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Fungal infections are a major global health problem, particularly in vulnerable populations such as immunocompromised individuals and those living in resource-limited settings. Traditional anti-fungal agents are often associated with high costs, side effects, and the emergence of drug-resistant strains. Therefore, there is a need for the development of new, safe, and effective anti-fungal agents. This study aimed to investigate the anti-fungal activity of mint essential oil against selected fungal strains. The leaves of mint were gathered, dried and powdered. After that, hydro distillation method was used for isolation of essential oil. The anti-fungal activity of the essential oil was evaluated against three fungal strains, including Aspergillus niger, Cryptococcus neoformans, and Trichophyton interdigitale, using a twofold dilution. The results showed that the mint essential oil had a significant effect on the growth of the fungi, with two of the three strains being highly sensitive. The concentrated essential oil was found to be most effective at preventing fungal growth. These findings suggest that Mint essential oil has potential as an anti-fungal agent, particularly against Cryptococcus neoformans. The use of different types of plant essential oil such as mint essential oil could provide a promising approach for the development of new anti-fungal agents, particularly in the context of drug-resistant infections.
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Eurasian J. Sci. Eng., 2023, 9(2), 162-169
https://doi.org/10.23918/eajse.v9i2p21
RESEARCH ARTICLE Open Access
Determination of Antifungal Activity of Essential Oil of Mentha piperita
(Mint)
Peshawa Y. Aziz 1 & Shokhan H. Azeez *2 & Nabaz Hamarashid Hama 3 & Hevar Nyaz
Abdulqadir 3 & Othman Mohammad 1
1 Department of Medical Laboratory Science, Technical College of Applied Science, Sulaimani
Polytechnic University, Sulaimani, Kurdistan Regional Government, Iraq
2* Department of Dental Nursing, Sulaimani Technical Institute, Sulaimani Polytechnic University,
Sulaimani, Kurdistan Regional Government, Iraq
3 Medical Laboratory Science Department, College of Science, Komar University of Science and
Technology, Sulaimani, Kurdistan Region, Iraq
Article History
Received: 25.03.2023
Revised: 28.05.2023
Accepted: 22.06.2023
Communicated by: Dr. Orhan Tug
*Email address:
shokhan.azeez@spu.edu.iq
*Corresponding Author
Copyright: © 2023 by the author.
Licensee Tishk International
University, Erbil, Iraq. This article is
an open access article distributed
under the terms and conditions of the
Creative Commons Attribution-
Noncommercial 2.0 Generic License
(CC BY-NC 2.0)
https://creativecommons.org/licenses/
by-nc/2.0/
Abstract:
Fungal infections are a major global health problem, particularly in vulnerable
populations such as immunocompromised individuals and those living in
resource-limited settings. Traditional anti-fungal agents are often associated
with high costs, side effects, and the emergence of drug-resistant strains.
Therefore, there is a need for the development of new, safe, and effective anti-
fungal agents. This study aimed to investigate the anti-fungal activity of mint
essential oil against selected fungal strains. The leaves of mint were gathered,
dried and powdered. After that, hydro distillation method was used for isolation
of essential oil. The anti-fungal activity of the essential oil was evaluated against
three fungal strains, including Aspergillus niger, Cryptococcus neoformans, and
Trichophyton interdigitale, using a two-fold dilution. The results showed that
the mint essential oil had a significant effect on the growth of the fungi, with
two of the three strains being highly sensitive. The concentrated essential oil
was found to be most effective at preventing fungal growth. These findings
suggest that Mint essential oil has potential as an anti-fungal agent, particularly
against Cryptococcus neoformans. The use of different types of plant essential
oil such as mint essential oil could provide a promising approach for the
development of new anti-fungal agents, particularly in the context of drug-
resistant infections.
Keywords: Antifungal, Hydro Distillation, Mentha Piperita, Multi Drug
Resistance
1. Introduction
Natural compounds from both plant and animal sources have traditionally been used in medicine due
to their broad therapeutic spectrum. Many scientists believe that our world's natural resources will be
the key to developing anti-fungal medicine. Although anti-fungal medications are generally effective
in treating fungal infections, there are instances where certain fungal infections may be resistant to
specific medications, requiring alternative treatment options, and as a result, people are suffering from
highly resistant fungal strains [1]. Because of advances in medicine, treating a fungal infection is far
more difficult than treating a bacterial infection. Fungi, like bacteria, can develop multi-drug
Eurasian J. Sci. Eng., 9(2) (2023), 162-169 P a g e | 163
resistance, making them considerably more hazardous when they cause infection. According to a
recent study on fungal infections, there are 150 million cases of severe fungal infections per year,
resulting in 1.7 million deaths, and the figure is still rising [1]. Scientists believe that Mentha piperita
can be employed in the medical area to treat microorganism infections because it is a natural resource
from our environment [2].
Cryptococcus neoformans, Candida albicans, and Trichophyton interdigitale are fungi that can cause
serious infections in humans. Cryptococcus neoformans can cause meningitis, particularly in
immunocompromised individuals, and is treated with anti-fungal agents such as fluconazole and
amphotericin B [3]. Candida albicans can cause a range of infections, including thrush, systemic
candidiasis, and invasive candidiasis, and is treated with anti-fungal agents such as fluconazole,
echinocandins, and amphotericin B [4]. Trichophyton interdigitale can cause various dermatophytosis
infections such as athlete's foot and nail infections, and is treated with topical anti-fungal agents such
as terbinafine and azoles [5].
Since ancient times Mentha has been used by different cultures for a variety of purposes. Mint has
numerous health advantages [6]. For example, it is a nutrient-dense food high in vitamin A, as well as
an antioxidant that aids the human body in eliminating cells harmed by free radicals [7]. Nowadays
one of the most frequent digestive system disorders is irritable bowel syndrome (IBS). According to
research, peppermint oil has a significant impact on the treatment of IBS. This is thought to be due to
methanol, a chemical in the oil that can help relieve IBS symptoms by relaxing the muscles of the
digestive tract [8]. A study by West et al, of the impact of peppermint oil in treatment of 700 IBS
patients, found that 75% of those who took peppermint oil showed signs of improvement in their IBS
symptoms [8].
Mint can also help with dyspepsia, Indigestion can occur when food remains in the stomach for an
extended period of time [9]. It has been found that using peppermint oil with meals helps food flow
through the stomach faster, which may alleviate symptoms of this form of indigestion [10]. Another
research project conducted a clinical experiment in which peppermint oil and caraway oil administered
as capsules demonstrated therapeutic effects similar to those seen with indigestion medication,
providing relief from stomach discomfort and other digestive issues [11].
Some researchers have stated that plant essential oil can increase brain function; however, the accuracy
of this claim has yet to be determined. In one of these researches, 144 adults who smelt peppermint oil
scent five minutes before testing found that it boosted their memory considerably [12]. Another study
discovered that sniffing peppermint oil while driving improved attention and reduce levels of
frustration, anxiety, and exhaustion among study participants [13].
Influenza is one of the most common winter ailments. Most flu medications contain methanol, which
is also present in peppermint oil. Hence, there are numerous studies indicating that mint can truly
benefit patients with cold symptoms [14]. Flu medication can be used as a nasal decongestant, to
remove congestion and improve airflow and breathing [15]. In addition, one recent study found that
mint helped participants to overcome cold symptoms by making them appear to go away [16].
Mint essential oil has been widely utilized as an anti-bacterial agent. As is well known, more bacteria
strains are developing resistance to drugs and because humanity is running out of options, scientists
are turning to natural medicines like mint oil to treat these illnesses. In one study, volatile oil extracted
from the leaves of Mentha piperita L. was used to treat multi drug resistance (MDR) strains from
hospitalized patients [17]. The researchers discovered that the minimum inhibitory concentration
Eurasian J. Sci. Eng., 9(2) (2023), 162-169 P a g e | 164
(MIC) was lower for Staphylococcus aureus, Escherichia coli, and Proteus mirabilis strains (20
mg/mL) and higher (40 mg/mL) for Klebsiella pneumoniae, Pseudomonas aeruginosa, and
Acinetobacter baumannii strains. The anti-bacterial qualities of mint essential oil are thought to be
derived from the components found in mint leaves, which are l-menthol, menthone, menthyl acetate,
and limonene [18]. These components within the leaf make it a good candidate for use as an anti-
bacterial, fungal, or viral treatment. More studies and investigations are now needed to determine the
true potencies of these medicinal aromatic herbs. The aim of the present study is to determine the
antifungal effect of the essential oil of Mentha piperita.
2. Methodology
2.1 Collection And Preparation Of Plant Essential Oil
The fresh mentha leaves were gathered in April 2022 from Chawg village, 4 kilometers from Halabja
city in the Kurdistan Region of Iraq. The University of Sulaimani's College of Agriculture supplied
voucher specimens that were used to identify the plant. Next, the leaves were dried and powdered
using a low-temperature (28°C) drying method. The drying process took place over a period of two
days and to minimize the loss of volatile compounds, the powdered leaves were sealed in an airtight
container. After that, hydro distillation procedure was used to isolate the essential oil. In accordance
with a documented protocol, 100g of powder was soaked in 350 ml of distilled water for 3 hours in a
conical flask [19]. After filtering, the essential oil was subsequently collected.
2.2 Microbial Strains
The Mentha essential oils were tested against Cryptococcus neoformans, Candida albicans and
Trichophyton interdigitale. These microorganisms were obtained from the Pasteur Institute of Iran in
Tehran. The fungal strains were stored at 4°C.
2.3 Agar Well Diffusion Method
In December 2022, at Komar University of Science and Technology's Microbiology lab, Department
of Medical Laboratory Science, antifungal activity against three different strains of fungi was
investigated. Each fungus was suspended in 100 microliters of water and adjusted to the McFarland
density standard of (0.7) [20]. Using sterilized cotton swabs, the suspensions were dispersed over
potato dextrose agar, and 75 microliters of mint essential oil at one concentration, 1/4, were added
directly to the wells using the agar well diffusion method. The essential oil was diluted with water
prior to the experiment. The plates were then incubated at a temperature of 25°C for a duration of six
days to allow for the diffusion of the essential oil components. After the incubation period, the diameter
of the inhibitory zone (mm) surrounding the wells was measured as an indicator of antimicrobial
activity [20].
3. Results
According to the findings, the Mint essential oil is highly sensitive to two of the three strains tested.
In the study, the mint essential oil was shown to be most effective in preventing fungal growth. The
effect of the essential oil differed based on the strain. Figure 1 shows that Trichophyton interdigitale
had the smallest inhibitory zone, while Cryptococcus neoformans had the biggest, measuring 24mm
for the 1/4 concentration, and Candida albicans had a medium sized inhibitory zone, measuring 17mm
(Figures 2 & 3 and Table 1).
Eurasian J. Sci. Eng., 9(2) (2023), 162-169 P a g e | 165
Figure 1: Shows anti-fungal activity of mint essential oil against Trichophyton interdigitale.
Figure 2: Shows anti-fungal activity of mint essential oil against Cryptococcus neoformans.
Figure 3: Shows anti-fungal activity of mint essential oil against Candida albicans.
Eurasian J. Sci. Eng., 9(2) (2023), 162-169 P a g e | 166
Table 1: shows the average of inhibition zone in (mm) for each strain of fungus
Fungus Name
Replicate 1
Inhibition
zone (mm)
Replicate 2
Inhibition zone
(mm)
Replicate 3
Inhibition
zone (mm)
Cryptococcus
neoformans
23
24
25
Candida albicans
16.5
17.5
17
Trichophyton interdigit
0
0
0
4. Conclusion and Discussion
Essential oils from the Mentha leaves were successfully isolated by hydro distillation. The different
types of essential oils are important natural materials. These are used in food production and in
industrial applications. Essential oils of various species of Mentha may contain antibacterial and
antifungal compounds which are important for preservation. The current study has demonstrated the
antifungal activity of Mentha essential oil against several human fungal pathogens.
The data of this study showed that Mentha oil exhibited sufficient antifungal activity against
Cryptococcus neoformans and Candida albicans but had no detectable effect on Trichophyton
interdigitale. It is the first study in iraqi kurdistan that used mint eesential oil on these specius of fungi.
In another study, some important plant extracts (mint, pomegranate and coriander) have been
investigated on Candida glabrata specius and all three plant extracts showed antifungal properties
[21]. In other research, the essential oil of pepperment showed significant antifungal activity against
Alternaria alternaria, Fusarium tabacinum, Penicillum spp., Fusarium oxyporum, and Aspergillus
fumigates [22].
The previous studies showed that the essential oils of mint possess antimicrobial activity [23,24]. The
Mentha oils have demonstrated a broad spectrum of effects against the fungal species [25]. In a
different study, essential oils of various herbs including Mentha spicata and Mentha piperita were
tested against microbial food poisoning and human pathogens, M. spicata oils contained carvone and
menthone, these compounds showed antifungal activity even more powerful than that of commercial
fungicide bifonazole [26]. It is concluded that the essential oils of Mentha have great antifungal
potential against some of the fungal species and could be employed as natural fungicides and
preservatives. This study provides an overview on the susceptibility of human fungi towards mint
essential oils and their constituents. In another study, the effect of Mentha oil on Salmonella enteritidis
and Listeria monocytogenes in a suitable medium was analyzed,    
activity basically relied on its concentration, pH, composition, temperature and type of microbe [27].
In research by Devkatte and his coworkers in 2005, data on peppermint oil against C. Albicans showed
an inhibition zone of approximately 18 mm [28]. Furthermore, Agarwal and his coworkers in 2010
found a 22.2 mm inhibition zone for Mentha oil against C. Albicans [29]. The well diffusion data
obtained in the current study showed similar or slightly different results in comparison with the above
studies. Thus, essential oils of Mentha might have potential antimicrobial characteristics.
In conclusion, the results of this study demonstrate the anti-fungal activity of Mint essential oil against
selected fungal strains. Regarding the use of Mint essential oil as a natural and safe anti-fungal agent,
Eurasian J. Sci. Eng., 9(2) (2023), 162-169 P a g e | 167
further investigation is needed to determine the mechanism of action of the essential oil and to evaluate
its safety and efficacy in vivo.
5. Author’s Contribution
“Authors confirm that the manuscript has been read and approved by all named authors. We also
confirm that each author has the same contribution to the paper. We further confirm that the order of
authors listed in the manuscript has been approved by all authors.”
6. Conflict of Interest

7. Acknowledgments
The authors would like to thank Komar University of Science and Technology for their help to perform
this study.
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... Particularly the essential oil's inhibition effect against C. utilis at a concentration of 62.5 µg/ml (MIC), which is consistent with other studies (Mkaddem et al., 2009). The antimicrobial activity of the essential oil, especially towards Candida sp., according to the literature, can be attributed to the presence of active compounds such as pulegone (Dhingra and Chopra, 2023), 1,8-Cineole (Shahina et al., 2022), and menthone (Aziz et al., 2023). However, the synergistic effect of minor components of the essential oil still needs to be considered. ...
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It is important to realize that guidelines cannot always account for individual variation among patients. They are not intended to supplant physician judgment with respect to particular patients or special clinical situations. IDSA considers adherence to these guidelines to be voluntary, with the ultimate determination regarding their application to be made by the physician in the light of each patient's individual circumstances. © 2015 The Author 2015. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail [email protected]/* */
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