Archives of

1. Introduction
Because of socio-economic growth and industrialization, the physical activity of mankind has remarkably decreased, entailing chronic, deteriorating consequences such as obesity, diabetes mellitus, and cardiovascular disorders [1-3]. Hence, nowadays, sports and fitness activities, whether for recreation or professionally, are more emphasized for improving body health status [4]. Wrestling has been a symbol of power and virility for people in a number of countries such as Iran, and many youth Iranians are involved in wrestling throughout the country. Due to the combating nature of this sports activity, profuse body sweating, and persistent contact with the fighting mat, wrestlers are mostly exposed to acquiring and/or transmitting fungal infections [5].
Nevertheless, poor hygienic conditions both for the sportsman himself/herself and/or the sporting environment, would not only endanger the health of the surrounding individuals from a public health perspective but also affect one’s ability to compete [6]. Primary preventive measures comprising routine showering and public awareness have been advocated, but still not adequately effective [7]. Thus, the causative agent must be identified to barricade infectious outbreaks [8]. Evidence-based data demonstrate that wrestling clubs if they are not controlled or examined, are considered suitable places for the growth of pathogenic fungi [9,10]. Accordingly, the surfaces of different devices such as mats, locker rooms, towels, floors, and W.C. walls are also suitable for the growth of dermatophyte species such as Trichophyton and Epidermophyton, as well as saprophytic fungi, including Mucor, Rhizopus, Aspergillus, Penicillium, and Scopulariopsis [11]. An important aspect of opportunistic fungi is the pathogenic potential for immune deficient and immune-compromised subjects. Furthermore, Saprophytes can cause nail dystrophy. According to previous studies, the most common etiologic agents of onychomycosis are Aspergillus spp., Acremonium spp., Scopulariopsis spp., Penicillium spp., and fusarium with an incidence rate of 1.43%-17.6% [12]. In sports with skin-to-skin contact, including wrestling ringworm, Tinea gladiatorum, has been found to be more common [13-17]. Trichophyton tonsurans was also isolated from wrestling mats in Iran [18,19].
Additionally, bodybuilding clubs are among extremely populated places, facilitating the risk of athletes’ exposure
Fungal Contamination of Wrestling and Bodybuilding Clubs in Hamedan Province, Western Iran
Zainab Sadeghi Dehkordi1*ID, Aso Hajiazizi1ID, Shahriar Keihani1ID, Bahareh Kordi2ID, Ali Sadeghinasab3ID
1Department of Pathobiology, Faculty of Veterinary Science, Buali Sina University, Hamedan, Iran
2PhD Student, Department of Parasitology, Faculty of Veterinary Medicine - Ferdowsi University of Mashhad. Mashhad, Iran
3Department of Clinical Sciences, Bu-Ali Sina University, Hamedan, Iran
*Corresponding Author: Zainab Sadeghi Dehkordi, Emails: z.sadeghidehkordi@basu.ac.ir and dehkordisz@gmail.com
Received: April 4, 2022, Accepted: September 11, 2022, ePublished: December 29, 2022
https://jhygiene.muq.ac.ir/
10.34172/AHS.11.4.208.2
Vol. 11, No. 4, 2022, 286-290
Original Article
Arch Hyg Sci. Volume 11, Number 4, 2022 287
Fungal Contamination of Clubs, Hamedan
to mycoses. Moreover, such perimeters possess several
fomites such as towels, practice clothing and wrestling
mats, occlusion, trauma, shower sharing, and sweating
with resultant maceration of the epidermis which may
carry the sources of infection [5]. Therefore, understanding
different transmission routes of fungal contamination
is crucial in planning effective interventions to break
the chain of transmission. Although extensive surface
contamination with fungi has been demonstrated, there
is no information available on the health status of the
devices, tools, and surfaces of wrestling and bodybuilding
clubs in Hamedan. Thus, this study was the first one to
report the fungal contamination of tools and surfaces in
wrestling and bodybuilding clubs in Hamedan province,
western Iran in 2018.
2. Materials and Methods
Nine sports centers in Hamedan province having wrestling
and bodybuilding clubs were enrolled in this crosssectional
study from September 2018 to May 2019. These
clubs were located in different areas such as Hamedan
(n = 4), Bahar (1), Malayer (1), Razan (1), Asadabad (1),
and Maryanaj (1) (Figure 1). Seven samples were taken
for each club. Initially, a number of 100 pieces of mats
(10 × 10 cm) were prepared and autoclaved at 121ºC for
15 minutes. For each wrestling club, sufficient pieces of
mats were placed in direct contact with various surfaces,
including the wrestling mat, parquet, wall, and floor,
WC bathroom, locker room, and the bodybuilding club.
Next, each piece was kept within a clean nylon, tagged,
and transferred to the laboratory. For air sampling, the
Sabouraud dextrose agar (SDA) culture medium, which
was placed 1 m higher than the earth’s surface, was
exposed to the environmental air for 30-60 minutes.
By tapping the back surface of each contaminated mat
piece, the culture mediums were inoculated by the mat
components. The media were sealed and incubated at
room temperature (SDA) or 37ºC (sabouraud dextrose
agar (SSD)) for the aim of fungal growth. Macroscopic
and microscopic examinations were performed following
2-3 weeks of fungal growth [5]. The slide culture method
was implicated in the case of difficulty in diagnosis. Fungi
were identified mostly by the close examination of their
morphology and characteristics. In slide cultures, the
fungi were grown directly on the slide on a thin film of
agar. Accordingly, there was no need to remove a portion
of the fungus from a culture plate and transfer it to the
slide. Briefly, a sterile U-shaped glass rod on the filter
paper was placed [8], and enough sterile water was poured
(about 4 mL) onto the filter paper to completely moisten
it. Then, a sterile slide was placed on the U-shaped rod
with forceps. Gently, a scalpel was flamed to sterilize, and
a 5-mm square block of the medium was cut from the
plate of Sabouraud’s agar. The block of agar was picked
up by inserting the scalpel, and this block was carefully
transferred aseptically to the center of the slide. Four sides
of the agar square were inoculated with spores or mycelial
fragments of the fungus for examination. Aseptically, a
sterile cover glass was placed on the upper surface of the
agar cube. Then, the cover was placed on the petri dish
and incubated at room temperature for 48 hours. After 48
hours, the slide was examined under low power. If growth
Figure 1. Hamedan Province Geographical Location
Sadeghi Dehkordi et al
288 Arch Hyg Sci. Volume 11, Number 4, 2022
had occurred, there would be the growth of hyphae and
the production of spores. If growth was inadequate and
spores were not evident, the mold was allowed to grow
for another 24-48 hours before making the stained slides.
Next, a drop of lactophenol cotton blue stain was placed
on a clean microscope slide [5,8]. The cover glass was
removed from the slide culture, and the block of agar
was discarded. A drop of 95% ethanol was added to the
hyphae on the cover glass. As soon as most of the alcohol
was evaporated, the cover glass was placed, and the mold
was placed side down on the drop of lactophenol cotton
blue stain on the slide. Finally, given that the microscopic
examination can help identify fungal infections using
morphological features, the slides were examined under a
microscope [20]. Eventually, the chi-square test was used
for the investigation of the relevance between variables.
3. Results
The results of the present investigation demonstrated
that out of 100 examined specimens from nine different
sports centers in Hamedan province, 68 specimens were
positive for fungal culture. The most frequently isolated
fungal genera included Rhizopus (24%), Penicillium
(24%), Aspergillus flavus (23%), Aspergillus niger (19%),
yeast (7%), Scopulariopsis (6%) Mucor, and Ulocladium
(1%), the details of which are presented in Table 1. No
dermatophyte contamination was observed in any of
the samples. The most contaminated surfaces (place of
sampling) were wrestling mats (95.7%), followed by the
floor and wall (89.5%), air (86.7%) WC (82.4%), and
locker room (50%). The findings revealed that there
was a statistically significant association between the
total fungal load and different kinds of sampling places
(P = 0.019).
No fungus was detected in the culture of 32 specimens.
The highest and lowest prevalence rates of fungal
contamination were related to Razan and Malayer (87.5
and 72.7%), respectively. No significant association was
found between the study area and fungal genera or species
(P = 0.868). The highest isolated fungi were Rhizopus
and Penicillium (24%) while the lowest ones were yeast,
Mucor, and Ulocladium (1%).
4. Discussion
To the best of our knowledge, this is the first study to
assess fungal contamination on inanimated objects (i.e.,
mat, floor and wall, locker room, bath and WC, parquet,
and environmental air). This study also provided insight
into the epidemiology and distribution of various fungal
contamination in nine wrestling and bodybuilding body
building clubs in Hamedan province, western Iran. In
the present study, 88.8% of the clubs were positive for at
Table 1. Distribution of isolated fungi species according to the place of sampling (surfaces) and area
Mat Bath-WC Floor-Wall Lock Room Parquet
Bodybuilding
Tool
Air
Club 1 - -
Penicilium
A. niger
A. flavus
Rhizopus
Penicilium
A. flavus
Yeast
Rhizopus
A. niger
Ulocladium
Rhizopus
Penicilium
Rhizopus
Penicilium
A. flavus
Club 2
Rhizopus
A. flavus
Yeast
A. niger
A. flavus
Yeast
Penicilium
A. flavus
Yeast - - Yeast
Club 3 Penicilium -
A. niger
A. flavus
- - -
A. niger
A. flavus
Club 4
Rhizopus
Yeast
- Rhizopus - - -
Rhizopus
Scopulariopsis
Club5
Rhizopus
A. flavus
Scopulariopsis
A. flavus
Penicilium
A. flavus
Scopulariopsis
-
Penicilium
A. flavus
-
Penicilium
A. flavus
Scopulariopsis
Club 6
Penicilium
A. niger
Rhizopus
Mucor
Penicilium
Yeast
- Penicilium
Penicilium
A. niger
A. flavus
Rhizopus
Penicilium
A. niger
Yeast
-
Club7
A. niger
Yeast
Rhizopus
Penicilium
Yeast
- -
Penicilium
Yeast
A. flavus
A. niger
Rhizopus
Rhizopus
Club 8
Penicilium
A. niger
A. flavus
Rhizopus
Penicilium
Yeast
A. flavus
Penicilium
A. niger
Rhizopus
Scopulariopsis
- - Yeast -
Club 9
Rhizopus
A. flavus
Rhizopus
Penicilium
A. flavus
Rhizopus
Penicilium
- - - Penicilium
Note. **Club 1-4: Hamedan; Club 5: Maryanaj; Club 6: Razan; Club 7: Asadabad; Club 8: Bahar; Club: Malayer; A. flavus: Aspergillus flavus; A. niger: Aspergillus niger.
Arch Hyg Sci. Volume 11, Number 4, 2022 289
Fungal Contamination of Clubs, Hamedan
least one saprophytic fungal agent. Saprobes are a group of
fungal species with saprophytic nutrition via a process of
chemoheterotrophic extracellular digestion, being involved
in the decaying organic matter. They may be implicated
in wrestlers’ mycoses through wrestling mats, WC and
bathroom, wall and floor, parquet, bodybuilding clubs, and
the environmental air. An important aspect of opportunistic
fungi is their pathogenic potential for immune deficient
and immune compromised subjects [12].
In sports with skin-to-skin contact such as wrestling,
T. tonsurans was isolated from wrestling mats in France
and Sweden [21,22]. In Iran, previous studies showed
that dermatophyte infection may vary among wrestlers
ranging from 24 to 47.5% [23,24]. T. tonsurans and
Trichophyton rubrum were also isolated from wrestling
mats in different regions of Iran [5,18,19,25,26].
No dermatophyte species were isolated in the current
study. Some reasons could illustrate why we were unable
to detect dermatophytes from the cultures of different
objects. The samples were taken after sterilizing the mats
and other objects. All tools and equipment were washed
or disinfected after the practice time. The disinfectant
used by each club contains an active fungicidal ingredient.
These would have not revealed the presence of ringworm
fungi. In addition, dermatophytes were probably there,
but we did miss our chance by sampling at the end of the
day. We may have also missed the opportunity to find any
dermatophyte by sampling twice a week. Further, had no
access to the specimens of wrestler skin or clothing the
wrestlers. Dermatophytes are fungi that require keratin
for growth and spread by direct contact from other
people. In this regard, our findings are in line with those
of previous studies by Kohl et al [27] and Ghasemi et al
[12].
5. Conclusion
In this study, the most contaminated places of sampling
were wrestling mats (95.7%) and the floor and wall
(89.5%). We would recommend that common-sense
hygiene measurements be continued, including showering
after every encounter, washing practice clothes daily, and
disinfecting mats daily [28]. Educating wrestlers, coaches,
parents, and members of the medical community about
skin infections and their prevention, recognition, and
treatment is crucial and a part of our continuing effort
[29]. The surface should then be cleaned with soap and
water or wiped with a clean paper towel lightly wetted
with a germicide registered with environmental surfaces.
These measures are applicable for most nonabsorbent
athletic surfaces. Surfaces should be allowed to dry
sufficiently to prevent possible injuries [30].
Acknowledgments
The authors would like to thank Somayeeh Moradi and Sakineh
Azami, the technicians of Parasitological Laboratory in Veterinary
Science Faculty, Bu-Ali Sina University.
Author Contributions
Conceptualization: Zainab Sadeghi Dehkordi.
Methodology: Zainab Sadeghi Dehkordi, Aso Hajiazizi, Shahriar
Keihani.
Validation: Zainab Sadeghi Dehkordi.
Formal Analysis: Ali Sadeghinasab.
Investigation: Zainab Sadeghi Dehkordi, Bahareh Kordi.
Resources: Zainab Sadeghi Dehkordi.
Data Curation: Zainab Sadeghi Dehkordi, Ali Sadeghinasab.
Writing—Original Draft Preparation: Zainab Sadeghi Dehkordi,
Bahareh Kordi.
Writing—Review and Editing: Zainab Sadeghi Dehkordi.
Visualization: Zainab Sadeghi Dehkordi, Shahriar Keihani, Aso
Hajiazizi.
Supervision: Zainab Sadeghi Dehkordi.
Project Administration: Zainab Sadeghi Dehkordi.
Funding Acquisition : all of Authors.
Conflict of Interests
The authors declare that they have no competing interests.
Funding
This research received no grant from Bu-Ali Sina University.
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