Extended Abstract

Introduction
Dermatophytosis is a significant dermatological condition affecting both humans and animals, caused by a group of fungi known as dermatophytes. These fungi invade keratinized tissues such as skin, hair, and nails, leading to various fungal infections named Tinea based on the infection site. Dermatophytosis is a contagious disease that can be transmitted directly or indirectly and also through contact with infected animals.
Allylamines, polyenes, and azoles are key drug classes used topically and systemically to treat dermatophytosis. However, treatment failures due to increasing drug resistance are becoming more common.
Artemisia spp. is a genus of plants in the Asteraceae family widely distributed across different regions of the world. Various bioactive compounds, including phenols, terpenoids, sterols, and polyacetylenes, have been reported in this genus, exhibiting antimicrobial, antiviral, anti-inflammatory, and antifungal properties. This study aimed to determine the in vitro antifungal activity of petroleum ether, dichloromethane, ethyl acetate, ethanol, and hydroethanol extracts of the aerial parts of Artemisia khorassanica, Artemisia scoparia, and Artemisia vulgaris against Trichophyton rubrum, Trichophyton verrucosum, Epidermophyton floccosum, and Microsporum canis.
Methods
This descriptive study was conducted on fungal isolates of Trichophyton rubrum, Trichophyton verrucosum, Microsporum canis, and Epidermophyton floccosum in the Mycology Group at Kermanshah University of Medical Sciences, Iran.
Preparation of Fungal Isolates and Culture Media
Three fungi, Microsporum canis (5069), Trichophyton verrucosum (5056), and Trichophyton rubrum (5143), were obtained from the Fungal Collection Center, and Epidermophyton floccosum was isolated from patients. The fungi were cultured on Sabouraud Dextrose Agar containing chloramphenicol and cycloheximide for 14 days at 25±2°C.
Preparation of Plant Species
Various species of the Artemisia genus, including Artemisia khorassanica, Artemisia scoparia, and Artemisia vulgaris, were collected and identified in Khorasan Razavi Province.
Extraction
For extraction, aerial parts of the plants were sequentially extracted using five solvents with different polarities to evaluate the antifungal effects of non-polar, semi-polar, and polar compounds.
Antifungal Activity by Agar Dilution Method
For initial screening, plant extracts were dissolved in dimethyl sulfoxide (DMSO) and mixed with sterile molten Sabouraud Dextrose Agar containing chloramphenicol and cycloheximide to a final concentration of 1.25 mg/mL. Then, 100 µL of each fungal conidia suspension (10^6 colonies/mL) was inoculated onto the mixed media or control and incubated at 25±2°C for 14 days. Antifungal effects were evaluated by observing complete growth inhibition of the fungi.
Minimum Inhibitory Concentration (MIC)
The minimum inhibitory concentration of the extracts was determined by agar dilution. Final concentrations of each extract in the medium ranged from 1250 ~ 39.06 µg/mL. MIC was determined after a 14-day incubation period at 25±2°C by observing no fungal growth. DMSO (31.25 µL/mL) and terbinafine (0.0039 ~ 2 µg/mL) in the medium were used as negative and positive controls, respectively.
Preliminary Phytochemical Screening
Petroleum ether and dichloromethane extracts were evaluated for the presence of terpenoids, sterols, and flavonoids using standard methods.
Sterol Detection: Liebermann–Burchard reaction, where a blue or green color indicates the presence of steroids.
Terpenoid Detection: Formation of a reddish-brown color at the interface indicates the presence of terpenoids.
Flavonoid Detection: Appearance of a red or orange color indicates the presence of flavonoids.
Results
Fifteen extracts from the aerial parts of Artemisia khorassanica, Artemisia scoparia, and Artemisia vulgaris showed varying degrees of inhibitory activity against Epidermophyton floccosum, Trichophyton rubrum, Microsporum canis, and Trichophyton verrucosum. Microsporum canis exhibited the highest sensitivity (86.66%) and Trichophyton verrucosum the highest resistance (100%) to the tested extracts. Both Epidermophyton floccosum and Trichophyton verrucosum showed higher resistance to terbinafine (MIC 0.25 µg/mL) compared to the other fungi (MIC 0.0039 µg/mL).
In initial screening, the extracts of Artemisia khorassanica, Artemisia scoparia, and Artemisia vulgaris showed fungal growth inhibition in 6, 12, and 9 cases, respectively. Despite the highest extraction yield, hydroethanol extracts showed the least antifungal activity. Petroleum ether and dichloromethane extracts from all three Artemisia species inhibited two or three fungal isolates, with dichloromethane extracts showing relatively higher extraction yields. The lowest MIC (78.12 µg/mL) was observed for petroleum ether and dichloromethane extracts of Artemisia scoparia against Trichophyton rubrum and Microsporum canis.
Preliminary phytochemical screening of petroleum ether and dichloromethane extracts mainly indicated the presence of terpenoids. The response to flavonoid detection tests in dichloromethane extracts decreased in Artemisia scoparia, Artemisia vulgaris, and Artemisia khorassanica, corresponding to a reduction in antifungal activity.
Conclusion
The highest extraction yield was observed in Artemisia scoparia (33%), followed by Artemisia vulgaris (32.5%), and Artemisia khorassanica (21.18%). All extracts showed weaker antifungal effects compared to the control (terbinafine). However, Epidermophyton floccosum and Trichophyton verrucosum exhibited higher resistance to terbinafine. Artemisia scoparia showed the highest and Artemisia khorassanica the lowest fungal growth inhibition among the tested extracts.
Ethical Statement
The present study was approved by the Research Ethics Committees of Kermanshah University of Medical Sciences (IR.KUMS.REC.1398.877).
Funding
This article is part of Mohammad Mahdi Hosseini's thesis for obtaining a professional doctorate in pharmacy from the Faculty of Pharmacy, Kermanshah University of Medical Sciences, Kermanshah, Iran.
Conflicts of Interest
The authors have no conflicts of interest.
Acknowledgement
We thank the Vice-Chancellor for Research and Technology at Kermanshah University of Medical Sciences for financial support.

Key Message
Some lipophilic compounds in various extracts, particularly petroleum ether and dichloromethane extracts of Artemisia scoparia, exhibit significant in vitro antifungal activity against dermatophytes.