Isolation, Identification and Antifungal Susceptibility of Dermatophytes Isolated from Clinically Suspected Cases of Tinea Infections in Pakistan

Main Article Content

Shumaila Shakir
Sidrah Saleem
Wajhiah Rizvi
Abdul Waheed
Javid Iqbal


Dermatophytosis or tinea is a type of cutaneous infection caused by keratinophilic fungi, infecting the skin, nails and hair. A correct diagnosis is important for epidemiological purposes and initiating appropriate treatment. An increase in the prevalence of fungal infection worldwide is due to abuse of antibiotics, immunosuppressive treatments and numerous medical conditions.

Aim: To isolate, identify, and examine the in-vitro antifungal susceptibility of dermatophytes in clinically suspected cases of tinea infections.

Methodology: After taking informed consent, we took 65 patients suspected of tinea infection and obtained samples from skin, hair and nail, under aseptic precautions, at Department of Microbiology, University of Health Sciences (UHS), Lahore, Pakistan. The identification of dermatophytes was performed using potassium hydroxide (KOH) mounts and culture on Sabouraud Dextrose Agar (SDA) and Dermatophyte Test Medium (DTM). The cultures were incubated at 30ºC for up to 4 weeks in case of SDA and 2 weeks in case of DTM. Lactophenol cotton blue (LCB) stain was used to identify the species morphologically. Susceptibility test was done by agar diffusion method using antifungal disks and zones of inhibition were measured.

Results: More females (55.38%) than males (44.61%) were observed in the study. Most of the cases belonged to the age categories of 1-10 years and 21-30 years. Tinea corporis was the most common clinical type found (27.69%) followed by Tinea capitis (21.53%) and Tinea cruris (12.30%).Trichophyton mentagrophytes was the commonest species isolated (32%)  followed by Trichophyton violaceum (28%) and Trichophyton rubrum (12%). Terbinafine was seen to be the most effective drug against the isolates, followed by clotrimazole. Fluconazole showed least activity.

Conclusion: Fungal culture remains the gold standard in identifying the causative species. Terbinafine promises to be a potent antifungal, whereas fluconazole has low efficacy against such organisms. Disk diffusion method adopted for antifungal susceptibility is cost effective and easily performable in small laboratories not having an established mycology bench.

Dermatophytes, disk diffusion method, antifungal drugs, susceptibility, tinea infections, Pakistan

Article Details

How to Cite
Shakir, S., Saleem, S., Rizvi, W., Waheed, A., & Iqbal, J. (2019). Isolation, Identification and Antifungal Susceptibility of Dermatophytes Isolated from Clinically Suspected Cases of Tinea Infections in Pakistan. Microbiology Research Journal International, 29(5), 1-11.
Original Research Article


Grumbt M, Monod M, Yamada T, Hertweck C, Kunert J, Staib P. Keratin degradation by dermatophytes relies on cysteine dioxygenase and a sulfite efflux pump. J Invest Dermatol. 2013;133(6):1550–5.
DOI: 10.1038/jid.2013.41

Shalaby MF, El-Din AN, El-Hamd MA. Isolation, identification and In vitro antifungal susceptibility testing of dermatophytes from clinical samples at Sohag University Hospital in Egypt. Electronic Physician. 2016;8(6): 2557–2567.
DOI: 10.19082/2557

Asticcioli S, Di Silverio A, Sacco L, Fusi I, Vincenti L, Romero E. Dermatophyte infections in patients attending a tertiary care hospital in northern Italy. New Microbiol. 2008;31(4):543-8.

Falahati M, Akhlagi L, Lari AR, Alaghehbandan R. Epidemiology of dermatophytoses in an area of south of Tehran, Iran. Mycopathologica. 2003;156 (4):279-287.

Sen SS, Rasul ES. Dermatophytosis in Assam. Indian Journal of Medical Microbiology. 2006;24(1):77.

Ekwealor CC, Oyeka CA. Cutaneous mycoses among rice farmers in Anambra State, Nigeria. Journal of Mycology; 2013.

Martin AG, Kobayashi GS. Superficial fungal infection: Dermatophytosis, tineanigra, piedra. In: Freedberg IM, Eisen AZ, Wolff K, Austen KF, Goldsmith LA, Katz SI, Fitzpatrick TB, editors. Dermatology in General Medicine,Fifth Edition, Mc Graw-Hill, New York. 1999; 2337–2357.

Starova A, Balabanova-Stefanova M, V’lckova-Lasckoska M. Dermatophytes in republic of macedonia. Contributions, Sec Biol Med Sci MASA. 2010;31:31.

Hanif F, Ikram A, Abbasi SA, Malik N. Pattern of dermatophytes among dermatological specimens at AFIP, Rawalpindi. Journal of Pakistan Association of Dermatologists. 2012;22(2).

Krishan Anand S, Ray R, Chatterjee M, Khandare M. A cross sectional descriptive study on clinical type and etiological agent of superficial dermatophytosis. JDA Indian Journal of Clinical Dermatology. 2018;1: 71-74.

Ghannoum MA, Isham NC. Dermatophytes and dermatophytoses. 2nd ed. Clinical Mycology. 2009;375-384.

Weitzman I, Summerbell RC. The dermatophytes. Clinical Microbiology Reviews. 1995;8(2):240-259.

Ananthanarayan R, Paniker CK. Medical mycology, Chapter 65 text book of microbiology. 8th edition. Hyderabad, India: Universities Press Private Limited. 2009;604–7.

Dhinakaran A, Kalaiselvam M, Sekar V, Sethubathi GVB. Isolation of Penicillium sp. and its antagonistic activity against dermatophytes from volcano soil of Baratang Island, Andaman. International Journal of Pharmaceutical Sciences and Research. 2012;3(2):564.

Achterman RR, White TC. Dermatophyte virulence factors: Identifying and analyzing genes that may contribute to chronic or acute skin infections. Int J Microbiol; 2012

Côbo EC, Silva JC, Cota UA, Machado JR, Castellano LR. Evaluation of a modified microscopic direct diagnosis of dermatophytosis. Journal of Microbiological Methods. 2010;81(2):205-207.

Robert R, Pihet M. Conventional methods for the diagnosis of dermatophytosis. Mycopathologia. 2008;166(5-6):295-306.

Shams-Ghahfarokhi M, Shokoohamiri MR, Amirrajab N, Moghadasi B, Ghajari A, Zeini F, et al. In vitro antifungal activities of Allium cepa, Allium sativum and ketoconazole against some pathogenic yeasts and dermatophytes. Fitoterapia. 2006;77:321– 323.

Nyilasi I, Kocsube S, Krizsán K, Galgo L, Papp T, Pesti M, Nagy K. Susceptibility of clinically importantdermatophytes against statins and different statin-antifungal combinations. Med Mycol. 2014;52:140– 148.

Esteban A, Abarca ML, Cabanes FJ. Comparison of disk diffusion method and broth microdilution method for antifungal susceptibility testing of dermatophytes. Medical Mycology. 2005;43(1):61-66.

Yadav A, Urhekar AD, Mane V, Danu MS, Goel N, Ajit KG. Optimization and isolation of dermatophytes from clinical samples and in vitro antifungal susceptibility testing by disc diffusion method. RRJMB. 2013; 2(3):19-34.

Jessup CJ, Warner J, Isham N, Hasan I, Ghannoum MA. Antifungal susceptibility testing of Dermatophytes: Establishing a medium for inducing conidial growth and evaluation of susceptibility of clinical isolates. Journal of Clinical Microbiology. 2000;38(1):341-344.

Nweze EI. Dermatophytoses in domesticated animals. Revista do Instituto de Medicina Tropical de São Paulo. 2011; 53(2):94-99.

Macura AB. Dermatophyte infections. International Journal of Dermatology. 1993;32(5):313-323.

Barry AL, Paller MA, Rennie RP, Fuchs PC, Brown SD. Precision and accuracy of fluconazole susceptibility testing by broth microdilution, Etest and disk diffusion methods. Antimicrobial Agents and Chemotherapy. 2002;46(6):1781-1784.

Meis J, Petrou M, Bille J, Ellis D, Gibbs D, Global Antifungal Surveillance Group. A global evaluation of the susceptibility of Candida species to fluconazole by disk diffusion. Diagnostic Microbiology and Infectious Disease. 2000;36(4):215-223.

Matar MJ, Ostrosky-Zeichner L, Paetznick VL, Rodriguez JR, Chen E, Rex JH. Correlation between Etest, disk diffusion, and micro dilution methods for antifungal susceptibility testing of fluconazole and voriconazole. Antimicrob Agents Chemother. 2003;47(5):1647–51.

Hazen Kevin. Davise H. Larone, ed. Medically important fungi: A guide to identification, 4th edn. Mycopathologia. 2003;156:383-384.

Winn Washington C, Elmer W Koneman. Koneman's color atlas and textbook of diagnostic microbiology. 6th ed. Philadelphia: Lippincott Williams & Wilkins; 2006.

Keyvan P, Leila B, Zahra R, Manuchehr S, Kamiar Z. In vitro activity of six antifungal drugs against clinically important dermatophytes. Jundishapur Journal of Microbiology. 2009;(4 Autumn):158-163.

Pakshir K, Akbarzadeh MA, Bonyadpour B, Mohagheghzadeh AA. In vitro activity and comparison of clotrimazol, fluconazol and nystatin against Candida vaginitis isolates in Shiraz, 2008. J Rafsanjan Univ Med Sci. 2010;9(3):210–20.

Farwa U, Abbasi SA, Mirza IA, Amjad A, Ikram A, Malik N, Hanif F. Non-dermatophyte moulds as pathogens of onychomycosis. J Coll Physicians Surg Pak. 2011;21(10):597-600.

Jahangir M, Hussain I, Khurshid K, Haroon TS. A clinico‐etiologic correlation in tinea capitis. International Journal of Dermatology. 1999;38(4):275-278.

Thakur R. Tinea capitis in Botswana. Clinical, Cosmetic and Investigational Dermatology. 2012;6:37-41.

Taylor RL, Kotrajaras R, Jotisankasa V. Occurrence of dermatophytes in Bangkok, Thailand. Sabouraudia: Journal of Medical and Veterinary Mycology. 1968;6(4):307-311.

Allah SS, Nada H, Mokhtar M. Yeast infections as a cause of nail disease in the Western province of Saudi Arabia. Egypt J Med Lab Sci. 2005;14:2.

Madhavi S, Rama Rao MV, Jyothsna K. Mycological study of dermatophytosis in rural population. Ann Biol Res. 2011;2(3): 88-93.

Santosh HK, Jithendra K, Rao AVM, Buchineni M, Pathapati RM. Clinico-mycological study of dermatophytosis our experience. Int. J. Curr. Microbiol. App. Sci. 2015;4(7):695-702.

Agrawalla A, Jacob M, Sethi M. A clinico-mycological study of dermatophytosis in Nepal J Dermatol. 2001;28:16-21.

Bindu V, Pavithran K. Clinico-mycological study of dermatophytosis in Calicut. Indian Journal of Dermatology, Venereology and Leprology. 2002;68(5):259.

Kumar S, Mallya SP, Shenoy SM. Trichophyton rubrum: The commonest isolate from dermatophytosis. Research Journal of Pharmaceutical, Biological and Chemical Sciences. 2012;3(1):101-106.

Abu-Elteen KH, Malek MA. Prevalence of dermatophytoses in the Zarqa district of Jordan. Mycopathologia. 1999;145(3):137-142.

Cetinkaya Z, Kiraz N, Karaca S, Kulac M, Ciftci IH, Aktepe OC, Altindis M, Kiyildi N, Piyade M. Antifungal susceptibilities of dermatophytic agents isolated from clinical specimens. European Journal of Dermatology. 2005;15(4):258-261.

Ghannoum M. Antifungal susceptibility testing of dermatophytes. Dermatology Online Journal. 2001;7(1).

Afshari MA, Shams-Ghahfarokhi M, Razzaghi-Abyaneh M. Antifungal susceptibility and virulence factors of clinically isolated dermatophytes in Tehran, Iran. Iranian Journal of Microbiology. 2016;8(1):36–6.
(Fluconazole Least Effective, Dd Method)

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;4:430-436.