KOH Preparation Research Articles

Clinically Cure with Oral Fluconazole of a Wide Spread Tinea Corporis in Immunocompromised Patient

Tinea Corporis is a commonly superficial fungal infection that mainly affects the skin either immunocompetent or immunocompromised host. Widespread Tinea corporis has been rarely reported. We report an extensive superficial fungal infection with a successful clinical cure with oral Fluconazole. A 38-year- old male presented generalized acuate and annular elevated erythematous scaly border with central clearing distribute on the trunk, both hand and leg. The blood test was confirmed positive for human immunodeficiency virus. KOH preparation was requested which was shown a positive result. The diagnosis was widespread tinea corporis. He was misdiagnosed and received inappropriate treatment with various topical steroid and systemic antihistamine and steroid for several months without any improvement. We emphasized the necessary mycological intervention that enables fast and provides the correct diagnosis and thus medical care. Furthermore, in extensive skin lesions immunocompromised comorbidity should be requested. Mycology detection such as fungal culture and PCR were important tools diagnoses to detect agent-causal, however, it was costly and required a period of time. KOH preparation is one of the in-office methods effective and fast in assisting in an appropriate diagnosis.

Molecular Detection of Microsporum Canis Infection in Cats in Baghdad Governorate

Microsporum canis is a keratinized skin fungus that causes diseases in all animals, especially in cats. It is able to infected human causes Tinea corporis and Tinea capitis. In this study,the aim was toward detection and confirmation of Microsporum canis infection in cats in Baghdad governorate using polymerase chain reaction (PCR).The diagnosis depend on sequencing of the PCR products obtained by using a universal primer to amplification of the primer DppV which was 895 bp.A total of two hundred and eighty three hair and skin scraping samples were collected from infected cats with and without skin lesions. Preliminarily examined of samples with 10% KOH preparation, and cultured for fungal identification . For the culture; Saboruad dextrose agar was used . Culture results showed that four isolates were positive for M. canis , the best growth of these isolates was confirmed by PCR and DPPV gene then sequencing of this gene. PCR technique showed that one isolate from the total of 283 isolates were assured for the DPPV gene and yielded a band at 895 bp, this finding is regarded as distinctive for the genus Microsporum canis ,which was dumped in the GenBank nucleotide sequence database under accession number OR126262. Results sequencing of this gene in Iraq revealed compatibility with global results in USA and Belgium.

Isolation and Identification of Microsporum Canis from cats in Baghdad governorate

Microsporum canis is a keratinized skin fungus that causes diseases in animals, especially cats. It is able to infect human causes Tinea corporis and Tinea capitis . The aim of this study is to isolate and identify the dermatophyte fungi Microsporum canis from cats in Baghdad / Iraq .This research focused on the epidemiology of M. canis found in cats.A total of two hundred and eighty three hair and skin scraping samples were collected from infected cats .preliminarily examined with 10% KOH preparation, and cultured for fungal identification . For the culture; Saboruad dextrose agar was used . The results of this study showed that four samples were positive for M.canis . The pathogen was found both at the site of the lesion and at other sites in the body. The pathogen can be found in the hair of cats with and without skin lesions.

Clinicomycological Study of Dermatophyte Infections in Patients Attending Dermatology Outpatient Department in a Tertiary Care Hospital

Context: Dermatophytosis is a fungal disease characterized by the infection of skin, hair, and nails. There is an upsurge of complicated superficial dermatophytosis and an increase in the incidence of chronic, relapsing, recurrent cases in India that are also often unresponsive to conventional doses of drugs. Aims: The aim of this study was to study the prevalence of dermatophyte species in the particular study area and it is susceptibility to itraconazole, terbinafine, and fluconazole. Settings and Design: All dermatophytes isolates obtained were subjected to 10% KOH mount, fungal culture, and antifungal sensitivity testing for fluconazole, itraconazole, and terbinafine. Materials and Methods: Newly diagnosed cases of dermatophytosis without treatment for the past 3 months were included in the study. KOH preparation was observed and subjected to fungal culture. Then, the susceptibility of the dermatophytes to oral antifungals (itraconazole, terbinafine, and fluconazole) was done. Results: Out of 28 patients with tinea, 14 males and 12 females were tested KOH positive. Out of them, 23 patients showed positive for dermatophyte growth. In them, 15 isolates were Trichophyton mentagrophytes and 8 isolates were Trichophyton rubrum. Antifungal drug susceptibility testing data indicated that, for the two species, MIC of itraconazole ranged between 0.03 and 16 μg/ml, terbinafine between 0.03 and 16 μg/ml, and fluconazole between 0.25 and 64 μg/ml. Conclusions: The study provided information regarding the prevalent species of dermatophyte in our study area and it is high sensitivity to itraconazole compared to terbinafine and the lowest for fluconazole. This gives us guidance as to which antifungal to use empirically in our clinical practice to treat tinea infection.

A STUDY ON CLINICO-ETIOLOGICAL PROFILE OF MYCOTIC KERATITIS IN TERTIARY CARE HOSPITAL AT GOVERNMENT MEDICAL COLLEGE, KOTA (RAJASTHAN)

Introduction: Keratitis is an inflammation of the cornea. Infective keratitis is an ocular emergency that requires prompt and appropriate management to ensure the best visual outcome for the patient. This study was planned to determine the fungal aetiological agents and predisposing risk factors causing corneal ulcer in patients. Methodology: This study was conducted in department of Microbiology at a tertiary medical facility This Analytical Cross-sectional study, was conducted for the study period of one year from May 2021 to April 2022. Total 100 patients of infective keratitis were enrolled in the study period. Cases who have received treatment previously either systemic or topical antifungal in the last six months duration were excluded from the study. Results: Trauma was found to be the most common predisposing factors present in almost two third (64%) of cases. The peak of the fungal kerartits cases were seen in September- October and Februrary- March. Sensitivity and specificity of 10% KOH wet mount was 93.75% and 98.5% respectively. Out of 31 culture positive the most common organisms isolated were Aspergillus spp in 16 samples, followed by Fusarium spp in 6 samples, and Candida spp in 6 samples. C.albicans was found 100% sensitive to Clotrimazole, Nystatin and Amphotericin-B, while 75% sensitive to Itraconazole and 50% sensitive to Fluconazole. Conclusion: Mycotic keratitis has a significant burden in corneal keratitis. The KOH preparation can be used as an ideal technique for revealing fungal elements. The awareness about the ocular hygiene and protection, from occupational hazard needs to be emphasized in case of any ocular trauma and disease.

A STUDY ON CLINICO-MICROBIOLOGICAL PROFILE OF CONJUNCTIVAL FLORA IN CONGENITAL DACROCYSTITIS

Dacryocystitis is an inammation of the lacrimal sac and duct. It may be congenital or acquired.It is an important cause of ocular morbidity both in children and adults. To isolate & identify the causati PURPOSE- ve bacteria from clinically diagnosed cases of congenital dacryocystitis & to study the organism in relation to clinical prole. -Gram฀s stain MATERIALS & METHODS & Giemsa stain ,Culture media,10 % KOH preparation , MacConkey฀s agar, 5% Sheep Blood agar, Chocolate agar .Biochemical tests on Isolated colonies from the plates were identied by Colony morphology, Gram฀s stain, Oxidase test, Catalase test, and Motility.For further identication isolated colony was inoculated into peptone water and Hiss serum for fastidious bacteria, which was incubated aerobically and inoculated into appropriate medias for biochemical tests like Indole production, Methyl red, Voges-proskauer test, Citrate utilization, Urease hydrolysis, Sugar fermentation, Triple Sugar Iron agar, Nitrate reduction test and Amino acids decarboxylase tests. The specic tests like Inulin fermentation, Optochin sensitivity test and Bile solubility test were done. Antimicrobial susceptibility tests: The standardized Kirby-Bauer disc diffusion test of the Clinical and Laboratory Standards institute (formerly NCCLS) was used for testing. The media used was Mueller-Hinton agar for non-fastidious organisms. 5% Sheep Blood agar added to Mueller-Hinton agar for fastidious organisms and Haemophilus Test Medium was used for testing Haemophilus inuenzae and it was read after 16-18 hours. The zone of inhibition was measured and reported as Susceptible or Resistant. For detection of methicillin resistant Staphylococcus aureus detection disc tests for methicillin resistance was done using 10μg oxacillin disc on Mueller-Hinton agar containing 4% sodium chloride. In our study , femal RESULTS- es are more (7) as compared to males (4)& the culture positive cases were 10 and in 1 case culture showed no bacterial growth. The most frequent Gram-positive isolates are Streptococcus pneumoniae 7 (63.63%) followed by Staphylococcus aureus 1 (9.09%) and the most frequent Gram-negative isolates are Pseudomonas aeruginosa 3 (27.27%). In our study the Gram-positive isolates were most sensitive to CONCLUSION- Vancomycin (100%), followed by Tobramycin and Linezolid (97.14 %). Least sensitive antibiotic against Gram-positive organisms was Penicillin (51.42 %). The Gram-negative organisms were most sensitive to Tobramycin and Gentamicin (100%), followed by Amikacin, Oaxacin (92.59 %), Cefepime (88.88 %). Least sensitive antibiotic against Gram-negative organisms was Ciprooxacin (77.77 %)

Fusarium as a Cause of Mycetoma: A Case Report and Review of the Literature

Mycetoma is a chronic, slow progressive granulomatous infection of skin and subcutaneous tissue; is classified into Eumycetoma (caused by fungi) and Actinomycetoma (caused by Actinomycetes). Fusarium, is a known cause of mycetoma. This case report describes the isolation and identification of Fusarium sp. from a lesion on the right foot of a female patient. This case report is about a 23-year-old female with complaints of gradual enlarging nodular swelling with multiple discharging sinuses along with granules on the right foot, on and off for last seven years. The granules obtained from the lesions are the best specimen for culture as they contain microcolonies of causative agents and it’s a hallmark of mycetoma. Granules were collected aseptically, crushed and examined microscopically on 20% KOH preparation. Plenty of thin hyaline, septate hyphae were seen. Granules were washed with sterile saline and inoculated on Sabouraud’s Dextrose Agar (SDA) and potato dextrose agar and incubated aerobically at 25°C and 37°C in two sets of each. Colonies grew at inoculated site, pinkish in the centre and whitish in the periphery. The isolate was identified as Fusarium sp. on the basis of morphology. The patient was started on Itraconazole 200 mg and is currently on the same treatment, improving and under follow-up. Mycetoma is a rare disease caused by both fungal and bacterial microorganisms. Definitive diagnosis should be made on the basis of culture or histopathological examination. Once the diagnosis is made, use of appropriate anti-fungal therapy with or without surgical interventions can result in complete cure without recurrence.

Study of Clinico-mycological profile of onychomycosis at a tertiary care hospital

Abstract Introduction: Onychomycosis (OM) is a common nail disorder caused by dermatophytes, non-dermatophyte moulds and yeasts. The increased prevalence of OM along with the wide range of associated fungi necessitates the accurate laboratory isolation and identification of the causative fungus for optimal management strategies. The aim of the study was to evaluate clinical, epidemiological and mycological characteristics of OM at a tertiary care hospital in Karimnagar. Materials and Methods: The study was conducted at Department of Microbiology for a period of one year. 109 patients clinically diagnosed as OM were subjected to detailed history, clinical examination and investigations. Nail clippings and subungual debris of OM patients were subjected to KOH preparation and culture. Isolates were identified by colony characters, pigment production, microscopic morphology. Results: The maximum number of OM cases belonged to the age group 21-30 years with a male preponderance. Higher incidence was noted in males (71.5%) than females (28.4%). Direct microscopy of the nail clippings in 40% KOH solution was positive in 56.8% and culture was positive in 75.2% cases. The common etiological agent was dermatophytes (45.12% cases) followed by yeasts (34.14% and non dermatophyte molds (20.7% cases). T.rubrum was the commonest among dermatophytes and Candida albicans among the yeasts. Conclusions: Although onychomycosis is not life threatening, it can cause a significant negative impact on the quality of life of infected patients. Early diagnosis and treatment is important to prevent distortion of the nails which can prevent loss of working efficiency of the commonly affected population.

P396 Utility of nasal swabs as samples for the diagnosis of Mucormycosis using molecular methods for detection

Poster session 3, September 23, 2022, 12:30 PM - 1:30 PM ObjectivesMucormycosis is an aggressive, life-threatening infection caused by fungi in the order Mucorales. There was an explosion of new cases of rhino-sino-orbital mucormycosis following the COVID pandemic in India, and the need for easy and rapid diagnostics was felt. The current diagnosis of mucormycosis relies on mycological cultures, radiology, and histopathology. These methods lack sensitivity and are most definitive later in the course of infection, resulting in the failure of timely intervention.A real-time multiplex PCR platform is commercially available for the detection of Rhizopus spp., Mucor spp. Rhizomucor spp., Lichtheimia spp., and Cunninghamella spp. (PN-700, MucorGenius®, PathoNostics®, Maastricht, The Netherlands) This real-time PCR has been validated to identify these fungal pathogens from bronchoalveolar lavage, tissue, and serum samples. This study aimed to validate this PCR-based system to detect Mucorales from nasal swab samples and evaluate its utility in the detection of Mucorales from nasal cavities of high-risk patients developing signs and symptoms of mucormycosis.MethodsA single-center cross-sectional observational study was conducted on 50 hospitalized adult patients with signs and symptoms of mucormycosis. Nasal swabs were taken for PCR analysis once there was a clinical suspicion and were compared with the results of the gold standard. The gold standard for the diagnosis of mucormycosis was the conventional method (KOH mounted microscopy/HPE). Demographic details and risk factors for these patients were recorded, and the RTPCR-based test was run on the nasal swab samples of all these 50 patients. The workflow is depicted graphically in Fig. 1 (Created with BioRender.com).ResultsThe study population mean (SD) age was 50 (16) years and consisted of 32 (64%) males. A total of 39 (78%) patients were known cases of diabetes mellitus, 48 (96%) patients had amphotericin B intake, and 20 (40%) had posaconazole intake. In all, 21 (42%) patients had a past history of COVID-19 infection; 14 patients had received steroids and 10 patients received oxygen support. PCR for Mucorales was positive in 15 (30%) patients while the KOH mount was positive in 4 (8%) patients.ConclusionThese results are not encouraging for the use of nasal swabs as the sample for diagnosis of mucormycosis. Though the PCR performed better on the swab samples than KOH preparation and culture techniques, it highlights the importance of using standard sampling methods.

P207 A rare case of disseminated Blastomycosis in an immunocompetent host: A report from North India

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PM ObjectivesBlastomycosis is a serious life-threatening systemic infection caused by dimorphic fungi Blastomyces dermatitidis. Infection is acquired via inhalation of airborne conidia or traumatic inoculation. It may produce a spectrum of infections ranging from asymptomatic infections in immunocompetent patients to disseminated disease, including skin, bone, genitourinary tract, and central nervous system (CNS) involvement in immunocompromised patients. This fungal infection is very rare in India. We report a unique case of disseminated blastomycosis in an immunocompetent patient.MethodsA 37-year-old male patient from Kanpur, Uttar Pradesh presented with chief complaints of hematuria associated with passage of blood clots for 3 months. There was no history of dysuria, fever, or pain abdomen. When he was evaluated for gross hematuria, his USG abdomen was suggestive of bilateral renal masses. To rule out bilateral renal cell carcinoma an FDG pet scan was done which was suggestive of FDG avid lesions in the bilateral kidneys. He also had enlarged left supraclavicular and left cervical lymph nodes. Multiple nodular and verrucous lesions were seen all over the face. He complained of pain in the left ankle; with a small wound opening on the medial malleolus of the left ankle.ResultsA skin biopsy was taken from one of the nodular and verrucous lesions over face. A total of 10% wet mount KOH preparation shows no epithelial cells, plenty of pus cells, no RBCs, and plenty of thick-walled round yeasts and some with broad-based budding yeasts (8-12 microns) suggestive of Blastomyces species. Renal abscess pus FNAC, left cervical lymph node, and CT-guided biopsy from the medial malleolus of the left ankle also showed similar round yeasts and some with broad-based budding suggestive of Blastomyces species. Hence, a preliminary diagnosis of disseminated blastomycosis was made.Cultures were put in SDA at 25°C and 37°C and after a week, colonies appeared as fluffy white glabrous with reverse cream-colored non-sporulating colony. On day 25, colony appears as tan glabrous fluffy with brownish reverse with sporulation. LPCB was done from the colony which showed thin septate hyphae with pedunculated and sessile spherical to pyriform and smooth walled microconidia suggestive of Blastomyces dermatitidis.He was started on IV liposomal amphotericin B for 20 days. HCV RNA was detected incidentally and he had transaminitis therefore, he could not be started on itraconazole, he was started on fluconazole. His facial lesions cleared dramatically; his left ankle pain and swelling had resolved. At present, he was symptomatically better and was discharged on 200 mg fluconazole once daily and advised for follow-up after 2 weeks.ConclusionBlastomycosis is a relatively uncommon geographically restricted chronic granulomatous disease that mainly occurs in the endemic regions. This fungus lives primarily in moist soil and decomposing matter, and inhabits the mid-west, south-central, southeastern United States as well as the boreal forests of Ontario and Quebec in Canada. Very little is known about the natural habitat and environmental distribution of Blastomyces species in India. This case is unique because the patient is immunocompetent and has no history of travel outside of India.

P266 A case of mixed fungal infection causing invasive fungal Rhinosinusitis in a post-COVID patient

Poster session 2, September 22, 2022, 12:30 PM - 1:30 PMObjectivesTo identify the fungal etiology of invasive nasal sinusitis in a patient of post-COVID.MethodsA 34-year-old non-diabetic man, who had mild coronavirus disease (COVID) infection 2 months back presented with left-sided nasal obstruction, headache accompanied by malodorous, thick, mucopurulent discharge for the last 2 weeks.A CT scan of sinuses revealed opacification of left posterior ethmoid and sphenoid sinus without bony erosion or calcification.The patient underwent unilateral functional endoscopic sinus surgery (FESS) and debridement of tissue from the affected sinuses.ResultsA KOH preparation of the debrided tissue showed thin septate hyphae. Gram-stained smear of the debrided material showed thin septate fungal hyphae with clamp-like connections (Fig. 1). Histopathological examination revealed features of the inflammatory polyp. In Sabouraud dextrose agar the fungal colony grew fast and its aerial mycelium is white and cotton-like. The fungus was phenotypically identified as Schizophyllum communae (identification was confirmed at the National Reference Centre).He continued to do well but about 2 months later he started experiencing headaches and pain behind the eyes. He also complained of nasal stuffiness (left greater than right) and yellowish nasal discharge. At this point, a CT scan revealed soft tissue density with interspersed hyperdensity in sphenoid sinus bilateral ethmoid, and bilateral maxillary sinuses with associated bony erosion and possible extension into the right cavernous sinus and extrachonal compartment of right orbit suggestive of invasive fungal sinusitis (Fig. 2).Bilateral FESS was done. Extensive fungal material was observed in the sphenoid sinus and thorough debridement was performed.Mycological studies of the debrided tissue showed thin septate hyphae. Schizophyllum communae was again isolated in culture.Histopathological section showed inflammatory cells and several slender, branching septate fungal hyphae.The invasive nature of the infection prompted reexamination of the histopathology slides and cultures for the presence of other fungi particularly of the Mucorales group but no other fungus could be identified.The patient was started on liposomal amphotericin B and the tissue blocks were sent for fungal identification to the National Reference Centre through sequencing following DNA extraction from the paraffin blocks. Amplification of the 18 S rDNA region (coding for the 18 S RNA) using ZM primers followed by sequencing revealed the presence of R. arrhizus.The patient was continued on amphotericin B. The patient was discharged after 2 weeks with oral isuvuconazole. At six months follow up he is doing well with no evidence of active infectionConclusionAs the novel COVID‐19 continues to rampage, an abrupt increase in the number of opportunistic fungal infections has been observed.Invasive and often fatal rhino-cerebral Mucormycosis is now been increasingly reported in patients who have had COVID-19 infection in the recent past. In this case, there was a dual fungal infection causing rhino-sinusitis which was established through conventional culture and PCR assays from paraffinized tissue sections. Increased awareness of the existence of dual mold infections in at-risk patients is necessary for optimum management. PCR methods in tissue sections increase the diagnosis of dual mold infections.

MICROBIOLOGICAL PROFILE IN WOMEN WITH SYMPTOMATIC VAGINAL DISCHARGE

The study's main goals are to determine the prevalence of symptomatic vaginal discharge in different age groups and the microbiological makeup of women who report with symptomatic vaginal discharge. The study was conducted in 200 cases. A detailed clinical history and examination of all cases were done. A speculum examination done to visualize cervix and vagina , swabs were collected from the posterior fornix of vagina and sent for microbiological examination. Following tests were done a) Gram staining and Nugent scoring system used, based on the morphology of the bacteria, score >/= to 7 is considered positive for bacterial vaginosis, b) Wet mount preparation done to look for motility of Trichomonas, c) KOH preparation, d) Swabs are inoculated in Blood agar, Mac Conkey's agar and Sabouraud's dextrose agar. Maximum patients had associated symptoms as foul smell (27%),followed by itching(26%) ,others like abdominal pain(11.5%),burning micturition(6%) , heavy menstrual bleeding(4.5%) and low backache(2.5%). Majority had homogeneous discharge(44.5%), followed by mucopurulent(19.5%),curdy white(18%) and yellowish (18%). Incidence of bacterial vaginosis is maximum(42.5%), followed by vaginal candidiasis (21.5%),other non specific (17%) and Trichomonas(8%). In around 11% of cases no cause was identified.

Alkali Niobate Powder Synthesis Using an Emerging Microwave-Assisted Hydrothermal Method.

For more than five decades, alkali niobate-based materials (KxNa1−xNbO3) have been one of the most promising lead-free piezoelectric materials researched to be used in electronics, photocatalysis, energy storage/conversion and medical applications, due to their important health and environmentally friendly nature. In this paper, our strategy was to synthetize the nearest reproductible composition to KxNa1−xNbO3 (KNN) with x = 0.5, placed at the limit of the morphotropic phase boundary (MPB) with the presence of both polymorphic phases, orthorhombic and tetragonal. The wet synthesis route was chosen to make the mix crystal powders, starting with the suspension preparation of Nb2O5 powder and KOH and NaOH alkaline solutions. Hydrothermal microwave-assisted maturation (HTMW), following the parameter variation T = 200–250 °C, p = 47–60 bar and dwelling time of 30–90 min, was performed. All powders therefore synthesized were entirely KxN1−xNbO3 solid solutions with x = 0.06–0.69, and the compositional, elemental, structural and morphological characterization highlighted polycrystalline particle assemblage with cubic and prismatic morphology, with sizes between 0.28 nm and 2.95 μm and polymorphic O-T phase coexistence, and a d33 piezoelectric constant under 1 pC/N of the compacted unsintered and unpoled discs were found.

Prevalence and Risk Factors of Zoonotic Dermatophyte Infection in Pet Rabbits in Northern Taiwan.

Dermatophytes are the group of keratinophilic fungi that cause superficial cutaneous infection, which traditionally belong to the genera Trichophyton, Microsporum, and Epidermophyton. Dermatophyte infection is not only a threat to the health of small animals, but also an important zoonotic and public health issue because of the potential transmission from animals to humans. Rabbit dermatophytosis is often clinically identified; however, limited information was found in Asia. The aims of this study are to investigate the prevalence and to evaluate the risk factors of dermatophytosis in pet rabbits in Northern Taiwan. Between March 2016 and October 2018, dander samples of pet rabbits were collected for fungal infection examination by Wood’s lamp, microscopic examination (KOH preparation), fungal culture, and PCR assay (molecular identification). Z test and Fisher’s exact test were performed to evaluate the potential risk factors, and logistic regression analysis was then performed to build the model of risk factors related to dermatophyte infection. Of the collected 250 dander samples of pet rabbits, 29 (11.6%) samples were positive for dermatophytes by molecular identification. In those samples, 28 samples were identified as the T. mentagrophytes complex and 1 sample was identified as M. canis. Based on the results of the Firth’s bias reduction logistic analyses, animal source (rabbits purchased from pet shops) and number of rearing rabbits (three rabbits or more) were shown as the main risks for dermatophyte infection in the pet rabbits in Taiwan. The results of the present study elucidate the prevalence of rabbit dermatophyte infection, pathogens, and risk factors in Taiwan, and provide an important reference for the prevention and control of rabbit dermatophytosis.

Evaluation of early detection of mucorales by rapid one step off- plate protein extraction using maldi -tof ms

Aim: To validate a simple one step method of protein extraction of moulds for routine early detection of fungi using automated instrument like MALDI- TOF MS. Methodology: A total of 123 clinical mould isolated were tested in Department of Microbiology, Indira Gandhi Institute of Medical Sciences, Patna, Bihar, India during period of 3 months (August 2021-October 2021) with the routine Laboratory and culture investigations. Out of these specimens, approximately 50 specimens were suspected of Mucorales based on their growth timing, colony morphology, history of patient, character of tissue or sample, and corresponding KOH mount findings having hyphae in tissues or samples were included in study. Samples were inoculated on routine fungal culture media like SDA. The plates were examined at earliest for appearance of any growth and the process of protein extraction was done to be analysed in MALDI- TOF MS and compared with KOH preparation findings of sample. The culture were again incubated further for appearance of mature colony and later on isolates were identified by LPCB mount examination. Results: Around 70% growth of mucorales could be identified by this method and 30% could not be identified.

Crush cytology smears – A novel approach to rapid diagnosis of rhino-orbital mucormycosis in post-COVID-19 patients: A single-center study

Background: With recent surge in SARS-COVID-19, cases of rhinocerebral mucormycosis are on rise. Rapid diagnosis of mucormycosis is essential as delay in the treatment is associated with dreadful outcome. Many studies have found that crush cytology smears can rapidly diagnose mucormycosis as compared to histopathology, thereby reducing the time for arriving at final diagnosis, here, we present a novel approach to the rapid diagnosis of mucormycosis by crush cytology and analyze whether it has a higher sensitivity in diagnosing cases of mucormycosis. Aims and Objectives: This study aims to analyze the efficacy of crush cytology smear in rapid diagnosis of mucormycosis. Materials and Methods: Biopsies received from all the clinically suspected cases of mucormycosis with a history or associated present COVID-19 infection. The biopsy was processed for KOH preparation, crush cytology smear and confirmation were done by histopathological examination. Special stains were done wherever necessary. Results: Out of 50 cases of clinically suspected mucormycosis, 43 confirmed on histopathology. Out of 43 confirmed cases of mucormycosis, 33 (76.74 %) were male and 10 (23.25%) were female with male: female ratio of 3.3:1. The mean age of the studied cases was found to be 52.32±12.59 years. The most common presenting complaint was fever (79.07%), followed by headache/retro-orbital pain (62.79%), rhinorrhea (58.14%), and facial swelling (53.49%). Out of 43 patients, 41 recovered from COVID-19 disease with mean duration of 12 days. Hospital stays of patients with for COVID-19 varied from 7 days to maximum of 35 days. Out of 43 cases of mucormycosis, 24 cases were diagnosed with KOH mount with sensitivity of 55.81% and 39 cases with crush cytology smears with sensitivity of 90.69%. Conclusion: The diagnosis of mucormycosis is done with the gold standard methods such as histopathological examination and culture studies but the present study emphasizes on use of crush cytological smears prepared from sinonasal biopsies in suspected cases of mucormycosis in rapid and accurate diagnosis of mucormycosis.

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.

Prevalence of Microsporum canis from Pet Cats in Small Animal Hospitals, Chiang Mai, Thailand.

Dermatophytosis is a disease caused by dermatophytes, a group of fungi that can cause disease both in humans and animals. The important genera that are pathogenic in animals include Trichophyton and Microsporum. Microsporum canis is an important species because it can cause zoonosis and is commonly found in domestic animals. Cats, which live very close to humans, may expose humans to this pathogen. This research focused on the epidemiology of M. canis found in cats. Hair samples were collected via the Mackenzie technique from cats with and without skin lesions, preliminarily examined with 10% KOH preparation, and cultured for fungal identification. Samples were confirmed with molecular techniques including polymerase chain reaction, gel electrophoresis, and sequencing. Samples were collected from 138 cats located in 93 households, 43 from cats with skin lesions (31.16%) and 95 from cats without skin lesions (68.84%). Eighteen cats with lesions (13.04%) and ten cats without lesions (7.2%) were found to carry M. canis. In eleven of the eighteen cats both with skin lesions and positive for M. canis (61.11%), the pathogen was found both at the site of the lesion and at other sites in the body. Because the pathogen can be found in the hair of cats with and without skin lesions, owners, keepers, veterinarians, and others who come into contact with these animals are at risk of infection if they are not aware or do not take precautions after contact with them.

Evaluation of Sensitivity and Specificity of Direct Microscopical Examination of Suspected Mucor Mycosis Samples by Potassium Hydroxide (KOH) during Covid-19 Pandemic Era

Introduction: Mucormycosis is a common fungal infection in India even before the COVID-19 pandemic. The incidence of Mucor mycosis has entered public consciousness in response to an outbreak of cases in India during COVID-19 pandemic. Mortality is unacceptably high, reaching 80% in some case series. Prompt diagnosis is crucial because treatment initiation is time-critical due to the rapid progression of the infection. Direct KOH mount is a simple, rapid, inexpensive point-of-care test, which requires minimum technical aids. Interpretation of KOH smears requires some amount of experience. The present study was to evaluate the sensitivity and specificity of Direct Microscopical examination by KOH preparation of suspected Mucor mycosis samples taking the fungal culture as gold standard. Material and Methods: 230 samples were received in Microbiology Department from May 2021 to August 2021 from the patients suspected of Mucor mycosis. Samples were processed for KOH preparation using different concentrations of KOH like 20% and 40% till all tissues get digested. Samples were cultured also in Sabourauds dextrose agar (SDA) medium. Results of microscopy and culture were statistically analyzed using SPSS software version 23. Result: Sensitivity of KOH preparation was 67%, specificity was 87%. Positive predictive value was 76% and negative predictive value was 81% and accuracy was 80% when compared with the fungal culture. Discussion: Direct microscopic examinations using KOH preparation are the simplest, cheapest method used for the diagnosis of mycotic infections. This study is valuable in establishing a reliable method for early information on diagnosis of Mucor mycosis that may be crucial for determining appropriate therapy for the successful treatment of Mucor mycosis patients. Conclusion: KOH examination plays an important role in the diagnosis of Mucor mycosis suspected patients during COVID-19 pandemic era when compared with fungal culture which is a gold standard test in diagnosis of Fungal infection. It is the simplest and cheapest method that can be developed easily with minimal infrastructure or human resources in resource-limited countries during the COVID-19 era.

Evaluating diagnostic utility of PAS stained skin scrape cytology smear in clinically suspected superficial cutaneous mycoses: A simple yet unpracticed technique.

Background and Objectives:For fungal dermatitis, a wet Potassium Hydroxide (KOH) preparation of the skin scrapings forms the routine practice for diagnosis. This study was carried out to determine the diagnostic accuracy and to evaluate the value-added information of PAS-stained scrape cytology smear in evaluating the presence of fungal elements.Methods:This prospective study was carried out on the patients clinically diagnosed with superficial cutaneous mycoses. For each patient, a wet KOH preparation and one PAS-stained skin scrape cytology (SSC) smear was prepared.Results:Out of the 52 suspected cases of superficial cutaneous mycoses, 50 showed fungal elements on either or both the techniques. The presentation was Tinea cruris together with Tinea corporis in 21 cases (42%), isolated tinea cruris in 19 cases (38%), tinea corporis in 8 cases (16%), and tinea corporis with tinea manuum and onychomycosis in 2 cases (4%). KOH preparation was positive in 45 out of 50 cases (90%) and SSC was positive in 49 out of 50 cases (98%). The sensitivity, specificity, positive predictive value, and negative predictive value for PAS-stained SSC smear was 98%, 100%, 100%, and 66% and that for KOH preparation was 90%, 100%, 100%, and 28%, respectively. Fungal elements quantity was graded on SSC smear as 1+(5), 2+(19), 3+(19), 4+(6), and in one case, it was negative.Conclusion:Incorporation of the PAS-stained SSC smear for fungal dermatitis ensures more efficient and confident diagnosis and the slides are available for archivable studies.