Comparative evaluation of histopathological analysis, KOH wet mount and fungal culture to diagnose fungal infections in post-COVID patients

Context and Aim: There is increasing prevalence of post-COVID fungal infection of rhinoorbitocerebral region especially mucormycosis and aspergillosis in India.[1] Early diagnosis of these fungal infections are of utmost importance, since it may improve outcome and survival.[2],[3],[4],[5],[6],[7],[8] The objective of this study was to evaluate and compare routine laboratory diagnostic methods, that is, histopathological examination, KOH wet mount and fungal culture in the diagnosis of post-COVID fungal infections. Materials and Methods: A total of 106 specimens of clinically suspected patients of post-COVID fungal infection of rhinoorbitocerebral region received in histopathology department were included in this study. The data of KOH wet mount and culture were acquired from the microbiology department after histopathological examination. Result: Approximately 88.68% of patients were diagnosed having fungal infections by one of the laboratory methods. The sensitivity of histopathological examination was highest (79.78%), followed by KOH wet mount (58.51%) and fungal culture (35.10%). Rhizopus species of zygomycetes group were the most common isolate (24.24%) on SDA culture. Overall 76% concordance was found between histopathological examination and fungal culture report for morphological identification of fungi. Conclusion: For the diagnosis of post-COVID fungal infection of Rhino-orbito-cerebral region, histopathological examination is was found to be more sensitive and rapid method to detect fungal hyphae. It leads to early treatment, prevents morbidity and mortality.

A retrospective clinicopathological analysis of rhino-orbital mucormycosis in second wave of COVID-19

Background: During the present surge of COVID-19 positive cases, concurrent multifold increase in the incidence of mucormycosis cases has resulted into significant morbidity and mortality. We retrospectively evaluated the clinicopathological features along with microbiological examination findings in histologically diagnosed cases of rhino-orbital mucormycosis. Material and Methods: All the H and E and special stained slides of included mucormycosis cases were retrieved from the records and were evaluated with microbiological findings including screening KOH mount examination and culture results. Results: Out of 16 cases with available details, 10 cases had the previous history of diabetes mellitus. The most frequent single site of involvement was maxillary sinus (7/25) followed by nasal cavity, orbit, ethmoid and sphenoid sinuses. While comparing the histological diagnosis with KOH mount findings and culture results, 15 cases revealed consistent results. Conclusion: A high clinical suspicion, monitoring, early diagnosis, and timely management can improve the morbidity and mortality of this life-threatening complication.

MYCOLOGICAL PROFILE OF DERMATOPHYTES

Aims: Dermatophytoses are commonest type of fungal infections seen in human beings, invading keratinised tissue skin, hair and nails by keratinophilic fungi. Dermatophytoses can be treated effectively by antifungals. Retrospective study A total of 200 patients which were Settings and Design: Methods and Material: referred from the department of dermatology to the department of microbiology were collected from July 2020 to June 2021. After cleaning with povidone iodine and 70% alcohol, the skin was scrapped, hair was plucked and nails were clipped. Skin, Hair and Nail were treated with 10%, 20% and 40% KOH respectively to identify septate hyphae and arthrospores. The specimen was cultured on Sabouraud's dextrose agar (SDA) with and without cycloheximide. Microscopic description of the growth was done by Lactophenol cotton blue (LPCB) staining. Antifungal susceptibility testing was done by E strip method using Rosewell Park memorial institute (RPMI) media for Ketoconazole, Itraconazole, Fluconazole, Voriconazole, Posaconazole, Griseofulvin and Terbina?ne. Out of 200 samples, the commonest age group affected was between (21-30 yrs) 39.5%. Female to Results: male ratio was 2.5:1.6. Tinea corporis was 63% and tinea cruris 27%. The commonest species isolated was Trichophyton rubrum 47.5%. Antifungal susceptibility test – Most susceptible to Itraconazole 81.4% and resistant to Griseofulvin 67.9%. Conclusions: Dermatophytosis is prevalent in high temperature zones. Most common isolates were T. rubrum and Trichophyton mentagrophytes in tinea cruris and tinea corporis respectively, they were most susceptible to Itraconazole and resistant to Griseofulvin. Resistance pattern of antifungal drugs helps in deciding empirical therapy for patient's better outcome.

Clinico-Mycological Study of Onychomycosis and Pattern of Drug Resistance

Introduction: Onychomycosis is a fungal infection of thetoenails or fingernails that may involve any component of thenail unit, including the matrix, bed, or plate. Onychomycosisis caused by dermatophyte fungi, non dermatophyte mouldsand yeasts. Prevalence rates for OM varying from 3 to 5%have been found in most studies; however, a few reportssuggest a higher prevalence of even up to 26% in the generalpopulation. Hence through this study an attempt was made toidentify the various clinical patterns of onychomycosis and toidentify pattern of drug resistance in onychomycosis.Material and methods: The proposed study was carriedout in Outpatient Department of Dermatology Venereologyand Leprosy, Rohilkhand Medical College and Hospital,Bareilly, a tertiary care hospital in western Uttar Pradeshfor one year (November 2017- October 2018) on patientshowing nail changes suggestive of onychomycosis, usingdirect microscopy using potassium hydroxide. The culturemedium SDA with chloramphenicol was used. Sensitivity toTerbinafine, Fluconazole, Itraconazole and Griseofulvine wasstudied.Result: The most common pattern of onychomycosis seenwas distal lateral subungual onychomycosis. Pattern of drugresistance is maximum with Griseofulvin (49%) and least withTerbinafine (20%).Conclusion: A total of 104 patients were enrolled, withmean age 35.93 years. Males were more commonly involvedcompared to females. The culture results were positive among55 patients and negative in 49 patients. The most commoncausative agent isolated was dermatophyte moulds followedby non-dermatophytes and yeasts.

Utility of Urine KOH in Detecting Candiduria in Infants

Background@#Candida species are common cause of urinary tract infection in infants requiring medical care. Candida fungal elements may be demonstrated in urine using microscopic examination with potassium hydroxide (KOH). However, detection of these elements does not always correlate with candiduria. @*Objectives@#To establish the utility of urine KOH in identifying candiduria and to determine the risk factors, as well as urinalysis and CBC parameters associated with candiduria. @*Methods@#This prospective cross-sectional study included admitted infants 1 year and below with urine culture and with any risk factor/s for candiduria. Additional urine KOH testing was done using clean catch or catheter method. Urine culture was used as the gold standard. @*Results@#Among the 90 study participants with both urine culture and urine KOH, 13 (14%) had candiduria. The use of indwelling catheter, presence of urinary tract anomalies, positive leukocyte esterase in urinalysis, and increased monocyte counts in CBC are all associated with candiduria. Urine KOH has sensitivity of 100%, (CI 75.2-100%), specificity 59.7%, (CI 47.9-70.7%), PPV 29.5%, (CI 17.7-45.2%), and NPV 100%, (CI 92.2-100%) in detecting candiduria.@*Conclusions@#Negative urine KOH has excellent negative predictive value, while positive urine KOH result may warrant further investigation. Urine KOH results should be interpreted with caution depending on patient’s risk factors, clinical status, and other laboratory results prior to initiation of empiric antifungal therapy. Positive urine KOH may not always require treatment.

Mixed dermatophytic infections: a learning perspective and report of ten cases

Background: Isolation of two or more than two pathogenic fungi from the same body site in a patient is considered as a rare entity and very few cases have been reported in literature. These types of infections are called as mixed/ combined fungal infections. Author are enumerating ten cases of superficial mycoses in which two different dermatophytes were grown from the same focus.Methods: From clinically suspected cases of dermatophytosis, skin and hair samples were collected from the affected sites and examined by standard mycological procedures. Microscopy was done by using 10% KOH wet mount. Culture was put on Sabouraud’s dextrose agar with cyclohexamide medium. Growth was identified by lactophenol cotton blue mount.Results: Mixed dermatophytes were obtained from tinea corporis (five cases), tinea capitis (four cases) and tinea cruris (one case) patients. Fungal combinations from given cases involved two different species of genus Trichophyton which were as follows: T. violaceum+T. tonsurans, T. verrucosum+T. tonsurans, T. violaceum (violet) and T. violaceum (white), T. mentagrophytes+T. Violaceum, T. rubrum+T. tonsurans, T. violaceum+T. rubrum, T. rubrum+T. mentagrophytes, T. verrucosum+T. mentagrophytes, T. mentagrophytes+T. tonsurans, Malassezia+T. mentagrophytes.Conclusions: Inspite of the frequent occurrence of dermatophytic infections worldwide, reports on mixed dermatophytes are very few. With proper sample collection and proper identification procedures, more cases can be identified and added to the existing literature.

Visual Outcome among Patients Undergone Manual Small Incision Cataract Surgery Following their Identification in Screening Eye Camp in Jammu Province.

Introduction: One of the documented leading cause ofblindness in India is cataract as many patients with cataract donot have access to hospitals and surgery and to avoid blindnessdue to cataract, the only remedy is to perform hospital basedcataract surgery on a large scale. There is an increase in anumber of cataract patients due to improved quality of life,health indices and increased life expectancy. Study aimedto see the visual outcome and complications among patientsundergone manual SICS with PC-IOL implantation.Material and Methods: The present prospectiveobservational study was conducted on 72 patients who wereselected in various screening eye camps to undergo cataractextraction surgery by manual small incision cataract surgerytechnique with posterior chamber IOL implantation (MSICSwith PC-IOL).Results: Total of 72 eyes of patients who underwent manualsmall incision cataract surgery were studied. Best correctedpreoperative visual acuity of ≥ 6/60 was found in 13 patientswhile postoperatively on day1 total of 60 patients had visualacuity of ≥ 6/60. The final 6th week postoperative bestcorrected visual acuity of 6/12-6/9 was found in 65 patients.Iris prolapse was seen in 3 patients followed by posteriorcapsule rent which was seen in 2 patients. Mild postoperativeuveitis was seen in 20 followed by striate keratopathy in 8patients while hyphaema was seen in 2 patients.Conclusion: MSICS is a safe and effective procedure, due toits low rates of intraoperative and postoperative complicationswhich are easily treatable. Visual acuity is only one measureof the functional success of cataract surgery and the goodpostoperative visual outcomes achieved by our patientsfurther adds on to its benefits as a good surgical technique.MSICS can be performed as procedure of choice for largevolume cataract surgeries

Comparison of fluorescent staining versus KOH wet-mount microscopy for detection of superficial fungal infection / 中华皮肤科杂志

Objective To compare the diagnostic value of fluorescent staining versus KOH wet-mount microscopy in detecting superficial fungal infection.Methods Totally,600 specimens from cases of clinically diagnosed superficial fungal infections and 102 from cases of clinically diagnosed Malassezia infection (including 54 cases of pityriasis versicolor and 48 cases of Malassezia folliculitis) were collected from the dermatology clinic of Tenth People's Hospital of Tongji University between July 2017 and February 2018.These specimens were subjected to fluorescent staining and KOH wet mount separately followed by direct microscopy,and the positive rate and average time for slide reading were compared between the two methods.Culture served as the gold standard method,and the missed diagnosis rate was compared between the two methods.Statistical analysis was carried out using chi-square test or Fisher's exact test for comparing enumeration data,and paired t test for comparing emeasurement data.Results Of the 600 specimens from clinically diagnosed superficial fungal infection cases,fungi were detected in 546 (91.00％) and 489 (81.50％) by fluorescent staining and KOH wet-mount microscopy respectively (x2 =22.83,P ＜ 0.05).Fluorescent staining showed significantly shorter average reading time (73.67 ± 13.56 s)compared with KOH wet-mount microscopy (87.12 ± 15.83 s,t =14.60,P ＜ 0.05).Among the 54 specimens from pityriasis versicolor cases,fluorescent staining and KOH wet-mount microscopy positive results in 51 (94.44％) and 50 (92.59％) specimens respectively (adjusted x2 =0,P ＞ 0.05),with the average reading time being 38.36 ± 8.79 s and 41.25 ± 15.67 s respectively (t =1.14,P ＞ 0.05).Of the 48 specimens from Malassezia infection cases,43 (89.58％) and 11 (22.92％) specimens were detected to be positive for fungi by fluorescent staining and KOH wet-mount microscopy respectively (x2 =43.34,P ＜ 0.05),and fluorescent staining showed shorter average reading time (42.14 ± 12.61 s) compared with KOH wet-mount microscopy (103.56 ± 9.48 s,t =17.83,P ＜ 0.05).Among the 600 specimens from superficial fungal infection cases,culture yielded fungi in 479.Moreover,476 specimens were found positive by fluorescent staining,and 3 were found negative (0.63％),while KOH wet-mount microscopy showed 465 positive results and 14 negative results (2.92％).There was a significant difference in the missed diagnosis rate between the two methods (x2 =7.25,P ＜ 0.05).Conclusion Compared with KOH wet-mount microscopy,fluorescent staining can increase the detection rate,reduce missed diagnosis rate and shorten reading time.

Métodos de diagnóstico en micología / Diagnostic methods in mycology

Resumen El diagnóstico de las micosis constituye un problema clínico y terapéutico de interés actual debido al incremento y gravedad de las infecciones diseminadas. Por esta razón, en los últimos años se ha priorizado la búsqueda de nuevos métodos para el diagnóstico rápido y preciso. En la actualidad contamos con métodos tradicionales como KOH, cultivo, histopatología y serología, y, de forma más reciente, con métodos moleculares que permiten mayor rapidez y sensibilidad. En esta revisión se hace énfasis en la importancia de los métodos de diagnóstico micológico para la dermatología y la medicina general, ya que debido al desconocimiento en este campo, la preparación del paciente no es la adecuada y, por otro lado, la dificultad en la interpretación de los resultados lleva a confusión y tratamientos incorrectos.

Abstract Diagnosis of fungal infections is a clinical and therapeutic problem of current interest due to the increase and severity of disseminated infections. For this reason in recent years have been mainly seeking new methods for fast and accurate diagnosis. At present we have traditional methods like KOH, culture, histopathology and serology, and more recently molecular methods that allow greater speed and sensitivity in diagnosis. This review emphasizes the importance of mycological diagnosis methods for dermatology and general medicine, as due to ignorance in this field, patient preparation is not adequate and secondly the difficulty in interpreting results and treatment choice are misleading.

The use of Proteinase K to access genitalia morphology, vouchering and DNA extraction in minute wasps

ABSTRACT Genitalia are rich source of characters in insect taxonomy. Usually, they are examined after dissection and cleaning with potassium hydroxide (KOH), procedure that may damage both genital morphological structures and intracellular molecular contents. Enzymatic procedure with Proteinase K has been used to clean muscle off the genitalia while extract DNA, but its damage to the genital structures has not been evaluated. Herein, we qualitatively compare the use of KOH and Proteinase K to prepare genital structures in minute insects (Hymenoptera: Bethylidae). We show that Proteinase K is better to preserve the genital structure and provides quality DNA.

Comparison of five methods for the detection of fungal infections in vaginal secretions / 国际检验医学杂志

Objective To compare the similarities and differences of the five detection methods used in the detection of fungi in vaginal secretions,and find the most sensitive、the most specific、the fastest、the most cost effective and the simplest method used in the detection of fungi in vaginal secretions.Methods A total of 442 patients were selected from the Department of Gynecology of Shenzhen OCT Hospital from May 2016 to August 2016.The vaginal secretion of 442 specimens was detected by using the methods of fungi culture、saline and KOH suspension method,Gram stain,Wright''s stain and Vaginitis Multi Test Kit.In these five methods,Fungi culture were using as gold standard to evaluate the specificity,sensitivity,negative predictive value,positive predictive value and accuracy of the other four methods.Results Using the fungus culture method to detect 442 cases of vaginal secretion,we found the positive rate of mycotic infection was 34.8%(154/442).Compared with the fungi culture method,the Specificity of saline and KOH suspension method was 97.9%,the sensitivity was 64.9%,the negative predictive value was 83.9%,the positive predictive value was 94.3% and the accuracy was 86.4%;the Specificity of Gram stain was 96.5%,the Sensitivity was 83.1%,the negative predictive value was 91.4%,the positive predictive value was 92.7% and the accuracy was 91.8%;the Specificity of Vaginitis Multi Test Kit was 84.7%,the Sensitivity was 46.8%,the negative predictive value was 74.8%,the positive predictive value was 62.0% and the accuracy was 71.5%;the Specificity of Wright''s stain was 96.9%,the Sensitivity was 78.6%,the negative predictive value was 89.4%,the positive predictive value was 93.1% and the accuracy was 90.5%.Conclusion Gram stain could be the most sensitive and specific method in the four methods,with highest accuracy,and the the fastest,the most cost effective and the simplest method for the detection of fungi in vaginal secretions.The accuracy of detecting fungi in vaginal secretions could be improved by the combination of Gram stain method in clinical work.

A clinicomycological study of onychomycosis in a rural hospital in Central India.

Background: Onychomycosis; fungal infection of nails account for about half of the nail diseases. Common site of involvement is toenails. Various etiological agents including dermatophytes, yeasts and non-dermatophytic moulds (NDM) are responsible. It is difficult to treat onychomycosis as compare to other dermatophytic infections because of the inherent slow growth of the nail. Aim: To diagnose etiological agents of onychomycosis on KOH, Calcofluor white (CFW), KOH treated Nail Clipping with Periodic Acid Schiff (KONCPA) and SDA culture. Objectives: 1) To determine the fungal etiological agents of onychomycosis. 1) To correlate clinical parameters with the mycological findings. Methods: The study was carried out in department of Microbiology, MGIMS, Sewagram, Wardha. A total of 44 cases with signs of onychomycosis were enrolled in the study which were subjected for microscopic examination by 20% KOH, CFW and KONCPA. Mycological culture was done on Sabouraud’s dextrose agar (with and without antibiotics). Results: On analysis, the positivity by 20% KOH and CFW was 45.5%, 63.4% respectively while by KONCPA it was found to be 25%. In 38.6% fungal cultures revealed growth. At present, the etiological agents were dermatophytes (12.5%), especially Trichophyton rubrum, nondermatophytic isolates (75%) include Aspergillus spp., Penicillium species, Rhizopus and Candida spp. (8.3%). In our study toenails were affected in 84% and distolateral subungual onychomycosis (DLSO) was the commonest clinical presentation. Conclusion: Along with dermatophytes, NDM and yeasts are also important etiological agents of onychomycosis in our set up.

Prevalence and Etiological Agents of Onychomycosis.

Onychomycosis is a common nail infection caused by dermatophytes, yeast or other non-dermatophyte molds and have been known to be associated with significant physical and psychological morbidity. This study was performed to determine the prevalence and the etiologic agents of onychomycosis in patients attending RIMS, Imphal, Manipur for a period of two years (January 2010 to December 2012). Nail samples of 347 clinically suspected cases of onycomycosis attending dermatology OPD RIMS, Manipur were collected and were subjected to potassium hydroxide (KOH) mounts for direct microscopy and fungal culture on Sabouraud’s dextrose agar (SDA). The male female ratio of the participants was 1:1.81 (165 male and 302 female). Out of 467, 399 (85.44.%) were positive by culture and KOH mount. Young adults in the age group of 21-30 years were commonly affected. In female patients finger nail infection was more common than toe nail infection. Both toe and finger nails involvement were noticed in nine patients of which two were male and seven were female. The most frequently isolated fungus was dermatophytes, in 107 (40.38%) patients followed by Aspergillus spp in 101 (38.11%) and Candida spp in 17 (6.4%). This study demonstrated that dermatophytes, as well as moulds especially Aspergillus spp are commonly isolated from onychomycosis patients in our region.

Clinico-epidemiological study of pityriasis versicolor in a rural tertiary care hospital.

Background: The study was conducted to know the incidence of pityriasis versicolor in relation to age, sex, seasons and occupation in a rural set up. Methods: A total of 105 patients of pityriasis versicolor were clinically evaluated and confirmed mycologically at central lab in a tertiary care hospital in South India. The obtained data was recorded and analysed accordingly. Results: Male preponderance was observed in the study. Disease was most pronounced in younger age group (21-30 years) especially students. 61.9% of patients sought medical advice on cosmetic grounds while 38% had pruritis. Chest was the commonest site of affection followed by neck and back. Conclusion: Early identification of the yeast by simple laboratory techniques would help prevent recurrences, systemic complications and cosmetological problems which are high especially among the students and younger age groups.

Duration Required for Negative Conversion of KOH Mount in Toenail Onychomycosis According to the Clinical Factors and Treatment Regimens / 대한의진균학회지

BACKGROUND: Onychomycosis is one of the most common diseases of the nails accounting for up to 50% of all nail problems. There are many literatures on the cure rate and duration for complete cure according to the various factors. However, little information is available regarding duration required for negative conversion of KOH mount. OBJECTIVE: The purpose of this study was to evaluate the duration required for negative conversion of KOH mount in patients with toenail onychomycosis according to the clinical factors and treatment regimens. METHODS: Medical records of 325 patients with onychomycosis between May 2001 and February 2012 were reviewed retrospectively to obtain the following information: age, sex, presence of diabetes mellitus (DM), extent of nail involvement, therapeutic methods, and result of KOH mount at each timepoint. RESULTS: Extent of nail involvement, old age (> or =60) and DM were negatively associated with the duration required for conversion of KOH mount. Extent of nail involvement also negatively affected the rate of conversion of KOH mount at week 48. Positive KOH mount at week 28 was negatively associated with complete cure. CONCLUSIONS: Extent of nail involvement, old age, and DM can prolong the duration required for negative conversion of KOH mount. Also, positive KOH mount at week 28 had negative effect on the cure of onychomycosis.

Fungal spectrum in otomycosis at tertiary care hospital.

Introduction: Otomycosis is a superficial mycotic infection of the external auditory canal that is caused by opportunistic fungi. It is a common fungal infection of the ear that is worldwide in distribution. The infection usually characterized by inflammation, pruritus, trauma to external auditory canal, scaling and severe discomfort such as suppuration and pain. We have tried to clarify the different causative factors & predisposing factors for otomycosis in our environment. Material & Methods: In this study we were including 100 patients (53 female and 47 male) with presumptive diagnosis of otomycosis. We performed mycological analysis on swab or fungal ball from external auditory canal of 100 patients. The diagnosis was confirmed micro biologically by direct microscopy (10% KOH mount) and fungal culture methods. Results: Otomycosis was more common among females as compared to males & also common in age groups of 20-35 yrs. The most common fungal pathogens isolated were Aspergillus niger (75.82%) followed by Aspergillus fumigatus( 13.19 %), Candida albicans ( 7.69 %) & Aspergillus flavus ( 3.3 %). while bacterial co infection/ super infection was detected in 6 cases. Conclusion: In this study we observe that otomycosis is unilateral condition more common in young age housewives and farmers particularly during summer and rainy season.

Various Nail Disorders Misdiagnosed and Treated as Onychomycosis / 대한피부과학회지

BACKGROUND: Treatment of onychomycosis should be commenced after mycological confirmation due to the side effects and long duration of oral antifungal agents. However, many doctors treat patients with nail disorders that resemble onychomycosis without any mycological examination (ME) in clinical practice. OBJECTIVE: To investigate the various nail disorders misdiagnosed as onychomycosis and to evaluate the performing ratios of appropriate diagnostic procedures of onychomycosis. METHODS: The study was performed in a total of 48 patients who had a history of being misdiagnosed and treated for onychomycosis. We investigated the characteristics of each patient, the specialty of their previous doctor, and their previous treatment modality and duration. Whether or not they had previously received proper diagnostic procedures was also investigated. All of the data were collected by questionnaires, and their final diagnoses were also recorded. RESULTS: Among the 48 patients, 31 (31/48, 64.7%) were female, and the mean age of the patients was 45.5+/-15.7. Eighteen patients (18/48, 37.5%) were referred from dermatologists and 30 (30/48, 65.2%) from non-dermatologists. The results of the questionnaires revealed that only 10 of the dermatologists (10/18, 55.6%) performed ME before initiating treatment for onychomycosis, and none of the non-dermatologists did it. The duration of incorrect previous systemic anti-fungal treatment was also significantly longer when administered by non-dermatologists as compared with dermatologists (p=0.042), and over half of the patients (30/46, 65.2%) were treated with fluconazole. Onycholysis (22/48, 45.8%) and trachyonychia (18/48, 37.5%) were the most common final diagnoses, but various nail disorders such as pitting nail, median nail dystrophy, and subungual melanoma were also included. CONCLUSION: To avoid misdiagnosis and incorrect anti-fungal treatment, people with nail disorders that resemble onychomycosis should be recommended to visit dermatologists, and the doctors should perform appropriate mycological examinations in these patients.

The Isolation of Malassezia Yeasts in the Comedones of Acne Vulgaris / 대한의진균학회지

BACKGROUND: Both Malassezia yeast and Propionibacterium acnes form part of the normal flora of the human skin and hair follicles. The former is the etiological agent of Malassezia(Pityrosporum) folliculitis and the latter is one of the major factors in the pathogenesis of acne vulgaris. These two follicular diseases can coexist on a certain area of the skin, but there have been few reports about their coexistence in the literature. OBJECTIVE: The aim of this work was to investigate the distribution of Malassezia yeasts in the comedones of patients clinically diagnosed as acne vulgaris for elucidation of the coexistence of the two diseases, and for information on the predominance of the colonized Malassezia species and on relationship between certain species and Malassezia folliculitis. METHODS: The spore load in the comedonal plugs of 32 patients with acne vulgaris was graded using direct microscopy of KOH/Parker ink mounts. The comedonal specimens were cultured on Looming & Notman's media and the isolated Malassezia yeasts were identified to species level by their colony morphologies, microscopic morphologies and physiological characteristics. RESULTS: On direct microscopy, 8 of 32 patients (25%) showed a 4+ spore load, which is considered as a diagnostic grading index of Malassezia folliculitis. The predominant Malassezia(M.) species from 32 patients with acne vulgaris were M. restricta, M. globosa, M. furfur in descending order. Three strains of M. restricta, 4 strains of M, globosa, 2 strains of M. furfur and 1 strain of M. obtusa were isolated from the comedones of the 8 patients with 4+ spore load. CONCLUSION: This study shows that Malassezia folliculitis might coexist with acne vulgaris on the face, but there was no relationship between certain species and Malassezia folliculitis. The results suggest that antibiotic resistant acne vulgaris should be examined by direct microscopy of KOH/Parker ink mounts to confirm the coexistence of Malassezia folliculitis and acne vulgaris.

Avaliação da lise celular pelo hidróxido de potássio a 3 e coloração de Gram / Evaluation of 3 KOH celular lysis technic and Gram stain

Cinco métodos para coloração de Gram foram comparados. O melhor resultado foi obtido com o método descrito em Lennette et al., 1974. A técnica da lise pelo KOH 3% foi comparada com a coloração de Gram em 164 cepas bacterianas. A técnica apresentou resultados compatíveis com os obtidos no método de Gram para todas as bactérias analisadas.

Comparative Study for METHODSfor Diagnosing Onychomycosis / 대한피부과학회지

BACKGROUND: Onychomycosis is very common nail problem, so an in xpensive, quick and sensitive test is essential for screening nail specimens. Recently, there is a report of new method for diagnosing onychomycosis u.;ing KOH treated nail clippings which vri then crushed and finally stained with periodic acid-Schifft(PAS) stain (KONCPA). OBJECTIVE: We evaluate the susefulness of the new methods using KOH treated nail clippings and nail debris which were then crushed and finally stained with FS (KONPA) or chlorazol black E(KONBE), for the diagriosis of onychomycosis. METHODS: We compare different methods for diagnosing onychonycisis such as KOH stains, fungal cultures, histologic evaluation, SEM, KONCPA, KONBE, and KOHJPA. RESULTS: KONPA was proved to be more effective indentifying uiigal hyphae in comparison with conventional KOH nail scraping preparation, fungal culture, and FONBE. The positive rates of each method were 74%, 46%, 43%, and 63%, respectively. Also, KENPA proved to be more rapid and easy to perform in cotnparison to the histologic evaluation of tiail clippings and SEM. CONCLUSION: KONPA is a sensitive, quick, and readily available teled for use in clinical settings in cases that are highy siispected to be onychomycosis clinicaly, but show negative results using conventional methods