Direct Mycological Examination Research Articles

Label-Free Optical Transmission Tomography for Direct Mycological Examination and Monitoring of Intracellular Dynamics

Live-cell imaging generally requires pretreatment with fluorophores to either monitor cellular functions or the dynamics of intracellular processes and structures. We have recently introduced full-field optical coherence tomography for the label-free live-cell imaging of fungi with potential clinical applications for the diagnosis of invasive fungal mold infections. While both the spatial resolution and technical set up of this technology are more likely designed for the histopathological analysis of tissue biopsies, there is to our knowledge no previous work reporting the use of a light interference-based optical technique for direct mycological examination and monitoring of intracellular processes. We describe the first application of dynamic full-field optical transmission tomography (D-FF-OTT) to achieve both high-resolution and live-cell imaging of fungi. First, D-FF-OTT allowed for the precise examination and identification of several elementary structures within a selection of fungal species commonly known to be responsible for invasive fungal infections such as Candida albicans, Aspergillus fumigatus, or Rhizopus arrhizus. Furthermore, D-FF-OTT revealed the intracellular trafficking of organelles and vesicles related to metabolic processes of living fungi, thus opening new perspectives in fast fungal infection diagnostics.

Favorable outcome of Lasiodiplodia theobromae keratomycosis : a clinical case and systematic review

BackroudKeratitis caused by Lasiodiplodia theobromae is rare and typically associated with a poor prognosis. Current literature lacks sufficient evidence on effective management of patients with this condition.Case presentationA 74-year-old former agricultural worker presented with a red right eye, discomfort, and decreased visual acuity, progressing over three days without treatment. Examination revealed type 2 diabetes and a non-perforating, spiculated corneal abscess with a hypopyon in the right eye. Initial treatment included a triple antibiotic therapy and supportive care. Direct mycological examination identified numerous septate mycelial filaments. Antifungal treatment with natamycin and voriconazole, both topically and orally, was initiated. Cultures confirmed Lasiodiplodia theobromae. The patient showed significant improvement. Treatment continued for eight weeks, with a final visual acuity of 20/50 due to a stromal scar.ConclusionAn extensive literature review conducted in November 2023, using databases such as PubMed and Google Scholar with the keywords “lasiodiplodia” and “keratitis” yielded no previous cases of this specific condition being managed solely with the combined use of natamycin and voriconazole. This antifungal combination is commonly included in most management protocols for fungal keratitis. Factors such as the use of corticosteroids and delayed diagnosis were noted to adversely affect the prognosis. This case and this systematic review underscores the potential for non-surgical management options in severe fungal keratitis.

Identification of primary metabolites in fungal species of Trichophyton mentagrophyte and Trichophyton rubrum by NMR spectroscopy.

Superficial mycoses are fungal infections limited to the outermost layers of the skin and its appendages. The chief causative agents of these mycoses are dermatophytes and yeasts. The diagnosis of dermatophytosis can be made by direct mycological examination with potassium hydroxide (10%-30%) of biological material obtained from patients with suspected mycosis, providing results more rapid than fungal cultures, which may take days or weeks. This information, together with clinical history and laboratory diagnosis, ensures that the appropriate treatment is initiated promptly. However, false negative results are obtained in 5%-15%, by conventional methods of diagnosis of dermatophytosis. To study the metabolic profiles of the commonly occurring dermatophytes by NMR spectroscopy. We have used 1D and 2D Nuclear Magnetic Resonance (NMR) experiments along with Human Metabolome Database (HMDB) and Chenomx database search for identification of primary metabolites in the methanol extract of two fungal species: Trichophyton mentagrophyte (T. mentagrophyte) and Trichophyton rubrum (T. rubrum). Both standard strains and representative number of clinical isolates of these two species were investigated. Further, metabolic profiles obtained were analysed using multivariate analysis. We have identified 23 metabolites in the T. mentagrophyte and another 23 metabolites in T. rubrum. Many important metabolites like trehalose, proline, mannitol, acetate, GABA and several other amino acids were detected, which provide the necessary components for fungal growth and metabolism. Altered metabolites were defined between Trichophyton mentagrophyte and T. rubrum strains. We have detected many metabolites in the two fungal species T. mentagrophyte and T. rubrum by using NMR spectroscopy. NMR spectroscopy provides a holistic snapshot of the metabolome of an organism. Key metabolic differences were identified between the two fungal strains. We need to perform more studies on metabolite profiling of the samples from these species for their rapid diagnosis and prompt treatment.

Deep cutaneous mycoses in kidney transplant recipients: Diagnostic and therapeutic challenges.

Deep cutaneous mycoses (DCMs) are rare infections that extend throughout the dermis and subcutis, often occurring after inoculation with pathogenic fungi. Trends toward a growing incidence have been observed that may be partially related to an increasing population of solid organ transplant patients. The aim of this study is to describe the diagnostics and the outcomes of DCM among kidney transplant recipients so as to optimize their management. We performed a retrospective review of cases of DCM occurring among kidney transplant recipients in our institution over 12 years. Twenty cases were included. Lesions were only located on the limbs and presented mainly as single (10/20, 50%) nodular lesions (15/20, 75%), with a mean size of 3cm. Direct mycological examination was positive for 17 patients (17/20, 85%) and the cultures were consistently positive. Thirteen different fungal species were observed, including phaehyphomycetes (n=8), hyalohyphomycetes (n=3), dermatophytes (n=1), and mucorale (n=1). The (1-3) beta-D-glucan antigen (BDG) was also consistently detected in the serum (20/20, 100%). Systematic imaging did not reveal any distant infectious lesions, but locoregional extension was present in 11 patients (11/14, 79%). Nineteen patients received antifungal treatment (19/20, 95%) for a median duration of 3 months, with surgery for 10 (10/20, 50%). There is a great diversity of fungal species responsible for DCMs in kidney transplant recipients. The mycological documentation is necessary to adapt the antifungal treatment according to the sensitivity of the species. Serum BDG positivity is a potentially reliable and useful tool for diagnosis and follow-up.

Rare Presentation of Chromoblastomycosis Due to Multiple Simultaneous Inoculations: A Case Report

Chronic infectious, granulomatous and suppurative dermatosis, classified among the subcutaneous mycoses, prevalent in tropical and subtropical regions and caused by the traumatic implantation of dematiaceous fungal species, where the presence of muriform bodies is an expression of the causal agent in the grafted tissue, are characteristic of chromoblastomycosis. Considered the second implantation mycosis in the world, it manifests itself with slow and progressive growth lesions of exophytic and verrucous plaques and black dots on the surface. The disease is considered a neglected and occupational disease, which occurs mainly among agricultural workers, coconut and babassu harvesters, lumberjacks, and traders of agricultural products. It is important to highlight that people at risk of contracting chromoblastomycosis work in tropical countries, where the temperature can be above 40 °C in summer, and generally refuse to wear protective equipment during the day (shoes, gloves, clothes, etc.), although they know that this type of prophylactic measure can prevent different types of diseases. These vulnerable people often live in low-income countries and sometimes live far from medical services and, once infected, do not seek medical attention. We report below an exuberant and unusual case due to multiple simultaneous inoculations on the back after the patient suffered an accidental fall while working on his farm. The diagnosis was established by direct mycological examination, histopathological examination, culture, microculture, and MALDI-TOF MS analysis that identified the agent of the Fonsecaea pedrosoi.

Report of terbinafine resistant Trichophyton spp. in Italy: Clinical presentations, molecular identification, antifungal susceptibility testing and mutations in the squalene epoxidase gene.

Numerous reports of resistance to terbinafine in Trichophyton spp. from all over the world are arousing justified attention and concern. Point mutations in the gene that encodes the squalene epoxidase (SQLE) enzyme are responsible for these therapeutic resistances. Primary objective of the study was to describe first isolates of Trichophyton spp. resistant to terbinafine among the patients treated between September 2019 and June 2022 at the Dermatology Units of Ospedale Maggiore Policlinico and San Bortolo Hospital. Secondary objective was to study the resistance mechanism. Patients with confirmed Trichophyton spp. infection has been treated with systemic and topical terbinafine. Patients were then re-evaluated 12 weeks after the therapy. Patients with incomplete or absent response to terbinafine underwent a new skin scraping for direct mycological examination, new identification of dermatophyte species from culture and MALDI-TOF, molecular species identification, antifungal susceptibility testing and molecular analysis of SQLE gene. We identified five patients without clinical response to treatment with terbinafine. The DNA sequencing of the ITS region identified one Trichophyton rubrum and four Trichophyton indotineae. The T. rubrum strain showed minimum inhibitory concentration (MIC) (90% growth inhibition) of 4 mg/L for terbinafine. The four T. indotineae strains showed a MICs range of 0.25-4 mg/L for terbinafine. The analysis of the SQLE gene in the T. rubrum strain showed a nucleotide substitution generating a missense mutation (L393F). The SQLE gene sequencing in the T. indotineae strains showed a nucleotide substitution generating a missense mutation (F397L) in two strains, a nucleotide substitution L393S in one strain and a nucleotide substitution F415C in another strain. We report the first cases of terbinafine-resistant Trichophyton isolates in the Italian population. Solid antifungal management programs will be needed to promote more responsible use of antimycotics and preserve their therapeutic efficacy to control antifungal resistance.

Development of a Multiplex qPCR Assay for Fast Detection and Differentiation of Paracoccidioidomycosis Agents.

Classic paracoccidioidomycosis (PCM) is a potentially deadly neglected tropical systemic mycosis caused by members of the Paracoccidioides brasiliensis complex (P. brasiliensis s. str., P. americana, P. restrepiensis, and P. venezuelensis) and P. lutzii. The laboratorial diagnosis of PCM relies on observing pathognomonic structures such as the "steering wheel" or "Mickey Mouse" shape in the direct mycological examination, fresh biopsied tissue in 10% KOH, histopathological analysis, and/or the isolation of the fungus in culture. However, these procedures are time-consuming and do not allow for the speciation of Paracoccidioides due to overlapping morphologies. Here, we propose a new one-tube multiplex probe-based qPCR assay to detect and recognize agents of the P. brasiliensis complex and P. lutzii. Primers (Paracoco-F and Paracoco-R) and TaqMan probes (PbraCx-Fam, Plu-Ned, and Paracoco-Vic) were developed to target the rDNA (ITS2/28S) in the Paracoccidioides genome. A panel of 77 Paracoccidioides isolates revealed a 100% specificity (AUC = 1.0, 95% CI 0.964-1.000, p < 0.0001) without cross-reacting with other medically relevant fungi or human and murine DNA. The lower limit of detection was 10 fg of gDNA and three copies of the partial rDNA amplicon. Speciation using qPCR was in perfect agreement with AFLP and TUB1-RFLP markers (kappa = 1.0). As a proof of concept, we assessed a panel of 16 formalin-fixed and paraffin-embedded specimens from histopathologically confirmed PCM patients to reveal a significant sensitivity of 81.25% and specificity of 100% (AUC = 0.906 ± 0.05, 95% CI = 0.756-0.979, p < 0.0001, Youden index J = 0.8125). Our assay achieved maximum sensitivity (100%) and specificity (100%) using fresh clinical samples (n = 9) such as sputum, bronchoalveolar lavage, and tissue fragments from PCM patients (AUC = 1.0, 95% CI 0.872-1.000, p < 0.0001, Youden index J = 1.0). Overall, our qPCR assay simplifies the molecular diagnosis of PCM and can be easily implemented in any routine laboratory, decreasing a critical bottleneck for the early treatment of PCM patients across a vast area of the Americas.

Survey on Dermatophytes Isolated from Animals in Switzerland in the Context of the Prevention of Zoonotic Dermatophytosis

Most inflammatory dermatophytoses in humans are caused by zoophilic and geophilic dermatophytes. Knowledge of the epidemiology of these fungi in animals facilitates the prevention of dermatophytosis of animal origin in humans. We studied the prevalence of dermatophyte species in domestic animals in Switzerland and examined the effectiveness of direct mycological examination (DME) for their detection compared to mycological cultures. In total, 3515 hair and skin samples, collected between 2008 and 2022 by practicing veterinarians, were subjected to direct fluorescence microscopy and fungal culture. Overall, 611 dermatophytes were isolated, of which 547 (89.5%) were from DME-positive samples. Cats and dogs were the main reservoirs of Trichophyton mentagrophytes and Microsporum canis, whereas Trichophyton benhamiae was predominantly found in guinea pigs. Cultures with M. canis significantly (p < 0.001) outnumbered those with T. mentagrophytes in DME-negative samples (19.3% versus 6.8%), possibly because M. canis can be asymptomatic in cats and dogs, unlike T. mentagrophytes, which is always infectious. Our data confirm DME as a reliable, quick, and easy method to identify the presence of dermatophytes in animals. A positive DME in an animal hair or skin sample should alert people in contact with the animal to the risk of contracting dermatophytosis.

Tinea capitis: new insights into an old disease

Tinea capitis is a superficial dermatophytic infection of the scalp. This common dermatosis occurs predominantly in children. The clinical manifestation of the disease is heterogeneous, and vary widely depending on the pathogenic fungal agent. Direct mycological examination and cultures are mandatory for an accurate diagnosis and species identification. Treatment should be both local and systemic, and ideally is tailored to the dermatophytic species identified by the laboratory diagnostic work up. Secondary prophylaxis through supplementary measures is crucial to avoid epidemic outbreak and patient reinfection.

Chromoblastomycosis: A case series from Eastern China.

Chromoblastomycosis (CBM) is a chronic fungal infection of the cutaneous and subcutaneous tissues caused by brown pigmented fungi. Fonsecaea monophora is one of the most common pathogens of CBM in China. Most formal cases have been reported from Southern China, however, the infection is not uncommon in Eastern China where very few case series are available. To describe the clinical aspects of CBM, we report a series of 11 cases between 2018 and 2021 at a single medical center in Eastern China. The patients were predominately male (n = 9) and the disease duration ranged from 3 months to 20 years. Plaque type lesions were the most common clinical manifestations. There were 7 cases of mild forms and 3 cases of severe forms. Among the 3 severe cases, one case gave up treatment due to economic poverty; one case did not respond to a 1-year systemic treatmen; one case was cured by combination therapy of 10 months. Other cases were cured by treatment with antifungal agents. All cases of direct mycological examination were positive. All isolates were identified by morphology and sequencing of the the ITS regions of ribosomal DNA, Ten were F. monophora and 1 was Cladophialophora carrionii. All cases had been evaluated at other clinics, where 8 cases were misdiagnosed as other diseases. As a neglected tropical disease (NTD), CBM is still a major challenge in the field of dermatology, especially in its severe clinical forms. As an effective and simple diagnostic method of CBM, direct microscopic examination should be further promoted in rural hospitals.

P456 Defungi: direct mycological examination of microscopic fungi images

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PMObjectiveTo classify five fungi types using two different deep learning approaches and three different convolutional neural network models, VGG16, Inception V3, and ResNet50.MethodA mycological laboratory in Colombia donated the images used for the development of this research work. They were manually labeled into five classes and curated with subject matter expert assistance. The images were later cropped and modified with automated coding routines to produce the final dataset.ResultsWe present experimental results classifying five types of fungi using two different deep learning approaches and three different convolutional neural network models, VGG16, Inception V3, and ResNet50. The first approach benchmarks the classification performance for the models trained from scratch, while the second approach benchmarks the classification performance using pre-trained models based on the ImageNet dataset. Using k-fold cross-validation testing on the 5-class dataset, the best performing model trained from scratch was Inception V3, reporting 73.2% accuracy. Likewise, the best performing model using transfer learning was VGG16, with 85.04% accuracy.ConclusionThe statistics provided by the two approaches create an initial benchmark to encourage future research work to improve classification performance. Furthermore, the dataset built is published on Kaggle and GitHub to encourage future research.

P308 Jorge lobo's disease in brazil: a new recognized reservoir?

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMLobomycosis (lacaziosis) is a chronic subcutaneous disease caused by the uncultivated fungus Lacazia loboi. The disease was first described in 1931 by Jorge Lobo in Brazil and has been reported in South and Central America. Lobomycosis is a rare disease that affects certain geographic regions, mainly in countries of low socioeconomic status, but its prevalence among indigenous people in Brazil is exceptionally high. The state of Maranhão is located in the northeast region of the country and there are rare clinical reports of imported cases of the disease in our region. This is the first published series of autochthonous cases of lacaziosis in Maranhão.ObjectiveTo describe seven cases of Lacazia loboi infections in immunocompetent patients in an endemic area for other subcutaneous mycoses.MethodsWe retrospectively reviewed the medical records of all patients who developed Jorge Lobo's diseases in a new area recognized as a potential environment reservoir. An incident case was defined as a patient who developed proven lacaziosis based on the presence of topical fungal elements in the tissue on histopathology or mycologic direct exam. Epidemiological and clinical data of all cases of lacaziosis were collected using a standard clinical form.ResultsIn the period from 2000 to 2021, seven patients were identified. All cases were diagnosed by histopathology and direct mycological examination of the lesions. A total of 6 out of 7 (85.7%) patients were men and the mean age was 57 years (52-70 years). The median time between onset of infection symptoms and diagnosis was ˃2 years in 6 patients, with 15 years of symptoms duration in patient 7. All patients reported having worked with agricultural activities before. A total of 50% of patients were diagnosed with localized lesions and the other half with disseminated lesions. Subcutaneous lesions involved the upper limbs (4/6; 66.6%), ears (2/6; 33.3%), lower limbs (1/6; 16.6%), and trunk (1/6; 16.6%). All patients exhibited nodules as a dominant pattern of skin lesion, sometimes coalescing with the tumor's appearance. Regarding treatment, a combination of itraconazole and clofazimine therapy was added to all patients. Surgical excision was possible only in 2 episodes due to the limited availability of this procedure at the site. Cure was documented in 2 cases, with no recurrence to date.ConclusionThe present series represents the first and largest collection of case studies of lacaziosis in Maranhão state, with all cases classified as autochthonous. Lacaziosis involves mainly adult males with different occupational risks such as agricultural labor, and other workers exposed to contaminated soil and plant materials. Few data are available to support suggested therapy with itraconazole and clofazime. Excision surgery is important for treatment due to the absence of clinical trials in the medical literature. Epidemiological surveillance studies of new cases of lacasiozis in Maranhão are important for mapping this new reservoir and developing public policies for diagnosis and treatment.

ATYPICAL ORAL MANIFESTATION ASSOCIATED WITH CANDIDA COLLICULOSA IN A PEDIATRIC HEMATOLOGICAL PATIENT

A 9-year-old boy under chemotherapeutic treatment for acute myeloid leukemia evolved with edema in the lower lip, with a crusted ulcer circumscribed by an erythematous and painless area, purulent secretion, and pseudomembrane, measuring approximately 1.5 cm at its largest diameter, with no history of trauma. The patient was being treated with cefepime, fluconazole, vancomycin, metronidazole, hydrocortisone, and tretinoin, as well as antiemetic and antihistamine drugs. Cell smears were collected for exfoliative cytology; direct mycologic, bacterial, and viral culture; and fungi investigation. No viruses or bacteria were identified. The direct mycologic examination showed yeast cells without pseudohyphae and the search for fungi was positive for <i>Candida colliculosa</i>. The technique used to identify the <i>Candida</i> strain was matrix-assisted laser desorption/ionization-time-of-flight. The dose of fluconazole was increased, and the improvement of the lesion accompanied the patient's immune recovery until hospital discharge.

Accuracy of direct examination and culture as compared to the anatomopathological examination for the diagnosis of chromoblastomycosis: a systematic review

BackgroundChromoblastomycosis is a skin infection caused by dematiaceous fungi that take the form of muriform cells in the tissue. It mainly manifests as verrucous plaques on the lower limbs of rural workers in tropical countries. ObjectivesThe primary objective of this review is to evaluate the accuracy of diagnostic methods for the identification of chromoblastomycosis, considering the histopathological examination as the reference test. MethodsMEDLINE, LILACS and Scielo databases were consulted using the terms “chromoblastomycosis” AND “diagnosis”. The eligibility criteria were: studies that evaluated the accuracy of tests for the diagnosis of chromoblastomycosis. Eleven studies were selected. Statistical analysis included the calculation of sensitivity and specificity of the diagnostic methods. ResultsConsidering the histopathological examination as the reference test, the culture showed a sensitivity (S) of 37.5% - 90.9% and a specificity (Sp) of 100%; while direct mycological examination showed S = 50% - 91.6% and Sp of 100% . Considering the culture as the reference test, the serology (precipitation techniques) showed S of 36% - 99%; and Sp of 80% - 100%; while the intradermal test showed S of 83.3% - 100% and Sp of 99.4% - 100%. Study limitationsThe small number of studies and very discrepant sensitivity results among them do not allow the calculation of summary measures through a meta-analysis. ConclusionsDirect mycological examination, culture, intradermal test and serology show sensitivity and specificity values ​​for the diagnosis of chromoblastomycosis with no significant difference between the studies.

Multiple ulcerated lesions in a patient from Amazon, Brazil

Multiple ulcerated lesions in a patient from Amazon, Brazil

Identification of Candida colliculosa in an oral ulcer of a pediatric hematological patient

INTRODUCTION: Candida colliculosa is a yeast of food origin classified as Non-Albicans Candida (NAC), with its pathogenic potential confirmed, but little known. Like other opportunistic infections, diseases caused by NAC species are most commonly found in immunosuppressed patients. OBJECTIVES: The objective of this work is to report the case of a pediatric patient who developed an atypical lesion associated with Candida colliculosa in the mouth. CASE REPORT: A 9-year-old boy under chemotherapeutic treatment for Acute Myeloid Leukemia developed edema in the lower lip, with a persistent crusted ulcer circumscribed by an erythematous, painless, and progressing negatively. He was using antibiotics, antifungals, antiemetics, and antihistamines. RESULTS: It was decided to perform complementary exams for final diagnosis. Cell smears were collected for exfoliative cytology, direct mycological, bacterial and viral culture, and fungi investigation. No viruses and bacteria were identified. A direct mycological examination showed yeast cells without pseudohyphae and a Maldi-TOF technique confirmed the diagnosis of Candida colliculosa. FINAL CONSIDERATIONS: The use of antifungal agents associated with the patient's immunological recovery led to the complete healing of the lesion and hospital discharge. Identification of the fungal species was important not only to establish the diagnosis and most appropriate treatment but also to avoid the fungus spreading in the hospital environment.

Coexistence of onychomycosis and nail psoriasis and its correlation with systemic treatment.

Epidemiological studies indicate that onychomycosis may affect up to 79% of psoriatic patients. Onychomycosis in psoriatic patients is more commonly caused by yeasts comparing with non-psoriatic. Evaluate the prevalence of fungi in nail psoriasis. Evaluate the association between direct mycological examination (DME) and mycological culture, Nail Psoriasis Severity Index (NAPSI) and systemic treatment for psoriasis. Of 133 nails from 20 patients with nail psoriasis were evaluated as follows: 9 patients were using topical treatment and 11 were on systemic treatment. The assessment of psoriasis severity using NAPSI was performed in all psoriatic nails. The presence of fungi was confirmed in DME and culture. DME showed the presence of fungal elements in 45 nails (33.83%) with a predominance of blastoconidia (95.5%) No septate hyphae were seen. Mycological culture was positive in 36 (27.06%) samples. Among them, Candida grew in 31 (86.1%): Candida parapsilosis in 15 (48.38%), Candida spp in 14 (45.16%). No growth of dermatophytes was observed. Patients with systemic treatment showed a higher frequency of positive test (DME and culture) for fungi when compared to topic treatment (p:.006). There was a positive correlation between NAPSI, mycological culture and systemic treatment (p:.0063); the risk was four times higher (OR:4.0). Sample size. These results are consistent with some previous reports, Candida was the fungus with higher frequency on the psoriatic nails, however, the role of these fungi is controversial (contamination x colonisation x infection). The fact that the immunosuppressive treatment increases the chance of fungal infection leads us to a greater attention to this patient profile, to prevent the intensification of nail psoriasis (Köbner phenomenon).

Patrones dermatoscópicos en pacientes con diagnóstico clínico de onicomicosis y examen micológico directo positivo

IntroductionOnychomycosis is one of the most common nail disorders, representing almost 50% of them. The sensitivity and specificity of direct examination and mycological culture for diagnosis can be as low as 15%. The objective of this study was to describe dermatoscopic patterns in cases of onychomycosis confirmed by direct examination and to correlate them with the type of causal fungal agent and the clinical subtype of onychomycosis. Material and methodA prospective observational and descriptive cross-sectional study of 45 patients with clinical suspicion of onychomycosis was conducted. All patients underwent clinical examination, dermatoscopy with HEINE mini 3000 dermatoscopy, direct mycological examination with KOH and culture analysis. The most frequent dermatoscopic patterns were identified and their correlation with the type of causal fungal agent and the clinical subtype of onychomycosis were analyzed. Results40 patients with clinical suspicion of onychomycosis and positive direct mycological examination were included. 22 (55%) patients had lateral distal subungual onychomycosis, 12 (30%) total dystrophic onychomycosis, and 6 (15%) superficial white onychomycosis. The dermatoscopic patterns of onychomycosis found were: focal macular in 6 (15%) cases, longitudinal striations in 12 (30%), jagged edge with spikes in 11 (27.5%) and distal irregular termination in 11 (27.5%) patients. Focal macular pattern was associated with lateral distal subungual onychomycosis (p = 0.002) and dermatophyte non mycelial fungus (p = 0.0001), the presence of longitudinal striations was associated with lateral distal subungual onychomycosis (p = 0.005) and dermatophyte fungus (p = 0.0001), jagged edge with spikes was associated with lateral distal subungual onychomycosis (p = 0.005) and dermatophyte fungus (p = 0.0001), finally distal irregular termination pattern was associated with total dystrophic onychomycosis (p = 0.0001), dermatophyte fungi (p = 0.0001) and yeasts (p = 0.004). ConclusionDermoscopy is a simple and fast technique that can provide immediate information to guide the diagnosis of onychomycosis.

Keloid-like lesions in a farmer from Amazonas

A 54-year-old male farmer from Eirunepé (Amazon, Brazil) presented with a 30-year history of pink nodules and plaques on his left thigh and knee. In 2016, excisions of the lesions was performed at his hometown, but recurrence was observed after 3 months. Physical examination revealed papules, plaques, and keloid-like nodules on his left thigh and knee and some scarring areas (Fig 1). Direct mycological examination using potassium hydroxide was performed (Fig 2). A punch biopsy was also performed and histopathological examination using Grocott stain was performed (Fig 3).

Conidiobolomycosis in Ovine in Southeast Bahia - Brazil

Background: Conidiobolomycosis is a highly lethal, granulomatous disease that primarily affects the respiratory system of sheep. The etiological agents are fungi of the genus Conidiobolus, including Conidiobolus coronatus, C. incongruus, and C. lamprageus. In Northeast Brazil, this disease is particularly important considering the significant impact sheep and goats have on the regional economy. The present report describes the occurrence of conidiobolomycosis in two sheep from the same property in the Itabuna-BA region that were referred to the Veterinary Hospital of the State University of Santa Cruz, Bahia, Brazil. Case: The primary complaint in both animals was bilateral bloody nasal discharge over a period of approximately 15 days and frequent coughing. On physical examination, the animals exhibited “goosebumps” and opaque hair, with a body score of 1 (scale, 1-5), mild dehydration (7%), apathy, frequent cough with putrid odor, bilateral serosanguinolent nasal discharge, craniofacial asymmetry, expiratory dyspnea, enlargement of the retropharyngeal lymph nodes, audible pulmonary rales, and pain on percussion of the pulmonary field. A therapeutic support protocol was established to stabilize the animals until the results of complete blood count, radiography, and microbiological evaluation of nasal content were available. Definitive diagnosis of disease was made by direct positive mycological examination, that revealed the presence of wide rarely septate hyphae, and isolation and cultivation of the fungus Conidiobolus sp., with microculture on slides and staining with lactophenol blue cotton. Due to disease diagnosis and the severity of injuries observed, the prognoses of both animals were considered to be unfavorable and led to euthanasia and necropsy. At necropsy, the most relevant findings were granulomatous rhinitis with ascending inflammatory processes to the meninges and adjacent structures, in addition to cranioventral areas of pulmonary consolidation with drainage of purulent exudate at the cut surface, suggestive of bronchopneumonia. Histopathology revealed intense pyogranulomatous inflammation associated with the presence of hyphae in negative images within the cytoplasm of multinucleated giant cells in the nasal cavity, lungs, meninges, and brain. Discussion: The association of necropsy findings, histopathological changes, and microbiological isolation of the fungus facilitated understanding of the changes observed in the respiratory system and other organs, and enabled correlation between the recorded lesions and clinical manifestations exhibited by the animals. Based on the lesions observed clinically and at necropsy, both cases were diagnosed with rhinopharyngeal conidiobolomycosis. The histopathological and macroscopic changes observed were similar to what are often described in severe cases of the disease. Considering the severity of the disease and its high lethality in these animals, conidiobolomycosis can lead to significant damage of the production chain; this was observed in the present property, where three animals in a herd of 20 died with signs of the disease, highlighting the importance of the disease, particularly in the Northeast region, where the largest flocks of sheep are found in Brazil. The implementation of prophylactic measures is particularly important considering the unfavorable prognosis and the absence of effective treatments.