Comparative Analysis of Putative Virulence-Associated Factors of Microsporum canis Isolates from Human and Animal Patients.

Microsporum canis is a zoophilic dermatophyte and the most common fungus isolated from dogs and cats worldwide. To invade skin, this pathogen uses different enzymes, which may be associated with virulence, that contribute to the fungal pathogenicity. The aim of this study is to compare the expression of enzymes that may be associated with virulence, and thermotolerance of M. canis strains isolated from dogs, cats, and humans. The in vitro expression of the enzymes keratinase, catalase, urease, hemolysin, and aspartic protease was evaluated in 52 M. canis strains recently isolated from 14 human patients, 12 dogs, 15 symptomatic, and 11 asymptomatic cats. In addition, thermotolerance was assessed by comparative analysis of fungal growth at 25 °C and 35 °C. Keratinase activity was low in 34 and moderate in 18 strains. Aspartic-protease activity was low in 7, moderate in 33, and high in 12 strains. Hemolysin activity was low in 44 and moderate in 8 strains. All strains were classified as low producers of catalase. All but three strains produced urease in vitro, with a broad range of activity. The strains presented in vitro growth at the two studied temperatures were classified as presenting low (36.5%), medium (44.3%), or high (19.2%) thermotolerance. There was no statistically significant difference in the new putative virulence-associated factors studied among the different hosts, which suggests that they may have a similar role on human, cat, and dog infection. Also, no difference was observed between strains isolated from symptomatic and asymptomatic cats. This suggests that these factors have a limited impact on the fate of feline dermatophytosis caused by M. canis.

Laser treatment of onychomycosis due to Neoscytalidium dimidiatum: An open prospective study.

Conventional systemic and topical treatments have proven ineffective for the treatment of onychomycosis caused by Neoscytalidium dimidiatum. Our aim was to evaluate the effectiveness and safety of laser monotherapy for the treatment of onychomycosis caused by this pathogen. Patients with clinical onychomycosis of the toenails and positive results both on direct mycological examination and N. dimidiatum culture underwent four 1064 nm Nd:YAG laser sessions with 6-week intervals between sessions. Participants were monitored by clinical examination supported by dermoscopy, measurement of diseased nail and the onychomycosis severity index (OSI), and by mycological examination for 12 months after completion of treatment. Treatment outcome was based on clinical and laboratory criteria and was divided in complete or partial cure, clinical improvement, treatment failure and relapse. No patient had complete or partial cure at any time during the study. Clinical improvement was observed in 40.6% of the patients at the end of the laser sessions; however, it did not persist during the follow-up. Treatment failure was observed in 64.7% of the patients at the end of 12 month follow-up period. Direct microscopy and culture results remained positive in most patients. Adverse events, in addition to treatment-related pain, were observed and considered severe in one case. The 1064 nm Nd:YAG laser was not able to cure onychomycosis caused by N. dimidiatum but temporarily improved the clinical appearance of the nail; however, adverse events may occur.

Performance of mycology and histopathology tests for the diagnosis of toenail onychomycosis due to filamentous fungi: Dermatophyte and non-dermatophyte moulds.

Improvement of laboratory diagnosis of onychomychosis is important so that adequate treatment can be safely implemented. To evaluate and compare the performance of mycological and histopathological examinations in onychomycoses caused by dermatophyte and non-dermatophyte moulds. Patients with lateral/distal subungual onychomycosis in at least one hallux were enrolled in the protocol and assessed via mycological and histopathological tests. The isolation of filamentous fungi was considered the gold standard. Test performance was evaluated through sensitivity, specificity and positive and negative predictive values. A total of 212 patients were enrolled in the study. Direct microscopy (DM) was positive in 57.5% patients, and cultures in 34.4%. Among these patients, 23.3% were positive for dermatophytes, with Trichophyton rubrum the most frequently isolated, and 86.3% were positive for non-dermatophytes, with Neoscytalidium dimidiatum predominance. Histopathology was positive in 41.0% samples. Direct microscopy showed better sensitivity for non-dermatophyte moulds (P=.000) and nail clipping was more specific for dermatophyte (P=.018). Histopathology of the distal nail plate is a valuable complementary tool for the diagnosis of onychomycosis caused by dermatophytes and direct microscopy is especially useful for non-dermatophyte molds.

Cutaneous murine model of infection caused by Neoscytalidium dimidiatum: a preliminary study of an emerging human pathogen.

Neoscytalidium dimidiatum is an emerging fungus that causes a skin infection similar to dermatophytosis; it affects both immunocompetent and immunosuppressed individuals, and it may invade deeper tissues and organs and cause systemic disease. Little is known about the etiopathogenesis of the infection caused by this fungus, and no standard effective treatment is available. The aim of the present experimental study was to develop an animal model of skin infection with N. dimidiatum. BALB/c mice were inoculated with two fungal strains, and different routes of infection were tested. When challenged intradermally, N. dimidiatum strain HUPE164165 caused skin infection in 67% of the animals whereas strain HUPE115669 did it in 49%. Neoscytalidium dimidiatum was isolated from the skin of 25% of the animals inoculated via epidermal scarification and from 100% of the animals challenged via subcutaneous injection. Mice inoculated intradermally were followed-up during four weeks, and clinical samples were collected on days 3, 8, 15, and 29 after inoculation, corresponding to different stages of infection. The cutaneous infection rate, as measured by the recovery of N. dimidiatum strain HUPE164165 from skin biopsies of animals inoculated intradermally, revealed the presence of infection in 90% of the animals sacrificed at 3 days post-inoculation, 71% at 8, 85% at 15, and 33% at 29. Conidia and hyphae were observed in PAS-stained sections as well as a mild to moderate inflammatory infiltrate in haematoxylin-eosin, although it did not differ from animals inoculated either with T. quinckeanum or PBS. The intradermal route of inoculation was considered to be suitable for the study of skin infection with N. dimidiatum The animal model developed in this preliminary study is the first to allow the study of cutaneous infection with N. dimidiatum and may contribute to further investigations of the aetiology, immunology, pathogenesis and treatment targeting this emerging mycosis.