases/drug therapy , India/epidemiology , Tinea/veterinary , Tinea/microbiology , Tinea/drug therapy , Tinea/epidemiology , Antifungal Agents/therapeutic use , Microsporum/isolation & purification , Microsporum/genetics , Male , Female , Arthrodermataceae/isolation & purification , Arthrodermataceae/genetics , Arthrodermataceae/classification , Arthrodermataceae/drug effects , Itraconazole/therapeutic use , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , DNA, Fungal/genetics , DNA, Ribosomal Spacer/genetics
4.
Int J Mol Sci ; 25(5)2024 Feb 22.
Article in English | MEDLINE | ID: mdl-38473782

ABSTRACT

Microsporum canis is a widely distributed dermatophyte, which is among the main etiological agents of dermatophytosis in humans and domestic animals. This fungus invades, colonizes and nourishes itself on the keratinized tissues of the host through various virulence factors. This review will bring together the known information about the mechanisms, enzymes and their associated genes relevant to the pathogenesis processes of the fungus and will provide an overview of those virulence factors that should be better studied to establish effective methods of prevention and control of the disease. Public databases using the MeSH terms "Microsporum canis", "virulence factors" and each individual virulence factor were reviewed to enlist a series of articles, from where only original works in English and Spanish that included relevant information on the subject were selected. Out of the 147 articles obtained in the review, 46 were selected that reported virulence factors for M. canis in a period between 1988 and 2023. The rest of the articles were discarded because they did not contain information on the topic (67), some were written in different languages (3), and others were repeated in two or more databases (24) or were not original articles (7). The main virulence factors in M. canis are keratinases, fungilisins and subtilisins. However, less commonly reported are biofilms or dipeptidylpeptidases, among others, which have been little researched because they vary in expression or activity between strains and are not considered essential for the infection and survival of the fungus. Although it is known that they are truly involved in resistance, infection and metabolism, we recognize that their study could strengthen the knowledge of the pathogenesis of M. canis with the aim of achieving effective treatments, as well as the prevention and control of infection.


Subject(s)
Microsporum , Virulence Factors , Humans , Animals , Virulence Factors/metabolism , Microsporum/genetics , Microsporum/metabolism , Animals, Domestic , Subtilisins/metabolism
5.
Med Mycol J ; 65(1): 1-5, 2024.
Article in English | MEDLINE | ID: mdl-38417881

ABSTRACT

Microsporum canis is a type of dermatophyte that causes zoonotic dermatophytosis in cats and dogs. We report three cases of tinea corporis due to M. canis from a single household with a domestic cat as a pet. The cases included a woman in her thirties (mother), a girl in her teens (older sister), and a girl in her teens (younger sister). Following sudden hair loss in the domestic cat, annular erythema with pruritus and scales appeared on the face, neck, and limbs of the older sister, younger sister, and mother, sequentially; they subsequently visited our hospital. Potassium hydroxide direct microscopy revealed filamentous fungi on all three women. In addition, short-haired colonies with a white to yellowish-white color and extending in a radial manner were found in cultures using a flat plate agar medium. A slide culture with the same medium indicated pointed spindle-shaped macroconidia with 7-8 septa. Therefore, the cases were diagnosed as tinea corporis due to M. canis. Genetic analysis of the cells of the cat and the mother, older sister, and younger sister using multilocus microsatellite typing (MLMT) indicated that all cases were classified into the same genotype, suggesting that the transmission route of these cases was familial. Here, we show that MLMT is useful in identifying the infection route in cases of tinea corporis due to M. canis.


Subject(s)
Dermatomycoses , Tinea , Humans , Adolescent , Female , Animals , Dogs , Cats , Tinea/diagnosis , Tinea/veterinary , Microsporum/genetics , Mothers , Microsatellite Repeats/genetics , Dermatomycoses/diagnosis , Dermatomycoses/microbiology
6.
Comp Immunol Microbiol Infect Dis ; 104: 102100, 2024 Jan.
Article in English | MEDLINE | ID: mdl-38043450

ABSTRACT

Microsporum canis is considered the common dermatophyte agent associated with ringworm in felines and canines. In the present study, we sampled n = 548 felines and canines for the probable isolation of M. canis. The rate of isolation from the cats and dogs was 70.27 % (52/74) and 1.68 % (8/474), respectively and Persian cats were found to be highly susceptible to M. canis infection. The strains were evaluated for their production of phospholipase, lipase, catalase, and hemolysis and their ability to grow at 35 â„ƒ. All the strains were identified as low producers of catalase and n = 17 strains exhibited high thermotolerance ability. Terbinafine was found to be the most effective antifungal drug and fluconazole was the least effective, in vitro. AFLP analysis revealed three genotypes of M. canis with 15 sub-clusters showing ≥ 90 % similarity and 7 sub-clusters exhibiting 100 % similarity. However, the phenotypic characters cannot be attributed based on the AFLP profiles.


Subject(s)
Cat Diseases , Dermatomycoses , Dog Diseases , Animals , Cats , Dogs , Antifungal Agents/pharmacology , Antifungal Agents/therapeutic use , Catalase/pharmacology , Dermatomycoses/drug therapy , Dermatomycoses/microbiology , Dermatomycoses/veterinary , DNA Fingerprinting/veterinary , Cat Diseases/microbiology , Amplified Fragment Length Polymorphism Analysis/veterinary , Dog Diseases/microbiology , Microsporum/genetics
7.
Pol J Vet Sci ; 26(4): 629-634, 2023 Dec 12.
Article in English | MEDLINE | ID: mdl-38088306

ABSTRACT

Dermatophytes from Microsporum, Trichophyton and Epidermophyton genera are divided into geophilic, zoophilic and anthropophilic species which cause skin infection in humans and wide group of animals, mainly mammals. Main species causing dermatophytosis in dogs and cats are Microsporum and Trichophyton. Conventional mycological diagnostic technique includes Saburaud Dextrose Agar (SAD) and others medium cultures, 10% KOH mount and direct microscopy of hairs and scraping. Molecular diagnostic become more frequent in veterinary practice due to shortening of waiting time. In this study we based on two PCR methods. The nested PCR amplified CHS1 gene for dermatophytes detection, and multiplex PCR coding ITS1 and ITS2 fragments for species identification of detected derpatophytes. Most frequently detected species was Microsporum canis, mainly in young cats. Geophilic Microsporum gypseum and anthropophilic Trichophyton rubrum was found primarily in dogs. Molecular methods in dermatophytosis identification are rapid in contrast to routinely, long lasting culture.


Subject(s)
Arthrodermataceae , Cat Diseases , Dermatomycoses , Dog Diseases , Tinea , Humans , Animals , Cats , Dogs , Arthrodermataceae/genetics , Dermatomycoses/diagnosis , Dermatomycoses/epidemiology , Dermatomycoses/veterinary , Poland/epidemiology , Cat Diseases/diagnosis , Cat Diseases/epidemiology , Dog Diseases/diagnosis , Dog Diseases/epidemiology , Microsporum/genetics , Trichophyton/genetics , Multiplex Polymerase Chain Reaction/veterinary , Tinea/diagnosis , Tinea/epidemiology , Tinea/veterinary , Mammals/genetics
8.
Med Mycol J ; 64(3): 63-72, 2023.
Article in English | MEDLINE | ID: mdl-37648500

ABSTRACT

Microsporum canis, a major causative agent of zoonotic dermatophytosis, has become prevalent in Japan. Molecular epidemiological surveys using multilocus microsatellite typing (MLMT), a sensitive genotyping tool for fungi, have been conducted to reveal intraspecies polymorphisms of M. canis.The present study utilized MLMT optimized for Japanese strains of M. canis to analyze epidemic trends of fungal infection. Six individual loci were targeted; namely, MS1, 2, 4, 5, 6, and 7. Analysis of data from 1974 through 2022 identified 416 strains, which were sorted into 60 genotypes by MLMT.The major genotypes showed changes in dominance during this period-changes that may reflect historical increases and decreases in the numbers of patients infected with M. canis patients. The main origins of infection included animal breeders and pet stores, as well as stray cat communities. Forty-nine episodes of familial outbreaks and cohabitant animal infections were recorded, and genotypes responsible for each episode were determined. MLMT analysis is not only a robust tool to understand population structures, but likely the most suitable method for tracking M. canis infections.


Subject(s)
Canidae , Microsporum , Animals , Cats , Japan/epidemiology , Molecular Epidemiology , Microsporum/genetics , Microsatellite Repeats
9.
Emerg Microbes Infect ; 12(1): 2219346, 2023 Dec.
Article in English | MEDLINE | ID: mdl-37288745

ABSTRACT

ABSTRACTGenomes of strains of the zoophilic dermatophyte Microsporum canis from invasive (disseminated and subcutaneous) and noninvasive (tinea capitis) infections were compared. Especially the disseminated strain showed significant syntenic rearrangements, including multiple translocations and inversions, and numerous SNPs and Indels in comparison to the noninvasive strain. In transcriptome analysis, both invasive strains were enriched for GO pathways related to components of the membrane, iron binding and heme binding, which possibly enables them to invade deeper into dermis and blood vessels. At 37 °C, invasive strains showed gene expression enriched for DNA replication, mismatch repair, N-glycan biosynthesis and ribosome biogenesis. The invasive strains were slightly less susceptible to multiple antifungal agents suggesting that acquired elevated drug resistance might be involved in the refractory disease courses. Patient with disseminated infection failed to respond to a combined antifungal treatment with itraconazole, terbinafine, fluconazole and posaconazole.


Subject(s)
Tinea Capitis , Transcriptome , Humans , Tinea Capitis/drug therapy , Tinea Capitis/microbiology , Microsporum/genetics , Antifungal Agents/pharmacology , Antifungal Agents/therapeutic use
10.
Med Mycol J ; 64(2): 37-43, 2023.
Article in English | MEDLINE | ID: mdl-37258133

ABSTRACT

A 64-year-old woman presented with a fist-sized, severely painful lesion with scales, crusts, pustules, erythema with subcutaneous abscess, and hair loss on the left temporal region. Direct microscopic examination revealed a large number of spores around the hair, which indicated ectothrix hair invasion, and some hyphae were also found. Histopathological examination showed significant inflammatory cell infiltration from the dermis to the subcutaneous tissues and into the hair follicles, destruction of the hair follicles with granulomatous reactions, and fungal masses along the hair within the hair follicles. Microsporum canis was identified based on morphological features via culture method and molecular biological analysis of the internal transcribed spacer region DNA sequence. The patient was diagnosed with kerion celsi caused by M. canis. For treatment of kerion celsi, we chose an oral antifungal agent, fosravuconazole (FRVCZ), which has been available since 2018 only in Japan. Clinical symptoms were cured in 12 weeks without scarring. No side effects were observed during oral administration of FRVCZ. The results of our case and several previous reports suggest that FRVCZ is effective in treating various types of dermatomycoses.


Subject(s)
Dermatologic Agents , Tinea Capitis , Female , Humans , Adult , Middle Aged , Antifungal Agents/therapeutic use , Tinea Capitis/diagnosis , Tinea Capitis/drug therapy , Tinea Capitis/microbiology , Microsporum/genetics , Hair/microbiology , Hair/pathology , Hair/ultrastructure , Dermatologic Agents/therapeutic use
11.
Med Mycol ; 61(5)2023 Apr 29.
Article in English | MEDLINE | ID: mdl-37120732

ABSTRACT

The classical dermatophytes diagnosis is based on mycological culture and microscopy observation both human and animal hair, skin, and nail samples. The aim of this work was to develop the new in-house real-time PCR with pan-dematophyte reaction for detection and identification of the main dermatophytes directly from hair samples, providing a simple and rapid diagnosis of dermatophytosis in dogs and cats. An in-house SYBR-Green real-time PCR was designed and used for detecting a DNA fragment encoding chitin synthase 1 (CHS1). A total of 287 samples were processed by culture, microscopic examination with KOH 10%, and real-time PCR (qPCR) analysis. Melting curve analysis of the CHS1 fragment revealed to be reproducible, showing a single distinct peak for each species of dermatophyte, namely Trichophyton mentagrophytes, T. verrucosum, Microsporum canis, and Nannizzia gypsea (formerly M. gypseum). Then, out of the 287 clinically suspected cases of dermatophytosis, 50% were positive for dermatophytes by qPCR, 44% by mycological culture, and 25% by microscopic examination. Microsporum canis was identified in 117 samples tested by culture and 134 samples tested by qPCR, followed by N. gypsea in 5 samples (either tested by culture or qPCR) and T. mentagrophytes detected in 4 and 5 samples when tested by culture or qPCR, respectively. Overall, qPCR allowed the diagnosis of dermatophytosis in clinical samples. The results suggest this newly proposed in-house real-time PCR assay can be used as alternative diagnosis and rapid identification of dermatophytes frequently associated to clinical hair samples of dogs and cats.


The aim of this work was to develop a molecular detection strategy for dermatophytes by SYBR-Green real-time PCR of hair samples from animals. The melting curve analysis of the CHS1 fragment revealed to be reproducible, showing a single distinct peak for distinct dermatophyte species and allowed the diagnosis of dermatophytosis in dogs and cats caused mainly by Trichophyton mentagrophytes, Microsporum sp., and Nannizzia gypsea).


Subject(s)
Arthrodermataceae , Cat Diseases , Dermatomycoses , Dog Diseases , Tinea , Cats , Animals , Dogs , Humans , Arthrodermataceae/genetics , Dermatomycoses/diagnosis , Dermatomycoses/veterinary , Real-Time Polymerase Chain Reaction/veterinary , Cat Diseases/diagnosis , Dog Diseases/diagnosis , Microsporum/genetics , Hair , Chitin Synthase/genetics , Tinea/veterinary , Trichophyton/genetics
12.
Vet Dermatol ; 33(6): 516-522, 2022 Dec.
Article in English | MEDLINE | ID: mdl-36000592

ABSTRACT

BACKGROUND: Dermatophytes are infectious zoonotic fungal agents that are common in animals worldwide. A new loop-mediated isothermal amplification (LAMP) method and quantitative (q)PCR can be used for identifying these agents. Both methods have high specificity and sensitivity, and are simple and quick to use. HYPOTHESIS/OBJECTIVES: To develop a LAMP and a rapid multiplex qPCR method for detecting Microsporum canis and Trichophyton mentagrophytes, which are the most common fungal species isolated from cats and dogs. MATERIAL AND METHODS: Both methods targeted the CHS-1 gene. Their specificity and sensitivity were tested using 64 M. canis and 44 T. mentagrophytes field strains. The validation of the methods was performed using 250 clinical fungal-positive hair samples. RESULTS: The specificity value was 100% for both methods. For LAMP, the sensitivity value was 96.9% for M. canis and 93.2% for T. mentagrophytes. For qPCR, the sensitivity values were 98.4% for M. canis and 97.7% for T. mentagrophytes. Similar specificity and sensitivity results were obtained from the validation study using 250 clinical hair samples. LAMP and multiplex qPCR took 30 and 45 min (respectively) for both targets. The limit of detection (LOD) assays for both targets were 10 and 1 spore/mL for LAMP and multiplex qPCR, respectively. CONCLUSION: These findings demonstrate that the LAMP and multiplex qPCR methods targeting CHS-1 gene developed in this study can be used both for point-of-care testing and in the laboratory for detecting M. canis and T. mentagrophytes with high specificity and sensitivity with an internal control.


Subject(s)
Arthrodermataceae , Dogs , Cats , Animals , Trichophyton/genetics , Microsporum/genetics , Real-Time Polymerase Chain Reaction/veterinary , Sensitivity and Specificity
13.
J Dermatol ; 49(7): 682-690, 2022 Jul.
Article in English | MEDLINE | ID: mdl-35411631

ABSTRACT

Multilocus microsatellite typing was performed on 124 strains of Microsporum canis, which is the most prevalent causative fungus of zoonotic dermatophytosis, isolated in Japan between 1974 and 1981. The strains had been enclosed in glass ampoules by freeze dried process. Genotypes were detected by capillary electrophoresis targeted on six microsatellite regions, and 19 genotypes were found among these 124 strains. The most frequent genotype, which had been reported as genotype L in a previous study, comprised 56 of the 124 strains (45.2%), although genotype L was reported to comprise only six of 165 strains (3.6%) isolated between 2010 and 2017. A decrease in the prevalence of genotype L may have led to the retreat of M. canis infection in 1996 to 2006. On the other hand, genotype A, the second most predominant genotype in the aforementioned studies of strains isolated between 2010 and 2017, comprised just four of 124 strains (3.2%) in the present study. Thus, these studies reveal that prevalences of some major genotypes have changed over the last 40 years. Genotype consistency of strains was proven in all of 12 familial cases, each of which was infected with a single genotype. We emphasize the importance of fungal culture collection for further studies with new techniques in the future.


Subject(s)
Dermatomycoses , Tinea , Dermatomycoses/epidemiology , Dermatomycoses/microbiology , Genotype , Humans , Japan/epidemiology , Microsporum/genetics , Tinea/epidemiology , Tinea/microbiology
14.
Jpn J Infect Dis ; 75(2): 105-113, 2022 Mar 24.
Article in English | MEDLINE | ID: mdl-34334533

ABSTRACT

Multilocus microsatellite typing (MLMT) was performed on 93 strains of Microsporum canis (M. canis) isolated between 2012 and 2017 from 75 cats, 8 dogs, and 10 pet owners. These strains were derived from 2 major reservoirs: commercial breeding facilities and pet shops (PS), and stray cats and pet cats that went outdoors and came in contact with stray cats (Outdoor). Six microsatellite markers were used for genotyping. These 93 strains included 22 genotypes; 11 had been previously detected in Japan while the other 11 were new. Strains belonging to the previously reported genotypes, P and A, were distributed widely throughout Japan. Genotype P was the most frequent, accounting for 37 (39.8%) of the 93 strains. Most were derived from Outdoor sources. Genotype A was the second most frequent (11 of 93 strains, 11.8%). Most of the genotype A strains were derived from the PS reservoir. All new genotypes were detected in isolates from cats. Many of these were derived from the Outdoor reservoir. Consistency of infection was observed in 18 of the 19 familial cases. These findings indicate that genotypes differ in strains derived from PS and Outdoor reservoirs. MLMT genotyping is useful for tracking the routes of spread and transmission of M. canis in Japan.


Subject(s)
Microsatellite Repeats , Microsporum , Animals , Cats , Dogs , Japan/epidemiology , Microsporum/genetics , Molecular Epidemiology
15.
Front Cell Infect Microbiol ; 11: 727665, 2021.
Article in English | MEDLINE | ID: mdl-34604111

ABSTRACT

Microsporum canis, a common pathogenic skin fungus, can cause dermatophytosis in humans and animals. Zinc is an important trace element and plays an important role in the growth and metabolism of fungi. Currently, the effects of zinc deficiency on growth, gene expression, and metabolic pathway have not been clarified in M. canis. Therefore, M. canis was cultured under zinc restriction, and RNA-Seq was conducted in this study. The growth of M. canis was severely inhibited, and many genes showed significant upregulation and downregulation in M. canis with zinc deficiency. Zinc deficiency could negatively affect the gene expression and biological metabolic pathway in M. canis. The zinc-responsiveness transcriptional activator (ZafA) gene was significantly upregulated and shared homology with Zap1. Thus, the ZafA gene might be the main transcription factor regulating M. canis zinc homeostasis. The ZafA gene knockout strain, ZafA-hph, was constructed via Agrobacterium tumefaciens-mediated transformation (ATMT) in M. canis for the first time to assess its function. In vitro growth ability, hair biodegradation ability, virulence test, and zinc absorption capacity in ZafA-hph and wild-type M. canis strains were compared. Results showed that the ZafA gene plays an important role in zinc absorption, expression of zinc transporter genes, and growth and pathogenicity in M. canis and can be used as a new drug target. Cutting off the zinc absorption pathway can be used as a way to prevent and control infection in M. canis.


Subject(s)
Microsporum , Zinc , Animals , Humans , Microsporum/genetics , RNA-Seq , Virulence
16.
Mycopathologia ; 186(4): 487-497, 2021 Aug.
Article in English | MEDLINE | ID: mdl-34164772

ABSTRACT

Epidermophyton floccosum is one of the most common agents of human superficial fungal infections, compared with genus Trichophyton and Microsporum, it possesses uniqueness in ecology traits and rarely causing hair infections. E. floccosum is so far the only representative species of genera Epidermophyton, and it is known as anthropophilic dermatophytes. To further reveal the genome sequences and clues of virulence factors, thus in this study, we sequenced the genome of E. floccosum (CGMCC (F) E1d), and performed comparative genomic analysis with other dermatophytes. It is revealed that E. floccosum owns the largest genome size and similar GC content compared with other dermatophytes. A total of 7565 genes are predicted. By comparing with the closest species N. gypseum, our study reveals that number and structure of adhesion factors, secreted proteases and LysM domain might contribute to the pathogenic and ecological traits of E. floccosum. Mating genes is also detected in genome data. Furthermore, we performed AFLP analysis trying to discuss intraspecific differences of E. floccosum, but no significant relationship is found between genotype and geographical distribution. Upon above, our study provides a deeper understanding and strong foundation for future researches about E. floccosum.


Subject(s)
Epidermophyton , Amplified Fragment Length Polymorphism Analysis , Genomics , Microsporum/genetics , Trichophyton/genetics
17.
J Mycol Med ; 31(3): 101157, 2021 Sep.
Article in English | MEDLINE | ID: mdl-34147757

ABSTRACT

BACKGROUND: Tinea capitis is a dermatophyte infection of the scalp and hair that affects a large number of people worldwide. The disease commonly affects children and manifests with varying degrees of hair loss, scalp inflammation, and psychosocial impact. In Nigeria, the burden of tinea capitis is worrisome affecting over 15,000,000 school-age children. Molecular techniques complement the conventional mycological examinations in laboratory diagnosis of tinea capitis. In this study, we identified dermatophytes species causing tinea capitis in Kano, Nigeria, using ITS-based nucleotide sequencing technique in addition to conventional mycological examination. METHODS: We collected 112 samples from the scalp of children with clinically diagnosed tinea capitis at the dermatology clinic of Murtala Muhammad Specialist Hospital, Kano, between April and September 2019. The samples were processed and subjected to direct microscopy and mycological culture to isolate dermatophytes species that were identified morphologically and using ITS sequencing. RESULTS: Out of the 112 patients investigated, the majority (59.8%) were between the ages 6 and 9 years with a mean age of 7.3 ± 1.9 years. Males (79.5%) were predominantly affected. Black dot (46.4%) was the most common clinical type of tinea capitis followed by gray patch (39.3%) and kerion (1.8%). Favus was not observed. Microsporum audouinii (45.7%) was the predominant etiologic agent followed by Trichophyton soudanense (28.6%), T. violaceum (22.9%), and T. tonsurans (2.9%). CONCLUSION: The prominence of anthropophilic dermatophytes as the main causes of tinea capitis in our localities suggests that public health interventions to promote health education and good hygiene practices would minimize the transmission rate of tinea capitis among children in the study area.


Subject(s)
Arthrodermataceae , Tinea Capitis , Arthrodermataceae/genetics , Child , Health Promotion , Humans , Male , Microsporum/genetics , Nigeria/epidemiology , Scalp , Tinea Capitis/epidemiology , Trichophyton/genetics
19.
J Vet Sci ; 22(1): e4, 2021 Jan.
Article in English | MEDLINE | ID: mdl-33522156

ABSTRACT

BACKGROUND: Microsporum canis is a zoonotic disease that can cause dermatophytosis in animals and humans. OBJECTIVES: In clinical practice, ketoconazole (KTZ) and other imidazole drugs are commonly used to treat M. canis infection, but its molecular mechanism is not completely understood. The antifungal mechanism of KTZ needs to be studied in detail. METHODS: In this study, one strain of fungi was isolated from a canine suffering with clinical dermatosis and confirmed as M. canis by morphological observation and sequencing analysis. The clinically isolated M. canis was treated with KTZ and transcriptome sequencing was performed to identify differentially expressed genes in M. canis exposed to KTZ compared with those unexposed thereto. RESULTS: At half-inhibitory concentration (½MIC), compared with the control group, 453 genes were significantly up-regulated and 326 genes were significantly down-regulated (p < 0.05). Quantitative reverse transcription polymerase chain reaction analysis verified the transcriptome results of RNA sequencing. Gene ontology enrichment analysis and Kyoto Encyclopedia of Genes and Genomes enrichment analysis revealed that the 3 pathways of RNA polymerase, steroid biosynthesis, and ribosome biogenesis in eukaryotes are closely related to the antifungal mechanism of KTZ. CONCLUSIONS: The results indicated that KTZ may change cell membrane permeability, destroy the cell wall, and inhibit mitosis and transcriptional regulation through CYP51, SQL, ERG6, ATM, ABCB1, SC, KER33, RPA1, and RNP genes in the 3 pathways. This study provides a new theoretical basis for the effective control of M. canis infection and the effect of KTZ on fungi.


Subject(s)
Antifungal Agents/pharmacology , Dermatomycoses/veterinary , Dog Diseases/drug therapy , Ketoconazole/pharmacology , Microsporum/drug effects , Transcriptome , Animals , Dermatomycoses/drug therapy , Dogs , Gene Expression Profiling/veterinary , Microsporum/genetics
20.
Med Mycol ; 59(4): 355-365, 2021 Apr 06.
Article in English | MEDLINE | ID: mdl-32640011

ABSTRACT

Arthroderma is the most diverse genus among dermatophytes encompassing species occurring in soil, caves, animal burrows, clinical material and other environments. In this study, we collected ex-type, reference and authentic strains of all currently accepted Arthroderma species and generated sequences of three highly variable loci (ITS rDNA, ß-tubulin, and translation elongation factor 1-α). The number of accepted species was expanded to 27. One novel species, A. melbournense (ex-type strain CCF 6162T = CBS 145858T), is described. This species was isolated from toenail dust collected by a podiatrist in Melbourne, during an epidemiological study of four geographical regions of Eastern Australia. Trichophyton terrestre, Chrysosporium magnisporum, and Chrysosporium oceanitis are transferred to Arthroderma. Typification is provided for T. terrestre that is not conspecific with any of the supposed biological species from the former T. terrestre complex, that is, A. insingulare, A. lenticulare and A. quadrifidum. A multi-gene phylogeny and reference sequences provided in this study should serve as a basis for future phylogenetic studies and facilitate species identification in practice. LAY ABSTRACT: The genus Arthroderma encompasses geophilic dermatophyte species that infrequently cause human and animal superficial infections. Reference sequences from three genetic loci were generated for all currently accepted Arthroderma species and phylogeny was constructed. Several taxonomic novelties are introduced. The newly provided data will facilitate species identification and future taxonomic studies.


Subject(s)
Arthrodermataceae/classification , Arthrodermataceae/genetics , DNA, Fungal/genetics , Genes, Fungal/genetics , Phylogeny , Australia , DNA, Ribosomal Spacer/genetics , Humans , Microsporum/classification , Microsporum/genetics , Peptide Elongation Factor 1/genetics , Trichophyton/classification , Trichophyton/genetics , Tubulin/genetics
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