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KSÜ Tarım ve Doğa Derg 25 (1): 83-87, 2022
KSU J. Agric Nat 25 (1): 83-87, 2022
https://doi.org/10.18016/ksutarimdoga.vi.880151
Biological Activities of Wild Poisonous Mushroom
Entoloma sinuatum
(Bull.) P. Kumm
(Boletales)
Celal BAL1 , Hayri BABA2, Ilgaz AKATA3, Mustafa SEVINDIK4, Zeliha SELAMOGLU5, Hasan AKGÜL6
1Gaziantep University, Oguzeli Vocational School, Gaziantep, Turkey, 2Hatay Mustafa Kemal University, Faculty of Science and Literature,
Department of Biology, Hatay, Turkey, 3Ankara University, Science Faculty Department, Biology, Ankara, Turkey, 4Department of Food
Processing, Bahçe Vocational School, Osmaniye Korkut Ata University, Osmaniye, Turkey, 5Nigde Ömer Halisdemir University, Faculty of
Medicine, Department of Medical Biology, Nigde, Turkey, 6Akdeniz University, Science Faculty, Biology Department, Antalya, Turkey.
1https://orcid.org/0000-0001-6856-3254, 2https://orcid.org/0000-0002-1837-4321, 3https://orcid.org/0000-0002-1731-1302
4https://orcid.org/0000-0001-7223-2220, 5https://orcid.org/0000-0001-9056-6435, 6https://orcid.org/0000-0001-8514-9776
: bal@gantep.edu.tr
ABSTRACT
In many studies conducted in recent years, it has been found that the
beneficial natural ingredients found in mushrooms are very important
in curing various diseases and eliminating disease factors. It has been
reported that fungi that spread in different countries have biological
activities. In this study, antioxidant level (TAS), oxidant level (TOS),
and antibacterial and antifungalactivity of
Entoloma sinuatum
(Bull.)
P. Kumm were determined. Ethanol extract of the mushroom was
extracted in a Soxhlet apparatus. Antioxidant and oxidant levels were
determined using Rel Assay TAS and TOS kits. Also, antimicrobial
activity was tested against bacterial and fungal strains using the agar
dilution method. As a result of the studies, the TAS value of the
mushroom was determined 2.64±0.15, the TOS value was 6.58±0.23,
and the OSI (Oxidative stress index) value was 0.25±0.02. In addition,
it was determined that the ethanol extract of the mushroom was
effective against bacteria at 200 and 400 μg/mL, and against fungi at
50 μg/mL concentrations. As a result, it was determined that
E.
sinuatum
is a natural antioxidant and antimicrobial agent.
Research Article
Yabani Zehirli Mantar
Entoloma sinuatum
(Bull.) P. Kumm 'un (Boletales) Biyolojik Aktiviteleri
ÖZET
Son yıllarda yapılan birçok çalışmada mantarlarda bulunan faydalı
doğal bileşenlerin çeşitli hastalıkları iyileştirmede ve hastalık
etkenini ortadan kaldırmada çok önemli olduğu bulunmuştur. Farklı
ülkelerde yayılış gösteren mantarların biyolojik aktivitelerinin olduğu
bildirilmiştir. Bu çalışmada Türkiye’de Entoloma sinuatum (Bull.) P.
Kumm’un antioksidan seviyesi, oksidan seviyesi ve antimikrobiyal
aktivitesi belirlendi. Mantarın etanol özütü soxhlet cihazında
çıkarıldı. Antioksidan ve oksidan seviyeleri Rel Assay TAS ve TOS
kitleri kullanılarak belirlendi. Ayrıca antibakteriyel ve antifungal
aktivite agar dilüsyon metodu kullanılarak bakteri ve fungus
suşlarına karşı test edildi. Yapılan çalışmalar sonucunda mantarın
TAS değeri 2.64±0.15, TOS değeri 6.58±0.23 ve OSI (Oksidatif stres
indeksi) değeri 0.25±0.02 olarak belirlendi. Ayrıca mantarın etanol
özütünün bakterilere karşı 200 ve 400 μg/mL, funguslara karşı 50
μg/mL konsantrasyonlarda etkili olduğu belirlendi. Sonuç olarak E.
sinuatum'un doğal antioksidan ve antimikrobiyal ajan olduğu
belirlendi.
Araştırma Makalesi
To Cite :
Bal C, Baba H, Akata I, Sevindik M, Selamoğlu Z, Akgül H 2022. Biological Activities of Wild Poisonous
Mushroom
Entoloma sinuatum
(Boletales). KSU J. Agric Nat 25 (1): 83-87. https://doi.org/10.18016/
ksutarimdoga.vi.880151.
INTRODUCTION
In terms of nutritional values of mushrooms, it has
been utilized in the treatment of cancer, cholesterol-
lowering, stress, insomnia, asthma, allergies, and
diabetes, and it has been included in studies that it is
effective. Besides, mushrooms contain various
KSÜ Tarım ve Doğa Derg 25 (1): 83-87, 2022
KSU J. Agric Nat 25 (1): 83-87, 2022
Araştırma Makalesi
Research Article
84
secondary metabolites, including phenolic compounds,
polyketides, terpenes, and steroids that act as
antioxidants (Ahmed et al., 2015; İnci and Kırbağ,
2018; Krupodorova and Sevindik, 2020). In studies on
natural mushrooms in different parts of the world, it
has been reported that mushrooms have different
biological activities such as antioxidant, antimicrobial,
anticancer, antitumor, antiproliferative, anti-
inflamatuvar, DNA protective, anti-aging, and
antiallergic (Bae et al., 2007; Bal et al., 2017; Garcia-
Lafuentea, et al., 2010; Osman and Toliba, 2019;
Sevindik et al., 2017; Zhang et al., 2019; Umaña et al.,
2020). In this context, the discovery of new natural
products has been inevitable, especially in recent
years, due to the interest of people in natural products
and the possible side effects of synthetic drugs. In this
study, wild poisonous mushroom
Entoloma sinuatum
(Bull.) P. Kumm was used as a material.
E. sinuatum
(livid entoloma, livid agaric, livid pinkgill,
leaden entoloma, and lead poisoner) is a poisonous
fungus that has been detected in many regions of the
world. Fruiting bodies of the mushroom appear in late
summer and autumn (Zeitlmayr, 1976). It is seen in
deciduous woodlands covered with clay and calcareous
soils, usually Oak and Beech, rarely in birch or in
nearby park areas. Sometimes it meets in the form of
a fairy ring. It can be seen alone or in groups and most
members of the genus are saprotrophic. In young form,
it can be mixed with the edible
Calocybe gambosa
or
Clitopilus prunulus
. Also, an ordinary observer might
confuse
E. sinuatum
with the meadow mushroom
Agaricus campestris
(Lamaison and Polese, 2005). For
this reason, it has played a leading role in many
mushroom poisoning cases in Europe.
E. sinuatum
poisoning is generally not fatal. Following
consumption of mushrooms, the gastrointestinal
syndrome is observed. Depression and delirium are
rare sequelae (Bastida et al., 1987). In this study, the
antioxidant, oxidant, and antimicrobial activities of
wild poisonous mushroom
E. sinuatum
were
determined.
MATERIALS and METHODS
Sample extraction
Mushroom samples were collected from the Belgrad
forest (41°10'56.63"N- 28°59'5.91"E, 145m)
(Istanbul/Turkey). After the samples were collected,
they were dried in a dryer (Profilo, PFD2350W) for
about 12 hours at 45 °C and 10 g of each mushroom
sample was taken and ground into powder
(WANXINGBO, Grinder). Powder samples were
extracted with 200 mL of ethanol for 6 h in the Soxhlet
apparatus then obtained by filtration through filter
paper (ISOLAB, 125mm) and the solvent was removed
on a rotary evaporator (Heidolph Laborota 4000
Rotary Evaporator) at 40 °C. Crude extracts were
stored at + 4 ° C until further conducting experiments.
Antibacterial activities
The antimicrobial activity tests of ethanol extract of
the mushroom sample were determined by the agar
dilution method. The lowest concentration of the
extract preventing the growth of microorganisms was
determined as the MIC (Minimum Inhibitor
Concentration) value. Extract concentrations were set
at 6.25 to800 µg/mL in distilled water (CLSI, 2002;
CLSI, 2003).
Staphylococcus aureus
(ATCC 29213),
Enterococcus faecalis
(ATCC 29212),
Escherichia coli
(ATCC 25922) and
Pseudomonas
aeruginosa
(ATCC
27853) were used as test bacteria. Bacteria were pre-
cultured in Muller Hinton Broth medium. Amikacin,
Ampicillin, and Ciprofloxacin were used as positive
controls (Hindler et al., 1992).
Antifungal activities
Candida
albicans
(ATCC 10231) and
C. tropicalis
(ATCC 13803) were used as test fungi. Fungi were pre-
cultured in RPMI 1640 Broth medium. Fluconazole
and Amphotericin B were used as positive controls
(Hindler et al., 1992).
Antioxidant activities
The total antioxidant status, total oxidant status, and
oxidative stress index of ethanol extracts of
mushrooms were evaluated using Rel Assay TAS and
TOS kits. Trolox was utilized as a calibrator in
antioxidant studies. Hydrogen peroxide was used as a
calibrator in oxidant studies (Erel, 2004; Erel, 2005).
The OSI (Oxidative stress index) value is determined
by proportioning the TAS value to the TOS value. The
following formula was used for the OSI value
(Sevindik, 2018).
RESULTS and DISCUSSION
Antibacterial and Antifungal activities
Antibacterial and antifungal drugs used today are
inadequate due to the antibiotic resistance that
microorganisms have gained in recent years.
Moreover, people turned to natural products due to the
possible side effects of antibacterial and antifungal
drugs (Liu et al., 2017; Abdalla et al., 2020). This trend
has made the discovery of new antibacterial and
antifungal natural products. In this study, the
antibacterial and antifungal activities of the ethanol
extract of
E. sinuatum
was determined. The findings
obtained were shown in Table1.
In previous studies on
Entoloma
species, the methanol
extract of
Entoloma speculum
was reported to be
effective against
Xanthomonas campestris,
Pseudomonas syringae, Agrobacterium tumefaciens,
Klebsiella pneumonia, Escherihia coli, Salmonella
typhi, P. aeruginosa, Staphylococcus aureus,
Streptomyces pneumoneae, Candida albicans,
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KSU J. Agric Nat 25 (1): 83-87, 2022
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Research Article
85
Chrysosporium merdarium, Trichophyton rubrum,
Chrysosporium keratinophilum, Fusarium solani,
Penicillium chrysogenum, Aspergillus flavus
and
A.
niger
in different concentrations (Kodiyalmath and
Krishnappa, 2018). In another study, ethanol extract
of
Entoloma
nubigenum
was reported to be effective
against
Bacillus subtilis, Escherichia faecalis,
Staphylococcus aureus, Penicillium notatum,
and
Ceratocistys pilifera
(Reinoso et al., 2013). In our
study, the antibacterial and antifungal activities of
ethanol extract of
E. sinuatum
was determined. As a
result of this, it has been determined to be effective
against
S. aureus
and
P. aeruginosa
in 400 µg/mL
concentration,
E. faecalis
and
E. coli
at 200 µg/mL
concentration,
C. albicans
and
C.
tropicalis
at 50
µg/mL concentration. In other words, the antifungal
activity of the mushroom extract appears to be higher.
It was also determined that the fungus can be used as
a natural antibacterial and antifungal source.
Table 1. Antibacterial and antifungal activities of ethanol extract of
E. sinuatum
Çizelge 1. E. sinuatum'un etanol ekstresinin antibakteriyel ve antifungal aktiviteleri
Sample
Antibacterial (µg/mL)
Antifungal (µg/mL)
S. aureus
E. faecalis
E. coli
P. aeruginosa
C. albicans
C. tropicalis
Entoloma sinuatum
400
200
200
400
50
50
Flukonazole
-
-
-
-
1.56
3.12
Ampicillin
3.12
1.56
3.12
-
-
-
Ciprofloxacin
0.78
0.78
1.56
3.12
-
-
Antioxidant activities
Antioxidant compounds are found in many natural
products including mushrooms that spread in many
different ecosystems. Mushrooms are very vital
sources of antioxidants with their numerous
antioxidant-effective enzymes and phenolic
compounds (Lu et al., 2018; İnci et al., 2019).
Table 2. TAS, TOS and OSI Values of ethanol extract of
E.
sinuatum
Çizelge 2. E. sinuatum'un etanol ekstresinin TAS, TOS ve
OSI Değerleri
Sample
TAS (mmol/L)
TOS (µmol/L)
OSI
E. sinuatum
2.64±0.15
6.58±0.23
0.25±0.02
Values are presented as mean±S.D
Many studies have shown that mushrooms have high
antioxidant activities. The total antioxidant and total
oxidant status of used in our study were determined
for the first time. In previous TAS and TOS studies on
wild mushrooms, TAS value of
Tricholoma
virgatum
was reported as 3.754, TOS value was 8.362 and OSI
value was 0.223 (Selamoğlu et al., 2020). The TAS
value of
Cerioporus varius
was reported as 2.312, TOS
value was 14.358 and OSI value was 0.627 (Sevindik,
2019). TAS value of
Suillus granulatus
was reported as
3.143, TOS value was 18.933 and OSI value was 0.603
(Mushtaq et al., 2020). The TAS value of
Lactifluus
rugatus
was reported as 3.237, the TOS value was
8.178 and the OSI value was 0.254 (Sevindik, 2020).
The TAS value of
Infundibulicybe geotropa
was
reported as 1.854, TOS value was 30.385 and OSI
value was 1.639 (Sevindik et al., 2020). The TAS value
of
Lepista nuda
was reported as 3.102, the TOS value
was 36.920, and the OSI value was 1.190 (Bal et al.,
2019). Compared to these studies, the TAS value of
E.
sinuatum
was found to be higher than
I. geotropa
and
C. varius
, lower than
L. rugatus, L. nuda, S.
granulatus
and
T. virgatum
. TAS value shows the
whole of the antioxidant compounds produced within
the mushroom (Krupodorova and Sevindik, 2020). The
difference in TAS values determined in studies on
different mushrooms draws attention. The difference
in TAS value between species is due to the difference
in the potential of fungi to produce compounds with
antioxidant properties.
When TOS values were examined,
E. sinuatum
was
determined to be lower than
C. varius, L. rugatus, I.
geotropa, L. nuda
and
T. virgatum
. The TOS value
indicates the whole of the oxidant compounds produced
by the fungus as a result of environmental effects
metabolic activities (Krupodorova and Sevindik, 2020).
TOS value of
E. sinuatum
used in our study was
generally found to be low. In addition, the OSI value
shows how much the oxidant compounds produced in
the mushroom's body are suppressed by the
endogenous antioxidants (Krupodorova and Sevindik,
2020).
E. sinuatum
suppressed better than
I. geotropa,
L. rugatus, S. granulatus, L. nuda,
and
C. varius
, less
than
T. virgatum
. Determining the oxidative stress
index is important in determining the antioxidant
potential of the mushroom. The OSI value shows the
success of suppressing the oxidant compounds
produced by the fungus by the antioxidant defence
system. In this context, it is seen that the antioxidant
defence system of
E. sinuatum
is successful in
suppressing oxidant compounds. As a result, it was
determined that
E. sinuatum
has antioxidant
potential.
CONCLUSION
In this study, the antioxidant, oxidant, and
antimicrobial potential of wild poisonous mushroom
E.
sinuatum
was determined. As a result, it was found
that the antioxidant potential of the mushroom was at
KSÜ Tarım ve Doğa Derg 25 (1): 83-87, 2022
KSU J. Agric Nat 25 (1): 83-87, 2022
Araştırma Makalesi
Research Article
86
normal levels. It was also found that the oxidant level
was low, yet, fungi were more effective against fungi.
Besides, it was determined that
E. sinuatum
could be
a natural antioxidant and antimicrobial source.
ACKNOWLEDGEMENTS
We thanks to Osmaniye Korkut Ata University,
Central Research Laboratory for their support.
Researchers Contribution Rate Declaration Summary
The authors declare that they have contributed equally
to the article.
Conflicts of Interest Statement
The article authors declare that they do not have any
conflict of interest.
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