SPECTRUM OF SUPERFICIAL MYCOSIS IN A TERTIARY CARE HOSPITAL- A MATTER OF CONCERN

BACKGROUND : Superficial mycosis has been recognized as a common fungal condition worldwide, including India. It refers to fungal infection of skin and its appendages. AIM & OBJECTIVES : 1) To determine the frequency of fungal agents isolated from clinically suspected cases of superficial mycoses, in a tertiary health care hospital. 2) To access the risk factors associated with it. 3) To analyse the demographic profile associated with superficial mycosis. MATERIAL & METHODS: The prospective study was carried out, over a period of 12 months (January 2021 to December 2021), samples such as skin scarping, hair plugs and nail clipping obtained from patients were submitted to Department of Microbiology for the fungal etiology. Samples were subjected to direct microscopy by KOH mount and fungal culture as per standard convectional technique. RESULTS: A total of 63 clinically suspected cases of superficial mycosis were enrolled in the study. Among the isolates recovered the most common were Dermatophytes(41.26%) followed by Candida(14.28%), Aspergillus(6.3%), Mucor(3.1%) and remaining 23.80% were sterile. Among the Dermatophytes, T.rubrum 42.30% was the predominant pathogen followed by T.mentagrophytes 34.61%. The most common age group affected was 21-40 years of age with male predominance of male to female ratio of 1.5:1. Out of total of 48 samples of superficial mycosis 26 were positive by direct microscopy and 48 samples were positive both by microscopy and culture. Most of the positive cases were recovered from immunocompromised individuals suffering from diabetes (37.5%) followed by prolonged antibiotic therapy (25.02%), long steroid therapy (12.5%) and chemotherapeutic agents (10.41%). The cases were mainly seen in the months between April to July which correlates the infection with the humid season. CONCLUSION: The study pinpointed dermatophytes as the most common clinical pattern of superficial mycosis with a male predominance. Dermatophytic infection is one of the emerging fungal disease along with non dermatophytic molds, especially in immunocompromised individuals prompt treatment and management can herald the onset of ensuing complications, thereby limiting the morbidity and thus improving the quality of life.

Antifungal susceptibility testing for dermatophytes isolated from clinical samples by microbroth dilution method

Background: The cases of dermatophytoses have increased over the past few decades. Dermatophytoses affect the outer layers of skin, nails and hairs without tissue invasion. These infections are mostly not dangerous but, are important as public health problem particularly in the immunocompromised. The increased use of antifungal drugs for prolonged periods may lead to acquired antifungal resistance among previously susceptible strains. With this background present study was conducted to know the susceptibility pattern of dermatophytes.Methods: A total 35 isolates of dermatophytes isolated from clinically suspected cases of dermatophytoses were examined. Broth microdilution method M38-A2 approved protocol of CLSI (2008) for filamentous fungi was followed for determining the susceptibility of dermatophyte species to antifungal agents- itraconaole, fluconazole and ketoconazole.Results: Itraconazole minimum inhibitory concentrations (MIC) varied from 0.0156 to 1 µg/ml for all dermatophytes. T. rubrum species showed higher MIC range for Ketoconazole than T. mentagrophytes and T. tonsurans. Fluconazole had poor susceptibility for all dermatophytes by having higher MIC values.Conclusions: The MIC values observed in present study will help clinician to select an appropriate antifungal agent with minimal side effects. The data from present study can be useful as reference for future studies covering large no. of isolates and more drugs.

Evaluación in vitro de la actividad antimicrobiana de un propóleo chileno sobre muestras clínicas de exudados bucofaríngeos y cepas ATCC / In vitro evaluation of the antimicrobial activity of a Chilean propolis on clinical samples of oropharyngeal exudates

Propolis is a substance manufactured by Apis mellifera and has been widely used in folk medicine due to its high concentration of bioactive compounds. The purpose of the following study was to characterize and evaluate in vitro the antimicrobial properties of propolis on clinical samples and ATCC strains. The chemical characterization of propolis presents a concentration of total polyphenols of 247 ± 9 mg EAG g-1 MS, flavones and flavonols 75± 4 mg EQ g-1 MS, flavanonones and flavanonols 118 ± 11 EP g-1 MS. HPLC-DAD identified apigenin, galangin, phenethyl ester of caffeic acid and pinocembrin, in addition to 16 compounds by HPLC MS/MS. Chilean propolis is a natural antimicrobial, showing effectiveness in strains ATCC Staphylococcus aureus, Candida albicans, Trichophyton rubrum and clinical samples of Staphylococcus aureus unlike Escherichia coli. These results demonstrate the antimicrobial effectiveness of the synergy of compounds present in propolis against different human pathogens.

El propóleos es una substancia fabricada por Apis mellifera y ha sido utilizado ampliamente en la medicina popular debido a su alta concentración de compuestos bioactivos. El propósito del siguiente estudio fue caracterizar y evaluar in vitro las propiedades antimicrobianas del propóleos sobre muestras clínicas y cepas ATCC. La caracterización química de propóleos presenta una concentración de polifenoles totales de 247 ± 9 mg EAG g-1 de MS, flavonas y flavonoles 75 ± 4 mg EQ g-1 de MS, flavanononas y flavanonoles 118 ± 11 EP g-1 de MS. Mediante HPLC-DAD se identificó apigenina, galangina, fenetil éster del ácido cafeico y pinocembrina, además de 16 compuestos mediante HPLC MS/MS. El propóleos chileno es un antimicrobiano natural, observándose efectividad en cepas ATCC Staphylococcus aureus, Candida albicans, Trichophyton rubrum y muestras clínicas de Staphylococcus aureus a diferencia de Escherichia coli. Estos resultados demuestran la efectividad antimicrobiana de la sinergia de compuestos presentes en el propóleos ante diferentes patógenos humanos.

Prevalence of Dermatophytosis in patients in a tertiary care centre in and around Cuddalore district

Background: The common cause of skin infections are dermatophytes and opportunistic fungi. Aim of this study was to isolate and identify the fungal agents from clinical samples from patients with different mycoses. Superficial mycoses are among the most frequent forms of human infections, affecting more than 20–25% of the world’ population. They are predominantly caused by a group of closely related keratinophilic mycelia fungi (dermato-phytes) in the genera of Trichophyton, Microsporum, and Epidermophyton. These groups of fungi invade the stratum corneum of the skin or other keratinized tissues derived from the epidermis such as hair and nails. Aim and objectives: The present work aims at evaluation of rate of incidences of dermatophytic infections (group wise), and etiological agents with associated symptoms. In addition, we also determine to test the sensitivity of present diagnostic procedures. Materials and methods: Samples were collected from 100 patients. A portion of each sample was examined microscopically and the remaining portion of each sample was cultured onto plates of Sabouraud’s dextrose agar containing chloramphenicol with and without cycloheximide. Dermatophyte isolates were identified by studying macroscopic and microscopic characteristics of their colonies .Preliminary identification was done by conventional methods and confirmed by Polymerase Chain Reaction – Restriction Fragment length Polymorphism (PCR -RFLP) technique. Results: All over 62/100 specimens were KOH positive and 48/62 (82.7%) samples were culture positive. Of these, highest isolation rate was obtained in opportunistic mycoses such as candidiasis (29/29, 100%). Dermatophytes were isolated in 41/62 (66.3%) specimens and Trichophyton rubrum was the commonest isolate in skin samples (44/62) among the patients suffering from dermatophytosis. Phaeoannellomyces Wernicke was isolated in a patient suffering from tinea nigra. P. Gunasekaran. Prevalence of Dermatophytosis in patients in a tertiary care centre in and around Cuddalore district. IAIM, 2017; 4(8): 91-95. Page 92 The study signifies the importance of mycological examination in the diagnosis of various mycoses for their effective management. Conclusion: Dermatophytoses are distributed worldwide with increased incidence especially in a tropical country like India. Several factors such as age, sex, illiteracy, poor hygiene and social economy influence the dermatophyte infection. In the present study we have attempted to understand the epidemological status of the dermatophytes in a tertiary care centre in Cuddalore, Tamil Nadu, India. Tinea corporis was the predominant clinical site from which dermatophytes were isolated. T. Rubrum and T. Mentagrophytes have been the major etiological agents and that has been evinced by our study.

Comparative Study of In Vitro Inhibitory Effects of UVC, Terbinafine Hydrochloride 1% and Paeonia Natural Extracts on Trichophyton rubrum / 대한의진균학회지

BACKGROUND: Trichophyton(T.) rubrum is most common fungal pathogen that causes tinea pedis and onychomycosis. It recurrently infects human and usually persists for very long time, provoking public health concern. Due to the limitation in current treatment options, alternative therapies are desirable. We investigated the inhibitory effect of UVC, terbinafine hydrochloride 1% and paeonia natural extracts on T. rubrum in vitro. OBJECTIVE AND METHOD: Total 25 T. rubrum strains were cultured for 10 days on Mycosel agar plate; 5 strains of T. rubrum and 5 copies for each strain. They were divided into 5 groups: control, UVC irradiation, terbinafine spray, paeonia natural extracts spray, UVC and paeonia natural extracts sprays. The cultured media were irradiated for 1 hour daily for 3 weeks in the germicidal lamp emitting 253.7 nm (UVC), power of 2.875 mW/cm2 at 10 cm distance. Terbinafine and paeonia extracts was sprayed twice on the surface to fully cover the colony area. The median diameter of each colony were measured every other day for 3 weeks. The change of colony diameter and the growth rate were analyzed. RESULTS: The UVC had virtually no effect on restraining the growth of T. rubrum, similar with the growth of the control group. However, both the terbinafine spray and paeonia extracts slowed down the growth rate remarkably and showed a similar effect. CONCLUSION: We could only figure out the fungistatic effect, and not the fungicidal effect of paeonia extract and terbinafine hydrochloride in vitro. UVC irradiation setting in this study was totally ineffective. More studies are needed on more variable wavelength and the fluence of UVC irradiation. In addition, further verification on the mechanism and the effect of anti-fungal activity by paeonia extracts are needed.

Isolation, Identification and Prevelance of Dermatophytes in Tertairy Care Hospital in Gulbarga District.

Dermatophytosis is one of the most commonly encountered cutaneous fungal infections worldwide. The higher prevalence in tropical and subtropical areas is supposedly due to the hot and humid climatic condition. Other factors like personal hygiene and habits, prevalence of virulent species in the soil are also known to influence the infection. A high prevalence of dermatophytoses has been reported from India. Hence, this study was undertaken to identify, speciate and study the sensitivity pattern of dermatophytosis from clinical samples received at Basveshwar Teaching & General Hospital, Gulbarga. A total of two hundred clinically diagnosed randomly selected patients of dermatophytosis attending the out patient department of Dermatology and Venereology, Basveshwar Teaching & General Hospital & M.R. Medical College, Gulbarga were studied. Skin, nail and hair specimens taken from the patients were processed by potassium hydroxide preparation (KOH) for direct microscopy and culture. Dermatophytosis was more common in the age group of 21-30 years (36%) and in males (73%). Fungi was demonstrated in 130 cases by KOH and in 96 cases by culture; 78 cases were positive by both direct examination and culture. Tinea corporis was the commonest clinical type encountered (54.5%) followed by tinea cruris (25.5%). Tricophyton rubrum (46.87%) was the commonest aetiological agent in majority of clinical types followed by Trichophyton mentagrophytes (36.46%), Microsporum gypseum (4.16%), Trichophyton tonsurans (1.04%) , Epidemophyton floccosum (8.33%). Microsporum audouinii (2.08%) and Trichophyton violecium (1.04%). Dermatophyte infections are very common in our country where hot and humid climate along with poor hygienic conditions play an important role in the growth of these fungi. By and large, Trichophyton species forms the commonest etiological agent of dermatophytosis. Tinea rubrum was the commonest isolate in tinea corporis, tinea cruris and Onychomycosis. Microporium audouinii was isolated from 1 case of tinea capitis.

A Case of Concomitant Tinea Infection Diagnosed with Molecular Biologic Technique / 대한의진균학회지

Trichophyton(T.) rubrum is known to be the most common causative agent of tinea cruris, tinea corporis, and tinea pedis, and hygiene, nutritional status, and high temperature/humidity are known to play an important role in pathogenesis. Although there has been a report on a case of co-infection in which tinea pedis subsequently developing tinea cruris with the foot lesion being a reservoir of the later lesion, there is of yet no report which confirmed co-infection of two different lesions through identification of definite causative organisms. This case concerns a 32 year-old patient who presented with erythematous annular scaly patches on the groin and axilla. Although no organism was identified on mycosel culture, DNA base sequence and RFLP analysis of the scales revealed T. rubrum in both of the lesions. Considering that this is a rare case of tinea cruris accompanied by tinea corporis in the axilla and groin confirmed by molecular biological techniques, the authors present it with review of literature.

Strain Differentiation of Trichophyton rubrum Isolated from 2 Sites of the Same Patients by Analysis of Ribosomal DNA Nontranscribed Spacer / 대한의진균학회지

BACKGROUND: In Korea, Trichophyton (T.) rubrum is the most common dermatophyte and occupied more than 80% of all isolates. Although strains differentiation is essential for epidemiologic studies, differentiation of T. rubrum is difficult. OBJECTIVE: The aim of this study was determined if 2 strains of T. rubrum, obtained from 2 different infection sites of the same patient, are the identical. METHOD: Amplification of tandemly repetitive subelements TRS-1 and TRS-2 of ribosomal DNA nontranscribed spacer (rDNA NTS) was performed on 20 strains of T. rubrum. They were isolated from skin lesions of tinea cruris and tinea pedis or tinea unguium in 10 patients. RESULTS: Twenty strains were cultured on Sabouraud dextrose agar slant plate and all strains showed the portwine strain. The amplification of TRS-1 from 20 strains resulted in 15 strains (75%) with type 1 (434 bp), 2 strains (10%) with type 2 (634 bp), 2 strains (10%) with type 3 (834 bp) and 1 strain (5%) with type 6 (434 bp + 634 bp). Each strains from 4 of 10 (40%) patients had different types of TRS-1: one patients had type 2 (groin) and type 1 (sole), one patient had type 6 (groin) and type1 (sole), one patient had type 3 (groin) and type 2 (sole), and one patient had type 1 (groin) and type 3 (sole). The infected sites or disease duration did not have noticeable difference in type of TRS-1. The amplification of TRS-2 resulted in all 20 strains with type II (502 bp), since differentiation was not possible. CONCLUSION: The patient with different type of strains present on infected sections of the body, indicated the possibility of different fungal transmission routes. Specific amplification of subrepeat elements in rDNA NTS was simple and reproducible method for typing of T. rubrum and was useful for epidemiologic studies.

Four Cases of Majocchi's granuloma / 대한의진균학회지

Dermatophytes usually do not invade beyond the epidermis. However mechanical breakage of the skin resulting from scratching or trauma and immunocompromised state, such as diabetes mellitus, lymphoma, and long-term steroid use may allow penetration of the fungi into reticular dermis. Cutaneous granulomas produced by infection with superficial fungi are infrequently recognized. We report four cases of dermatophytic granuloma on the lower extremities. Histopathologic examinations of the skin lesions of four patients showed chronic granulomatous inflammation with fungal elements. Cultures of Sabouraud's media with excised tissue revealed Trichophyton(T.) rubrum in two patients and T. mentagrophytes in one patient. The patients were treated with oral administration of terbinafine or itraconazole for 2-4 weeks.

The Effect of Incubation Temperature on Susceptibility Test of Dermatophytes to Oral Antifungal Agents / 대한피부과학회지

BACKGROUND: thermotherapy has been shown to be effective in the treatment of some fungal infections. Dermatophyte are well grown at 25degrees C rather than 37degrees C or high temperature. OBJECTIVE: An vitro test was done to assess the complemental effect and optimal conditions of local heating on the susceptability of t. rubrum to systemic antifungal agents. METHODS: Microdilution susceptability test to ketoconazole and itraconazole was done using 96 well microplate. Eight strains of T. rubrum were isolated from patients withtinea pedis and were cultured at 25degrees C, 37degrees C and 42degrees C for 1, 8 or 24 hours per day. MIC were checked at 4th, 7th, 9th day after inoculution. RESULTS: The growth without antifungal agents at 37degrees C and 42degrees C were decreased by 805 and 50% of the growth at 25degrees C respectively. Seven day after inoculation was the proper time to check the MIC. MIC50 of ketoconazole was the lowest at 42degrees C for 24hours per day in value of 0.006microgram/ml, and 0.09microgram/ml at 37degrees C for 24hours per day, 0.37microgram/mlat 42degrees C, for 8hours per day and 37degrees C for 8hours per day. MIC at 42degrees C for 1 hours er day, 37degrees C for 1 hyours and 25degrees C for 24hours per ady MIC were the same in value of 0.05microgram/ml. MIC50 of itraconazole was the lowest at 42degrees C for 24hours per day in value of 0.006microgram/ml, 0.01microgram/ml at 37degrees C for 24hours per day, 0.02microgram/ml at 37degrees C for 8hours per day. MIC at 42degrees C for 8hours per day, 42degrees C for 1hours per day, 37degrees C for 24hours per day MIC were the same in value of 0.05microgram/ml. CONCLUSION: Incubation at 37degrees C for 24 hours per day or 42degrees C for 24 hours per day increased the susceptability of T. rubrum to ketoconazole and itraconazole.

A Case of White Superficial Onychomycosis / 대한피부과학회지

Clinical types of onychorpycosis, consist of distal subungual orychomycosis, white superficial onychomycosis, proximal subongual onychomycosis and candidal onychomycosis. White superficial onychomycosis aippears as white, sharply outlined areas on the surfaces of toenails. The fingernails are not affected. Trichophyton mentogrophytes, rarely Trichophyton rubrm, is the causa ive dermatophytes. The auther reports a case of white superficial onychomycosis of the fingernail caused by T. rubrm.