Subtilisin 3 production from Microsporum canis is independent of keratin substrate availability.

Dermatophytes are highly infectious fungi that cause superficial infections in keratinized tissues in humans and animals. This group of fungi is defined by their ability to digest keratin and encompasses a wide range of species. We investigated a critical adhesion protein, subtilisin 3, utilized by Microsporum canis during initial stages of infection, analyzing its production and expression under varying growth conditions. Additionally, as this protein must be expressed and produced for dermatophyte infections to occur, we developed and optimized a diagnostic antibody assay targeting this protein. Subtilisin 3 levels were increased in culture when grown in baffled flasks and supplemented with either l-cysteine or cat hair. As subtilisin 3 was also produced in cultures not supplemented with keratin or cysteine, this study demonstrated that subtilisin 3 production is not reliant on the presence of keratin or its derivatives. These findings could help direct future metabolic studies of dermatophytes, particularly during the adherence phase of infections.

Dermatopathology and the Diagnosis of Fungal Infections.

Diagnosis of superficial/cutaneous fungal infections from skin, hair and nail samples is generally achieved using microscopy and culture in a microbiology laboratory, however, any presentation that is unusual or subcutaneous is sampled by taking a biopsy. Using histological techniques a tissue biopsy enables a pathologist to perform a full examination of the skin structure, detect any inflammatory processes or the presence of an infectious agent or foreign body. Histopathological examination can give a presumptive diagnosis while a culture result is pending, and may provide valuable diagnostic information if culture fails. This review demonstrates how histopathology contributes to the diagnosis of fungal infections from the superficial to the life threatening.

Dermatofitosis por Microsporum gypseum en un niño con inmunodeficiencia primaria / Microsporum gypseum dermatophytosis in a child with primary immunodeficiency

El Microsporum gypseum es un hongo geofílico que puede producir lesiones cutáneas inflamatorias en personas sanas. Se han descripto lesiones más extensas en pacientes inmunocomprometidos. Se presenta el caso de un paciente con dermatofitosis, con exámenes micológicos positivos para Candida sp, Epidermophytom floccosum y Trichophyton tonsurans, al que, ante la mala respuesta al tratamiento con griseofulvina e itraconazol a dosis habituales, se le realizó biopsia cutánea para cultivo que evidenció la presencia de M. gypseum. Debido a la extensión y a la mala respuesta al tratamiento, se realizó evaluación inmunológica y se diagnosticó un defecto en STAT1 con ganancia de función (STAT1-GOF). Los pacientes que tienen esta inmunodeficiencia primaria son susceptibles a las infecciones micóticas, especialmente por Candida, pero también, aunque en menor medida, a virus y bacterias. El paciente aquí presentado recibió tratamiento prolongado con antimicóticos imidazólicos sistémicos, con resolución de las lesiones.

Microsporum gypseum is a geophilic fungus that can cause inflammatory skin lesions in heathy people. More extensive lesions have been described in immunocompromised patients. We present a patient with extensive dermatophytosis, which mycological examination led the identification of Candida sp, Epidermophyton Floccosum and Trichophyton tonsurans and showed poor response to treatment with griseofulvina and itraconazol at usual doses. When skin biopsy was performed, it had positive culture for M. gypseum. Due to the extension and poor response to treatment, immunological assessment was performed and it showed a defect of STAT1 with gain of function (STAT 1-GOF). Patients with primary immunodeficiency are susceptible to fungal infections, especially Candida but also virus and bacteria, although to a lesser extent. The patient received long-term treatment with systemic imidazole antifungal recovering for the lesions.

[Microsporum gypseum dermatophytosis in a child with primary immunodeficiency]. / Dermatofitosis por Microsporum gypseum en un niño con inmunodeficiencia primaria.

Microsporum gypseum is a geophilic fungus that can cause inflammatory skin lesions in heathy people. More extensive lesions have been described in immunocompromised patients. We present a patient with extensive dermatophytosis, which mycological examination led the identification of Candida sp, Epidermophyton Floccosum and Trichophyton tonsurans and showed poor response to treatment with griseofulvina and itraconazol at usual doses. When skin biopsy was performed, it had positive culture for M. gypseum. Due to the extension and poor response to treatment, immunological assessment was performed and it showed a defect of STAT1 with gain of function (STAT 1-GOF). Patients with primary immunodeficiency are susceptible to fungal infections, especially Candida but also virus and bacteria, although to a lesser extent. The patient received long-term treatment with systemic imidazole antifungal recovering for the lesions.

El Microsporum gypseum es un hongo geofílico que puede producir lesiones cutáneas inflamatorias en personas sanas. Se han descripto lesiones más extensas en pacientes inmunocomprometidos. Se presenta el caso de un paciente con dermatofitosis, con exámenes micológicos positivos para Candida sp, Epidermophytom floccosum y Trichophyton tonsurans, al que, ante la mala respuesta al tratamiento con griseofulvina e itraconazol a dosis habituales, se le realizó biopsia cutánea para cultivo que evidenció la presencia de M. gypseum. Debido a la extensión y a la mala respuesta al tratamiento, se realizó evaluación inmunológica y se diagnosticó un defecto en STAT1 con ganancia de función (STAT1-GOF). Los pacientes que tienen esta inmunodeficiencia primaria son susceptibles a las infecciones micóticas, especialmente por Candida, pero también, aunque en menor medida, a virus y bacterias. El paciente aquí presentado recibió tratamiento prolongado con antimicóticos imidazólicos sistémicos, con resolución de las lesiones.

Gout-like hands.

A 65-year-old man presented to our hospital with a 1-year history of multiple nodules on his arms and hands without itching or pain.

Histoplasma capsulatum var. duboisii: A KwaZulu-Natal, South Africa public sector perspective.

BACKGROUND: Histoplasma capsulatum var. duboisii (H. duboisii) causes potentially fatal disease in the immunocompromised patient, and the literature on these cases is limited. The study was undertaken to describe the cases of H. duboisii seen in a resource-limited setting. METHODS: A 5-year retrospective, laboratory-based histopathologic review of all H. duboisii cases. RESULTS: A total of 24 patients were diagnosed with cutaneous H. duboisii infection. The male-to-female ratio was 1.4. All the patients were human immunodeficiency virus (HIV) positive, and 70.83% (17 of 24 patients) were on antiretroviral therapy. The clinically identified lesions were as follows: plaques, nodular plaques, papules, papulo-pustules, and umbilicated papules. Histopathological appraisal revealed organisms typical of H. duboisii. The commonest epidermal alterations were interface change, parakeratosis, spongiosis, ulceration, acanthosis, hyperkeratosis, transepidermal elimination, and exocytosis in decreasing order. The dermal changes included histiocytic pattern (n = 14), neutrophil-rich (n = 13), non-necrotizing granulomatous inflammation (n = 9), and karyorrhexis (n = 6). Histoplasma was confirmed in 53% (8 of 15 samples) of the tissue samples submitted for routine fungal culture. CONCLUSION: This study showed the demographics, clinical and histopathology features of H. duboisii infection in a resource-limited setting. Further research on histopathological features of this rare infection is essential to expand on the knowledge base and support findings in this study.

Novel approach of dermatophytosis eradication in shelters: effect of Pythium oligandrum on Microsporum canis in FIV or FeLV positive cats.

BACKGROUND: Shelters and similar facilities with a high concentration and fluctuation of animals often have problems with various infections, which are usually difficult to solve in such environments and are very expensive to treat. This study investigated the eradication of Microsporum canis, the widespread cause of zoonotic dermatophytosis in shelters, even in immunosuppressed feline leukaemia virus or feline immunodeficiency virus positive cats. RESULTS: Our study showed the increased effectiveness of an alternative topical therapy for affected animals using the mycoparasitic fungus Pythium oligandrum, which is gentler and cheaper than the standard systemic treatment with itraconazole, and which can also be easily used as a preventative treatment. A decrease in the number of M. canis colonies was observed in cats treated with a preparation containing P. oligandrum 2 weeks after the start of therapy (2 cats with P-1 score, 2 cats with P-2 score, 5 cats with P-3 score) compared with the beginning of the study (9 cats with P-3 score = massive infection). The alternative topical therapy with a preparation containing P. oligandrum was significantly more effective compared with the commonly used systemic treatment using itraconazole 5 mg/kg in a 6-week pulse. After 16 weeks of application of the alternative topical therapy, the clinical signs of dermatophytosis were eliminated throughout the whole shelter. CONCLUSION: The complete elimination of the clinical signs of dermatophytosis in all cats indicates that this therapy will be useful for the management and prevention of zoonotic dermatophytosis in animal shelters.

[Cutaneous complications following hematopoietic stem cell transplantation]. / Complications cutanées post-allogreffe de cellules souches hématopoïétiques.

BACKGROUND: Hematopoietic stem cell allograft is a treatment for patients with severe constitutional or acquired hematopoietic system diseases. This act is always linked to complications requiring multidisciplinary care. Our study describes the post-allograft cutaneous complications. METHODS: A prospective study was conducted at the Hematology department of "20 Août Hospital" in Casablanca during a period going from January 2018 to December 2020; including all patients who presented acute or chronic cutaneous complications post-allograft. RESULTS: Twenty-five patients were included. All patients received induction chemotherapy (Busulfan/Fludarabine or Busulfan/Melphalan). A skin infection was found in 8 patients : four cases of Malassezia folliculitis, one case of perineal zona, one case of genital herpes, one case of varicella and one case of Candida sepsis. The acute graft versus host reaction was found in 3 patients, revealed by an erythematous rash all over the body. The chronic graft versus host reaction was found in five patients on a lichenoid form. Nine patients had a hyperpigmentation of the folds followed by detachment in the same areas, concluding to a Busulfan toxidermy. DISCUSSION: Hematopoietic stem cell allograft has many complications. The literature mainly specifies hematological and digestive complications, while skin complications are little described. Our series is special by reporting different types and mechanisms of skin complications that can occur; with a predominance of skin graft-on-host reactions and infections. It also reports an unusual Busulfan toxidermy.

Ultraviolet Fluorescence as a Field-Applicable Screening Tool for Lesions Consistent with Ophidiomycosis in Lake Erie Watersnakes (Nerodia sipedon insularum).

Ophidiomycosis, commonly called snake fungal disease, has been linked to significant morbidity of free-ranging snakes in North America and Europe. Diagnosis of ophidiomycosis currently requires detection of skin lesions via physical exam or characteristic histopathology as well as detection of the causative agent, Ophidiomyces ophidiicola, through quantitative (q)PCR or fungal culture of a skin swab or tissue sample. While reliable, these methods require specialized training, invasive procedures (e.g., biopsy), and several days or weeks to receive results. Additionally, screening entire populations can quickly become costly. A fast, easy-to-use, cost-efficient, and sensitive screening tool is needed to optimize conservation strategies and treatment intervention. Our objective was to investigate the association between skin fluorescence under long-wave ultraviolet (UV) light (365 nm) and the detection of Ophidiomyces ophidiicola DNA using qPCR. Fifty-eight Lake Erie watersnakes (Nerodia sipedon insularum) collected in June of 2018 and 2019 from islands in western Lake Erie, Ottawa County, Ohio, US were visually inspected for skin lesions, photographed under natural light and UV light, and swabbed for qPCR analysis. Fluorescence was highly associated with the presence of skin lesions, and the presence of at least one fluorescent skin lesion was 86% sensitive and 100% specific for identifying animals with apparent ophidiomycosis, with a positive predictive value of 100%. While we recommend performing standard diagnostics along with fluorescence, our study supports the use of visual UV fluorescence identification as a preliminary, affordable, noninvasive, and field-applicable method to screen populations for ophidiomycosis.

Cutaneous Histoplasmosis in HIV Seronegative Patients: A Clinicopathological Analysis.

BACKGROUND: Diagnosis of extrapulmonary histoplasmosis in HIV seronegative and immunocompetent patients is often challenging, so a high index of suspicion is required. Cutaneous manifestation of infection shows a wide spectrum of lesions including erythematous plaques; maculopapules; crusted, verrucous, or desquamative papules and nodules; abscesses; and mucocutaneous ulcers among others. Due to the variations in its clinical presentation, histopathology plays a very important role in the detection of spores and the confirmation of diagnosis. OBJECTIVES: The aim of our study was to analyze clinicopathological characteristics of cutaneous manifestations of biopsy-proven histoplasmosis in HIV seronegative individuals. We also examined the utility of Fite stain for the diagnosis of Histoplasma capsulatum on tissue biopsy sections. METHODS: This was a retrospective, observational study on 7 patients who were HIV seronegative and clinically manifested with isolated cutaneous lesions or disseminated disease. Skin biopsy from the lesions was performed on all 7 patients. In addition to H&E staining and special stains for detecting fungus, Fite staining was performed on all of the cases to study its utility in detecting H. capsulatum spores. RESULTS: The skin lesions were widely disseminated in all patients and the most common cutaneous lesions were papules, present in all 7 patients. On review of the H&E-stained slides, the most common pattern was histiocytic lobular panniculitis-like infiltrate observed in 4 cases. Fite stain highlighted the yeast as magenta-colored spores on a blue background in all cases, except for 1 with a granulomatous pattern. CONCLUSION: A primary cutaneous manifestation of H. capsulatum infection in non-HIV-infected individuals is extremely rare. Fite stain could aid in differentiating the spores of H. capsulatum from those of other fungi, Cryptococcus and Candida in particular.

Metabolomic analysis of Trichophyton rubrum and Microsporum canis during keratin degradation.

Keratin is important and needed for the growth of dermatophytes in the host tissue. In turn, the ability to invade keratinised tissues is defined as a pivotal virulence attribute of this group of medically important fungi. The host-dermatophyte interaction is accompanied by an adaptation of fungal metabolism that allows them to adhere to the host tissue as well as utilize the available nutrients necessary for their survival and growth. Dermatophyte infections pose a significant epidemiological and clinical problem. Trichophyton rubrum is the most common anthropophilic dermatophyte worldwide and its typical infection areas include skin of hands or feet and nail plate. In turn, Microsporum canis is a zoophilic pathogen, and mostly well known for ringworm in pets, it is also known to infect humans. The aim of the study was to compare the intracellular metabolite content in the T. rubrum and M. canis during keratin degradation using liquid chromatography system coupled with tandem mass spectrometer (LC-MS/MS). The metabolite "fingerprints" revealed compounds associated with amino acids metabolism, carbohydrate metabolism related to the glycolysis and the tricarboxylic acid cycle (TCA), as well as nucleotide and energy metabolism. The metabolites such as kynurenic acid, L-alanine and cysteine in case of T. rubrum as well as cysteine and riboflavin in case of M. canis were detected only during keratin degradation what may suggest that these compounds may play a key role in the interactions of T. rubrum and M. canis with the host tissue. The metabolomic results were completed by qPCR gene expression assay. Our findings suggest that metabolomic analysis of T. rubrum and M. canis growing in culture media that mimic the dermatophyte infection could allow the understanding of processes involved in the pathogenesis of dermatophytes.

Synergistic antifungal evaluation of over-the-counter antifungal creams with turmeric essential oil or Aloe vera gel against pathogenic fungi.

BACKGROUND: The frequent incidence of fungal infection and widespread of antibiotic resistance are emergent concerns in public health. Hence, there is a need to harness the potential of natural bioactive compounds from plant towards treatment of fungal infection. Combination effect of antibiotic creams with natural products from plants is prospective strategy to produce new antifungal agent. This study therefore, revealed antifungal effect of combined Antifungal Creams (AFCs) with Turmeric Essential Oil (TEO) or Aloe vera Gel (AVG). METHODS: Phytochemicals and bioactive compounds in TEO and AVG were revealed using GC-MS. Bioactive compounds in plant extracts were compared to known compounds in database library of National Institute of Standards and Technology (U.S.). Antifungal activity and synergistic effect of AFCs with TEO or AVG were carried out using agar well diffusion method. RESULTS: Phenol, flavonoids, saponins, alkaloids, steroids, terpenoids and cardiac glycosides were present in TEO and AVG. GCMS revealed thirty-six (36) and eighteen (18) bioactive compounds in TEO and AVG, respectively. AFCs displayed zones of inhibition with values ranged from 5.0 to 14.3 mm, TEO was 5.0 to 11.0 mm and AVG was 8.0 to 11.7 mm against tested fungi. Minimum Inhibitory Concentration (MIC) by AFCs, TEO and AVG ranged from 1.25 to 10.0 mg/ml. Combinatory effects of AFCs with TEO or AVG revealed synergistic and indifferent properties. CONCLUSION: Development of novel products using bioactive ingredients from plants with commercially available AFCs will serve as potential alternative therapy to cure dermatological infections with no side effects.

Diagnostic Utility of Biplex/Multiplex Polymerase Chain Reaction in Infectious Granulomatous Dermatitis in North Indian Population.

BACKGROUND: A definite diagnosis of infectious granulomatous dermatitis (IGD) is difficult for both practicing dermatologists and dermatopathologists due to overlapping clinical and histomorphological features. We aimed to explore the role of multiplex polymerase chain reaction (PCR) for identifying a definite etiological agent for diagnosis and appropriate treatment in IGD in formalin-fixed paraffin-embedded tissue. MATERIALS AND METHODS: Sixty-two cases of IGD were included, excluding leprosy. The histochemical stains including Ziehl-Neelsen, periodic acid-Schiff, and Giemsa were performed in all cases. A multiplex PCR was designed for detection of tuberculosis (TB) (IS6110 and mpt64), fungal infections (ITS1, ITS2; ZM1, and ZM3), and leishmaniasis (kDNA). The results of histomorphology, histochemical stains, and multiplex PCR were compared. RESULTS: Among 62 cases, the sensitivity rate of PCR detection for organisms was 16.7%, 0%, 100%, 72%, 75%, and 66.7% in patients with TB, suggestive of TB, leishmaniasis, fungal infections, and granulomatous dermatitis not otherwise specified and granulomatous dermatitis suggestive of fungus, respectively. The TB PCR using IS6110 primers was negative in all cases; however, PCR using mpt64 primers was positive in 33.33% cases of scrofuloderma. The histochemical stains including Ziehl-Neelsen for acid-fast bacilli, periodic acid-Schiff for fungus, and Giemsa for Leishman-Donovan bodies showed positivity in 11.3%, 43.5%, and 3.2%, respectively. CONCLUSION: A multiplex PCR (Mycobacterium tuberculosis, Leishmania, and panfungal) is highly recommended in all cases of IGD where an etiological agent is difficult to establish by skin biopsy and histochemical stains along with a clinicopathological correlation. This will augment in appropriate treatment and will reduce empirical treatment and morbidity in such patients.

A rare case of cutaneous Papiliotrema (Cryptococcus) laurentii infection in a 23-year-old Caucasian woman affected by an autoimmune thyroid disorder with hypothyroidism.

In recent years, the frequency of infections due to saprophytic fungi has increased. Cryptococcus laurentii, recently classified as Papiliotrema laurentii, is responsible for fungemia, meningitis, and superficial infections. Here, we report the first case of cutaneous Papiliotrema (Cryptococcus) laurentii infection in a 23-year-old Caucasian woman affected by an autoimmune thyroiditis with hypothyroidism. Impairments of the immune system are often associated with unusual fungal infections, which cannot be neglected. The isolate strain was susceptible to Amphotericin B while resistant to fluconazole, itraconazole, voriconazole, and terbinafine. The patient was successfully treated with Amphotericin B.

Saccharomyces cerevisiae as a skin physiology, pathology, and treatment model.

Saccharomyces cerevisiae serves as a useful model in experimental biology. Within dermatology research, several studies have examined this organism's role in skin physiology, pathology, and treatment. Saccharomyces cerevisiae has been used to explore the mechanisms of melanogenesis as its extract inhibits key enzymes involved in melanogenesis and melanosome transfer. Additionally, the lack of probiotic intestinal Saccharomyces cerevisiae has been associated with psoriasis, potentially related to the anti-inflammatory effects of the yeast. Furthermore, antibodies against Saccharomyces cerevisiae have been observed in skin conditions, including atopic dermatitis. Saccharomyces cerevisiae may even cause skin infections, such as septic emboli in a patient with acute myelogenous leukemia. Lastly, Saccharomyces cerevisiae has potential use in vaccine development against melanoma and is utilized to study various treatment modalities such as zinc pyrithione, an ingredient often used in anti-dandruff shampoo.

[Subcutaneous infection by Curvularia pallescens in a lung transplant recipient: First report in Spain]. / Micosis cutánea por Curvularia pallescens en un paciente trasplantado pulmonar: primer caso descrito en España.

BACKGROUND: Curvularia is a filamentous dematiaceous fungus increasingly recognized as a pathogen in immunocompromised patients. The most common clinical entities associated with this fungus are allergic sinusitis, cutaneous infection and keratitis. In this article, a report on the first clinical case of Curvularia pallescens cutaneous infection in Spain and its treatment is described. CASE REPORT: A 68 year-old man with a history of lung transplantation presented to Dermatology Unit due to a skin lesion in the knee that had been evolving for 6 months. A skin biopsy was performed for its study. In the histopathological study, an intense and non-specific inflammatory reaction in the dermis was observed, and with Grocott stain and periodic acid Schiff abundant septate hyphae and spores were found in the dermis. The culture of the sample revealed a filamentous fungus whose microscopic examination allowed to identify the genus as Curvularia. Using MALDI-TOF mass spectrometry and molecular identification, the fungus was finally identified as Curvularia pallescens. The patient underwent surgical resection of the lesion and was treated with posaconazole, evolving favorably. CONCLUSIONS: The species of Curvularia should be considered causal agents of fungal skin infections in immunosuppressed patients. This clinical case, which showed good clinical response after surgical resection and treatment with posaconazole, is the first described in Spain due to this species.

Pseudomycetoma of the scalp caused by Microsporum canis

Abstract Pseudomycetoma is an extremely rare deep mycosis, caused by dermatophytic fungi that penetrate the tissue from infected follicles of tinea capitis. Both clinically and histopathology are similar to eumycetoma, being distinguished through the isolation of the fungus, which in the case of pseudomycetoma can be Microsporum spp. or Trichophyton spp. genre. We present a 24-year-old man with an exuberant tumor in the occipital region with fistula, whose histopathological examination evidenced grains composed of hyaline hyphae and the culture for fungi isolated the agent Microsporum canis. Combined treatment of surgical excision followed by oral griseofulvin for two years was performed, with resolution of the condition.

Pseudomycetoma of the scalp caused by Microsporum canis.

Pseudomycetoma is an extremely rare deep mycosis, caused by dermatophytic fungi that penetrate the tissue from infected follicles of tinea capitis. Both clinically and histopathology are similar to eumycetoma, being distinguished through the isolation of the fungus, which in the case of pseudomycetoma can be Microsporum spp. or Trichophyton spp. genre. We present a 24-year-old man with an exuberant tumor in the occipital region with fistula, whose histopathological examination evidenced grains composed of hyaline hyphae and the culture for fungi isolated the agent Microsporum canis. Combined treatment of surgical excision followed by oral griseofulvin for two years was performed, with resolution of the condition.

Effect of chlorogenic and gallic acids combined with azoles on antifungal susceptibility and virulence of multidrug-resistant Candida spp. and Malassezia furfur isolates.

Chlorogenic acid (CHA) and gallic acid (GA) are safe natural phenolic compounds that are used as enhancers of some drugs in influencing antioxidant, anticancer, and antibacterial activities. Among fungi, Candida spp. and Malassezia spp. are characterized by an increasing prevalence of multidrug resistance phenomena and by a high morbidity and mortality of their infections. No data are available about the efficacy of CHA and GA combined with azoles on the antifungal susceptibility and on the virulence of both fungi. Therefore, their antifungal and antivirulence effects have been tested in combination with fluconazole (FLZ) or ketoconazole (KTZ) on 23 Candida spp. and 8 M. furfur isolates. Broth microdilution chequerboard, time-kill studies, and extracellular enzymes (phospholipase and hemolytic) activities were evaluated, displaying a synergistic antifungal action between CHA or GA and FLZ or KTZ on C. albicans, C. bovina, and C. parapsilosis, and antagonistic antifungal effects on M. furfur and Pichia kudriavzevii (Candida krusei) isolates. The time-kill studies confirmed the chequerboard findings, showing fungicidal inhibitory effect only when the GA was combined with azoles on Candida strains. However, the combination of phenolics with azoles had no effect on the virulence of the tested isolates. Our study indicates that the combination between natural products and conventional drugs could be an efficient strategy for combating azole resistance and for controlling fungistatic effects of azole drugs.