Molecular study and antifungal susceptibility profile of Trichophyton rubrum and Trichophyton mentagrophytes strains isolated from lesions of humans and cattle
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Abstract
Background and Objectives: Monitoring of contagious diseases is important to advance our knowledge of their epidemiology and to enable more impressive investigation and prevention efforts. This study aimed to examine antifungal drug susceptibility and molecular analysis of clinical isolates of Trichophyton rubrum and Trichophyton mentagrophytes in humans and cattle.
Materials and Methods: A total of 400 patients and 500 cattle were evaluated in this study. Dermatophytosis was confirmed in cases by direct microscopy and culture methods. Antifungal drug susceptibility profiles, MIC50, and MIC90 of isolates were determined using the broth microdilution method. Multiplex-PCR, RAPD PCR, and sequencing methods were used for the genetic analysis of virulence genes and the ITS1 and ITS2 regions, respectively.
Results: A total of 175 patients and 120 cattle were diagnosed with dermatophytosis. Dermatophytes showed a remarkable rate (30%) of terbinafine resistance. T. mentagrophytes showed lower susceptibility than T. rubrum (MIC50=16 μg/mL). Strains harboring Mep1, Mep2, and Mep4 genes had the highest frequency among all genotypes. A RAPD-PCR dendrogram divided T. mentagrophytes and T. rubrum strains into three and six groups, respectively.
Conclusion: A notable rate of resistance to terbinafine in isolated dermatophytes was reported in this study. Examination of RAPD-PCR results showed that T. rubrum strains had higher genetic diversity than T. mentagrophytes. Genetic monitoring of dermatophytes must be considered an important factor in providing fungal infection prevention and treatment approaches.
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4. Celestrino GA, Verrinder Veasey J, Benard G, Sousa MGT. Host immune responses in dermatophytes infection. Mycoses 2021; 64: 477-483.
5. Cowen LE, Sanglard D, Howard SJ, Rogers PD, Perlin DS. Mechanisms of antifungal drug resistance. Cold Spring Harb Perspect Med 2014; 5: a019752.
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7. Xiao C, Wang J, Liao Z, Huang Y, Ji Q, Liu Y, et al. Assessment of the mechanism of drug resistance in Trichophyton mentagrophytes in response to various substances. BMC Genomics 2021; 22: 250.
8. Verrier J, Monod M. Diagnosis of dermatophytosis using molecular biology. Mycopathologia 2017; 182: 193-202.
9. Yang R-H, Su J-H, Shang J-J, Wu Y-Y, Li Y, Bao D-P, et al. Evaluation of the ribosomal DNA internal transcribed spacer (ITS), specifically ITS1 and ITS2, for the analysis of fungal diversity by deep sequencing. PLoS One 2018; 13(10): e0206428.
10. Dalis JS, Kazeem HM, Kwaga JKP, Kwanashie CN. Prevalence and distribution of dermatophytosis lesions on cattle in Plateau State, Nigeria. Vet World 2019; 12: 1484-1490.
11. Fattahi A, Shirvani F, Ayatollahi A, Rezaei‐Matehkolaei A, Badali H, Lotfali E, et al. Multidrug‐resistant Trichophyton mentagrophytes genotype VIII in an Iranian family with generalized dermatophytosis: report of four cases and review of literature. Int J Dermatol 2021; 60: 686-692.
12. Baeza LC, Giannini MJSM. Strain differentiation of Trichophyton rubrum by random amplification of polymorphic DNA (RAPD). Rev Inst Med Trop Sao Paulo 2004; 46: 339-341.
13. Shaw D, Singh S, Dogra S, Jayaraman J, Bhat R, Panda S, et al. MIC and upper limit of wild-type distribution for 13 antifungal agents against a Trichophyton mentagrophytes-Trichophyton interdigitale complex of Indian origin. Antimicrob Agents Chemother 2020; 64(4): e01964-19.
14. Ansari S, Hedayati MT, Zomorodian K, Pakshir K, Badali H, Rafiei A, et al. Molecular characterization and in vitro antifungal susceptibility of 316 clinical isolates of dermatophytes in Iran. Mycopathologia 2016; 181: 89-95.
15. Didehdar M, Shokohi T, Khansarinejad B, Sefidgar SAA, Abastabar M, Haghani I, et al. Characterization of clinically important dermatophytes in North of Iran using PCR-RFLP on ITS region. J Mycol Med 2016; 26: 345-350.
16. Frías-De-León MG, Martínez-Herrera E, Atoche-Diéguez CE, González-Cespón JL, Uribe B, Arenas R, et al. Molecular identification of isolates of the Trichophyton mentagrophytes complex. Int J Med Sci 2020; 17: 45-52.
17. Guo Y, Ge S, Luo H, Rehman A, Li Y, He S. Occurrence of Trichophyton verrucosum in cattle in the Ningxia Hui autonomous region, China. BMC Vet Res 2020; 16: 187.
18. Singh A, Masih A, Monroy-Nieto J, Singh PK, Bowers J, Travis J, et al. A unique multidrug-resistant clonal Trichophyton population distinct from Trichophyton mentagrophytes/Trichophyton interdigitale complex causing an ongoing alarming dermatophytosis outbreak in India: Genomic insights and resistance profile. Fungal Genet Biol 2019; 133: 103266.
19. Salehi Z, Shams-Ghahfarokhi M, Razzaghi-Abyaneh M. Antifungal drug susceptibility profile of clinically important dermatophytes and determination of point mutations in terbinafine-resistant isolates. Eur J Clin Microbiol Infect Dis 2018; 37: 1841-1846.
20. Nowrozi H, Nazeri G, Adimi P, Bashashati M, Emami M. Comparison of the activities of four antifungal agents in an in vitro model of dermatophyte nail infection. Indian J Dermatol 2008; 53: 125-128.
21. Taghipour S, Shamsizadeh F, Pchelin IM, Rezaei-Matehhkolaei A, Mahmoudabadi AZ, Valadan R, et al. Emergence of terbinafine resistant Trichophyton mentagrophytes in Iran, harboring mutations in the squalene epoxidase (SQLE) gene. Infect Drug Resist 2020; 13: 845-850.
22. Singh A, Masih A, Khurana A, Singh PK, Gupta M, Hagen F, et al. High terbinafine resistance in Trichophyton interdigitale isolates in Delhi, India harbouring mutations in the squalene epoxidase gene. Mycoses 2018; 61: 477-484.
23. Hiruma J, Noguchi H, Hase M, Tokuhisa Y, Shimizu T, Ogawa T, et al. Epidemiological study of terbinafine‐resistant dermatophytes isolated from Japanese patients. J Dermatol 2021; 48: 564-467.
24. Wiederhold NP, Fothergill AW, McCarthy DI, Tavakkol A. Luliconazole demonstrates potent in vitro activity against dermatophytes recovered from patients with onychomycosis. Antimicrob Agents Chemother 2014; 58: 3553-3555.
25. Baghi N, Shokohi T, Badali H, Makimura K, Rezaei-Matehkolaei A, Abdollahi M, et al. In vitro activity of new azoles luliconazole and lanoconazole compared with ten other antifungal drugs against clinical dermatophyte isolates. Med Mycol 2016; 54: 757-763.
26. Chowdhary A, Singh A, Singh PK, Khurana A, Meis JF. Perspectives on misidentification of Trichophyton interdigitale/Trichophyton mentagrophytes using internal transcribed spacer region sequencing: urgent need to update the sequence database. Mycoses 2019; 62: 11-15.
27. Leng W, Liu T, Wang J, Li R, Jin Q. Expression dynamics of secreted protease genes in Trichophyton rubrum induced by key host's proteinaceous components. Med Mycol 2009; 47: 759-765.
28. Gnat S, Łagowski D, Dyląg M, Ptaszyńska A, Nowakiewicz A. Modulation of ERG gene expression in fluconazole-resistant human and animal isolates of Trichophyton verrucosum. Braz J Microbiol 2021; 52: 2439-2446.
29. Angiolella L. Virulence Regulation and Drug-Resistance Mechanism of Fungal Infection. Microorganisms 2022; 10: 409.
30. Khosravi A, Behzad F, Sabokbar A, Shokri H, Haddadi S, Masoudi‐Nejad A. Molecular typing of Epidermophyton floccosum isolated from patients with dermatophytosis by RAPD‐PCR. J Basic Microbiol 2010; 50 Suppl 1: S68-73.
31. Nenoff P, Verma SB, Uhrlaß S, Burmester A, Gräser Y. A clarion call for preventing taxonomical errors of dermatophytes using the example of the novel Trichophyton mentagrophytes genotype VIII uniformly isolated in the Indian epidemic of superficial dermatophytosis. Mycoses 2019; 62: 6-10.
2. Sen S, Borah SN, Bora A, Deka S. Rhamnolipid exhibits anti-biofilm activity against the dermatophytic fungi Trichophyton rubrum and Trichophyton mentagrophytes. Biotechnol Rep (Amst) 2020;27:e005166.
3. Klinger M, Theiler M, Bosshard PP. Epidemiological and clinical aspects of Trichophyton mentagrophytes/Trichophyton interdigitale infections in the Zurich area: a retrospective study using genotyping. J Eur Acad Dermatol Venereol 2021; 35: 1017-1025.
4. Celestrino GA, Verrinder Veasey J, Benard G, Sousa MGT. Host immune responses in dermatophytes infection. Mycoses 2021; 64: 477-483.
5. Cowen LE, Sanglard D, Howard SJ, Rogers PD, Perlin DS. Mechanisms of antifungal drug resistance. Cold Spring Harb Perspect Med 2014; 5: a019752.
6. Taghipour S, Pchelin IM, Zarei Mahmoudabadi A, Ansari S, Katiraee F, Rafiei A, et al. Trichophyton mentagrophytes and T. interdigitale genotypes are associated with particular geographic areas and clinical manifestations. Mycoses 2019; 62: 1084-1091.
7. Xiao C, Wang J, Liao Z, Huang Y, Ji Q, Liu Y, et al. Assessment of the mechanism of drug resistance in Trichophyton mentagrophytes in response to various substances. BMC Genomics 2021; 22: 250.
8. Verrier J, Monod M. Diagnosis of dermatophytosis using molecular biology. Mycopathologia 2017; 182: 193-202.
9. Yang R-H, Su J-H, Shang J-J, Wu Y-Y, Li Y, Bao D-P, et al. Evaluation of the ribosomal DNA internal transcribed spacer (ITS), specifically ITS1 and ITS2, for the analysis of fungal diversity by deep sequencing. PLoS One 2018; 13(10): e0206428.
10. Dalis JS, Kazeem HM, Kwaga JKP, Kwanashie CN. Prevalence and distribution of dermatophytosis lesions on cattle in Plateau State, Nigeria. Vet World 2019; 12: 1484-1490.
11. Fattahi A, Shirvani F, Ayatollahi A, Rezaei‐Matehkolaei A, Badali H, Lotfali E, et al. Multidrug‐resistant Trichophyton mentagrophytes genotype VIII in an Iranian family with generalized dermatophytosis: report of four cases and review of literature. Int J Dermatol 2021; 60: 686-692.
12. Baeza LC, Giannini MJSM. Strain differentiation of Trichophyton rubrum by random amplification of polymorphic DNA (RAPD). Rev Inst Med Trop Sao Paulo 2004; 46: 339-341.
13. Shaw D, Singh S, Dogra S, Jayaraman J, Bhat R, Panda S, et al. MIC and upper limit of wild-type distribution for 13 antifungal agents against a Trichophyton mentagrophytes-Trichophyton interdigitale complex of Indian origin. Antimicrob Agents Chemother 2020; 64(4): e01964-19.
14. Ansari S, Hedayati MT, Zomorodian K, Pakshir K, Badali H, Rafiei A, et al. Molecular characterization and in vitro antifungal susceptibility of 316 clinical isolates of dermatophytes in Iran. Mycopathologia 2016; 181: 89-95.
15. Didehdar M, Shokohi T, Khansarinejad B, Sefidgar SAA, Abastabar M, Haghani I, et al. Characterization of clinically important dermatophytes in North of Iran using PCR-RFLP on ITS region. J Mycol Med 2016; 26: 345-350.
16. Frías-De-León MG, Martínez-Herrera E, Atoche-Diéguez CE, González-Cespón JL, Uribe B, Arenas R, et al. Molecular identification of isolates of the Trichophyton mentagrophytes complex. Int J Med Sci 2020; 17: 45-52.
17. Guo Y, Ge S, Luo H, Rehman A, Li Y, He S. Occurrence of Trichophyton verrucosum in cattle in the Ningxia Hui autonomous region, China. BMC Vet Res 2020; 16: 187.
18. Singh A, Masih A, Monroy-Nieto J, Singh PK, Bowers J, Travis J, et al. A unique multidrug-resistant clonal Trichophyton population distinct from Trichophyton mentagrophytes/Trichophyton interdigitale complex causing an ongoing alarming dermatophytosis outbreak in India: Genomic insights and resistance profile. Fungal Genet Biol 2019; 133: 103266.
19. Salehi Z, Shams-Ghahfarokhi M, Razzaghi-Abyaneh M. Antifungal drug susceptibility profile of clinically important dermatophytes and determination of point mutations in terbinafine-resistant isolates. Eur J Clin Microbiol Infect Dis 2018; 37: 1841-1846.
20. Nowrozi H, Nazeri G, Adimi P, Bashashati M, Emami M. Comparison of the activities of four antifungal agents in an in vitro model of dermatophyte nail infection. Indian J Dermatol 2008; 53: 125-128.
21. Taghipour S, Shamsizadeh F, Pchelin IM, Rezaei-Matehhkolaei A, Mahmoudabadi AZ, Valadan R, et al. Emergence of terbinafine resistant Trichophyton mentagrophytes in Iran, harboring mutations in the squalene epoxidase (SQLE) gene. Infect Drug Resist 2020; 13: 845-850.
22. Singh A, Masih A, Khurana A, Singh PK, Gupta M, Hagen F, et al. High terbinafine resistance in Trichophyton interdigitale isolates in Delhi, India harbouring mutations in the squalene epoxidase gene. Mycoses 2018; 61: 477-484.
23. Hiruma J, Noguchi H, Hase M, Tokuhisa Y, Shimizu T, Ogawa T, et al. Epidemiological study of terbinafine‐resistant dermatophytes isolated from Japanese patients. J Dermatol 2021; 48: 564-467.
24. Wiederhold NP, Fothergill AW, McCarthy DI, Tavakkol A. Luliconazole demonstrates potent in vitro activity against dermatophytes recovered from patients with onychomycosis. Antimicrob Agents Chemother 2014; 58: 3553-3555.
25. Baghi N, Shokohi T, Badali H, Makimura K, Rezaei-Matehkolaei A, Abdollahi M, et al. In vitro activity of new azoles luliconazole and lanoconazole compared with ten other antifungal drugs against clinical dermatophyte isolates. Med Mycol 2016; 54: 757-763.
26. Chowdhary A, Singh A, Singh PK, Khurana A, Meis JF. Perspectives on misidentification of Trichophyton interdigitale/Trichophyton mentagrophytes using internal transcribed spacer region sequencing: urgent need to update the sequence database. Mycoses 2019; 62: 11-15.
27. Leng W, Liu T, Wang J, Li R, Jin Q. Expression dynamics of secreted protease genes in Trichophyton rubrum induced by key host's proteinaceous components. Med Mycol 2009; 47: 759-765.
28. Gnat S, Łagowski D, Dyląg M, Ptaszyńska A, Nowakiewicz A. Modulation of ERG gene expression in fluconazole-resistant human and animal isolates of Trichophyton verrucosum. Braz J Microbiol 2021; 52: 2439-2446.
29. Angiolella L. Virulence Regulation and Drug-Resistance Mechanism of Fungal Infection. Microorganisms 2022; 10: 409.
30. Khosravi A, Behzad F, Sabokbar A, Shokri H, Haddadi S, Masoudi‐Nejad A. Molecular typing of Epidermophyton floccosum isolated from patients with dermatophytosis by RAPD‐PCR. J Basic Microbiol 2010; 50 Suppl 1: S68-73.
31. Nenoff P, Verma SB, Uhrlaß S, Burmester A, Gräser Y. A clarion call for preventing taxonomical errors of dermatophytes using the example of the novel Trichophyton mentagrophytes genotype VIII uniformly isolated in the Indian epidemic of superficial dermatophytosis. Mycoses 2019; 62: 6-10.
| Files | ||
| Issue | Vol 14 No 4 (2022) | |
| Section | Original Article(s) | |
| DOI | https://doi.org/10.18502/ijm.v14i4.10246 | |
| Keywords | ||
| Trichophyton rubrum; Trichophyton mentagrophytes; Dermatophytosis; Random amplified polymorphic DNA-PCR | ||
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How to Cite
1.
Mohammadifard H, Amini K, Bayat M, Hashemi SJ, Noorbakhsh F. Molecular study and antifungal susceptibility profile of Trichophyton rubrum and Trichophyton mentagrophytes strains isolated from lesions of humans and cattle. Iran J Microbiol. 2022;14(4):587-597.
