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 Table of Contents  
ORIGINAL ARTICLES
Year : 2022  |  Volume : 9  |  Issue : 2  |  Page : 54-58

A clinico-mycological study of dermatophytoses and their in-vitro sensitivity to antifungal drugs


1 Department of Dermatology, Venereology, and Leprosy, Vydehi Institute of Medical Sciences and Research Centre, Whitefield, Bangalore 560066, Karnataka, India
2 Department of Microbiology, Vydehi Institute of Medical Sciences and Research Centre, Whitefield, Bangalore 560066, Karnataka, India

Date of Submission05-Jun-2021
Date of Decision25-May-2022
Date of Acceptance05-Jul-2021
Date of Web Publication27-Oct-2022

Correspondence Address:
Shishira R Jartarkar
Department of Dermatology, Venereology, and Leprosy, No. 272, 8th Cross, 29th Main, HSR Layout, 1ST Sector, Bangalore 560102, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijdpdd.ijdpdd_41_21

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  Abstract 

Background: There is an increase in the frequency of resistant and recurrent dermatophyte infections in India. We undertook a study to isolate the organisms in different clinical presentations and to study their sensitivity to clotrimazole, miconazole, fluconazole, and griseofulvin. Aims: The aim of this article was to study the clinico-mycological pattern of dermatophytes and their in-vitro sensitivity to commonly used antifungals. Objectives: The objectives were as follows: (1) to study the clinico-mycological patterns of dermatophytosis and (2) to study the in-vitro sensitivity patterns of culture isolates to clotrimazole, miconazole, fluconazole, and griseofulvin. Materials and Methods: A total number of 48 patients were included in the study. The study period was from August 2019 to February 2021. Skin scrapings and nail clippings were inoculated in Sabouraud dextrose agar. Isolates were identified based on gross colony characteristics and microscopic morphology of their micro- and macroconidia and accessory structures. The isolates were then transferred to sterile distilled water in vials and stored. The clinical isolates stored in the stock solution were subcultured on to Potato dextrose agar. Then, the antifungal disks and Griseofulvin E strip were applied on to the plates. The zones of inhibition around the disks were measured and recorded. In the case of E strip, the minimum inhibitory concentration value is read from the scale in terms of µg/mL, where the ellipse edge intersects the strip. Results: The total number of cases included were 48. Commonest clinical presentation was co-occurrence of Tinea cruris and corporis (33%). Trichophyton mentagrophytes was isolated in 56% of cases. Isolates showed highest sensitivity to clotrimazole (87.5%) followed by miconazole (60.4%). Only 8.3% were sensitive to fluconazole and 37.5% were sensitive to griseofulvin. Conclusion: T. mentagrophytes is the causative organism in significant numbers of dermatophytic infections. Though broth dilution is the standard method for sensitivity assay of dermatophytes, disk diffusion method could become a more simple alternative.

Keywords: Clotrimazole, disk diffusion, fluconazole, griseofulvin, miconazole, Trichophyton mentagrophytes


How to cite this article:
Kadnur M, Jartarkar SR, Narayanaswamy G, Manoj Kumar AS, Arora S, Balakrishnan T. A clinico-mycological study of dermatophytoses and their in-vitro sensitivity to antifungal drugs. Indian J Dermatopathol Diagn Dermatol 2022;9:54-8

How to cite this URL:
Kadnur M, Jartarkar SR, Narayanaswamy G, Manoj Kumar AS, Arora S, Balakrishnan T. A clinico-mycological study of dermatophytoses and their in-vitro sensitivity to antifungal drugs. Indian J Dermatopathol Diagn Dermatol [serial online] 2022 [cited 2022 Dec 9];9:54-8. Available from: https://www.ijdpdd.com/text.asp?2022/9/2/54/359770




  Introduction Top


Dermatophytoses is one of the most common dermatoses. Though not a serious disease, it significantly affects quality of life.[1],[2] A simple and easily treatable condition a few years ago has become one of the most commonly discussed topics for its recent increase in prevalence, recurrence, and resistance to treatment. These changes in the behavior of the disease could be due to the host factors, environmental factors, change in the species, and resistance to antifungal agents. In our study, we attempted to make a correlation of clinical features with culture pattern and also to identify the sensitivity pattern of the culture isolates to clotrimazole, miconazole, fluconazole, and itraconazole.


  Materials and Methods Top


All clinically suspected cases of dermatophytoses visiting dermatology OPD in our institute whose KOH mount was positive were included in the study. Those on immunosuppressants and topical and systemic antifungals in the last 1 month were excluded. A total number of 48 patients were included.

A detailed history including demographics, recurrences, previous treatment, etc. was taken. A detailed physical examination was done. Sample like scrapings of skin, nail clippings, and hair clippings was taken for KOH mount.

All samples (skin scrapings and nail and hair clippings) which showed fungal elements on KOH mount were inoculated on to Sabouraud dextrose agar (SDA) plate under aseptic precautions and incubated at 25°C for 2–3 weeks. Isolates were identified based on gross colony characteristics and microscopic morphology of their micro- and macroconidia and accessory structures. The isolates were then transferred to sterile distilled water (DW) in vials and stored.

Antifungal susceptibility testing

Antifungal susceptibility testing was done for the clinical isolates using disc diffusion method and E strip method. Paper disks containing clotrimazole (10 mcg), miconazole (30 mcg), and fluconazole (10 mcg) from Hi Media were used in the disk diffusion method to evaluate the in-vitro activity of the antifungal agents by measuring the mean diameter of inhibition around the disks. Griseofulvin E strip (0.002–32 mcg/mL, Hi Media) is used for calculating the minimum inhibitory concentration (MIC) of the drug [Table 1].
Table 1: Cut-off value for resistant, intermediate, and resistance

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The clinical isolates stored in the stock solution were subcultured on to Potato dextrose agar to enhance sporulation, and inoculum was prepared using colonies containing mixture of mycelia and conidia. The inoculum was spread evenly on the surface of 10 cm Petri dishes containing SDA medium and exposed to air dry. Then, the antifungal disks and Griseofulvin E strip were applied on to the plates, after which the plates were incubated at 25°C for 5–10 days. The zones of inhibition around the disks were measured and recorded. In the case of E strip, the MIC value is read from the scale in terms of µg/mL, where the ellipse edge intersects the strip. Criteria of susceptibility and resistance of antifungal disks were measured according to manufacturers’ guidelines.

Statistical analysis

Data were entered in MS Excel and analyzed using SPSS version 19.


  Results Top


The total number of cases included were 48. Males were 34 and females 14.

Mean age of the patients was 35 years. Mean duration of the disease was 9 months.

The commonest clinical presentation was co-occurrence of Tinea cruris and T. corporis (33.3%), followed by T. cruris (20.8%) and by T. corporis (18.7%).

Trichophyton mentagrophytes was isolated in 56.2% of the cases. T. rubrum was isolated in 16.6% of the cases. Microsporum species accounted for 12.5%. [Table 2], [graph 1]
Table 2: Fungal isolate in different clinical presentations

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Graph 1: Fungal isolates in different clinical presentations

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Drug sensitivity results

All dermatophytes combined showed highest sensitivity to clotrimazole (87.5%), followed by miconazole (60.4%).

Only 8.3% were sensitive to fluconazole and 37.5% were sensitive to griseofulvin. [Table 3], [graph 2]
Table 3: In-vitro sensitivity pattern of dermatophytes to antifungals

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Graph 2: In-vitro sensitivity pattern of dermatophytes to antifungals

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Among T. mentagrophytes which was the major species causing infection, 88% were sensitive to clotrimazole, 8% to fluconazole, 60% to miconazole, and 38% to griseofulvin.


  Discussion Top


Dermatophytoses is one of the most common infections of skin. Recently, there have been reports about the increase in the number of cases with extensive infection, resistant infections, and recurrent infections.[3],[4]

Most of the patients in our study were males (70.8%). The studies by Ghuse et al.[5] and Poojary et al.[6] also showed male preponderance. The reason for the male preponderance in our study could be due to the fact that most of our patients were immigrant manual laborers which was a male dominant group.

In our study, co-occurrence of T. cruris and T. corporis was the most common clinical presentation constituting 33%, followed by T. cruris 21% and T. corporis 19%. This is similar to the study by Poojary et al.,[6] in which co-occurrence of T. cruris and T. corporis was the most common presentation.

T. mentagrophytes was the most common organism which was isolated in 56% of the patients and T. rubrum in 17% of the patients. T. mentagrophytes was the most common organism in most of the clinical types.

Many studies from India show T. rubrum to be the dominant species.[7],[8],[9] However, of late studies are showing more contribution from T. mentagrophytes,[6],[10],[11] many a times contributing for more than 50% of the cases.[5],[12]

The shift in the species causing dermatophytoses could be one of the reasons for resistant and recurrent infections.

Among topical antifungals tested, 88% of the isolates were sensitive to clotrimazole and 60% of the isolates were sensitive to miconazole. Among systemic drugs tested, 38% of the isolates were sensitive to griseofulvin and 8% of the isolates were sensitive to fluconazole [Table 4] and [Table 5], [graph 3].
Table 4: Comparison between topical and systemic antifungals

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Table 5: A sensitivity pattern of our study could be compared with that in a study by Pakshir et al.[13]

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Graph 3: Comparison between topical (clotrimazole and miconazole) and systemic (fluconazole and griseofulvin) antifungals

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These findings could not be compared with more studies as most of the studies used broth method for sensitivity testing.

Limitations

  1. Small sample size


  2. Other commonly used drugs such as terbinafine, itraconazole, luliconazole etc. could not be tested in our study.



  Conclusion Top


T. mentagrophytes is the causative organism in a significant number of dermatophytic infections. Most of the isolated dermatophytes are still sensitive to clotrimazole in vitro. Though broth dilution is the standard method for sensitivity assay of dermatophytes, disc diffusion method can become a more simple alternative. In this regard, further studies on disc diffusion method are needed.[13]

Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

1.
Bashir S, Hassan I, Wani RT Influence of dermatophytosis on quality of life: A cross sectional study from Kashmir Valley in North India. Int J Community Med Public Health 2020;7:1711-6.  Back to cited text no. 1
    
2.
Patro N, Panda M, Jena AK The menace of superficial dermatophytosis on the quality of life of patients attending referral hospital in eastern India: A cross-sectional observational study. Indian Dermatol Online J 2019;10:262-6.  Back to cited text no. 2
    
3.
Bishnoi A, Vinay K, Dogra S Emergence of recalcitrant dermatophytosis in India. Lancet Infect Dis 2018;18:250-1.  Back to cited text no. 3
    
4.
Verma S, Madhu R The great Indian epidemic of superficial dermatophytosis: An appraisal. Indian J Dermatol 2017;62: 227-36.  Back to cited text no. 4
    
5.
Ghuse V, Someshwar S, Jerajani H Patterns of culture positivity and antifungal sensitivity in dermatophytosis. MGM J Med Sci 2019;6:105-12.  Back to cited text no. 5
    
6.
Poojary S, Miskeen A, Bagadia J, Jaiswal S, Uppuluri P A study of in vitro antifungal susceptibility patterns of dermatophytic fungi at a tertiary care center in western India. Indian J Dermatol 2019;64:277-84.  Back to cited text no. 6
    
7.
Chaudhary JK, Kumar A A clinico-mycological profile of dermatophytosis at a tertiary care hospital in Bihar. Int J Curr Microbiol App Sci 2016;5:181-9.  Back to cited text no. 7
    
8.
Hosthota A, Gowda T, Manikonda R Clinical profile and risk factors of dermatophytoses: A hospital based study. Int J Res Dermatol 2018;4:3860.  Back to cited text no. 8
    
9.
Sabtharishi V, Katragadda R, Ravinder T A study on the antifungal susceptibility pattern of dermatophytes isolated in a tertiary care hospital. Int J Bioassay 2017;6:5379-82.  Back to cited text no. 9
    
10.
Maity PP, Nandan K, Dey S Clinico-mycological profile of dermatophytosis in patients attending a tertiary care hospital in Eastern Bihar, India. J Evol Medical Dent Sci 2014;3:8263-9.  Back to cited text no. 10
    
11.
Agarwal RK, Gupta S, Mittal G, Khan F, Roy S, Agarwal A Antifungal susceptibility testing of dermatophytes by agar based disk diffusion method. Int J Curr Microbiol Appl Sci 2015;4:430-6.  Back to cited text no. 11
    
12.
Nenoff P, Verma SB, Vasani R, Burmester A, Hipler U, Wittig F et al. The current Indian epidemic of superficial dermatophytosis due to Trichophyton mentagrophytes—A molecular study. Mycoses 2018;62:336-56.  Back to cited text no. 12
    
13.
Pakshir K, Bahaedinie L, Rezaei Z, Sodaifi M, Zomorodian K In vitro activity of six antifungal drugs against clinically important dermatophytes. Jundishapur J Microbiol 2009;2: 158-63.  Back to cited text no. 13
    


    Figures

  [Figure 1], [Figure 2], [Figure 3]
 
 
    Tables

  [Table 1], [Table 2], [Table 3], [Table 4], [Table 5]



 

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