Azole resistance using agar screening in environmental isolates of Aspergillus species from Pakistan
DOI:
https://doi.org/10.61529/idjp.v34i3.377Abstract
Background: Azole resistance in clinical and environmental Aspergillus isolates has been reported globally. The environmental route of resistance follows the use of fungicides in agricultural practices and is known to be more common than the clinical route of resistance. We aimed to screen for azole resistance in Aspergillus species from environmental sources in Pakistan.
Material and Methods: This was a cross-sectional, observational study conducted from November 2021 to June 2023 at Aga Khan Aga Khan University Hospital, Karachi. Environmental Aspergillus isolates from across Pakistan were screened for azole resistance using previously described methods. Soil samples were collected from different towns and cities of Pakistan using convenience sampling. A suspension of soil in normal saline was cultured on SDA to yield fungal colonies, and onto media containing posaconazole, voriconazole and itraconazole in defined concentrations to screen for resistance. Growth on antifungal containing media was considered to be resistant.
Results: Soil was collected from 171 different sites across Pakistan. A total of 249 Aspergillus isolates were recovered from these soil samples. These included 99 (39.7%) A. niger, 84 (33.7%) A. flavus, 31 (12.4%) A. fumigatus, and other species. None of the isolates grew on any of the antifungal containing media, indicating no resistance.
Conclusion: Our study finds no indication of azole resistance in environmental isolates from Pakistan. However, methodologies such as MIC determination and genotypic sequencing are needed for a more robust analysis.
Keywords: Aspergillus species, Agar screening, Azole resistance, Environmental, Pakistan
References
Pérez-Cantero A, López-Fernández L, Guarro J, Capilla J. Azole resistance mechanisms in Aspergillus: update and recent advances. Int J Antimicrob Agents. 2020; 55(1): 105807. DOI: https://doi.org/10.1016/j.ijantimicag.2019.09.011
Denning DW, Venkateswarlu K, Oakley KL, Anderson MJ, Manning NJ, Stevens DA, et al. Itraconazole resistance in Aspergillus fumigatus. Antimicrob Agents Chemother. 1997;41(6):1364-8. DOI: https://doi.org/10.1128/aac.41.6.1364
van der Linden JW, Arendrup MC, Warris A, Lagrou K, Pelloux H, Hauser PM, et al. Prospective multicenter international surveillance of azole resistance in Aspergillus fumigatus. Emerg Infect Dis. 2015; 21(6): 1041-4. DOI: https://doi.org/10.3201/eid2106.140717
Latgé JP, Chamilos G. Aspergillus fumigatus and Aspergillosis in 2019. Clin Microbiol Rev. 2019; 33(1): e00140. DOI: https://doi.org/10.1128/cmr.00140-18
Nywening AV, Rybak JM, Rogers PD, Fortwendel JR. Mechanisms of triazole resistance in Aspergillus fumigatus. Environ Microbiol. 2020; 22(12): 4934-52. DOI: https://doi.org/10.1111/1462-2920.15274
Verweij PE, Ananda-Rajah M, Andes D, Arendrup MC, Brüggemann RJ, Chowdhary A, et al. International expert opinion on the management of infection caused by azole-resistant Aspergillus fumigatus. Drug Resist Updat. 2015; 21-22: 30-40. DOI: https://doi.org/10.1016/j.drup.2015.08.001
Chen Y, Dong F, Zhao J, Fan H, Qin C, Li R, et al. High azole resistance in Aspergillus fumigatus isolates from Strawberry Fields, China, 2018. Emerg Infect Dis. 2020; 26(1): 81-9. DOI: https://doi.org/10.3201/eid2601.190885
Ahangarkani F, Puts Y, Nabili M, Khodavaisy S, Moazeni M, Salehi Z, et al. First azole-resistant Aspergillus fumigatus isolates with the environmental TR(46) /Y121F/T289A mutation in Iran. Mycoses. 2020; 63(5): 430-6. DOI: https://doi.org/10.1111/myc.13064
Chowdhary A, Sharma C, Kathuria S, Hagen F, Meis JF. Prevalence and mechanism of triazole resistance in Aspergillus fumigatus in a referral chest hospital in Delhi, India and an update of the situation in Asia. Front Microbiol. 2015; 6: 428. DOI: https://doi.org/10.3389/fmicb.2015.00428
Mortensen KL, Mellado E, Lass-Flörl C, Rodriguez-Tudela JL, Johansen HK, Arendrup MC. Environmental study of azole-resistant Aspergillus fumigatus and other aspergilli in Austria, Denmark, and Spain. Antimicrob Agents Chemother. 2010; 54(11): 4545-9. DOI: https://doi.org/10.1128/aac.00692-10
Guinea J, Verweij PE, Meletiadis J, Mouton JW, Barchiesi F, Arendrup MC. How to: EUCAST recommendations on the screening procedure E.Def 10.1 for the detection of azole resistance in Aspergillus fumigatus isolates using four-well azole-containing agar plates. Clin Microbiol Infect. 2019; 25(6): 681-7. DOI: https://doi.org/10.1016/j.cmi.2018.09.008
Snelders E, Huis In 't Veld RA, Rijs AJ, Kema GH, Melchers WJ, Verweij PE. Possible environmental origin of resistance of Aspergillus fumigatus to medical triazoles. Appl Environ Microbiol. 2009; 75(12): 4053-7. DOI: https://doi.org/10.1128/aem.00231-09
Moin S, Manzar Abbas, Sidra Laiq, Joveria Farooqi, Afia Zafar, Kauser Jabeen. Triazole Resistance screening in environmental Aspergillus Species from Pakistan. Infect Dis J. 2020; 29 (3): 54-6. Available from: https://ojs.idj.org.pk/index.php/Files/article/view/30
Moin S, Farooqi J, Jabeen K, Laiq S, Zafar A. Screening for triazole resistance in clinically significant Aspergillus species; report from Pakistan. Antimicrob Resist Infect Control. 2020; 9(1): 62. DOI: https://doi.org/10.1186/s13756-020-00731-8
Iqbal N, Irfan M, Mushtaq A, Jabeen K. Underlying conditions and clinical spectrum of chronic pulmonary aspergillosis (CPA): An experience from a tertiary care hospital in Karachi, Pakistan. J Fungi (Basel, Switzerland). 2020; 6(2): 41. DOI: https://doi.org/10.3390/jof6020041
WHO fungal priority pathogens list to guide research, development and public health action. Geneva: World Health Organization; 2022. Availble from: https://iris.who.int/bitstream/handle/10665/363682/9789240060241-eng.pdf?s
Arendrup MC, Verweij PE, Mouton JW, Lagrou K, Meletiadis J. Multicentre validation of 4-well azole agar plates as a screening method for detection of clinically relevant azole-resistant Aspergillus fumigatus. J Antimicrob Chemother. 2017; 72(12): 3325-33. DOI: https://doi.org/10.1093/jac/dkx319
Nabili M, Shokohi T, Moazeni M, Khodavaisy S, Aliyali M, Badiee P, et al. High prevalence of clinical and environmental triazole-resistant Aspergillus fumigatus in Iran: is it a challenging issue? J Med Microbiol. 2016; 65(6): 468-75. DOI: https://doi.org/10.1099/jmm.0.000255
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2025 Syed Ali Raza Nasir, Sadaf Zaka, Joveria Farooqi, Adan Zubair, Kausar Jabeen
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
