KOH Microscopy Research Articles

Improvement of direct microscopy method for differential diagnosis of skin mycoses

Background and Aim.In the practice of veterinary doctors, a number of methods for diagnosing dermatomycoses are used, methods of direct and luminescent microscopy are undeservedly ignored. Сomparative analysis of staining methods for improving the method of direct microscopy, rapid and reliable detection of spores and mycelium in biomaterial, identification of pathogens and differential diagnosis of skin mycoses is of crucial importance for the effective treatment of animalsMaterials and Methods. 47 clinical samples of wool and skin taken from farm animals were examined for dermatomycetes. Diagnosis was performed using direct microscopy, fluorescence microscopy with calcofluor, and culture medium culture Saburo. Results. During the study of 47 samples of biological material, it was found that during microscopy the pathogen was detected in 10 cases (21.3%), during KOH microscopy - in 15 cases (29.9%), during KOH microscopy with other dyes in 11 cases (23.4%), during KOH microscopy with color calcofluor in 25 cases (53.2%). The efficiency of the KOH microscopy method with white calcofluor in comparison with direct microscopy was 60% higher, in comparison with KOH microscopy with other dyes - by 56%, classical KOH microscopy by 43.8%. By the culture method, pathogens were detected in 28 cases (59.6%), of which 7 Trichophyton spp., 1 Microsporum spp., 14 Aspergillius spp., 6 yeast. Growth was absent in 19 samples (40.4%), bacteria were detected in 3 samples (6.3%). In comparison with КOH microscopy with calcofluor white, positive results were confirmed in 89.3%.Conclusion. The use of coloring with various dyes of morphological elements of micromycetes in pure cultures allows you to identify mycelial hyphae of various thicknesses, spores, conidia, other morphological structures and carry out a quick preliminary identification of dermatomycetes and opportunistic mold fungi in smears.

Tinea capitis with kerion complicated by keloid formation in a 34-year-old man: A case report

A kerion, also known as a dermatophyte, is a type of inflammatory tinea capitis caused by an exaggerated immune response to a fungal ringworm infection primarily affecting the scalp. A keloid is an abnormal scar tissue proliferation occurring at the site of tissue injury or spontaneously. It occurs more commonly in darker-skinned individuals. The coexistence of kerion and keloid is rare. This case involves a 34-year-old man who presented with a four-year history of scalp swelling, itching, and occasional fluid discharge. There is a history of keloids in multiple parts of the body. Examination revealed multiple keloids on the scalp alongside a purulent, oozing rash with satellite nodules. Skin scraping for KOH microscopy was positive for fungal spores and hyphae. This case highlights the rare complication of keloid formation in kerion which can hinder effective treatment of the underlying fungal infection.

Diagnosis of mucormycosis from nasal swabs using commercial PCR platforms; a feasible alternative?

Rhino-orbital-cerebral mucormycosis (ROCM) is linked to uncontrolled diabetes, diabetic ketoacidosis, iron overload, corticosteroid therapy, and neutropenia. This study evaluated a commercial real-time PCR system's effectiveness in detecting Mucorales from nasal swabs in 50 high-risk patients. Nasal swab PCR showed 30% positivity, compared to 8% with KOH microscopy. Despite its improved sensitivity, nasal swab PCR has limitations, highlighting the importance of established sampling methods in mucormycosis diagnosis. Participants were predominantly male (64%), with diabetes (78%) and amphotericin B use (96%). Prior COVID-19 was 42%, with 30% positive for Mucorales by PCR, compared to 8% with KOH microscopy.

Comprative Study of two Different Microscopic Techniqes and Fungal Culture for Isolation of Dermatophytes

Introduction: Fungal infections are very common in man. They are assuming greater significance both in developed & developing countries due to advent of immunosuppressive drugs & disease hot & humid climate in tropical & subtropical countries like India makes dermatophytosis or ringworm a very common superficial fungal skin infection. Dermatophytosis is caused by dermatophytes, a group of keratinophilic fungi that require long incubation period to grow. The clinical presentation, though very typical of ringworm infection, is very often confused with other skin disorder particularly due to rampant application of broad – spectrum steroid containing skin ointments and cream leading to further misdiagnosis and mismanagement. Materials and Methods: Cross sectional lab based investigational study was conducted and skin, hair, nail sample from 110 clinically suspected cases of dermatophytosis were taken from OPD of district hospital of central India and were screened by direct microscopicexamination using 10% potassium hydroxide (KOH) with and without 40% dimethyl sulphoxide (DMSO) mount. 10% potassium hydroxide (KOH) with 40% Dimethyl sulphoxide mount (DMSO) mixed in equal proportion. Results: Direct microscopy using KOH mount is simple, rapid and easy method to visualize fungal elements. Many modification have been evolved to increase the specificity & sensitivity of KOH microscopy results like use of 5% glycerol, addition of 36% DMSO, and addition of parker’s blue(4) in this study we compered the results of conventional KOH mount and a modified technique (40%KOH with DMSO)(5). When doing direct microscopic. Conclusion: The modified KOH with DMSO mount had allowed fastest and better visualization of fungal elements at 10 minutes instead of routine 30 minutes.

A Comparison Between Potassium Hydroxide (KOH) Microscopy and Culture for the Detection of Post-COVID-19 Rhino-Orbital-Cerebral Mucormycosis.

Background and objective Mucormycosis is an emerging and serious angioinvasive infection caused by filamentous fungi related to the order of Mucorales and the class of Mucormycetes. There was a marked increase in the number of cases of mucormycosis in India following the second wave of the coronavirus disease 2019 (COVID-19) pandemic in the year 2021. In this study, we aimed to compare potassium hydroxide (KOH) microscopy with culture for the detection of post-COVID-19 rhino-orbital-cerebral mucormycosis (ROCM). Materials and methods The KOH microscopy was performed with a 10% or 20% KOH-mounted slide with specimens collected from suspected cases of post-COVID-19 ROCM. Simultaneously, the culture was done on Sabouraud dextrose agar (SDA). These were incubated at 37 ℃ and 25℃for 28 days. Diagnostic parameters were calculated by comparing KOH with gold standard culture. Results KOH mount was positive for broad aseptate fungal hyphae in 322 (54.1%) cases, while it was negative in 244 (41.0%) cases. KOH mount was positive for other fungi in 29 (4.8%) samples. The diagnostic accuracy of the KOH mount for Mucorales was 70.3%. KOH mount had asensitivity of 84.9%, specificity of 61.5%, positive predictive value (PPV) of 56.9%, and negative predictive value (NPV) of 87.2%. Conclusions Based on our findings, the KOH microscopy positivity rate was higher in tissue samples compared to nasal swabs, with a sensitivity of 84.9%, specificity of 61.5%, PPV of 56.9%, and NPV of 87.2%. The overall diagnostic accuracy of the KOH mount for Mucorales was 70.3%.

Clinicomycological Profile and Risk Factors for Dermatophytosis at a Teaching Tertiary Care Centre in North India: A Cross-sectional Study.

Dermatophytosis has gained significant importance in recent years owing to increased incidence, more atypical lesions, changing mycological profile and growing antifungal resistance. Therefore, this study was planned to know the clinicomycological profile of dermatophytic infections in patients attending our tertiary care centre. A total of 700 patients with superficial fungal infections belonging to all age groups and both sexes were taken up for this cross-sectional study. Sociodemographic and clinical details were noted on a prestructured proforma. Superficial lesions were clinically examined and the sample was collected by appropriate collection methods. Direct microscopy by potassium hydroxide wet mount was done to see hyphae. For culture Sabouraud's dextrose agar (SDA) with chloramphenicol and cyclohexamide was used. Dermatophytic infections were detected in 75.8% (531/700) patients. Young people belonging to age group of 21-30 years were commonly affected. Tinea corporis was the commonest clinical picture seen in 20% of the cases. Oral antifungals were taken by 33.1% and topical creams were used by 74.2% of the patients. Direct microscopy was positive in 91.3% and culture was positive for dermatophytes in 61% of the study subjects. T. mentagrophytes was the commonest dermatophyte isolated. Irrational use of topical steroids needs to be controlled. KOH microscopy can be useful as a point of care test for rapid screening of dermatophytic infections. Culture is necessary to differentiate various dermatophytes and to guide the antifungal treatment.

Is Serum Ferritin an Early Marker for COVID-19-Associated Mucormycosis?

Background Coronavirus disease 2019 (COVID-19)is a hyperinflammatory disease caused by severe acute respiratory syndrome coronavirus 2, which makes critically ill patients susceptible to invasive fungal infections. Invasive fungal infections such as mucormycosis are associated with high morbidity and mortality. This study aimed to determine theserum ferritin levels in COVID-19-associated mucormycosis (CAM) patients and isolate and identify the fungi causing secondary fungal infections in patients with suspected CAM. Methodology This cross-sectional study was conducted from June to September 2021 among CAM patients admitted to Bowring and Lady Curzon Hospital. After obtaining approval from the institutional ethics committee and valid consent, data regarding demographic details, past medical history, and serum ferritin levels, along with other blood investigations, were carefully collected from patients presenting with clinical features of mucormycosis and a history of COVID-19. Samples were examined under a bright field microscope using wet mounts of samples in KOH, cultured on Sabouraud's dextrose agar, and examined under a microscope after staining with lactophenol cotton blue for the isolation and correct identification of fungi. Statistical analysis was done using Microsoft Excel (Microsoft Corp., Seattle, WA, USA) and SPSS version 26.0 (IBM Corp., Armonk, NY, USA). A p-value less than <0.05 was considered statistically significant. Results A total of 95 patients with CAM were included in this study, comprising 70 males and 25 females. The mean age of presentation was 49.83 ± 12.41 years, with 73% males and 26% females. Type 2 diabetes mellitus was noted in 69% of patients, hypertension in 29%, and steroid use in 42%. The mean serum ferritin level was 537.38 ± 468.88 ng/mL. We found a significant association between increased serum ferritin and a history of diabetes. Serum ferritin levels had a statistically significant correlation with samples of patients who were positive for Mucorales under KOH microscopy. The fungal culture showed the growth of Aspergillus, Mucor, Rhizopus, and Candida. The mean value of serum ferritin in patients who showed mucor growth was 842.09 ng/mL. Conclusions We found a significant increase in serum ferritin levels in CAM patients. Ferritin can be used as an early marker for screening mucormycosis in COVID-19 patients. Monitoring patients with elevated serum ferritin levels in severe COVID-19, glycemic control, judicious use of corticosteroids, early diagnosis, and appropriate treatment can aid in better management of the disease.

Polarity Inversion of GaN via AlN Oxidation Interlayer Using Metal–Organic Vapor Phase Epitaxy

The polarity inversion of GaN films grown on vicinal sapphire substrates is demonstrated using the metal–organic vapor phase epitaxy (MOVPE) method via an AlN oxidation interlayer for application in wavelength conversion devices. By annealing the N‐polar AlN surface at 900 °C in air, the pit‐free Ga‐polar GaN layer can be regrown. Anisotropic etching in KOH and transmission electron microscopy confirms the formation of a GaN polarity‐inverted structure. X‐ray diffraction measurements reveal an increase in threading dislocations due to the nucleation island formation by low‐temperature GaN layer and strain relaxation at the regrown interface. The simple fabrication process of the GaN polarity‐inverted structure using an AlN oxidation interlayer will pave the way for applications in new devices that require polarity‐controlled and/or polarity‐inverted stacking structures.

MALDI-TOF-MS for Rapid Detection of Fungi causing Mucormycosis during COVID-19 Pandemic from a Tertiary Care Hospital in Southern India: A Retrospective Cohort Study

Introduction: Mucormycosis is an opportunistic fungal infection that became a public health emergency during the second wave of Coronavirus Disease-2019 (COVID-19). It is an acute, angioinvasive, opportunistic, and emerging mycosis caused by mucormycetes, resulting in a significantly fatal fungal infection among immunocompromised and/or immunosuppressed individuals. Mucormycosis has a specific predilection for blood vessels (angioinvasive) and tissues, leading to extensive necrosis and thromboembolic events. Globally, the incidence rate of mucormycosis varies from 0.005 to 1.7 per million people. Early diagnosis and appropriate therapy are crucial for clinical outcomes. Aim: To evaluate the mycological profile of mucormycosis during the COVID-19 pandemic using automated Matrix Assisted Laser Desorption Ionisation Time of Flight Mass Spectroscopy (MALDI-ToF-MS) in comparison with conventional fungal identification methods. Materials and Methods: A retrospective cohort study was conducted, collecting data from July 2020 to June 2022, involving 1,176 patients from various clinical departments who attended PSG Hospitals affiliated with PSG Institute of Medical Sciences and Research, Coimbatore, Tamil Nadu, India. Data analysis was performed from July 2022 to August 2022. Clinical specimens, including Bronchoalveolar Lavage (BAL), Sputum, Cerebrospinal Fluid (CSF), nasal swabs, tracheal aspirates, pus, blood, corneal scraping, eye discharge, and necrotised tissues, were subjected to Potassium Hydroxide (KOH) mount, conventional culture coupled with micrometry-aided microscopy, and MALDI-ToF-MS among suspected mucormycosis cases. The results were collected in Microsoft Excel, and data analysis was conducted using the International Business Machines Statistical Package for the Social Sciences statistical software (IBM SPSS) version 24.0. The study variables were expressed as descriptive statistics: frequency (N) and percentage (%). The incidence of mucormycosis, mycological profile, and diagnostic utility of conventional micrometry-aided microscopy-assisted culture were compared with the automated MALDI-ToF-MS protein signature method of testing. Results: Fungal elements were positive in 130 (11.05%) out of a total of 1,176 clinical samples, such as necrotised tissue, biopsy, and sputum, in the KOH mount. Fungal culture was positive in 258 (21.94%) out of 1176 samples, with mucormycosis detected in 73 (28.29%) of the culture-positive samples. The incidence of mucormycosis in this study was 73 (6.21%) out of 1176. A total of 101 (8.59%) samples showed positive results for mucormycetes through KOH, culture, and MALDI-ToF-MS. MALDI-ToF-MS could rapidly and specifically identify and confirm the causative agents of mucormycosis at their genus and species levels compared to KOH or culture. Conclusion: Simple, low-cost, and less laborious conventional culture coupled with micrometry-aided microscopy can detect Mucorales at the species level, where KOH microscopy could not differentiate between species. MALDI-ToF-MS can be employed for the rapid detection and confirmation of mucormycosis agents at the genus or species level

Relevance of Conventional Microscopy in the Diagnosis of Mucormycosis during COVID-19 Pandemic.

Mucormycosis is a lethal disease which bewildered the health-care community of India during the ongoing second wave of the COVID-19 pandemic. The diagnosis is challenging considering the poor isolation in culture. The aim of the study was to emphasize the utility of potassium hydroxide (KOH) mount examination using conventional light microscopy for early diagnosis of mucormycosis in resource-limited settings. A retrospective analysis of results for all the samples including tissue biopsies, swabs, and pus received in the laboratory for KOH microscopy was done, and results were recorded. The clinical and demographic details of the patients were collected from the hospital information system. A total of 75 samples from 50 patients were received in the laboratory. Out of these, 43 samples from 35 patients showed fungal hyphae (38 patients with only nonseptate hyaline hyphae, 2 with septate hyaline hyphae, and 3 samples with mixed infections). All patients except one were positive for severe acute respiratory syndrome coronavirus 2 infection. The most common age group was 45-59 years (40%), followed by 30-44 years (34.28%) with a male predominance. There was a significant difference in hemoglobin A1C (P = 0.005) and ferritin (P = 0.017) levels between laboratory-confirmed mucormycosis patients and clinically suspected mucormycosis patients without confirmation. Early diagnosis and initiation of targeted therapy is the cornerstone for treating mucormycosis patients. Hence, a rapid and reliable mode of diagnosis is the need of the hour. Conventional microscopy is such a tool that may be used, especially in resource-limited settings.

Tinea Gladiatorum Detection With a Dermatophyte Test Strip.

Determine sensitivity, specificity, positive predictive value (PPV), negative predictive value (NPV), and concordance of diafactory hs-TP (DTS) to detect tinea gladiatorum using direct potassium hydroxide (KOH) microscopy as the reference standard. Prospective, comparative study. Seventeen Minnesota high schools during the winter wrestling season. Seventy-one consecutive high school wrestlers identified with a suspicious rash during skin inspection. Samples were obtained from each rash for both DTS and direct KOH microscopy. Readings were recorded as positive or negative. Direct KOH microscopy identified tinea gladiatorum in 35 of the 71 samples (46%). DTS sensitivity was 80% (95% confidence interval 63%-92%), and specificity was 82% (66%-92%). PPV was 85% (68%-95%), and NPV was 86% (72 %-91%). The DTS result was 83% concordant (72%-91%) with direct KOH microscopy. Similar to rapid Covid antigen tests, DTS required brief, basic training to perform and gave onsite results in 5 to 30 minutes. Although DTS is not approved for use in the United States by the FDA, concordance compared with direct KOH microscopy in diagnosing tinea gladiatorum was similar to results reported for DTS-TU in tinea unguium and tinea pedis. Further study comparing DTS to a reference standard using PCR plus direct microscopy is warranted.

P262 Clinico- microbiological profile of post-COVID pulmonary fungal infections encountered during the second wave of COVID-19 pandemic at a tertiary care teaching hospital in the Himalayas

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM ObjectiveThe study aims to generate preliminary data about post-COVID pulmonary fungal infections in the Himalayas and analyze patients’ micro-radio-clinical profiles and outcomes.MethodologyWe conducted a retrospective study at a tertiary care teaching hospital in the Himalayas to generate preliminary post-COVID pulmonary fungal infection data. Sputum, Endotracheal Tube (ET), and Bronchoalveolar lavage (BAL) samples of patients sent to the Mycology laboratory were subjected to KOH mount and aerobic inoculation on Sabouraud dextrose agar plates at 37°C. The patients’ symptoms, diagnosis, clinical-radiological profile, and outcome were collected from the hospital database.ResultsAmong n = 16 cases of post-COVID pulmonary fungal infections aged 53 +/- 13.38 years, n = 7 (43.75%) had Pulmonary Aspergillosis (n = 5 A. fumigatus, n = 1 A. flavus, n = 1 A. niger), n = 5 (31.25%) had Pulmonary Mucormycosis (Rhizopus arrhizus), and n = 4 (25%) had mixed infection. In 2 of 4 mixed infection patients, R. arrhizus was identified on KOH microscopy and A. fumigatus on SDA Agar. Both A. fumigatus and R. arrhizus were identified on KOH Microscopy of the third patient, while only A. fumigatus was cultivated on his SDA Agar. Aspergillus flavus and R. arrhizus were isolated simultaneously from the sample of the last patient, but only R. arrhizus was identified on KOH Microscopy.Clinical symptoms were similar among Pulmonary Aspergillosis and Mucormycosis patients, but hemoptysis was reported only among Pulmonary Aspergillosis patients. Pre-existing co-morbid end-organ damage, AKI, CKD, CLD, COPD, and CAD was more common among Pulmonary Mucormycosis patients and rare among Pulmonary Aspergillosis patients. Treatment requirements and clinical outcomes of patients infected with either mold were similar. The clinical profile of mixed infection patients was notably different from the others. All the patients were males, none complained of chest pain or expectoration, and none had a history of PTB, AKI, CKD, CLD, COPD, or CAD. Only 2 (50%) mixed infection patients needed supplemental high flow oxygen, unlike all (100%) patients diagnosed with single mold infection. None of the mixed infection patients required steroids. Moreover, none of the mixed infection patients died, unlike 60% mortality in cases of single-species infections.On radiological investigation, n = 6 had typical thick-walled cavitary lesions with air-fluid levels and multiple centrilobular nodules giving a tree in bud appearance, of which n = 4 had bilateral lung involvement, and n = 2 had only one lung involved. n = 1 patient had a well-circumscribed lung abscess.ConclusionCOVID patients from the Himalayas had a higher prevalence of invasive pulmonary fungal infections, probably due to the dense surrounding vegetation. The immuno-compromised state following COVID-19 infection/treatment might be responsible for the progression of regular exposure to invasive pulmonary infection.

P242 Autochthonous cases of Madura foot from Jodhpur, Western Rajasthan, India

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMMycetoma has been classified as a neglected tropical disease by the WHO. ‘Madura foot’ is a prominent presentation of phaeoid fungi Madurella complex in India. Maduromycotic mycetoma is frequently encountered in Western Rajasthan, where it is endemic. A high prevalence in this area is due to the desert climate of prolonged hot sunshine and scanty annual rainfall. The classic clinical ‘triad of Mycetoma’ involves a painless hard swelling, multiple fistulas, and discharge of grains.We report two cases of eumycetoma from Western Rajasthan:Case Report 1: A 41-year-old male, farmer presented a history of progressive swelling of the left foot in the last 4 years following trauma to the left sole 5 years ago. Patient took multiple treatments with no improvement. General examination showed a pallor and inguinal lymphadenopathy. Local examination revealed tumefaction with multiple sinuses discharging serosanguinous fluid with occasional black granules. Laboratory tests revealed hemoglobin of 6.2 g/dL. Serology was positive for Hepatitis B virus. Direct 40% KOH microscopy of the crushed grains shows septate branching phaeoid hyphae. Whitish brown dry colonies with diffusible brown pigment were seen on Sabouraud Dextrose Agar at 25°C. Lactophenol Cotton Blue Stain showed dark septate hyphae with chlamydoconidia. Histopathological studies confirmed Eumycetoma. X-rays of his foot showed large destructive bony lesions. Initial therapy given to patient was to correct anemia followed by specific antifungal treatment. A below knee amputation of left leg was performed once the anemia was corrected.Case Report 2: A 55-year-old borderline diabetic male daily-wage worker presented with multiple nodular discharging lesions over the anterior shin of the right leg for past 7 years. The lesion started in leg as a small nodule that ruptured discharging black grains intermittently. He could not recall any significant trauma to his foot. Patient took multiple treatments with temporary relief but the disease recurred. Direct microscopy of 40% potassium hydroxide mount of the crushed grains showing thin septate branching phaeoid hyphae and on Sabourad Dextrose Agar at 25°C. White to brown color colony growth with diffusible brown pigment was seen after 2 weeks of incubation. X-rays indicated no bony involvement. Ultrasonographic study of lesions was suggestive of mycetoma. Patient was treated with itraconazole 400 mg/day for 4 days in the hospital and discharged subsequently.DiscussionThe foot is commonly involved in those with outdoor occupations. The combination of the clinical specific lesions, typical grains, microscopy, and histopathological studies are characteristic of the diagnosis. Radiodiagnosis is useful to determine the extension of the lesions in bone and other tissues. Combined medical and surgical treatment is recommended. Amputation is indicated in advanced mycetoma not responding to medical treatment with a severe secondary bacterial infection.ConclusionWe hope to spread awareness regarding fungal eumycetoma due to Madurella spp, an endemic but neglected tropical disease in Western Rajasthan. Clinical suspicion coupled to direct KOH microscopy, fungal culture, and histopathology can yield a definitive diagnosis. Commencement of medical and/or surgical treatment at an early stage is necessary to prevent complications.

P071 Isolate profiling and antifungal susceptibility determination for terbinafine and itraconazole among dermatophytes in a tertiary care hospital of western rajasthan

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM ObjectiveTo determine the species distribution of causative agents causing dermatophytosis, and their antifungal susceptibility pattern for terbinafine and itraconazole in trichophyton spp among samples collected in patients with dermatophytosis clinical suspicion during the period of December 1, 2020 to January 31, 2022.Materials and MethodsThis is a prospective study conducted in the Department of Microbiology of a tertiary care super specialty and referral Centre of western Rajasthan from December 1, 2020 to January 31, 2022.Skin scraping, nail clipping, and hair pluckings were collected in mycology lab from clinically suspected cases of dermatophytosis presenting to the department of dermatology for conventional identification, and antifungal susceptibility testing.The specimens were subjected to direct KOH and calcofluor white microscopy and conventional fungal culture on SDA at 25 ͦ C and 37 ͦ C.The cultures positive for dermatophytes were speciated by microslide culture lactophenol cotton blue mount, hair perforation test, and urease test.The isolates identified as Trichophyton spp were taken up for antifungal susceptibility testing against terbinafine and itraconazole by microbroth dilution according to CLSI-M38 A2 guidelines. Further terbinafine resistance gene evaluation for detection of C1191A and T1189C single nucleotide polymorphism in Squalene epoxide by Amplified Refractory Mutation System-Polymerase chain Reaction (ARMS-PCR) is undergoing for trichophyton spp.ResultsOver the 14-month study period, the laboratory processed total of 174 specimens: 134 skin scraping, 36 nail clipping, and 4 hair pluckings. Of them, 106 (61.62%) specimens were microscopy positive and 111 (63.79%) were culture positive. Out of the 111 culture-positive agents isolated, 94 (84.68%) were found to be dermatophytes. On isolate profiling of 94 dermatophytes T. mentagrophyte was found to be most common 45 (48%) followed by T. rubrum 27 (29%), T. tonsurans 20 (21%), T. verrucosum 1 (1.1%), and Microsporum spp 1(1.1%). Antifungal susceptibility of 93 Trichophyton spp against terbinafine showed resistance among 58.06% isolates with 83.33% isolates among terbinafine resistant cases showing ≥4 μg/ml minimum inhibitory concentration. There was no resistance detected for itraconazole with microbroth dilution.ConclusionA total of (54.02%) skin, hair, nail infections were found to be caused by dermatophytes.On isolate profiling, T. mentagrophyte, T. rubrum, and T. tonsurans were found to be predominant species among our isolates showing altered trend of local isolates from T. tonsurans being second most common spp isolated in past.On antifungal susceptibility >55% isolates showed resistance for Terbinafine with >80% having higher MIC of ≥4 μg/ml on the contrary there was no observed resistance for itraconazole.There is a need for encouraging dermatologists for prescribing routine fungal microscopy, culture, and AFST for dermatophytes in Western Rajasthan, to reduce the indiscriminate use of antifungals.

P320 Ten cases of tinea incognito due to Trichophyton rubrum, and literature review on the pathogenic characteristics

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMObjectiveWe present ten tinea incognito (TI) patients caused by Trichophyton rubrum combined with a review of the characteristics of tinea incognito infections reported worldwide over the past 20 years, leading to clinical requirements for diagnosis and treatment.Patients and MethodsPatients were scraped for direct KOH microscopy of the skin lesions at the initial visit, isolates were cultured in SDA medium for 14 days and species were identified by morphology and molecular analysis using ITS regions. Literature review by searching articles on ‘PubMed’ and ‘Web of Science’, using the keywords ‘Tinea incognito or Incognito’, Set the year as ‘2002-2022’, and screened the literature for cases with clear types of TI infecting pathogens. Clinical, mycological, and treatment data of all cases were collected and analyzed.ResultsWe report 10 cases of tinea incognito caused by T. rubrum, ranging in age from 5 to 70 years, with clinical manifestations of eczema-like (Figs. 1a, b, and f), SLE-like (Fig. 1d) and pustular psoriasis-like (Figure 1c, e, g, h, i and j), seen in multiple sites. All of our patients had either tinea pedis or onychomycosis themselves or in their family members.A total of 660 cases reported in the last 20 years, T. rubrum (279/660) was the main anthropophilic dermatophytes causing TI, T. mentagrophytes (162/660) and Microsporum canis (135/660) were the common zoophilic pathgens. There were median age differences in patients infected with T. rubrum, T. mentagrophytes, M. canis, and Nannizzia gypsea, at 56, 22.5, 23, and 5.5 years old, respectively (Fig. 2a). The TI patients infected by T. rubrum often had cutaneous or non-cutaneous underlying diseases (27/41), TI patients with zoophilic or geophilic dermatophyte infections had a definite animal or soil contact history (114/136). The face and trunk are the most frequently affected areas, followed by the extremities and the whole body (Fig. 2b). TI appears on the face and is often thought of as eczema, rosacea, DLE, photosensitive rash, and when it occurs on the trunk is misdiagnosed as eczema, pyoderma, psoriasis (Fig. 2c). In Asia and the Americas, anthropophilic dermatophytes are by far the most reported pathogens, especially T. rubrum. In Europe, zoonotic pathogens have been reported in excess of the anthropophilic (Fig. 2d). For patients in whom the nature of the lesions cannot be determined and with negative direct microscopic examination, dermoscopy can be used to aid in the diagnosis. Our review of all cases found that topical or systemic antifungal treatment always gave good efficacy.ConclusionTI has a broad spectrum of mimics, and an unrestricted range of target audiences, zoonotic pathogens are adapting to the human host, so the history of illness and contact is essential at the time of the initial visit. A definitive diagnosis must be obtained in a mycological laboratory, multiple modalities can be used to diagnose TI, and systemic antifungal treatment is often necessary.

P233 COVID-19-a ssociated Mucormycosis: an experience of invasive fungal sinusitis from a tertiary care referral center in Northern India

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMObjectivesThe study was aimed to determine the patient demographics, risk factors, which include comorbidities, presenting symptoms and signs, medications used to treat COVID-19, species of Mucorales isolated, and the management outcome of COVID-19-associated mucormycosis.MethodsIt was a 6-month retrospective, propensity score-matched, comparative study conducted at a tertiary care center, involving 124 patients with COVID-19 associated mucormycosis admitted between April and September 2021, who were suffering from or had a history of COVID-19 infection.ResultsAmong the 124 patients, 87 were male, and 37 were female. A total of 72.6% of patients received steroids, while 73.4% received antibiotics, and 55.6% received oxygen during COVID-19 management. The most common comorbidities were diabetes mellitus (83.9%) and hypertension (30.6%). A total of 92.2% had mucor, 16.9% had Aspergillus, 12.9% had both, and one patient had hyalohyphomycosis on fungal smear and culture. The comparative study showed the significant role of serum ferritin, glycemic control, steroid use, and duration in COVID-19-associated invasive fungal disease (P <.001). Headache and facial pain (68, 54.8%) were the most common symptoms. The most involved sinonasal site was the maxillary sinus (90, 72.6%). Direct KOH microscopy for fungus was positive in 100% of the cases, 82.2% were Mucorales; 12.9% showed mixed fungal hyphae (mucormycetes and Aspergillus spp.), 4% Aspergillus and hyaline septate non-Aspergillus fungus hyphae were seen in 0.8%. Mucoraceous fungi isolated were R. arrhizus, R. microsporus, R. homothallicus, Rhizomucor pusillus, and Leithemia corymbifera. The overall survival rate at the 3-month follow-up was 79.9%.ConclusionCOVID-19-related aggressive inflammatory response, uncontrolled glycemic level, and rampant use of steroids are the most important predisposing factors in developing COVID-19-associated invasive fungal sinusitis caused by Mucorales. A high index of suspicion, aggressive surgical and antifungal therapy resulted in the successful outcomes for most of the patients.

Real‐life applicability of the Euroarray dermatomycosis kit in the diagnosis of onychomycosis

Traditionally, KOH microscopy and fungal culture are the two preferred tests as gold standard for diagnosis of onychomycosis. Recently, other diagnostic methods have been developed to improve the microbiological diagnosis. The EUROArray dermatomycosis kit is a PCR-based microarray test system for the detection and direct identification of species that are most frequently involved in skin and nail infections. Our primary aim was to evaluate the real-life applicability of the EUROArray dermatomycosis kit in the diagnosis of onychomycoses. In addition, we compared the aetiology of onychomycoses found in our patients with those described in the literature. We prospectively studied consecutive 100 patients with suspected onychomycoses. Samples of suspect toenails were taken as part of routine medical management. Nail specimens were evaluated by means of three diagnostic methods: KOH preparation, culture and EUROArray dermatomycosis kit. Onychomycosis was diagnosed in 47/100 patients who proved positive on at least one reference diagnostic test and in 49/100 patients who proved positive on PCR. The combination of microscopy and PCR had better sensitivity than microscopy (p=.0397), fungal culture (p=.0061) and PCR alone (p=.0117). Moulds were more frequently positive in culture than in PCR (p=.033). Dermatophytes proved positive more frequent than moulds and yeasts in both culture and PCR; in particular, Trichophytoninterdigitale was the most frequent pathogen. In conclusion, introducing EUROArray dermatomycosis kit into the diagnostic algorithm of onychomycosis increases the sensitivity of direct microscopy and yields more rapid results than culture.

Crusted scabies in AIDS patient, a clinical challenge to be sorted out with a simple bedside test.

Norwegian or crusted scabies is a highly contagious severe variant of scabies described first among leprosy patients in Norway in 1848 by Boeck and Danielsen. Herein, we report a case of crusted scabies in an AIDS patient with large hyperpigmented macules covered with thick crusts present over the axilla, inguinal region, and gluteal region. Treatment started immediately with ivermectin, permethrin, and keratolytics after doing KOH microscopy. Mite population may exceed 1 million/person. Hence, it is highly infectious and can set off epidemics of scabies in home or institutions.

The clinical type and etiological agents of superficial dermatophytosis: A cross sectional study

Dermatophytosis are fungal infections caused by three genera of fungi that have the unique ability to invade and multiply within keratinized tissue (hair, skin, and nails). Although dermatomycoses are globally distributed, the endemic and most prevalent species of dermatophytosis differ strikingly from one geographic locality to another. Changing trend has been noticed in last few years with dermatophytic infections presenting as chronic, treatment unresponsive and recurrent. Also various microscopic and fungal culture studies have shown shift in identification of causative fungal species in recent years. Numerous studies have been done on the occurrence of dermatophytes in various parts of our country illustrating the range and changing pattern of fungal infection as well as causative fungal species. Total number of 150 patients attending outpatient department of our hospital who were clinically diagnosed as having superficial dermatophytosis were enrolled into the study. Patients were carefully screened as per inclusion and exclusion criteria and then enrolled in the study. Samples were taken from all the patients and examined for KOH direct microscopy and sent for fungal culture on Sabouraud’s Dextrose Agar as well as on Dermtophyte Test Medium. Results were then analyzed using standard statistical methods. Out of total 150 patients, 101 were males and 49 were females. Most common age group was 21-30 years (37.3%). 58 patients (38.7%) showed positivity to KOH microscopy as well as fungal culture. Additionally 25 more samples demonstrated positivity to KOH microscopy (total 83 patients) but negativity to culture, while 9 patient samples were positive to culture but negative to direct microscopy. Predominant fungal species isolated on culture was Trichophyton mentagrophytes (50.7%) while next common species isolated was T. tonsurans (29.9%). No significant association was found between dermatophyte isolate on culture and clinical type. Trichophyton mentagrophytes and Trichophyton tonsurans were the most common species isolated among subjects with Tinea faciei, Tinea cruris and Tinea corporis. The study showed a male preponderance and T. corporis was the commonest clinical type found. Majority of patients were in the 3rd decade and came within a duration of 1 month to 6 months of getting an infection. In patients diagnosed with tinea corporis, tinea cruris, tinea pedis and tinea manuum, T. mentagrophytes was the most predominant species isolated.

Microbiological and Radiological Perspective of Fungal Rhinosinusitis during the COVID-19 Pandemic: A Prospective Observational Study in A Tertiary Care Centre in Northern India

Radiological investigations are essential for the diagnosis and classification of fungal rhinosinusitis; however, radiological findings might occasionally be misleading. Computed tomography (CT) scan and magnetic resonance imaging (MRI) complement each other, facilitating clinicians to arrive at a diagnosis. Hence, even with the best radiological modalities, correlation between clinical and microbiological findings is crucial for the accurate diagnosis of fungal rhinosinusitis. In addition, the role of traditional methods such as KOH microscopy and culture should be accurately evaluated. To this end, we aimed to diagnose rhinosinusitis with a fungal etiology based on radiological findings and subsequently correlate these findings with those of microbiological techniques, namely culture and KOH microscopy. A total of 57 clinically suspected fungal rhinosinusitis cases were included in the study. Radiological investigations were performed using either CT or MRI. Tissues samples were processed and analyzed using KOH microscopy and culture. The results of the study suggest that using a single method for the diagnosis of fungal rhinosinusitis is inadequate. Rather, the diagnosis should be based on radiological as well as microbiological findings, especially for cases that are clinically ambiguous.