Mycological Culture Research Articles

The impact of silver and chitosan nanoparticles on A. fumigatus isolates from Ras cheese and sputum of respiratory patients

This work was designed to study the incidence of A.fumigatus in Ras cheese sold in different markets at Sohag city, Egypt and also in sputum samples of respiratory diseases patients admitted to Sohag University Hospital, as well as study the effect of silver (SNPs) and chitosan (CNPs) nanoparticles against the A. fumigatus isolates. Potato dextrose agar (PDA) was used for mycological culture of samples and the antifungal effect of nanoparticles (with different concentrations) using well diffusion method. High resolution transmission electron microscope (HRTEM) was used for detection of nanoparticles size, and scanning electron microscope (SEM) was used for investigation of nanoparticles effect on A. fumigatus morphology. The isolation rate of A. fumigatus were 22.11 % (21/95), 32.17 % (37/115) from Ras cheese and sputum samples, respectively. SNPs had inhibitory effect at concentrations 1 and 2 µg/ml, and complete growth inhibition with concentrations 4, 8, and 16 µg/ ml. While CNPs had no effect in concentrations 1 and 2 µg/ ml, but at concentrations 4, 8 and 16 µg/ ml represented inhibitory effect with mean inhibition zones 5±0.21, 9±0.41 mm and 11 ±0.77mm, respectively. SNPs and CNPs by SEM patently damaged the A.fumigatus cell structure. Using nanoparticles against A. fumigatus from food and human sources is important to stop food spoilage and protect human health. So, further studies are necessary for the investigation of suitable application methods, and the expected hazards which may occur after nanoparticles usage.

A clinico-mycological spectrum of superficial mycoses of scalp in pediatric population

Objective: A clinico-mycological study was conducted to assess the profile of dermatophytes and non-dermatophytes in patients with superficial mycoses of scalp.
 Methods: This Descriptive cross sectional study was conducted at Department of Microbiology, Basic Medical Sciences Institute, in collaboration with Department of Dermatology, Jinnah Postgraduate Medical Centre, Karachi. The study was carried out from May 2019 to September 2019. A total of 114 children were enrolled in the study having a clinical diagnosis of superficial mycoses of scalp. The hair specimens and scales from the scalp were collected. The specimens were processed for wet mounts (KOH and CFW staining) and mycological culture. .
 Results: The most common clinical presentation was found to be dermatophytoses (28%). Non-dermatophytoses was seen in 16.1% cases, while 55.2% cases were culture negative. The most common isolated dermatophyte was Trichophyton violaceum (35.4%), while among non-dermatophyte molds, Aspergillus spp accounted for 45% of cases.
 Conclusion: The unambiguous diagnosis of scalp mycoses can be made by mycological culture and conventional techniques like KOH mounts and CFW staining.
 Keywords: Calcoflour white stain, Dermatophytes, KOH mount, Tinea capitis

Enhancement in serum (1–3)-β-D-glucan level by cutaneous alternariosis: A case report and literature review

Contamination with the fungus Alternaria spp. is often considered to have originated from laboratory sources, which occasionally causes infection in immunocompromised patients, termed as phaeohyphomycosis. Here, we have reported a case of cutaneous alternariosis caused by Alternaria alternata. This diagnosis was based on microscopic examination and mycological culturing of patient's vesicular lesions, with the use of 5 molecular markers (namely, ITS, ATPase, Actin, rpb2, and tef1) for strain identification. We noted that Alternaria infection caused an increase in the serum level of (1–3)-β-D-glucan (BG) in the patients. To the best of our knowledge, no such finding has been reported in previously in the literature.

Molecular identification of Actinomadura madurae isolated from a patient originally from Algeria; observations from a case report

BackgroundMycetoma is a chronic granulomatous subcutaneous infection caused by anaerobic pseudofilamentous bacteria or fungi. It is commonly prevalent in tropical and subtropical countries. Men are more susceptible to the disease due to greater participation in agricultural works. Mycetoma commonly involves lower extremities, wherein untreated cases lead to aggressive therapeutic choices, such as amputation of the affected body organs and consequently lifelong disability.Case presentationIn this report, we present the rare case of a 58-year-old man, originally from Algeria with a left foot chronic tumefaction of 5 years. In the initial clinical examination, mycetoma was diagnosed based on tumefaction and the presence of multiple sinuses with the emission of white grains. The latter was observed via direct examination. The histopathological analysis demonstrated an actinomycetoma caused by bacteria, as the etiological agent. Imaging showed a bone involvement with osteolysis at the levels of 2nd to 4th metatarsal diaphysis. The mycological and bacterial cultures were both negative. For an accurate diagnosis, the obtained grains were subjected to molecular analysis, targeting the 16S-rDNA gene. Molecular identification yielded Actinomadura madurae as the causal agent, and 800/160 mg of trimethoprim/sulfamethoxazole was prescribed twice a day for 1 year, as a treatment.ConclusionConsidering low information about this disease, especially in non-endemic areas, it is of high importance to enhance the knowledge and awareness of clinicians and healthcare providers, in particular in the countries with immigration issues.

A cross sectional study to observe the diversity of fungal species in Onychomycosis isolated from a tertiary care hospital in Karachi.

To study the frequency and diversity of fungi involved in onychomycosis. The cross-sectional study was conducted from February 2018 to February 2019 at the Department of Microbiology, Basic Medical Sciences Institute, Jinnah Postgraduate Medical Centre, Karachi, and the University of Karachi. Specimens were taken from patients suffering with onychomycosis attending the Dermatology Department of Jinnah Postgraduate Medical Centre and were processed for the isolation of fungi. Specimens were processed for the diagnosis by potassium hydroxide mount for microscopic evaluation and mycological culture to determine the type of fungus involved in onychomycosis. Fungi were isolated using Sabouraud Dextrose Agar with antibiotics cycloheximide and chloramphenicol, and without antibiotics. Plates were observed for the growth periodically until 4th week. Appearance of any colony was studied further for identification on the basis of cultural characteristics and microscopy. Precise identification of Candida species was done. Of the 230 samples, 85(36.96%) were from males and 145(63.04%) from females. Potassium hydroxide mount showed fungal element in 134(58.26%) specimens, while in 111(48.26%) patients the fungal culture was positive. Nine (3.9%) cases were culture-positive but potassium hydroxide-negative, while among the 134(58.26%) potassium hydroxide-positive cases, 32(23.9%) were culture-negative.

Diphtheric aspergillosis tracheitis with gastrointestinal dissemination secondary to viral infections in a dairy calf

Diphtheric aspergillosis tracheitis is an uncommon syndrome described in human pathology, usually associated with immunosuppression in the affected individuals. Interestingly, no comparative/equivalent cases were found in domestic animals. This report describes the pathological and mycological findings associated with diphtheric aspergillosis tracheitis in an immunocompromised calf. The main pathological findings were diphtheric tracheitis and rhinitis, and necrotizing ruminitis associated with intralesional septate, acute branching fungal hyphae consistent with Aspergillus spp. Mycological culture and isolation confirmed the fungal hyphae as A. fumigatus due to characteristic features. Immunohistochemistry (IHC) assays identified intralesional antigens of bovine viral diarrhea virus (BVDV) and malignant catarrhal fever virus (MCFV) at the trachea and small intestine; IHC detected intralesional antigens of bovine alphaherpesvirus 1 (BoHV-1) only at the trachea. These findings confirmed the simultaneous occurrence of A. fumigatus with concomitant infections due to BVDV, MCFV, and BoHV-1 in this calf. Since ovine gammaherpesvirus-2 (OvHV-2) is the cause of MCF in Brail, it is likely that the intralesional MCFV antigens identified were those of OvHV-2. In this case, disseminated aspergillosis was probably associated with the undeveloped immunological status of the calf that was further impaired due to the combined immunodepressive effects of BVDV and BoHV-1 infections. Although BVDV and BoHV-1 are infectious disease pathogens frequently associated with the development of bovine respiratory disease (BRD) in feedlot and dairy cattle, the identification of intralesional OvHV-2-like antigens in several parts of the lungs suggest that this MCFV also played a role in the BRD-associated lesions identified in this calf.

Concordancia entre la técnica de cultivo micológico frente a la técnica citopatológica en el diagnóstico de dermatofitosis en cuyes de crianza intensiva

La dermatofitosis es una enfermedad que afecta al estrato córneo de la piel, pelo y uñas de los cuyes, causando un mal aspecto en la carcasa, afectando su comercialización, y generando pérdidas económicas. Se colectaron 189 muestras de cuyes con lesiones dermatológicas en granjas de crianza intensiva; las cuales fueron analizadas mediante cultivo micológico y citopatología en el Laboratorio de Microbiología y Microscopía de la Universidad Científica del Sur. Se halló una frecuencia de dermatofitosis de 18,5+-5.5% por cultivo micológico y 43 +-7,1% por citopatología; según el estrato etario la frecuencia de dermatofitosis fue de 0% / 0% en lactantes, 25,6% / 62% en recría y 4,8% / 6% en reproductores, por cultivo micológico y citopatología, respectivamente. Según la ubicación de la lesión la frecuencia de dermatofitosis fue mayor en las regiones frontal y nasal, con 41,7% / 70% y 28,1% / 67%, por cultivo micológico y citopatología, respectivamente; en cuanto al tipo de instalación, se presentó una frecuencia de 0% / 0% en animalescriados en jaulas, y 26,5% / 61% en animales de crianza en poza, por la técnica de cultivo micológico y citopatología, respectivamente. Al evaluar el grado de concordancia entre ambas técnicas se halló un valor de Kappa (k) igual a 0,46, considerada moderada.

Incidence and seasonal variation of pet dermatophytosis in Moscow region

In the current one-year study 2477 samples from pets clinically suspected for dermatophytosis (1010 from dogs and 1467 from cats) were examined by direct microscopy, conventional mycological culture and DTM culture. Direct microscopy revealed 1,28% positive samples, conventional culture – 8,0% and DTM culture – 27,7% positive samples. The average incidence of dermatophytosis in Moscow region was 14% (9,5% in cats and 4,5% in dogs), most of cases were diagnosed in autumn. DTM culture proved to be the most effective method for diagnosing dermatophytosis in pets.

Clinical significance of Candida isolation from dystrophic fingernails.

Candida onychomycosis mostly involves fingernails. Yet, in contrast to dermatophytes, Candida isolation from dystrophic fingernails does not prove casualty, as sample contamination and non-pathogenic Candida growth occur. Characterising treatment outcome of Candida-positive dystrophic nails is crucial to avoid unnecessary treatment. To investigate predicators associated with treatment outcome among Candida-positive dystrophic fingernails. A retrospective cohort study was carried out among 108 adults with Candida-positive dystrophic fingernails not cured with adequate systemic anti-fungal course. Diagnosis was based on a single mycological culture. Patients with treatment failure (n=85; 78.7% of the cases) were compared to patients with partial response (mild to almost cure; n=23; 21.3% of the cases) at 9 to 12months following treatment initiation. Treatment failure was significantly associated with primary onycholysis (odds ratio [OR] 2.8; 95% confidence interval [CI] 1.1-7.4) and prolonged dystrophy (12.8 vs. 3.7years in average), compared to partial treatment response. Non-responders had lower odds to present with distal lateral subungual onychomycosis, compared to partial responders (OR 0.3; 95% CI 0.1-0.7). Demographic and mycological characteristics, as well as number of nails affected, co-presence of paronychia, and treatment regime were not found to be associated with treatment response. Candida-positive primary onycholysis was shown to be non-responsive to systemic anti-fungal treatment, suggesting that anti-fungal treatment is not indicated. For other clinical scenarios, high proportions of treatment non-response suggest that determining causality of Candida should not be based on a single positive mycological culture.

Diagnosis of dermatophytoses: Comparison of mycological techniques

Background: Dermatophytes are a group of closely related fungi that have the capacity to invade keratinized tissue (skin, hair, and nails) of humans and other animals to produce an infection. Infection by this fungi are restricted to the non-living cornified layers as they lack the ability to penetrate the deeper tissues of immuno competent hosts. The great variation in clinical presentation is related to the involved species of fungus, size of the inoculum, involved site and immune status of the host. There are three genera of dermatophyte fungi: Trichophyton, Microsporum and Epidermophyton based on microscopic features. Each genus is characterized by a specific pattern of growth in culture and by the production of macroconidia or microconidia. The Trichophyton species usually infect the skin, hair and nails. Microsporum species infect the skin and hair but not the nails. Epidermophyton species infect the skin as well as nails but not hair. This study was carried out to determine the etiological agents in different forms of dermatophytosis and to compare the various laboratory techniques in the diagnosis of dermatophytosis. Materials and method: The study was conducted at the department of Microbiology, Srinivas Institute of Medical Sciences and Research Centre, Mangalore, India. 270 specimens obtained from clinically suspected cases of dermatophytoses between December 2017 to December 2019 were included in the study. The samples included skin scraping, nail clipping and plucked hair. The specimen was then subjected to Microscopic examinations like KOH, PAS, and Calcofluor white stain and Culture on SDA, SDA with Chloramphenicol and DTM. Results: Male patients infected were found to be 48.88% and female patients were 51.11%. The most common affected age group was 21-30 years (25.1%). More than 92.59% of samples were from Tinea unguium. Fungi was demonstrated in 31.85% of cases, by KOH wet mount, 41.48% by calcofluor white stain and 49.62% by PAS. Out of 135 specimens, 48 specimen yielded growth giving 35.55% culture positivity on SDA with chloramphenicol. Conclusion: A success rate of 41.48% was achieved for detecting fungal elements by microscopy as compared to culture. PAS Stain shows high reliability and sensitivity for detecting onychomycosis compared to KOH and mycological culture. PAS and the culture combination is superior to combined KOH and culture for detection of dermatophyte infection. Ninety six isolates were obtained from 270 patients which shows the prevalence of the isolate in and around Mangalore

Isolation and Molecular Characterization of Aspergillus fumigatus and Aspergillus flavus Isolated from Poultry Birds in Ado- Ekiti, Nigeria

Aim: The study was carried out to isolate and identify Aspergillus species from commercial birds with suspected aspergillosis in the poultry farms within Ado Ekiti metropolis Nigeria. 
 Place and Period of Study: The study was carried out in the Department of Microbiology, Faculty of Science, Ekiti State University Ado Ekiti, Nigeria in August 2016.
 Methodology: A total of 35 sick/suspected birds were collected randomly from three poultry farms. At Ago-Aduloju poultry farms, 15 samples were randomly collected from 1000 birds while at Ekiti State University poultry farms, 10 samples were randomly collected from 500 birds. At Federal Polytechnic Ado Ekiti poultry farms, 10 samples were randomly collected from 700 birds. The bird’s selection was on the basis of clinical signs and symptoms such as difficulty in breathing, weight loss, drooping of wings and exercise intolerance. Swab samples were collected from each suspected/sick bird for mycological culture and molecular characterization of the isolates from each bird was carried out. The isolates were identified based on the color of the culture on Potato Dextrose Agar and microscopic examination. Molecular identification was done using 23S Ribosomal RNA Gene and Partial Sequence.
 Results: Six fungal strains that showed similar morphological and cultural characteristics of Aspergillus species were isolated. The isolates were coded ASP 1, ASP 2, ASP 3, ASP 4, ASP 5, and ASP 6. The identified organisms were; Aspergillus fumigatus qH 107 (ASP 1), Aspergillus fumigatus qH 107 (ASP 2), Aspergillus flavus M09 (ASP 3), Aspergillus flavus UOMS6 (ASP 4), Aspergillus fumigatus qH 107 (ASP 5), Aspergillus flavus qH 107 (ASP 6).
 Conclusion: It is evident that Aspergillus species were predominant in poultry farms selected in this study. Necessary precaution should be put in place to prevent the spread of aspergillosis. Poultry farmers are advised to avoid damp environments, moldy feeds, dry and dusty litters. Adequate ventilation should always be provided in poultry farms to prevent Aspergillosis.

Microbiological picture of onychopathy in psoriatic patients

Objective.To evaluate the incidence of onychomycosis and bacterial contamination of onychopathy in patients with psoriasis.Material and methods.The study included 86 patients with skin psoriasis and abnormal nail plates or isolated nail psoriasis. Patients nail plates examined in laboratory using direct microscopy with 20 % KOH, mycological culture Sabourauds Dextrose Agar with chloramphenicol and сycloheximide, and bacteriological culture with indetification using the MALDI-TOF mass spectrometer.Results.Out of 86 patients, 27 (31.4 %) had onychomycosis (KOH-positive or KOH-negative with a positive result for dermatophytes in a culture study). Of the 27 patients with onychomycosis, 9 caused by pathogenic fungi, and 18 caused by opportunistic fungi. Of the 54 patients with nail psoriasis, 9 (16.7 %) had onychomycosis, 3 had dermatophytes, and 6 had opportunistic micromycetes. A total of 97 microbiological studies were conducted in 86 patients, in which the following microorganisms were detected: Staphylococcus caprae – 28 strains, Staphylococcus lugdunensis – 26, Staphylococcus epidermidis – 26, Staphylococcus haemolyticus – 15, Staphylococcus pettenkoferi – 13, Staphylococcus simulans – 11, Staphylococcus warneri – 8, Staphylococcus aureus – 5, Staphylococcus piscifermentans – 4, corinebacteria spp. – 3, Staphylococcus hominis – 3, Staphylococcus capitis – 3, Pseudomonas aeruginosa – 3, Staphylococcus pasteuri – 1, Staphylococcus cohnii – 1, Kocuria spp. – 1, Klebsiella pneumonia – 1.Conclusion.In our study, onychomycosis was detected in 31.4 % of patients with psoriasis who have onychodystrophy. In psoriatic onychia, onychomycosis occurred in 16.7 % cases. Pseudomonas nail infection was observed in two patients, one in combination with nail psoriasis.

Toward High-Throughput Fungal Electroculturomics and New Omics Methodologies in 21st-Century Microbiology and Ecology.

Modern microbiology and drug development are in a watershed moment with the advent of electroceuticals. In addition to genomics, electrical impulses in an organism are believed to contribute to tissue and cellular plasticity. Hence, electroceuticals or bioelectronics offers the promise to identify innovative approaches to treat human diseases. However, applications toward electromicrobiology are still limited and rare, despite the high potential to innovate the fields of both microbiology and therapeutics. For example, electric modalities for manipulating microbial growth are highly sustainable; can be combined with biopharmaceuticals, probiotics, and pharmacobiotics; and, thus, are well poised for use in medicine, public health, and ecology and diverse industries. We report here the introduction of a new research framework and technology platform for electroculturomics, by coupling standard solid-state mycological cultures with conductive treatment using a conformité Européene (CE-)-certified medical ionophoresis device. We share our experience with a diverse range of fungi that have been treated with the electroculturomics approach reported herein. We suggest that this line of inquiry can be extended to electrotranscriptomics and electrometabolomics by deploying electroculturomics in tandem with multi-omics approaches in the future. This article makes a specific contribution to fungal microbiology, and a broader contribution to advance the theory and practice of the field of electroculturomics emerging in 21st-century microbiology and ecology research.

Evaluation of MucorGenius® mucorales PCR assay for the diagnosis of pulmonary mucormycosis

ObjectivesWe aimed to assess the clinical relevance of the marketed pan-mucorales real-time PCR assay MucorGenius® (Pathonostics) on pulmonary specimens relative to that of in-house PCR assays and conventional mycology for the diagnosis of mucormycosis. MethodsIn total, 319 pulmonary samples from severely immunosuppressed patients at risk for invasive mold disease (IMD) were retrospectively included. Direct examination, mycological culture, and PCR testing were performed using three genus-specific in-house mucorales real-time PCR assays and MucorGenius®PCR. Results from Aspergillus testing, including galactomannan and PCR, were also collected. ResultsThe 319 patients were graded according to modified EORTC-MSG criteria as proven/probable mucormycosis (n=6), proven/probable invasive aspergillosis (IA) (n=63), Aspergillus–mucorales co-infections (n=4), possible IMD (n=152), and excluded IMD (n=94). The in-house and MucorGenius®PCR assays were positive for 33 (10.3%) and 27 (8.5%) samples, respectively, whereas culture was positive for only 10 (3.1%). The in-house and MucorGenius®PCR assays showed a sensitivity of 100% (10/10) and 90% (9/10) and a specificity of 95.7% and 97.9%, respectively. Both PCR assays allowed the detection of mucorales DNA in samples from 10 possible cases and six IA, all missed by culture. ConclusionsMucorGenius® showed good performance, despite missing some low fungal burden. Combining mucorales PCR with EORTC-MSG criteria greatly improved the diagnosis of mucormycosis.

Acral melanoma mimicking mycotic intertrigo

Dear Editors, The interdigital location of melanoma is the rarest presentation of the acral subtype.1 Acral lentiginous melanoma (ALM) accounts for 2% to 10% of melanoma cases according to data from the American Cancer Society (2017).2, 3 Plantar regions represent up to 73% of cases, and the incidence is mainly after the seventh decade of life.2, 4 A 73-year-old female patient, phototype III, presented with a complaint of a long-term brownish patch between the third and fourth left toes, with concomitant maceration (Figure 1A). Direct mycological and fungal cultures came back negative. An initial biopsy was performed, which did not reveal any malignancy. However, because of high clinical suspicion, the patient underwent a second ellipse biopsy (Figure 1B) in an attempt to remove as many parts of lesion as possible for a comprehensive analysis. Microscopic examination of this second sample revealed in situ acral lentiginous melanoma (Figure 2A,B), and the patient was referred to the Cancer Surgery Service. ALM was first described in 1976 by Reed.3, 4 Compared with other subtypes, it is less common in Caucasians and more frequent in higher phototype patients, as demonstrated in our case.1, 4 Interdigital location of ALM is the less common presentation of acral subtype.3, 4 Risk factors general and traditionally described for melanoma, such as sun exposure, phototype, childhood sunburn, and family or personal history of previous melanoma, appear to be less important in acral lesions.2 Some authors suggest recurrent trauma may be a potential risk factor, but incidence of melanoma is low when compared with the high frequency of trauma in these locations.2-4 A French series of 126 cases found that Breslow thickness, male gender, and amelanosis were significantly associated with worse prognosis.2 This seems to be worse in ALM because of often-delayed diagnosis. However, it is still controversial whether only the histological subtype could be a worse outcome variable.3 Differential diagnoses that should be considered include warts, callosities, eccrine poroma, fungal infections, foreign body, pyogenic granuloma, inflammatory lesions, bullous disorders, diabetic ulcer, nevi, keratoacanthoma, subungual bruising, and onychocryptosis.1, 3, 4 It is worth noting that up to 10% of ALM cases can simulate mycotic intertrigo,3 reaching diagnostic mistakes up to 20% to 34% in some cases series.4 We highlight that ALM located in the feet frequently has a delayed diagnosis and may simulate other benign pathologies, such as fungal infection primarily considered in the present case. We emphasise the need of routine examination of patients' feet skin in order to allow earlier diagnosis and thus a better prognosis. The authors declare no conflicts of interest.

Coinfection of domestic felines by distinct Sporothrix brasiliensis in the Brazilian sporotrichosis hyperendemic area

Microbial interactions may impact patient’s diagnosis, prognosis and treatment. Sporotrichosis is a hyperendemic neglected zoonosis in Brazil, caused by Sporothrix brasiliensis. Four pairs of clinical isolates of Sporothrix were recovered from four diseased cats (CIM01-CIM04, two isolates per animal) raising the possibility of coinfection in a sporotrichosis hyperendemic area, Brazil. Each isolate of the pair had distinct pigmentation in mycological culture, and was designated as “Light” or “Dark”, for low and high pigmentation, respectively. Dark isolates reacted strongly with monoclonal antibodies to melanin (p ≤ 0.05) by both ELISA and FACS quantitation, and displayed a ring pattern with some regions exhibiting higher punctuated labeling at cell wall by immunofluorescence. In turn, Light isolates reacted less intensely, with few and discrete punctuated labeling at the cell wall. PCR identified all isolates as S. brasiliensis, MAT1-2 idiomorph. Sequencing of β-tubulin and calmodulin genes followed by phylogenetic analysis placed all eight isolates within the same cluster as others from the Brazilian hyperendemic area. The ability of these strains to stimulate cytokine production by human PBMCs (Peripheral blood mononuclear cells) was also analyzed. CIM01 and CIM03 Light and Dark isolates showed similar cytokine profiles to the control strain, while CIM02 and CIM04 behaved differently (p < 0.001), suggesting that differences in the surface of the isolates can influence host-fungus interaction. MICs for amphotericin B, terbinafine, caspofungin, micafungin, itraconazole, fluconazole, and voriconazole were obtained (CLSI M38-A2/M27-A3). Pairwise comparisons showed distinct MICs between Sporothrix Light and Dark isolates, higher than at least two-fold dilutions, to at least one of the antifungals tested. Isolates from the same pair displayed discrepancies in relation to fungistatic or fungicidal drug activity, notably after itraconazole exposure. Since S. brasiliensis Light and Dark isolates show disparate phenotypic parameters it is quite possible that coinfection represents a common occurrence in the hyperendemic area, with potential clinical implications on feline sporotrichosis dynamics. Alternatively, future studies will address if this specie may have, as reported for other fungi, broad phenotypic plasticity.

Metagenomics of Black Grains: New Highlights in the Understanding Eumycetoma

Background: Eumycetoma is a debilitating chronic subcutaneous granulomatous disease, endemic in Sudan and many other tropical and subtropical countries. It can be caused by eight different fungal orders and Madurella mycetomatis as the main causative species. The gold standard diagnostic test is culture, however culture-independent methods as imaging, histopathological and molecular techniques can support diagnosis, especially in case of negative cultures. Methods: Amplicon-based Internal Transcribed Spacer 2 (ITS2) metagenomic technique was used to study black grains isolated from 14 tissue biopsies from patients with mycetoma. Furthermore, grains mycological culture and surgical biopsies histopathological examination were done. Results: M. mycetomatis (n=5) and Falciformispora spp. (n=4) organisms were identified by culture and confirmed by metagenomics. Metagenomics recognised, at the species level, Falciformispora as F. tompkinsii (n=3) and F. senegalensis (n=1), while in culture-negative cases (n=5), M. mycetomatis (n=3), F. senegalensis (n=1) and Fusarium spp. (n=1) were identified. Interestingly, the metagenomics results showed a “consortium” of different fungi in each sample, mainly Ascomycota phylum, including various species associated with eumycetoma. The microbial co-occurrence in eumycetoma showed the co-presence of Madurella with Trichoderma, Chaetomium, Malasseziales and Sordariales spp. While, Falciformispora was co-presented with Inocybe and Alternaria, and mutual exclusion with Subramaniula, Aspergillus and Trichothecium. Interpretation: Metagenomics had provided new insight into the aetiology of eumycetoma in samples with negative culture. Furthermore, into diversity and complexity of grains mycobiota, and it questioned the accuracy of traditional culture for the identification of causative agents. Funding Information: This study was supported by the Italian Agency for Development 62 Cooperation (AICS) (Grants AID10821 and AID10861). Declaration of Interests: The research work of Dr. Aleksandra Barac was supported by the Ministry of Education, Science and Technology, the Republic of Serbia (No. III45005). Ethical Approval Statement: The study was approved by the Mycetoma Research Center IRB, No. 12/2018. Informed consent was obtained from each adult patient, while an assent was taken from minors (age less than 18 years) with approval of their guardian.

Necrotizing Tracheobronchitis Caused by Aspergillus fumigatus in a Cow

This report describes a case of tracheobronchitis caused by Aspergillus fumigatus in a cow. A 4-year-old Jersey cow was submitted for necropsy examination. Grossly, the mucosa of the trachea and bronchi was diffusely reddened and was covered by a moderate amount of yellow, caseous material, sometimes with small foci of grey 'cotton-like' appearance, suggestive of filamentous fungus. Microscopical analysis of the trachea and bronchi showed marked diffuse mucosal and submucosal necrosis with a large amount of cellular debris, fibrinous exudation and inflammatory infiltration dominated by macrophages and neutrophils. There were large numbers of septate fungal hyphae with acute bifurcation angles and conidia and vesicles typical of Aspergillus spp. that were stained by the Grocott method. A. fumigatus growth in mycological culture from the trachea and bronchi was confirmed by molecular identification. Tracheobronchitis caused by A. fumigatus infection in cattle is poorly described and requires further research to improve the therapeutic management of the disorder.

Associated factors and spatial patterns of the epidemic sporotrichosis in a high density human populated area: A cross-sectional study from 2016 to 2018

An epidemiological characterization of human and feline sporotrichosis was carried out between 2016 and 2018, in a high density-populated area in Brazil. Professionals were trained to identify suspect cats and notify vets to interview the owners and collect swabs of the wounds from these animals. Mycological cultures were performed, and colonies identified as Sporothrix spp. Subsequently, data regarding the outcome from suspect animals were collected. Confirmed cases of human sporotrichosis (56) were also counted and analyzed for spatial distribution. 118 positive animals were observed. The prevalence of feline sporotrichosis was 8.36 ‰ (CI 95 %, 5.38–9.55 ‰). The odds for being positive in animals that lived only partially at home were 3.02 times greater than for those cats without access to the street (OR 3.02, CI 95 % 1,96-10,43). There was no statistically significant association between environmental variables and positive diagnosis, corroborating the hypothesis that direct transmission by infected cats plays a greater role in the occurrence and continuous outbreaks of sporotrichosis in Brazil. Among the positive animals, 61.90 % (CI 95 % 58.95–64.96) died, and they had an odds to die in the next six months 6.30 times greater than negative animals (p < 0.05, OR 6.30, CI 95 % 2,79-14,42). The case fatality rate was 55.08 % in cats (CI 95 % 49.20–51.15). The cause-specific death rate was 4.6 ‰ in cats (CI 95 % 3.4–6 ‰). Only 7.62 % (CI 95 % 7.12–8.16) positive cats were treated and cured. Among dead positive animals, 29.23 % were inappropriately discarded. In the study period 56 human cases were recorded in the Barreiro region. Regions with highest prevalence of feline sporotrichosis, had greater frequencies of both human and feline cases. This is the first report on the epidemic of sporotrichosis in Minas Gerais, Brazil. The free offer for treatment and veterinary care for these animals should be taken into consideration, as well as the collection and incineration of the dead ones, as measures of public health, followed by the guidance and care for the human patient.

Efficacy of 1% terbinafine and 1% luliconazole in the management of tinea corporis and tinea cruris

Background: Tinea or dermatophytosis is a fungal infection that commonly affects stratum corneum of the skin and its other appendages. Topical antifungal agents are the first-line treatment choice for dermatophytosis among them terbinafine, luliconazole, amorolfine, and sertaconazole are more effective. Aim and Objective: The aim of the study was to assess the efficacy and safety of topical luliconazole with topical terbinafine in tinea corporis and tinea cruris management. Materials and Methods: Study conducted on 180 cases diagnosed clinically and confirmed mycologically with tinea cruris and tinea corpis between age group of 21 and 60 years were recruited. Study consists of two groups, that is, Group 1 administered with 1% terbinafine and Group 2 with 1% luliconazole. Clinical signs such as pruritus, erythema, scaling and mycological culture, and KOH mount were assessed at the beginning, end of 1st, 2nd, and 4th weeks. Results: The comparison of mean pruritus score at the beginning, at the end of 1st week and 2nd week was statistically significant (P < 0.05). The mean erythema score was statistically significant between two study groups from the beginning to the 4th week. The reduction of mean scaling score was statistically significant at the end of 1st week. The KOH values between two study groups showed significant difference at the end of the 1st week (P = 0.028). Conclusion: The status of clinical signs, mycological culture, and KOH values was better in Group 1 than Group 2. Hence, 1% luliconazole is more efficacious than 1% terbinafine in the management of tinea corporis and tinea cruris for 2 weeks.