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1.
Microb Ecol ; 84(2): 363-375, 2022 Aug.
Article in English | MEDLINE | ID: mdl-34536095

ABSTRACT

The European hedgehog (Erinaceus europaeus Linnaeus) frequently colonises areas located close to human life in cities, as these are more suitable nest sites offering an abundance of food and allowing avoidance of predators. However, urbanisation has a significant impact on the epidemiology of infectious diseases, including dermatophytoses, the primary source of which are wild animals. In this study, we determined the spectrum of dermatophytes isolated from the European hedgehog and assessed their susceptibility profile to antifungal drugs. Symptomatic and asymptomatic dermatophyte infections were observed in 7.7% and 8% of the 182 examined free-living hedgehogs, respectively. In the pool of the isolated dermatophyte strains, Trichophyton erinacei was dominant (29.9%), followed by Trichophyton mentagrophytes (17.9%), Trichophyton benhamiae (13.4%), Nannizzia gypsea (11.9%), Microsporum canis (10.4%), Nannizzia nana (7.5%), Paraphyton cookei (6.0%), and Nannizzia fulva (3.0%). Susceptibility tests revealed the highest activity of luliconazole and the lowest of activity fluconazole among the azole drugs applied. Although terbinafine generally exhibited high efficacy, two Trichophyton mentagrophytes isolates showed resistance to this drug (MIC = 2 µg/ml) resulting from missense mutations in the SQLE gene corresponding to the amino acid substitution Leu393Phe. Summarising, our study has also revealed that such wildlife animals as hedgehogs can be a reservoir of pathogenic human dermatophytes, including harmful strains resistant to commonly used antifungal drugs.


Subject(s)
Arthrodermataceae , Tinea , Animals , Animals, Wild , Antifungal Agents/pharmacology , Arthrodermataceae/genetics , Hedgehogs/microbiology , Humans , Microbial Sensitivity Tests , Poland/epidemiology , Tinea/epidemiology , Tinea/microbiology , Tinea/veterinary , Trichophyton/genetics
2.
J Appl Microbiol ; 133(2): 544-554, 2022 Aug.
Article in English | MEDLINE | ID: mdl-35429221

ABSTRACT

AIMS: Indoor air quality in stables, cowsheds or henhouses has recently become in interest due to the potential risks of zoonotic infections. Cowsheds are commonly known to have high fungal loads, particularly if insufficient attention is paid to the monitoring and control of the indoor microclimate around three elements, that is heating, ventilation and lighting. The aim of this study was to determine the concentrations and spectrum of dermatophyte propagules in the indoor air of cowsheds. METHODS AND RESULTS: Air samples were collected on five farms, and the dermatophyte species were identified using MALDI-TOF MS analysis. The quantitative analysis of the fungal pollutants showed an average of 0.084 dermatophyte propagules (CFU) per m3 of flowing air in spring and 0.0239 CFU/m3 in the summer. Dermatophyte species were identified in case of 64.6% of the obtained colonies. Trichophyton verrucosum as dominant species was isolated on all five farms. In turn, Nannizzia gypsea was isolated on four farms and Trichophyton mentagrophytes as well as Paraphyton cookei were isolated on two farms. CONCLUSIONS: This study demonstrated that indoor aerosol appears to be one of the underestimated risks of dermatophyte infections. Moreover, the risk of zoonotic infections is posed by airborne zoophilic dermatophytes, especially T. verrucosum, whose prevalence of infections has been increasing in recent years. SIGNIFICANCE AND IMPACT OF THE STUDY: The ability of dermatophytes to infect animals and humans is thought to be a consequence of not only their adaptation to new ecological niches but also occurring as an aerosol component, which we demonstrate for the first time in this study. The microclimate of the cowshed may be an underestimated reservoir of zoophilic dermatophytes, which pose a zoonotic threat to farmers, animal breeders and veterinarians.


Subject(s)
Arthrodermataceae , Dermatomycoses , Animals , Attention , Dermatomycoses/microbiology , Humans , Microsporum , Trichophyton , Zoonoses/microbiology
3.
Infection ; 49(6): 1233-1240, 2021 Dec.
Article in English | MEDLINE | ID: mdl-34499324

ABSTRACT

PURPOSE: Anthropophilic dermatophytes as etiological factors of onychomycoses are more common than zoophilic fungi. In the case of the latter, reverse zoonoses are possible, which poses a threat to the persistence of dermatophytes in the environment. Nevertheless, without treatment, both types of tinea unguium may lead to complete nail plate destruction and secondary mixed infections with fungi and bacteria. One of the zoophilic dermatophytes that cause onychomycosis is Trichophyton verrucosum, whose prevalence has been increasing in recent years. Such infections are usually treated with allylamines and/or azoles, but such a conventional treatment of infections caused by T. verrucosum often fails or is discontinued by patients. METHODS: Herein, we reveal the results of our in vitro studies related to direct application of cold atmospheric pressure plasma (CAPP) on Trichophyton verrucosum growth, germination and adherence to nail as a new alternative treatment method of such types of dermatomycoses. RESULTS: Our in vitro studies showed that, while exposure to CAPP for 10 min delays germination of conidia and clearly impairs the fitness of the fungal structures, 15 min is enough to kill all fungal elements exposed to plasma. Moreover, the SEM images revealed that T. verrucosum cultures exposed to CAPP for 10 and 15 min were not able to invade the nail fragments. CONCLUSION: The results revealed that single exposure to CAPP was able to inhibit T. verrucosum growth and infection capacity. Hence, cold atmospheric pressure plasma should be considered as a promising alternative treatment of onychomycoses.


Subject(s)
Dermatomycoses , Onychomycosis , Plasma Gases , Animals , Arthrodermataceae , Atmospheric Pressure , Humans , Onychomycosis/drug therapy , Plasma Gases/therapeutic use , Zoonoses
4.
Mycoses ; 64(8): 967-975, 2021 Aug.
Article in English | MEDLINE | ID: mdl-33884673

ABSTRACT

OBJECTIVES: Dermatomycoses of zoophilic origin, especially those caused by Trichophyton mentagrophytes, often pose considerable therapeutic problems. This is reflected in the growing number of strains of this species with resistance to terbinafine caused by a mutation in the squalene epoxidase (SQLE) gene. Therefore, it is reasonable to look for alternative therapies to the commonly used terbinafine. The aim of the present study was to assess the in vivo effectiveness of topical therapy with luliconazole or terbinafine 1% cream. METHODS: Therapeutic efficacy was assessed using direct examination in KOH with DMSO, qPCR analysis with pan-dermatophyte primers and culturing. Moreover, in vitro susceptibility tests for luliconazole and terbinafine were performed. RESULTS: The results demonstrated significantly higher antifungal activity of luliconazole than terbinafine against dermatomycoses caused by T. mentagrophytes. The geometric mean of the MIC value for luliconazole against all T. mentagrophytes strains was 0.002 µg/ml, while this value for terbinafine was 0.004 µg/ml. In all studied cases, 28-day local therapy with luliconazole contributed to complete eradication of the aetiological agent of infection. CONCLUSIONS: Given the increasingly frequent reports of difficult-to-treat dermatophytoses caused by zoophilic terbinafine-resistant strains, the 1% luliconazole cream can be alternative solution in topical therapy.


Subject(s)
Antifungal Agents/therapeutic use , Arthrodermataceae/drug effects , Dermatomycoses/drug therapy , Imidazoles/therapeutic use , Terbinafine/pharmacology , Terbinafine/therapeutic use , Administration, Topical , Antifungal Agents/administration & dosage , Arthrodermataceae/classification , Arthrodermataceae/genetics , Drug Resistance, Fungal , Genotype , Humans , Imidazoles/administration & dosage , Imidazoles/pharmacology , Microbial Sensitivity Tests , Terbinafine/administration & dosage
5.
Mycoses ; 64(5): 537-549, 2021 May.
Article in English | MEDLINE | ID: mdl-33448025

ABSTRACT

BACKGROUND: Dermatophytoses have gained interest worldwide due to the increased resistance to terbinafine and azoles and difficulty in management of these refractory diseases. OBJECTIVES: In this study, we identified and analysed Trichophyton mentagrophytes clinical isolates obtained from humans with infections of animal origin. METHODS: We used quantitative real-time PCR (qRT-PCR) to examine the transcriptional modulation of three MDR genes (PDR1, MDR2 and MDR4) and analysed squalene epoxidase (SQLE) gene sequences from multidrug-resistant Trichophyton mentagrophytes isolates. RESULTS: The expression profile revealed a 2- to 12-fold increase in mRNA accumulation in the presence of any of the antifungals, compared to cells incubated without drugs. A statistically significant relationship between the isolates exposed to itraconazole and increased expression of the tested genes was revealed. Substantially lower transcription levels were noted for cells exposed to luliconazole, that is, a third-generation azole. Additionally, in the case of 50% of terbinafine-resistant strains, Leu397Phe substitution in the SQLE gene was detected. Furthermore, the reduced susceptibility to itraconazole and voriconazole was overcome by milbemycin oxime. CONCLUSIONS: In conclusion, our study shed more light on the role of the ABC transporter family in T. mentagrophytes, which, if overexpressed, can confer resistance to single azole drugs and even cross-resistance. Finally, milbemycin oxime could be an interesting compound supporting treatment with azole drugs in the case of refractory dermatomycoses.


Subject(s)
Arthrodermataceae , Drug Resistance, Fungal/genetics , Tinea/drug therapy , ATP-Binding Cassette Transporters/genetics , Animals , Antifungal Agents/pharmacology , Antifungal Agents/therapeutic use , Arthrodermataceae/drug effects , Arthrodermataceae/genetics , Arthrodermataceae/isolation & purification , Azoles/pharmacology , Azoles/therapeutic use , Drug Resistance, Multiple/genetics , Genes, Fungal , Humans , Macrolides/pharmacology , Macrolides/therapeutic use , Microbial Sensitivity Tests , Mutation, Missense , Real-Time Polymerase Chain Reaction/methods , Squalene Monooxygenase/genetics , Terbinafine/pharmacology , Terbinafine/therapeutic use , Trichophyton/drug effects , Trichophyton/genetics , Trichophyton/isolation & purification , Zoonoses
6.
Infection ; 48(6): 889-897, 2020 Dec.
Article in English | MEDLINE | ID: mdl-32770418

ABSTRACT

BACKGROUND: Dermatomycoses are the most common fungal infections in the world affecting a significant part of the human and animal population. The majority of zoophilic infections in humans are caused by Trichophyton mentagrophytes. Currently, the first-line drug for both oral and topical therapy is terbinafine. However, an increasing number of cases that are difficult to be cured with this drug have been noted in Europe and Asia. Resistance to terbinafine and other allylamines is very rare and usually correlated with point mutations in the squalene epoxidase gene resulting in single amino acid substitutions in the enzyme, which is crucial in the ergosterol synthesis pathway. PURPOSE: Here, we report terbinafine-resistant T. mentagrophytes isolates among which one was an etiological factor of tinea capitis in a man and three were obtained from asymptomatic foxes in Poland. METHODS: We used the CLSI protocol to determine antifungal susceptibility profiles of naftifine, amphotericin B, griseofulvin, ketoconazole, miconazole, itraconazole, voriconazole, and ciclopirox. Moreover, the squalene epoxidase gene of the terbinafine-resistant strains was sequenced and analysed. RESULTS: In the genomes of all four resistant strains exhibiting elevated MICs to terbinafine (16 to 32 µg/ml), single-point mutations leading to Leu393Phe substitution in the squalene epoxidase enzyme were revealed. Among the other tested substances, a MIC50 value of 1 µg/ml was shown only for griseofulvin. CONCLUSION: Finally, our study revealed that the terbinafine resistance phenomenon might not be acquired by exposure to the drug but can be intrinsic. This is evidenced by the description of the terbinafine-resistant strains isolated from the asymptomatic animals.


Subject(s)
Amino Acid Substitution , Antifungal Agents/pharmacology , Arthrodermataceae/drug effects , Drug Resistance, Fungal , Fungal Proteins/genetics , Squalene Monooxygenase/genetics , Terbinafine/pharmacology , Animals , Arthrodermataceae/genetics , Foxes/microbiology , Fungal Proteins/metabolism , Humans , Poland , Squalene Monooxygenase/metabolism
7.
Infection ; 48(3): 429-434, 2020 Jun.
Article in English | MEDLINE | ID: mdl-32232786

ABSTRACT

BACKGROUND: Fungal infections of the skin, hair, and nails are the largest and most widespread group of all mycoses. Nannizzia nana is a relatively rare etiological factor of dermatomycosis in humans, as it usually affects animals, e.g. pigs and boars. In addition to the zoophilic nature, there are also reports of the geophilic reservoir of this dermatophyte species. OBJECTIVE: In this study, we present symptomatic infections with N. nana aetiology in humans reported recently in Poland. Interestingly, these cases had a non-specific clinical picture and occurred as skin lesions on the neck and foot as well as onychomycosis of the toenails. From the medical history, the patients had no contact with pigs. METHODS: Diagnostics of these infections was performed with a combination of classical phenotypic and molecular genomic methods. The genomic diversity of the isolates was determined using the MP-PCR method. In vitro antifungal susceptibility tests against itraconazole, ketoconazole, terbinafine and naftifine hydrochloride were also performed. RESULTS: Nannizzia nana has been identified as an etiological factor of dermatomycosis. Moreover, heterogeneity of the genomes was revealed for the obtained strains. In vitro activities of antifungal agents showed that isolates were susceptible to all tested drugs. The patients were treated with oral terbinafine and topical ketoconazole cream, which led to a complete recovery. CONCLUSIONS: In conclusion, the cases studied by us may indicate that the infrequency of N. nana infections may not necessarily be related to the low infectivity of this fungal agent, but they are rather associated with misdiagnosis. Furthermore, N. nana reservoirs should also be sought in soil.


Subject(s)
Antifungal Agents/therapeutic use , Arthrodermataceae/isolation & purification , Dermatomycoses/diagnosis , Ketoconazole/therapeutic use , Administration, Oral , Administration, Topical , Adult , Aged , Arthrodermataceae/classification , Dermatomycoses/drug therapy , Dermatomycoses/microbiology , Diagnosis, Differential , Female , Humans , Male , Onychomycosis/diagnosis , Onychomycosis/drug therapy , Onychomycosis/microbiology , Soil Microbiology , Terbinafine/therapeutic use , Treatment Outcome
8.
BMC Dermatol ; 20(1): 9, 2020 09 29.
Article in English | MEDLINE | ID: mdl-32993612

ABSTRACT

BACKGROUND: The impact of Malassezia yeasts on skin mycobiome and health has received considerable attention recently. Pityriasis versicolor (PV), a common dermatosis caused by Malassezia genus worldwide, is a manifestation of dysbiosis. PV can be associated with hyper- and/or hypopigmented skin lesions. This disease entity is characterized by high percentage of relapses, which demands a proper antifungal therapy that is based on unambiguous species identification and drug susceptibility testing. CASE PRESENTATION: Comprehensive analysis of PV case in man presenting simultaneously hyper- and hypopigmented skin lesions was performed. Conventional and molecular diagnostic procedures revealed Malassezia furfur and Malassezia sympodialis, respectively as etiological agents of skin lesions observed. Susceptibility tests showed significantly lowered sensitivity of M. furfur cells to fluconazole. Based on susceptibility profiles local antifungal therapy with drugs characterized by entirely different mechanism of action was included. CONCLUSIONS: Our study indicates that cases of PV represented by two types of skin lesions in one patient may be associated with distinct Malassezia species. Moreover, as observed in this case, each of the isolated etiological agents of PV may differ significantly in susceptibility to antifungals. This can significantly complicate the treatment of dermatosis, which by definition is associated with a significant percentage of relapses. In the presented case localized topical treatment was sufficient and successful while allowing maintaining the physiological mycobiome.


Subject(s)
Antifungal Agents/therapeutic use , Ciclopirox/administration & dosage , Malassezia/isolation & purification , Mycobiome/drug effects , Skin/microbiology , Terbinafine/administration & dosage , Tinea Versicolor/drug therapy , Administration, Topical , Antifungal Agents/pharmacology , Drug Therapy, Combination , Humans , Male , Middle Aged , Pigmentation Disorders/etiology , Tinea Versicolor/complications
9.
Extremophiles ; 20(5): 641-52, 2016 Sep.
Article in English | MEDLINE | ID: mdl-27315167

ABSTRACT

Mycobiota are important in underground ecology. In 2014, we discovered dark stains on clayey sediments on the walls of Driny Cave, Slovakia. Our description is based on the morphology of the fungus and the phylogenetic relationships of the internal transcribed spacer (ITS) region. In addition, data on its capacity for the production of extracellular enzymes, growth, and survival in vitro at different temperatures are reported. Our analyses revealed that this dark stains on the wall was produced by Penicillium glandicola. The fungus was able to synthesize amylases, proteases and cellulases, but not pectinases and keratinases. The vegetative structures of mycelium of this fungus are viable in vitro after storage at cool temperatures (from -72 to 5 °C), and show active growth at temperatures from 5 to 25 °C, but without spore germination, and without active growth at 30 and 37 °C. Penicillium glandicola is a psychrotolerant species and belong to var. glandicola.


Subject(s)
Caves/microbiology , Geologic Sediments/microbiology , Microbiota , Penicillium/isolation & purification , Acclimatization , Cold Temperature , DNA, Intergenic , Fungal Proteins/metabolism , Penicillium/genetics , Penicillium/metabolism
10.
Molecules ; 21(12)2016 Dec 15.
Article in English | MEDLINE | ID: mdl-27983708

ABSTRACT

This review summarizes the current state of knowledge about the metabolism of cancer cells, especially with respect to the "Warburg" and "Crabtree" effects. This work also summarizes two key discoveries, one of which relates to hexokinase-2 (HK2), a major player in both the "Warburg effect" and cancer cell immortalization. The second discovery relates to the finding that cancer cells, unlike normal cells, derive as much as 60% of their ATP from glycolysis via the "Warburg effect", and the remaining 40% is derived from mitochondrial oxidative phosphorylation. Also described are selected anticancer agents which generally act as strong energy blockers inside cancer cells. Among them, much attention has focused on 3-bromopyruvate (3BP). This small alkylating compound targets both the "Warburg effect", i.e., elevated glycolysis even in the presence oxygen, as well as mitochondrial oxidative phosphorylation in cancer cells. Normal cells remain unharmed. 3BP rapidly kills cancer cells growing in tissue culture, eradicates tumors in animals, and prevents metastasis. In addition, properly formulated 3BP shows promise also as an effective anti-liver cancer agent in humans and is effective also toward cancers known as "multiple myeloma". Finally, 3BP has been shown to significantly extend the life of a human patient for which no other options were available. Thus, it can be stated that 3BP is a very promising new anti-cancer agent in the process of undergoing clinical development.


Subject(s)
Antineoplastic Agents/therapeutic use , Energy Metabolism/drug effects , Glycolysis/drug effects , Hexokinase/metabolism , Oxidative Phosphorylation/drug effects , Pyruvates/therapeutic use , Humans , Liver Neoplasms/drug therapy , Mitochondria/metabolism , Multiple Myeloma/drug therapy
11.
Med Dosw Mikrobiol ; 67(3-4): 221-31, 2015.
Article in English | MEDLINE | ID: mdl-27019916

ABSTRACT

A significant participation in the growing prevalence of fungal infections have basidiomycete yeasts of the genus of Cryptococcus. Among about seventy actually known species belonging to Cryptococcus genus there are only two: Cryptococcus neoformans and Cryptococcus gattii, which as anamorphs in parasitic stage of its life fully evolved the mechanisms predisposing them to be a human and animal pathogens. Specific attributes important in the pathogenesis of cryptococcosis called virulence factors allow them to colonize the mammalian tissues. Undoubtedly, the most important role in the pathogenesis play: the presence of polysaccharide capsule, synthesis of melanin, and the ability to grow in the range of temperatures 35-40°C.


Subject(s)
Cryptococcosis/microbiology , Cryptococcus gattii/pathogenicity , Cryptococcus neoformans/pathogenicity , Virulence Factors , Animals , Cryptococcus gattii/cytology , Cryptococcus gattii/metabolism , Cryptococcus neoformans/cytology , Cryptococcus neoformans/metabolism , Fungal Capsules , Humans , Melanins/biosynthesis
12.
Med Dosw Mikrobiol ; 67(2): 133-9, 2015.
Article in Polish | MEDLINE | ID: mdl-26591665

ABSTRACT

Systemic fungal infections are becoming an increasingly important problem. Exploring the development of mechanisms of pathogenesis, immune response of the human organism and the search for new potential antifungal agents requires in vivo testing. Mice, rats and rabbits are indispensable model organisms for this type of study. Unfortunately, such a kinds of studies carried out on a large scale are associated with high costs as well as with logistical and ethical problems. This paper reports proposal for the use of insects as model organisms to study the development of systemic fungal infections and analysis of biological activities of antifungal agents.


Subject(s)
Antifungal Agents/pharmacology , Disease Models, Animal , Insecta/microbiology , Mycoses/drug therapy , Mycoses/microbiology , Animals , Blattellidae/microbiology , Culex/microbiology , Drosophila melanogaster/microbiology , Mycoses/immunology
13.
Biochem Biophys Res Commun ; 432(1): 169-74, 2013 Mar 01.
Article in English | MEDLINE | ID: mdl-23337499

ABSTRACT

Candida glabrata, a human opportunistic pathogen is characterized by intrinsic, low susceptibility to fluconazole and a high capacity for acquiring high-level azole resistance. This is related to the elevated expression of genes belonging to the CgPdr1-governed regulon, comprising numerous genes, of which the multidrug ABC transporter-encoding CgCDR1, CgCDR2, CgSNQ2 are the best characterized. The function of certain PDR loci, such as CgRTA1 and CgRSB1 is poorly understood. These are homologs of ScRTA1 and ScRSB1 from Saccharomyces cerevisiae, members of the LTE family of plasma membrane proteins characteristic of fungi. While overproduced, they are involved in tolerance to 7-aminocholesterol or phytosphingosine, respectively. In this report we shed light on the differential regulation of CgRTA1 and CgRSB1 in C. glabrata. CgRTA1 expression positively correlated with intrinsic azole tolerance in clinical isolates. In contrast to CgRSB1, a high induction of CgRTA1 was observed upon fluconazole exposure, which was accompanied by a parallel up-regulation of its transcriptional activator CgPDR1. Hypoxia or presence of ketoconazole, both leading to ergosterol depletion, resulted in increased level of CgRTA1 transcript, whereas CgRSB1 was highly responsive to mitochondrial dysfunction. On the other hand, the expression of CgRTA1 was suppressed during growth in pseudohyphae formation promoting media. Our results are the first report linking the divergent regulation of LTE family members and azole sensitivity in C. glabrata.


Subject(s)
Candida glabrata/genetics , Drug Resistance, Fungal/drug effects , Fungal Proteins/genetics , Gene Expression Regulation, Fungal , Genes, Fungal , Membrane Proteins/genetics , Stress, Physiological/genetics , Azoles/pharmacology , Candida glabrata/drug effects , Candidiasis/microbiology , Humans
14.
Biochem Biophys Res Commun ; 434(2): 322-7, 2013 May 03.
Article in English | MEDLINE | ID: mdl-23541578

ABSTRACT

We have investigated the antifungal activity of the pyruvic acid analogue: 3-bromopyruvate (3-BP). Growth inhibition by 3-BP of 110 strains of yeast-like and filamentous fungi was tested by standard spot tests or microdilution method. The human pathogen Cryptococcus neoformans exhibited a low Minimal Inhibitory Concentration (MIC) of 0.12-0.15 mM 3-BP. The high toxicity of 3-BP toward C. neoformans correlated with high intracellular accumulation of 3-BP and also with low levels of intracellular ATP and glutathione. Weak cytotoxicity towards mammalian cells and lack of resistance conferred by the PDR (Pleiotropic Drug Resistance) network in the yeast Saccharomyces cerevisiae, are other properties of 3-BP that makes it a novel promising anticryptococcal drug.


Subject(s)
Antifungal Agents/pharmacology , Cryptococcus neoformans/drug effects , Pyruvates/pharmacology , Adenosine Triphosphate/antagonists & inhibitors , Adenosine Triphosphate/metabolism , Amphotericin B/pharmacology , Biological Transport , Cryptococcus neoformans/pathogenicity , Fluconazole/pharmacology , Glutathione/metabolism , Microbial Sensitivity Tests , Microbial Viability/drug effects , Time Factors
16.
Antibiotics (Basel) ; 11(6)2022 May 30.
Article in English | MEDLINE | ID: mdl-35740146

ABSTRACT

The "One Health" concept increasingly demonstrates the global spread of pathogenic (also eukaryotic) microorganisms and their zoonotic potential. Dermatophytes can cause superficial mycoses in humans and animals. Furthermore, the number of transmissions from asymptomatic carriers to humans has been on the rise over the last few years. This study was focused on the detailed characterisation of clinical isolates of Trichophyton quinckeanum with epidemiological analyses and characterisation of their in vitro antifungal susceptibility patterns. The isolated dermatophytes were identified with a combination of conventional and molecular methods. In turn, their susceptibility in vitro was tested according to the Clinical and Laboratory Standards Institute (CLSI) M38 ed.3 protocol. A total of 36 strains were isolated, with 21 cases of T. quinckeanum zoonoses resulting from direct contact with symptomatic cats (58.3%). The other 15 strains (41.7%) were isolated simultaneously from healthy cats and their owners. All strains showed high susceptibility to allylamine, pyridinone, and phenyl morpholine derivatives but were resistant to fluconazole and ketoconazole. In conclusion, our study shows the frequency of zoonoses contracted from asymptomatic cats. Moreover, the antifungal susceptibility profiles indicate the serious risk posed to animal owners by resistant strains of T. quinckeanum, which are often responsible for recalcitrant-to-treatment cases.

17.
Pathogens ; 11(7)2022 Jul 05.
Article in English | MEDLINE | ID: mdl-35890013

ABSTRACT

Opportunistic pathogens of the anamorphic genus Cryptococcus are unique considering their virulence factors that in the context of pathogenesis allowed them to achieve evolutionary success. Morphological transformation into giant (Titan) cells is one of the factors contributing to cryptococcosis. Recently established in vitro protocols demonstrate that 5 or 10% fetal bovine serum (FBS) combined with 5% CO2, 37 °C, and sufficiently low cell density, triggers cellular enlargement (Serum protocols). However, the FBS components that promote this morphological transition remain incompletely characterized. In search of minimal conditions necessary for stimulating the formation of Titan cells, we performed a study where we eliminated serum from the protocol (Serum-free protocol) and instead systematically adjusted the amount of glucose, source of nitrogen (ammonium sulfate), and the pH. We found that exposing cells to PBS with adjusted pH to 7.3, and supplemented with 0.05% glucose, 0.025% ammonium sulfate, 0.004% K2HPO4, 0.0035% MgSO4, in the presence of 5% CO2 at 37 °C triggers Titan-like cell formation to the same degree as the previously established protocol that utilized 10% FBS as the sole nutrient source. Titan-like cells obtained according to this Serum-free protocol were characterized by cell body size over ten microns, a single enlarged vacuole, thick cell wall, extensive polysaccharide capsule, and changes in the level of cell ploidy, all currently known hallmarks of Titan cells found in vivo. Strikingly, we found that in both, Serum and Serum-free protocols, an optimal pH for Titan-like cell development is ~7.3 whereas relatively acidic pH (5.5) prevents this morphological transition and promotes robust proliferation, while alkaline pH (~8.0) has a profound growth inhibitory effect. Our study demonstrates a critical role of pH response to the formation of Titan cells and indicates that conditions that allow restricted proliferation in the presence of 5% CO2 are sufficient for this morphological transition to form enlarged cells in Cryptococcus neoformans and Cryptococcus gattii species complex.

18.
Biology (Basel) ; 11(3)2022 Feb 23.
Article in English | MEDLINE | ID: mdl-35336726

ABSTRACT

In nature, there are many species of fungi known to produce various mycotoxins, allergens and volatile organic compounds (VOCs), as well as the commonly known etiological agents of various types of mycoses. So far, none of them have provoked so much emotion among homeowners, builders, conservators, mycologists and clinicians as Stachybotrys chartarum. This species compared to fungi of the genera Fusarium and Aspergillus is not as frequently described to be a micromycete that is toxigenic and hazardous to human and animal health, but interest in it has been growing consistently for three decades. Depending on the authors of any given review article, attention is focused either on the clinical aspects alongside the role of this fungus in deterioration of biomaterials, or aspects related to its biology, ecology and taxonomic position. On the one hand, it is well established that inhalation of conidia, containing the highest concentrations of toxic metabolites, may cause serious damage to the mammalian lung, particularly with repeated exposure. On the other hand, we can find articles in which authors demonstrate that S. chartarum conidia can germinate and form hyphae in lungs but are not able to establish an effective infection. Finally, we can find case reports that suggest that S. chartarum infection is linked with acute pulmonary hemorrhage, based on fungal structures recovered from patient lung tissue. New scientific reports have verified the current state of knowledge and note that clinical significance of this fungus is exceedingly controversial. For these reasons, understanding S. chartarum requires reviewing the well-known toxigenic features and harmful factors associated with this fungus, by gathering the newest ones into a coherent whole. The research problem related to this fungus seems to be not overly publicized, and there is still a demand to truthfully define the real threats of S. chartarum and phylogenetically related species. The most important problem, which should be fully elucidated as soon as possible, remains the clarification of the pathogenicity of S. chartarum and related species. Maybe it is urgent time to ask a critical question, namely what exactly do we know 28 years after the outbreak of pulmonary hemorrhage in infants in Cleveland, Ohio, USA most likely caused by S. chartarum?

19.
Pharmaceutics ; 14(6)2022 May 28.
Article in English | MEDLINE | ID: mdl-35745731

ABSTRACT

BACKGROUND: Dermatophytoses are one of the most prevalent infectious diseases in the world for which the pace of developing new drugs has not kept pace with the observed therapeutic problems. Thus, searching for new antifungals with an alternative and novel mechanism of action is necessary. OBJECTIVE: This study aimed to evaluate the antifungal activity of ebselen and diphenyl diselenide against Trichophyton mentagrophytes clinical isolates. METHODS: In vitro antifungal susceptibility was assessed for organoselenium compounds used alone or in combination with allylamines and azoles according to the 3rd edition of the CLSI M38 protocol. RESULTS: Ebselen demonstrated high antifungal activity with MICGM equal to 0.442 µg/mL and 0.518 µg/mL in the case of human and animal origin strains, respectively. The values of MICGM of diphenyl diselenide were higher: 17.36 µg/mL and 13.45 µg/mL for the human and animal isolates, respectively. Synergistic or additive effects between terbinafine and ebselen or diphenyl diselenide were observed in the case of 12% and 20% strains, respectively. In turn, the combination of itraconazole with diphenyl diselenide showed a synergistic effect only in the case of 6% of the tested strains, whereas no synergism was shown in the combination with ebselen. CONCLUSIONS: The results highlight the promising activity of organoselenium compounds against Trichophyton mentagrophytes. However, their use in combinational therapy with antifungal drugs seems to be unjustified due to the weak synergistic effect observed.

20.
Mycoses ; 54(2): 162-7, 2011 Mar.
Article in English | MEDLINE | ID: mdl-19925570

ABSTRACT

The study was performed to analyse the spectrum of dermatomycoses in southwest Poland during the period 2003-2007. A total of 10,486 patients were investigated for fungal skin infections by means of native specimen and cultivating procedures. Skin scrapings, plucked hairs and nail clippings were examined and identified by direct microscopy and culture. From 2468 patients, 2753 fungi were identified including dermatophytes, yeast and moulds. Among the dermatophytes, the most common pathogen isolated was Trichophyton rubrum (59.4%), followed in descending order by: Trichophyton mentagrophytes var. interdigitale (16.6%), Trichophyton mentagrophytes var. mentagrophytes (9.0%), Trichophyton tonsurans (6.8%), Microsporum canis (5.1%) and Epidermophyton floccosum (2.7%). Among the yeast-like fungi, a marked predominance of Candida species was observed (86.3%). Scopulariopsis brevicaulis was the most commonly isolated mould (25.2%). The most frequently affected body sites were the toenails (53.9%), followed by the fingernails (19.0%). In children under 15 years of age, glabrous skin was the most commonly affected body site with M. canis as the most frequent causative agent.


Subject(s)
Arthrodermataceae/isolation & purification , Dermatomycoses/epidemiology , Adolescent , Adult , Age Distribution , Aged , Arthrodermataceae/classification , Child , Child, Preschool , Dermatomycoses/microbiology , Female , Humans , Male , Middle Aged , Poland/epidemiology , Young Adult
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