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Journal of Microbiological Methods Mar 2017Malassezia spp. are part of the normal human and animal mycobiota but are also associated with a variety of dermatological diseases. The absence of a transformation...
Malassezia spp. are part of the normal human and animal mycobiota but are also associated with a variety of dermatological diseases. The absence of a transformation system hampered studies to reveal mechanisms underlying the switch from the non-pathogenic to pathogenic life style. Here we describe, a highly efficient Agrobacterium-mediated genetic transformation system for Malassezia furfur and M. pachydermatis. A binary T-DNA vector with the hygromycin B phosphotransferase (hpt) selection marker and the green fluorescent protein gene (gfp) was introduced in M. furfur and M. pachydermatis by combining the transformation protocols of Agaricus bisporus and Cryptococcus neoformans. Optimal temperature and co-cultivation time for transformation were 5 and 7days at 19°C and 24°C, respectively. Transformation efficiency was 0.75-1.5% for M. furfur and 0.6-7.5% for M. pachydermatis. Integration of the hpt resistance cassette and gfp was verified using PCR and fluorescence microscopy, respectively. The T-DNA was mitotically stable in approximately 80% of the transformants after 10 times sub-culturing in the absence of hygromycin. Improving transformation protocols contribute to study the biology and pathophysiology of Malassezia.
Topics: Agaricus; Agrobacterium tumefaciens; Coculture Techniques; Cryptococcus neoformans; DNA, Bacterial; Dermatomycoses; Genetic Vectors; Green Fluorescent Proteins; Humans; Malassezia; Microscopy, Fluorescence; Phosphotransferases (Alcohol Group Acceptor); Polymerase Chain Reaction; Transformation, Genetic
PubMed: 28064034
DOI: 10.1016/j.mimet.2017.01.001 -
Medical Mycology Nov 2018Cytochrome P450 CYP1A1 and CYP1B1 enzymes are regulated by the aryl hydrocarbon receptor (AhR), a transcription factor activated by a variety of ligands among which...
Cytochrome P450 CYP1A1 and CYP1B1 enzymes are regulated by the aryl hydrocarbon receptor (AhR), a transcription factor activated by a variety of ligands among which Malassezia metabolites. In this study, we analyzed the modulation of CYP1A1, CYP1B1, and AhR in human keratinocytes infected with different strains of Malassezia pachydermatis, as well as the upregulation of some genes involved in the epidermal homeostasis. We demonstrated that all the strains induced AhR activation and its nuclear translocation in HaCaT cells infected for 24 h, compared to untreated cells. The expression of CYP1A1 and CYP1B1, prototypical markers of the AhR signaling pathway, were upregulated with the level of CYP1A1 mRNA approximately 100-fold greater than that for CYP1B1. Filaggrin, involucrin, and TGaseI, proteins involved in epidermal differentiation, were all modulated by Malassezia pachydermatis strains, with the strongest induction observed for filaggrin. By contrast, quinone oxidoreductase 1 (NQO1), which is part of the antioxidant defense system involved in detoxification, was not modulated in our experimental model. In conclusions, our findings suggest that Malassezia pachydermatis infection of human keratinocytes induces activation of the AhR, and increases the expression of its responsive genes and markers of epidermal differentiation, paving the way for occurrence/exacerbation of pathological skin conditions.
Topics: Basic Helix-Loop-Helix Transcription Factors; Cell Line; Cytochrome P-450 CYP1A1; Cytochrome P-450 CYP1B1; Filaggrin Proteins; Gene Expression Profiling; Humans; Keratinocytes; Malassezia; RNA, Messenger; Receptors, Aryl Hydrocarbon; Transcription, Genetic
PubMed: 29462476
DOI: 10.1093/mmy/myy004 -
Revista Chilena de Infectologia :... Feb 2015
Topics: Animals; Humans; Malassezia
PubMed: 25860048
DOI: 10.4067/S0716-10182015000200014 -
International Journal of Dermatology May 2016The genus Malassezia comprises lipophilic species, the natural habitat of which is the skin of humans and other warm-blooded animals. However, these species have been... (Review)
Review
The genus Malassezia comprises lipophilic species, the natural habitat of which is the skin of humans and other warm-blooded animals. However, these species have been associated with a diversity of dermatological disorders and even systemic infections. Pityriasis versicolor is the only cutaneous disease etiologically connected to Malassezia yeasts. In the other dermatoses, such as Malassezia folliculitis, seborrheic dermatitis, atopic dermatitis, and psoriasis, these yeasts have been suggested to play pathogenic roles either as direct agents of infection or as trigger factors because there is no evidence that the organisms invade the skin. Malassezia yeasts have been classified into at least 14 species, of which eight have been isolated from human skin, including Malassezia furfur, Malassezia pachydermatis, Malassezia sympodialis, Malassezia slooffiae, Malassezia globosa, Malassezia obtusa, Malassezia restricta, Malassezia dermatis, Malassezia japonica, and Malassezia yamatoensis. Distributions of Malassezia species in the healthy body and in skin diseases have been investigated using culture-based and molecular techniques, and variable results have been reported from different geographical regions. This article reviews and discusses the latest available data on the pathogenicity of Malassezia spp., their distributions in dermatological conditions and in healthy skin, discrepancies in the two methods of identification, and the susceptibility of Malassezia spp. to antifungals.
Topics: Antifungal Agents; Dermatitis, Atopic; Dermatitis, Seborrheic; Dermatomycoses; Folliculitis; Humans; Malassezia; Psoriasis; Skin; Tinea Versicolor
PubMed: 26710919
DOI: 10.1111/ijd.13116 -
Medical Mycology Apr 2019The yeast Malassezia pachydermatis is a common commensal and occasional opportunistic pathogen of theskin microbiota of animals and humans. In this study, the...
The yeast Malassezia pachydermatis is a common commensal and occasional opportunistic pathogen of theskin microbiota of animals and humans. In this study, the susceptibility of M. pachydermatis isolates to fluconazole (FLC), itraconazole (ITZ), ketoconazole (KTZ), clotrimazole (CLZ), and miconazole (MCZ) alone and in combination with terbinafine (TRB), nystatin (NYS), and caspofungin (CSP) was evaluated in vitro based on the M27-A3 technique and the checkerboard microdilution method using Sabouraud dextrose broth with 1% tween 80 (SDB). Based on the mean FICI values, the main synergies observed were combinations of ITZ+CSP and CLZ+CSP (55.17%). The most significant combinations deserve in vivo evaluations because might provide effective alternative treatments against M. pachydermatis due to their synergistic interactions.
Topics: Antifungal Agents; Drug Combinations; Drug Resistance, Fungal; Drug Synergism; Fluconazole; Itraconazole; Malassezia; Miconazole; Microbial Sensitivity Tests
PubMed: 29924372
DOI: 10.1093/mmy/myy043 -
Annals of Laboratory Medicine Jan 2019
Topics: Adenocarcinoma; Catheter-Related Infections; Dermatomycoses; Gastrectomy; Humans; Malassezia; Male; Middle Aged; RNA, Ribosomal; Sequence Analysis, DNA; Skin Diseases; Stomach Neoplasms
PubMed: 30215238
DOI: 10.3343/alm.2019.39.1.99 -
Veterinary Dermatology Apr 2020Tolerance of Malassezia pachydermatis to azole drugs has been reported worldwide, from strains isolated from dogs. Canine Malassezia dermatitis often is treated with...
BACKGROUND
Tolerance of Malassezia pachydermatis to azole drugs has been reported worldwide, from strains isolated from dogs. Canine Malassezia dermatitis often is treated with shampoos containing 2% miconazole (MCZ) or other topical MCZ products.
OBJECTIVES
In the in vitro study herein, it was investigated whether MCZ-induced amino acid substitutions in the lanosterol 14-alpha-demethylase (ERG11) gene 1 lead to azole tolerance in M. pachydermatis.
METHODS AND MATERIALS
Toleranced to MCZ was induced in an azole-susceptible strain of M. pachydermatis (CBS1879 ) by culture in medium containing MCZ. Antifungal susceptibility to MCZ, clotrimazole (CTZ) and itraconazole (ITZ) was assessed using the modified broth microdilution (BM) method. To assess the potential mechanism of tolerance in the three MCZ-resistant strains, ERG11 was sequenced. The interaction between the calcineurin inhibitor tacrolimus and MCZ in the azole-tolerant isolates also was examined.
RESULTS
Three strains (NUBS19001 to NUBS19003) from CBS1879 cultured in medium containing MCZ exhibited minimum inhibitory concentrations (MICs) of 40 mg/L to MCZ, 5 mg/L to ITZ and >32 mg/L to CTZ, meaning that the isolates were tolerant. The combination of MCZ and tacrolimus exerted an indifferent effect against the MCZ-tolerant strain. BLAST analysis using the NCBI database showed mutations in the cytochrome p450 encoded by ERG11 in the MCZ-tolerant strains.
CONCLUSIONS
In the present in vitro study, it was shown that MCZ exposure can induce amino acid substitutions in ERG11 and subsequent tolerance of M. pachydermatis to several azoles. Whether topical therapy with azole-containing products can exert a similar effect in vivo is a question that requires further research.
Topics: Amino Acid Substitution; Antifungal Agents; Azoles; Culture Media; Cytochrome P-450 Enzyme System; Drug Resistance, Fungal; Fungal Proteins; Malassezia; Miconazole; Microbial Sensitivity Tests; Mutation
PubMed: 31729813
DOI: 10.1111/vde.12805 -
Medical Mycology Jan 2018The non-lipid-dependent yeast Malassezia pachydermatis is predominantly zoophilic but occasionally colonizes the human skin. This yeast caused an outbreak in a neonatal...
The non-lipid-dependent yeast Malassezia pachydermatis is predominantly zoophilic but occasionally colonizes the human skin. This yeast caused an outbreak in a neonatal iIntensive care unit (NICU). This study aimed to describe the molecular epidemiology of this M. pachydermatis outbreak. All the M. pachydermatis isolates collected at a French University Hospital from January 2012 to April 2013 were included in the study. M. pachydermatis isolates, sampled from various biological samples sites in 25 patients, were identified via MALDI-TOF mass spectrometry and typed using intergenic-spacer 1 (IGS1) nucleotide sequence polymorphisms analysis. By analyzing 90 IGS1 sequences (including 43 deposited in GenBank), we found that of the 186 M. pachydermatis isolates, 47 were viable for typing and all of them clustered within type 3; 78.7% clustered within the 3D subtype; the remaining clustered within three newly described subtypes: 3E (4.3%), 3F (8.5%) and 3 G (8.5%). No particular subtype was associated with a collection site or a particular time period. This first molecular investigation of a M. pachydermatis outbreak in neonates showed that multiple genotypes can colonize the same neonate patient by. The source of this polyclonal outbreak could not be identified. It stopped after infection control measures, including the prohibition of a lipid-rich moisturizing hand cream used by the health care staff, had been implemented.
Topics: Adult; Cluster Analysis; Cross Infection; DNA, Fungal; DNA, Ribosomal Spacer; Dermatomycoses; Disease Outbreaks; Female; France; Hospitals, University; Humans; Infant; Infant, Newborn; Infection Control; Intensive Care Units, Neonatal; Malassezia; Male; Molecular Epidemiology; Phylogeny; Sequence Analysis, DNA; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
PubMed: 28371911
DOI: 10.1093/mmy/myx022 -
Veterinary Dermatology Sep 1996Abstract Skin disease associated with the cutaneous commensal organisms Staphylococcus intermedius, Malassezia pachydermatis and Demodex canis is frequently... (Review)
Review
Abstract Skin disease associated with the cutaneous commensal organisms Staphylococcus intermedius, Malassezia pachydermatis and Demodex canis is frequently encountered in veterinary medicine. In treatment the aim is elimination of the commensal, but recurrence of skin disease is not unusual. In this review, these potentially pathogenic commensals and their ecology are discussed with particular reference to skin biology and the surface ecosystem. The strategies employed by the micro-organisms for survival and the defence mechanisms of the host are considered. Disease occurs when the virulence of the commensal overwhelms the resistance of the host. It is hoped that an understanding of the complex nature of the skin and its commensals will lead to a better understanding of those diseases associated with commensals and in consequence more effective treatment. Résumé- Les dermatoses associées aux organismes commensaux cutanés Staphylococcus intermedius, Malassezia pachydermatis et Demodex canis sont fréquentes en médecine vétérinaire. Le traitement vise àéliminer le commensal, mais les récidives sont fréquentes. Dans cet article, les commensaux potentiellement pathogènes et leur écologie sont discutés avec une référence particulière à la biologie cutanée et à l'écosystème de surface. Les stratégies employées par les microorganismes pour survivre et les mécanismes de défense de l'hote sont présentés. La pathologie se développe lorsque la virulence du commensal outrepasse la résistance de l'hote. Il faut espérer que la compréhension de la nature complexe de la peau et de ses commensaux, permettent une meilleure compréhension des pathologies associées à ces commensaux et par conséquent un traitement plus efficace. [Mason, I. S., Mason, K. V., Lloyd, D. H. A review of the biology of canine skin with respect to the commensals Staphylococcus intermedius, Demodex canis and Malassezia pachydermatis (Une revue de la biologie de la peau concernant les commensaux Staphylococcus intermedius, Demodex canis et Malassezia pachydermatis). Veterinary Dermatology 1996; 7: 119-132.] Resumen En medicina veterinaria son frecuentes las dermatopatías asociadas a los microorganismos comensales Staphylococcus intermedius, Demodex canis y Malassezia pachydermatis. El tratamiento busca la eliminatión del comensal, aunque no son raras las recidivas. En esta revisión se discute la ecología de estos comensales potencialmente patógenos, con especial énfasis en la biologia cutánea y el ecosistema superficial. Se tienen en cuenta las estrategias para la supervivencia utilizadas por los microorganismos y los mecanismos de defensa del huésped. La enfernedad se desarrolla cuando la virulencia del comensal supera la resistencia del huésped. Se espera que el mejor conocimiento de la naturaleza compleja de la piel y de sus comensales llevará a un mejor conocimiento de las enfermedades asociadas a los comensales y, consecuentemente, a un tratamiento más efectivo. [Mason, I. S., Mason, K. V., Lloyd, D. H. A review of the biology of canine skin with respect to the commensals Staphylococcus intermedius, Demodex canis and Malassezia pachydermatis (Revision de la biologia cutánea con respecto a los comensales Staphylococcus intermedius, Demodex canis y Malassezia pachydermatis). Veterinary Dermatology 1996; 7: 119-132.] Zusammenfassung- Hauterkrankungen in Verbindung mit den kutanen, kommensalen Organismen Staphylococcu intermedius, Malassezia pachydermatis und Demodex canis werden in der Veterinärmedizin häufig angetroffen. Das Ziel der Behandlung ist die Elimination der Kommensalen, aber die Rezidivierung der Hauterkrankung ist nicht ungewöhnlich. In dieser Übersicht werden diese potentiell pathogenen Kommensalen und ihre Ökologie mit speziellem Bezug zur Biologie der Haut und des Ober-flächenökosystems diskutiert. Die Überlebensstrategien dieser Mikroorganismen und die Abwehrmechanismen des Wirtes werden berücksichtigt. Eine Erkrankung tritt auf, wenn die Virulenz des Kommensal die Widerstandskraft des Wirtes übertrifft. Man hofft, daß das Verstehen der komplexen Natur der Haut und ihrer Kommensalen zum besserem Verständnis dieser mit den Kommensalen verbundenen Krankheiten und damit in der Folge zu wirksamerer Behandlung führt. [Mason, I.S., Mason, K.V., Lloyd, D.H. A review of the biology of canine skin with respect to the commensals Staphylococcus intermedius, Demodex canis and Malassezia pachydermatis (Übersicht über die Biologie der Haut des Hundes mit Berücksichtigung der Kommensalen Staphylococcus intermedius, Demodex canis und Malassezia pachydermatis). Veterinary Dermatology 1996; 7: 119-132.].
PubMed: 34644987
DOI: 10.1111/j.1365-3164.1996.tb00237.x -
Mycoses Dec 2022Malassezia (M.) pachydermatis as a frequent reason for dermatological consultation in dogs and cats was recently shown to be lipid-dependent, too. Lipolytic activity is...
BACKGROUND
Malassezia (M.) pachydermatis as a frequent reason for dermatological consultation in dogs and cats was recently shown to be lipid-dependent, too. Lipolytic activity is a prerequisite for activating antimicrobial effectivity of fatty acid esters.
OBJECTIVES
It was therefore of interest whether it is possible to induce this mechanism in M. pachydermatis and to identify possible differences between minimal and strong lipid-dependent strains.
METHODS
In an agar dilution test, the minimal inhibitory concentrations of six fatty acid esters were determined for seventeen M. pachydermatis strains. GC analysis of parent compounds and liberated fatty acids was used to quantify ester cleavage.
RESULTS
Hydrolysis was observed in all test strains in a homogenous manner but was dependent on the chemical structure. Lowest MICs (500 ppm after 14 days of incubation) were obtained applying glyceryl monocaprylate and 3-hydroxylpropyl caprylate, while the corresponding esters of undecylenic acid showed nearly twice the value. As shown by GC analysis with the reference strains CBS 1879 and CBS 1892 and 3-hydroxypropyl caprylate, hydrolysis and caprylic acid formation starts immediately and was dependent on yeast density. Furthermore, nine azole-resistant strains isolated from dogs with treatment failures showed MIC values comparable to the other strains and no resistance to monohydric fatty acid esters.
CONCLUSIONS
Medium-chain fatty acid esters may represent a new therapeutic option for veterinary use even in azole-resistant strains. The in vivo verification in M. pachydermatis-associated dermatitis in dogs and cats will be the next step for the successful development of new therapeutics.
Topics: Dogs; Cats; Animals; Malassezia; Azoles; Caprylates; Esters; Cat Diseases; Antifungal Agents; Dog Diseases; Fatty Acids; Dermatomycoses
PubMed: 35923130
DOI: 10.1111/myc.13512