Dermatophytosis Patients Research Articles

Automated grading system for quantifying KOH microscopic images in dermatophytosis

Concerning the progression of dermatophytosis and its prognosis, quantification studies play a significant role. Present work aims to develop an automated grading system for quantifying fungal loads in KOH microscopic images of skin scrapings collected from dermatophytosis patients. Fungal filaments in the images were segmented using a U-Net model to obtain the pixel counts. In the absence of any threshold value for pixel counts to grade these images as low, moderate, or high, experts were assigned the task of manual grading. Grades and corresponding pixel counts were subjected to statistical procedures involving cumulative receiver operating characteristic curve analysis for developing an automated grading system. The model’s specificity, accuracy, precision, and sensitivity metrics crossed 92%, 86%, 82%, and 76%, respectively. ’Almost perfect agreement’ with Fleiss kappa of 0.847 was obtained between automated and manual gradings. This pixel count-based grading of KOH images offers a novel, cost-effective solution for quantifying fungal load.

Effect of leech therapy in dermatophytosis: A randomized standard controlled trial

Background and purposeIn Unani medicine leech therapy is used as analgesic and resolvent purposes. The saliva of leech contains various biological substances like Hirudin, hyaluronidase, vasodilators, anesthetics, antibacterial, fibrinases, collagenase etc. To compare the efficacy of Leech therapy with local antifungal clotrimazole (1 %) cream in dermatophytosis on long term basis. Material and methodsA randomized, standard controlled, open labeled, clinical study was conducted on 40 patients of dermatophytosis, aged 18–60 years, and were randomly assigned into two groups; 20 patients in test group and 20 in control group. Patients were diagnosed clinically and confirmed through KOH examination. Complete haemogram, bleeding and clotting times, random blood sugar, HIV I & II, and HBsAg tests were carried out before starting the therapy, while haemogram was repeated after completion of treatment too. In test group, five sittings of leech therapy were applied, with intervals of one week. The control group was given clotrimazole (1 %) cream for topical application twice daily, continuously for a period of 28 days. Post-treatment follow-ups were done on 35th day in person, and then over the phone on 45th, 60th, and 90th days. Data were collected using Signs and Symptoms Severity Score for study, and Visual Analogue Scale (VAS). After completion of trial, appropriate tests were applied for statistical analysis of data. ResultsThe study shows, in most of the parameters, statically significant therapeutic effect in both the groups (p<0.001). No patients of either group reported any adverse event throughout the trial. Conclusionit appeared that leech therapy can contribute significantly in treating dermatophytosis (Qūbā) without any adverse effect, and with prolonged efficacy compared to standard treatment.

Study the effect itraconazole and luliconazole in dermatophytosis patients with associated atopic dermatitis: A prospective observational

Study the effect itraconazole and luliconazole in dermatophytosis patients with associated atopic dermatitis: A prospective observational

Efficacy and Safety of Amorolfine Lotion 0.25% w/v Compared to Amorolfine Cream 0.25% w/w in Patients With Superficial Fungal Infections of the Skin: A Multi-center, Randomized, Open-Label, Active-Controlled, Non-inferiority Phase III Clinical Trial.

Dermatophytosis, a major cause of superficial fungal infections, requires topical and systemic antifungals. Amorolfine, a morpholine derivative, is a new topical antifungal available in cream and lotion formulations. To evaluate the efficacy and safety of amorolfine lotion 0.25% compared to amorolfine cream 0.25% in patients with dermatophytosis. A multi-center randomized, two-arm, active-controlled, parallel, non-inferiority phase III clinical trial involving 284 dermatophytosis patients was conducted, with the test arm using amorolfine lotion and the reference arm using amorolfine cream. The study drugs were applied once daily in the evening for four weeks and patients were followed up for another two weeks. The primary endpoint was clinical cure, while secondary endpoints included mycological cure, composite cure, global efficacy assessment, and post-treatment relapse. Safety and tolerability were assessed. Amongst the enrolled patients, 69.9% and 68.1% of patients had tinea corporis, while 30.1% and 31.9% had tinea cruris. The majority of patients in both groups (99.3% test and 97% reference) achieved a clinical cure at the end of treatment. Mycological cure was achieved by 98.6% and 96.3% respectively. A composite cure was achieved by 98.6% in the test arm versus 96.3% in the reference arm. A total of two AEs were reported in two (1.4%) patients in the test group and three AEs were reported in three (2.1%) patients in the reference group, all of the AEs were mild and resolved within three days without supportive medication. No severe adverse effects were reported in any of the study subjects. Amorolfine lotion 0.25% w/v showed a non-inferior clinical, mycological, and composite cure in dermatophytosis patients, was well-tolerated, and had a similar safety profile to amorolfine cream 0.25% w/w.

Hypothalamus-pituitary-adrenal axis (HPA axis) suppression with inappropriate use of steroids in recalcitrant dermatophytosis - A cross-sectional study.

Improper use of over-the-counter (OTC) steroid medication has been linked to recalcitrant dermatophytosis. There is proven evidence of HPA axis suppression by the use of long-term oral steroids. This study aims to determine the prevalence and pattern of inappropriate OTC steroid use and its effects on the hypothalamus-pituitary-adrenal (HPA) axis in adults with recalcitrant dermatophytosis. This cross-sectional study of 2 months was conducted in a hospital setting and included patients of recalcitrant dermatophytosis with a history of OTC steroid use. Clinico-demographic details and basal serum cortisol levels were recorded in all and analyzed. Of a total of 103 patients, 59.22% (n = 61/103) were males, and the mean duration of steroid abuse was 17.78 months. About 48.54% (n = 50/103), 3.88% (n = 4/103), and 47.57% (n = 49/103) patients reported the use of topical steroids, oral steroids, and both oral and topical steroids, respectively. Among all the topical steroid users (n = 99), clobetasol propionate 48.48% (n = 48/99), while among oral steroid users (n = 53), prednisolone 45.28% (n = 24/53) were the most commonly used agents, respectively. The morning serum cortisol levels (8-9 AM) were found to be decreased in 42.7% (n = 44/103), with a mean value of 44.28 ± 17.34 μg/dL. Improper OTC steroid use in recalcitrant dermatophytosis leads to HPA axis suppression. This highlights the need for intervention from apex health officials.

Immunohistochemical analysis of chronic and recurrent dermatophytosis.

Dermatophytosis has assumed epidemic proportions with rising resistance, recalcitrance and recurrence, especially in tropical regions. While various factors contribute to high prevalence worldwide, yet little is known about the interactions between host defence mechanisms and dermatophytes, particularly in chronic and recalcitrant dermatophytosis. We aimed to compare the population of various immune cells in specimens of chronic recurrent dermatophytosis and those with acute superficial dermatophytosis. We investigated the density of various immune cells-Langerhans cells (CD1a+), macrophages (CD68+), dermal dendrocytes (Factor XIIIa+) in the skin of chronic dermatophytosis patients and those with successfully resolved lesions (controls). Langerhans cells were significantly decreased in the epidermis of patients, both in affected and unaffected areas in comparison with controls. In the dermis, however, no differences in the density of immune cells (macrophages and fibroblasts) were observed. The limited sample size and immune cells evaluated could be expanded further in future research. These results indicate that the decreased number of Langerhans cells could be a potential risk factor for the development of chronic and recurrent dermatophytosis.

Diagnosis of a recurred lesion in dermatophytosis patients after 2 weeks of antifungal therapy: A prospective observational study.

A prospective observational study was conducted on 157 cases of dermatophytosis with positive fungal test results. A fixed dose of 100 mg of oral itraconazole once daily was administered to all patients for 2 weeks. At the end of 2 weeks, patients were assessed for clinical cure and recurrence. Recurred cases were assessed for mycological profile using a fungal test (potassium hydroxide mount and/or fungal culture) for identifying fungal infection. Only eight (5.36%) patients showed clinical cure, and 141 (94.63%) patients developed recurrence after therapy. Of the 141 cases with recurrence, only 47 (33.33%) patients were positive for fungus. Eight (5.09%) patients were lost to follow-up. Frequently encountered risk factors in the study were topical steroid use, disease in family, associated atopic dermatitis and contact with pets. This is the first study that described the clinical diagnosis and mycological profile of the various lesions recurring at the previous tinea infection site in patients with dermatophytosis. Such patients presented not only with recurrent lesions of fungal infection but also developed various dermatoses unrelated to fungal infection at the sites of previous tinea infection. Various factors, which could have resulted in the observed changes, are reinfection by dermatophytes at the sites of previous tinea infection, inadequate antifungal therapy or antifungal resistance; or due to the effects of various topical steroid formulations used by the patients, such as anti-inflammatory or immunosuppressive effects or shift in immunity. Hence, diagnosis of the recurrent lesion at the site of previous dermatophytosis must be individualized and should be based on 1) duration of antifungal therapy received, 2) associated risk factors, 3) response to antifungal therapy, 4) evolution of the recurrent lesion, and/or 5) fungal tests.

A clinical prevalence of dermatophytic mycoses with an assessment of its clinical manifestations in a tertiary care hospital at Salem, South India.

Dermatophytosis is very common among all age groups throughout the world. The incidence of the same is increasing on a steady basis. Estimating the clinical prevalence of dermatophytes mycoses among the patients visiting the outpatient unit and assessing its distinct manifestations. A prospective observational study was conducted with the patients attending the Skin and STD outpatient unit of a tertiary care teaching hospital in Salem. A total of 3068 outpatients attended the department, of which 420 patients were diagnosed with dermatophytic mycoses and were taken for investigating the prevalence. A total of 420 dermatophytosis patients were included giving a percentage prevalence of 13.69%. There were more female patients (n = 213, 50.71%) than males (n = 207, 49.29%). The most common afflicted age group was 31-40 years (n = 99, 50.71%). Most of the patients had an atypical lesion called tinea incognita (n = 265, 63.09%) where there was no typical classic appearance of dermatophytic infections. The most prevalent clinical manifestation was tinea corporis (n = 73, 17.38%) followed by tinea cruris (n = 69, 16.43%). There were more newly diagnosed dermatophytosis cases (n = 326) than the previously diagnosed cases (n = 94). This study concludes that dermatophytic mycoses were more prevalent among females than males and among the age groups of 31-40 years. The most common clinical presentation was tinea incognita followed by tinea corporis.

Transcriptional analysis of Metalloprotease, Metallocarboxypeptidase, Isocitrate lyase, Citrate synthase, Malate synthase and Dipeptidylpeptidase V of Trichophyton rubrum isolates from dermatophytosis patients.

Introduction: Dermatophytes are pathogenic fungi that cause cutaneousinfection of human and animal and grow exclusively on the stratumcorneum, nail and hair. Methods: The present study was conducted on 160 samples from clinicallydiagnosed onycomycosis patients, further subjected to culture from nailsamples of patients attending dermatology OPD of a tertiary care hospital,Delhi from January 2016 to December 2018 Results: In a soy protein culture, a substantial proteolytic activity was seen, which was secreted by Trichophyton rubrum, Trichophyton mentagrophytes and Microsporum canis. This proteolytic activity was 55–75% inhibited by O-phenanthroline, which authenticate that metalloproteases were secreted by all three species. A consensus probe was constructed on previously characterized genes, which encodes metalloproteases (MEP) of the M36 fungalysin family in Aspergillus fumigatus, Aspergillus oryzae and M. canis. From genomic libraries of T. rubrum, T. mentagrophytes and M. canis, T. rubrum &amp; T. Mentagrophytesa five-member MEP family was isolated and also secretes MCP A &amp; MCPB, MEP 3, MEP4, IL, CS, MS AND DPP V of the M14 family accordingto the MEROPS proteolytic enzyme database. Conclusion: This study shows that Metalloprotease, Metallocarboxypeptidase, Isocitrate Lyase, Citrate Synthase, Malate Synthase and Dipeptidylpeptidase V are expressed in Trichophyton Rubrum Isolates from Dermatophytosis Patients.

EVALUATION OF IMMUNOLOGICAL MARKER INTERLEUKIN-17 AMONG PATIENTS WITH DERMATOPHYTOSIS INFECTION

This study was conducted to evaluates the immunological parameter IL-17) associated with development of dermatophyte infection. The study was carried out in the period beginning of December 2022 to end of June 2023.This study has been done at the Al-Hilla Teaching Hospital, Dermatology Unit, Babylon Government; ages range between 1 to 50 years in male and female. During the study a total 90 clinical specimens (Skin scraping and 5ml of blood) were obtained from all participants (45 dermatophytosis cases and 45 controls). Blood samples (serum introduced for evaluation of concentration level of IL-17 by ELISA. The ELISA was used to measure two types of cytokines (IL-17 and TNF-), and it was observed a significant increase ( P 0.0001 ) of serum IL-17 in dermatophyte infected patients (35.015 0.837 pg/ml) when compared with control subjects (6.481 0.848 pg/ml). The levels of IL-17 concentration in patients Infected with Dermatophyte Infection according to age groups appear non significant difference between age groups (P.value 0.8780) and the highest concentration level of IL-17 appears with age group (31-40 years) and (41-50 years) and were (37.191 2.806 Pg/ml) and (37.006 0.934 Pg/ml) respectively while the lowest concentration level of IL-17 with ( 10 years) was (33.784 4.756 Pg/ml). while in control group, appear non significant difference between age groups (P.value 00.8991) and the highest concentration level of IL-17 appears with age group (51-60 years) and ( 10 years) and were (8.636 2.300 Pg/ml) and (8.281 1.352 Pg/ml) respectively while the lowest concentration level of IL-17 with (31-40 years) was (5.869 1.002 Pg/ml). In depending on Gender, in dermatophyte infected patients group, the results have shown the high concentration level of IL-17 appear in male was (35.950 ± 0.916 Pg/ml) other than female (33.255 1.643 Pg/ml) with non significant difference (P.value 0.1266) also in control group , the results have shown the high concentration level of IL-17 appears in male was (7.383 0.911 Pg/ml) and the low concentration level appears in female and was (6.544 0.957 Pg/ml) , and this height also non significance (P.value 0.5381). The concentration level of IL-17 according to duration of infection in dermatophytosis patients, appear significant difference between groups (P.value 0.0001) and the highest concentration level of IL-17 appears with groups ( 2 years) and was (74.083 9.807 Pg/ml), while the lowest concentration level of IL-17 with (6 month) and was (33.892 2.890 Pg/ml).

Incidence of Dermatophytosis Based on Age and Gender at The Regional General Hospital in Gianyar District Hospitals

Background: Dermatophytosis is a superficial fungal infection caused by filamentous fungi that attack the keratinized tissue of the skin, nails, and hair. The clinical manifestations of dermatophytosis, in addition to depending on the source host factors, such as age, sex, and race, are significant epidemiological factors. However, the relationship of infection susceptibility remains unclear. Purposes: This study aimed to determine the relationship between age and sex and dermatophytosis incidence in Gianyar Regency. Methods: This study was observational with a cross-sectional design using 200 samples divided into 100 dermatophytosis patients and 100 non-dermatophytosis patients. Statistical analysis of the data used was a chi-square test with 95%CI. Results: The majority of patients with dermatophyte infections were &gt;19 years old (74%), female (57%), and tinea corporis (36%). The chi-square test showed that there was a significant relationship between age and sex with the incidence of dermatophytosis (p = 0.025; OR = 1.978; 95% CI = 1.087-3.599 and p = 0.003; OR = 2.357; 95% CI = 1.334-4.162). Conclusion: It can be concluded that older age and female tend to suffer dermatophytosis.

KAP survey in patients of dermatophytosis in a tertiary care hospital: a cross-sectional study

Background: Dermatophytes are a group of fungi with the capacity to invade keratinized tissue. Along with etiological factors, the clinical and epidemiological patterns has shown changing trend, leading to epidemic state of dermatophytosis. This could be attributed to inadequate knowledge and substandard practices followed by patients of dermatophytosis. Methods: A cross-sectional observational survey of patients with dermatophytosis who visited the outpatient department of dermatology at a tertiary healthcare facility over a period of two months to assess knowledge, attitude and practices regarding dermatophytosis management was performed. Results: A total of two hundred and twenty-seven (227) patients were recruited in during 2-month study period. The male:female ratio was 0.82:1. Fifty two percent respondents were from a rural background. Fifteen percent of respondents had cattles as pets. Majority of the respondents preferred wearing loose clothes (79%), 89% wore cotton as the material of cloth and 35% preferred wearing closed footwear. Seventy two percent were unaware of the infective nature of disease and 84% of the treatment of this condition 11.45% were hesitant to take oral treatment for the same. More than half of the patients gave a history of self-medication, and 27% of the patients gave a history of receiving steroid injections. Conclusions: This study concluded that there is an extensive knowledge gap in this subset of the population, which lead to faulty and substandard practices, such as self-treating, steroid abuse and visiting quacks before considering registered dermatologists for treatment purposes.

RISK OF DERMATOPHYTOSIS IN ADULT POPULATION WITH DIABETES MELLITUS TYPE 2 IN WESTERN UTTAR PRADESH: A CASE CONTROL STUDY

INTRODUCTION : Dermatophytosis is a common supercial fungal infection which has taken epidemic proportions in many pockets of India. Several risk factors have been implicated including environmental factors, treatment, host factor, Diabetes mellitus etc. for chronic and recurrent infections. The association of Diabetes and dermatophytosis is still ambiguous with contrasting data over years of research. Thus, there is need for more data collection and analysis to establish a clear association. · To assess OBJECTIVES: recurrent and atypical infections in diabetic and non-diabetic individuals. This w METHODOLOGY : as a case control study with 90 patients 45 diabetic and 45 non-diabetics having dermatophytosis attending skin OPD at Teerthankar Mahaveer Medical College, Moradabad were included in the study over a period of 12 months from July 2021 to June 2022. Atypical difcult to trea CONCLUSION : t infections were observed more in diabetic group. Therefore, these patients should undergo diabetic status analysis. Patients of dermatophytosis with DM-II should maintain strict glycaemic control. Treating physicians should be aware of all these aspects and comprehensively treat with pharmacological therapy and counselling

“A CROSS-SECTIONAL STUDY OF CLINICO-MYCOLOGICAL PROFILE OF DERMATOPHYTES AND ANTIFUNGAL SUSCEPTIBILITY AMONG PATIENTS ATTENDING DERMATOLOGY OPD AT A TERTIARY CARE HOSPITAL IN TELANGANA”

Background: Dermatophytosis is the most common supercial fungal infection worldwide, caused by dermatophytes that cause infections of the skin, hair, and nails due to their ability to invade keratin. Belong to three closely related genera: Trichophyton, Microsporum, and Epidermophyton. It is common in the tropics &amp; subtropical regions and in areas of high humidity. Recent years have seen an alarming increase in chronic, recurrent, and recalcitrant dermatophytosis in India. Despite the availability of a wide range of antifungals, treatment failure is often observed. This is often attributed to the probable emergence of drug-resistant strains. To identify t Objectives: he dermatophyte species causing Dermatophytosis and to determine antifungal susceptibility patterns to Fluconazole and Griseofulvin by Broth microdilution. A cross-sectional study was conducte Method: d over a period of 18 months duration i.e., from June 2021 to November 2022. Skin scrapings, hair plucking, and nail clippings were collected from 200 clinically diagnosed cases of dermatophytosis who attended the outpatient department of Dermatology (DVL) and samples were processed at the upgraded Department of Microbiology at Osmania General Hospital, Afzalgunj, Hyderabad. A total of 2 Results: 00 clinically diagnosed dermatophytosis patients were included in our study following inclusion and exclusion criteria. Their demographic details and clinical examination of cutaneous lesions were recorded. In our study, most patients affected were males 57% and the most common age group was 21-30 years seen in 25.5% of patients. Tinea corporis (40%) was the most common dermatophyte lesion followed by Tinea cruris (19.5%) seen in our study. KOH positivity was seen in 42% and culture was positive in 69.5% of cases. Dermatophytes isolated are 42% and non-dermatophytes isolated are 27.5% of cases in our study. The most common dermatophyte isolated in culture was Trichophyton rubrum (45%), followed by Trichophyton mentagrophytes 28 (33.3%). Among isolates of dermatophytes, Fluconazole is sensitive to 33.3% of isolates, whereas Griseofulvin is sensitive to 52.3% of isolates. Therapeutic failure is alarmingly common in t Conclusion: he current scenario of dermatophytosis in India. Failure is probably seen with all common isolates, younger patients, high contagious nature. This study will help to standardize care, provide guidance on management, and assist in clinical decision-making for healthcare professionals

Human-pet Relationship, Pet Abandonment, and Clinical Correlation for Patients Infected with Dermatophytosis of the Glabrous Skin

Objective: The study on human-pet relationship and pet abandonment among dermatophytosis patients is limited. This study aims to review these correlations. Materials &amp; Methods: A two-year retrospective cross-sectional study was conducted. Case record forms were reviewed for clinical manifestations, fungal identification, human-pet relationships, and changes in the relationships after dermatophytosis diagnosis. Results: A total of 230 dermatophytosis patients from the Dermatology outpatient clinic, Siriraj Hospital, were included. The mean age was 41.9 ± 19.1 years and 51.3% were female. Among 170 cases with positive fungal culture, zoophilic dermatophytosis from M. canis infection was identified in 15.9% which was predominately found in females and manifested as shorter duration of onset, and higher involvement on exposed areas when compared to anthropophilic dermatophytosis. Most (71%%) of patients with M. canis infection classified themselves as pet-lovers. The relationship with pets had changed after the dermatophytosis diagnosis in 41% of them which was statistically different from 8.8% in non-pet lovers (P = 0.001). The overall pet abandonment rate was 26.6%. The abandonment rate was 40.9% among non-pet lovers, while 30.6% was reported among pet lovers. Conclusion: Zoophilic M. canis infection was associated with rapid onset and on predominant-exposed areas. Some pets could be asymptomatic, so identification of the reservoirs of dermatophytosis is important in the treatment process and helps prevent future recurrence. Paying attention to human-pet relationships and pet abandonment is critical. Knowledge about dermatophytosis transmission, and appropriate pet management should be advised to decrease abandonment.

A Clinical-Mycological Study of Dermatophytosis in Western India with Focus on Antifungal Drug Resistance as a Factor in Recalcitrance.

Dermatophytosis have assumed epidemic proportions in India. Antifungal drug resistance solely cannot explain disease magnitude and changing epidemiology. Aim of this study was to analyse clinical-mycological aspects of dermatophytosis, and estimate contribution of drug resistance in clinical recalcitrance. This single-centre observational, cross-sectional, descriptive study was done in tertiary centre of western India after ethical approval, enrolling dermatophytosis patients of all ages and sex. After history and examination, KOH mount and culture in modified SDA medium was done. Culture positive isolates were subjected to E-strip antifungal susceptibility method to test MIC for Terbinafine, Itraconazole, Fluconazole and Griseofulvin. Total 300 patients were included, with mean age of 33.83±27.5 years and male-to-female ratio of 1.22:1; tinea corporis et cruris being commonest, 39.33% (n=118). Only 11.67% (n=35) were treatment naïve, having classical annular morphology. History of topical steroid abuse was found in 81.67% (n=245), with pseudoimbricate lesions in 70.61% (n=173). 86.67% (n=260) had KOH positivity while 83.33% (n=250) had culture positivity: Trichophyton mentagrophytes 45.6% (n=114), followed by Trichophyton rubrum in 34.4% (n=86). A total of 265 patients fit into definition of recalcitrance, from which 12.45%, i.e., 33 isolates showed in-vitro fluconazole resistance. 14.33% (n=43) cases were chronic, 37% (n=111) persistent, 46% (n=138) recurrent while 17% (n=51) had relapse in their disease course. Steroid abuse was the commonest denominator. Role of antifungal resistance in recalcitrant dermatophytosis remains debatable. Stopping steroid abuse, which is often the commonest culprit, with adherence to standard antifungal therapy remains the paradigm in management.

Study of pattern of superficial dermatophytic infection in paediatric patients in tertiary health care centre

Background: It is difficult to ascertain reliably the overall incidence and prevalence of the various cutaneous infections in different parts of the world because studies of one region of the country may not be a true representation of the overall disease pattern of that country; furthermore, incidence and prevalence figures may only be representative of the population sampling, which may have associated risk factors for infection. Most of the studies done in this field are dealt with adults or with a specific clinical form of dermatophytosis. Only few surveys were performed in pediatric age group.Methods: It is an observational type of study. The present study was conducted in Department of Dermatology, Rajshree Medical &amp; Research Institute, Bareilly, from 1st march 2021 to 28th February 2022.Results: Majority of the study population belonged to 11-15 years (48.7%) followed by 15-18 years (24.1%). The most commonly affected sites were Groin (47.2%) followed by trunk (29.9%), buttocks (29.1%), and upper limbs (24.0%). The predominant lesion type was Plaque (43.5%). The most common lesion feature found in most of the study group was discoloration (94.1%). The most common diagnosis was T. corporis (29.1%) followed by T. corporis + T. cruris (28.3%), T. cruris (22.4%).Conclusions: Total 237 patients of superficial dermatophytosis were included in the study, out of which mean age was 9.16±4.241 with female predominance (52.3%). Common presenting complaints were itching (95.3%), affected site was groin (47.2%) and annular plaque was predominant type of lesion. The most common made in our study was T. Cruris (47.2%).

P115 Reversed-phase high-performance liquid chromatography as rapid and simple method for quantifications of terbinafine drug in human serum from dermatophytosis cases

Poster session 1, September 21, 2022, 12:30 PM - 1:30 PM ObjectivesThe alarming situation of treatment failure cases in dermatophytosis and resistance to the first-line drug (terbinafine) is a worrisome condition for the management of tinea cases. However, studies also reported non-responders to terbinafine treatment even when the isolates are susceptible to this drug in vitro. Thus, evaluating the pharmacokinetic profile of terbinafine might help better manage dermatophytosis. This study was conducted to standardize and validate a rapid and simple reversed-phase high-performance liquid chromatography (HPLC)-based protocol for terbinafine in serum/plasma of dermatophytosis cases.MethodsHPLC analysis was standardized for terbinafine drug on an Agilent 1290 infinity system (Agilent Technologies InC., USA). Chromatographic parameters including mobile phase [acetonitrile (A), methanol (M), and water (W)], flow rate (0.7-1.5 ml/min), injection volume (20 μl), and various wavelengths ranging from 220 to 265 nm under isocratic conditions were assessed and optimized. The mobile phase consisted of a filtered and degassed mix of A: W and M: W with various ratios of 85:15, 60:40, 50:50, and M-100%, respectively. Quality control samples were prepared in drug-free serum by spiking with the terbinafine at 0.0312-100 μg/mL concentrations. An equal volume of serum and acetonitrile (A) were mixed. The mixture was vigorously vortexed for 30 s, followed by high-speed centrifugation at 13 000 rpm at 40°C for 10 min. The supernatant was transferred into the chromatographic vials and placed in the autosampler of HPLC for injection. The standardized method was tested in 6 dermatophytosis patients’ serum/plasma samples collected at 3-time points (first, second, and third week of start of antifungal).ResultsLinearity of calibration standard for terbinafine was optimized at 250C at a flow rate of 1.0 ml/min, injection volume 20 μl, 8 minutes run time with the standardized wavelength at 245 nm under isocratic conditions. The best suitable graph was determined by plotting the area under the curve (AUC) and peak height separately against the drug concentrations measured by reversed-phase- HPLC for terbinafine drugs (Fig. 1a and b). The standardized mobile phase consisted of filtered and degassed Methanol (100, v/v). The chromatographic separation was achieved on an Agilent C18 column, and 4.3 ± 1 time represents the peak for terbinafine drug. Based on the standardized protocol, six tinea cases were included for validation, and the therapeutic range achieved for terbinafine in clinical samples was 0.6 to 1.13 μg/ml.ConclusionsThe standardization of HPLC method was successfully applied to quantify terbinafine in spiked samples with terbinafine drug and showed no observable interferences at the standardized parameter. Further evaluation with larger number of samples is warranted.

Nannizzia incurvata in Hue city - Viet Nam: Molecular identification and antifungal susceptibility testing.

Nannizzia incurvata, a species belonging to the Nannizzia gypsea complex, is considered a neglected pathogen. To detected N. incurvata isolates from dermatophytosis patients in Hue city - Viet Nam, and test the antifungal susceptibility of this species. Moreover, fungal capability to produce hydrolytic enzymes was evaluated. Patients' samples were collected and cultured on Sabouraud-chloramphenicol-cycloheximide medium. Dermatophytes isolates were initially macroscopically and microscopically identified. ITS PCR-RFLP and ITS rDNA sequences were performed to determine and confirm species. An ITS Neighbor-Joining phylogenetic tree evaluated the genetic relationship among isolates. Fungal hydrolytic enzymes were examined, including lipase, phospholipase and protease. Antifungal susceptibility testing was carried out by the disk diffusion method. MICs of itraconazole, voriconazole, and terbinafine against these isolates were determined by the broth microdilution method. Twelve isolates of N. gypsea complex were preliminary morphologically identified. PCR-RFLP and ITS-rDNA sequencing identified and confirmed dermatophytes as N. incurvata strains, respectively. An evident polymorphism among isolates was highlighted in the phylogenetic tree. All isolates showed the activity of lipase, phospholipase, and protease production. Overall, all N. incurvata isolates were susceptible to itraconazole, voriconazole, clotrimazole, miconazole, and terbinafine. Few isolates were susceptible to griseofulvin, and none of them were susceptible to fluconazole. There was a presence of polyclonal N. incurvata isolates in dermatophytosis patients from Hue city, identified by PCR-RLFP and confirmed by ITS sequencing. We confirmed PCR-RLFP as a reliable technique to identify this species. Azole and terbinafine are the optimal choices for N. incurvata treatment except for fluconazole.

ENZYMES CHARACTERISTICS OF TRICHOPHYTON SPECIES ISOLATED FROM PATIENTS ATTENDING HUE UNIVERSITY OF MEDICINE AND PHARMACY HOSPITAL

Abstract: Background: Trichophyton is the common genus in dermatophytes fungi in the world. These fungi secreted virulence enzymes that help them take nutrition from the host. Objectives: 1. To determinate enzymes secretion ability of Trichophyton species isolated from patients, including lipase, phospholipase, and protease. 2. To evaluate the correlation between enzyme secretion ability and dermatomycoses clinical forms. Material and methods: Trichophyton species were isolated from the cultivation samples of dermatophytosis patients. Enzyme production experiments were checked by cultivating fungi into suitable solid agar media. Results: The prevalence of Trichophyton spp. producing protease, lipase, and phospholipase were 96.1%%, 86.2%, and 21.6%, respectively. There were 78.4% of fungal strains that produced both lipase and protease, and 17.6% of fungal strains produced three enzymes. The ability to produce protease of T. rubrum was higher than T. mentagrophytes, and there was no statistically significant difference between lipase and phospholipase activities of these two fungal species. There was no difference between the fungal enzymatic activities and dermatophytosis clinical forms. Conclusion: Trichophyton species isolated from patients attending Hue University of Medicine and Pharmacy Hospital produced more protease and lipase than phospholipase. T. rubrum strains showed higher ability protease produce than T. mentagrophytes. Key words: dermatophytes, Trichophyton spp., fungal enzyme