Hindawi

Evidence-Based Complementary and Alternative Medicine

Volume 2021, Article ID 3025848, 10 pages
https://doi.org/10.1155/2021/3025848

Research Article
Phytochemical Profile and Antimicrobial Activities of Edible
Mushroom Termitomyces striatus

Concepta N. W. Sitati ,1 Kenneth O. Ogila,2 Rebecca W. Waihenya,2 and Lucy A. Ochola3

1
Department of Biological Sciences, School of Pure and Applied Sciences, Mount Kenya University, P.O. Box 1869-30200,
Kitale, Kenya
2
Zoology Department, School of Biological Sciences, Jomo Kenyatta University of Agriculture and Technology,
P.O. Box 62000-00200, Nairobi, Kenya
3
Institute of Primate Research, P.O. Box 24481-00502, Karen, Nairobi, Kenya

Correspondence should be addressed to Concepta N. W. Sitati; [email protected]

Received 6 April 2021; Revised 4 June 2021; Accepted 16 September 2021; Published 19 October 2021

Academic Editor: Shih-Chao Lin

Copyright © 2021 Concepta N. W. Sitati et al. This is an open access article distributed under the Creative Commons Attribution
License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is
properly cited.
The mushroom Termitomyces striatus is an edible mushroom that grows wildly and belongs to the family Lyophyllaceae. Studies in
the last few decades have demonstrated that mushrooms and their active components have beneficial effects on a variety of
biological systems. Some mushrooms do exhibit antibacterial properties. Qualitative phytochemical profile was done on the
mushroom Termitomyces striatus to establish the presence of compounds responsible for important biological activities. This
study also investigated the effect of Termitomyces striatus extracts on certain bacterial strains that included Escherichia coli and
Pseudomonas aeruginosa representing the Gram-negative bacteria and Bacillus subtilis and Staphylococcus aureus representing
Gram-positive bacteria. The fungi were represented by Candida albicans and Saccharomyces cerevisiae. The mushroom was
collected in western Kenya, air-dried, and crushed into powder, followed by extraction using water, methanol, and dichloro-
methane (DCM) solvents. Antibacterial and antifungal activities were evaluated using the disc-diffusion method. Qualitative
phytochemical screening of the aqueous extract revealed the presence of alkaloids, flavonoids, steroids, sterols, saponins, phenols,
carbohydrates, and proteins. The three extracts exhibited antibacterial against tested bacterial strains. The DCM extract revealed
higher effects among the bacterial strains tested. The three extracts showed antifungal effects against C. albicans. However, both
methanol and aqueous extracts did not inhibit growth of S. cerevisiae. In conclusion, T. striatus extracts are a promising source of
novel antimicrobial and antifungal agents.

1. Introduction kingdom Fungi, and family Lyophyllaceae. There are about

30 species within this genus.
Since ancient times, wild mushrooms have been used as a Mushrooms have a history of medicinal use spanning
source of food [1]. Before the introduction of exotic over a millennia. Studies in the last few decades have
mushrooms, native mushrooms were highly consumed as demonstrated that mushrooms and their constituting active
vegetables in Kenya [2, 3]. Out of the 42 tribes in Kenya, 38 components have beneficial effects on a variety of biological
tribes are known to consume native mushrooms [4]. Wild systems [6, 7]. A study in [8] indicated the presence of
edible mushrooms have been collected and consumed in alkaloids in the extracts of wild edible termitophilous
Uganda during the rainy season and valued as a traditionally mushrooms. In this study, 0.077 mg/g of alkaloids was found
nutritious food. Some of these are Polyporus tenuiculus, in T. mammiformis followed by T. microcarpus (0.056 mg/g),
Termitomyces tyleranus, Termitomyces clypeatus, Volvariella T. medius (0.053 mg/g), T. badius (0.052 mg/g), and
speciosa, and Termitomyces microcarpus [5]. Termitomyces is T. striatus (0.050 mg/g), while the least was in T. heimii
a genus of mushrooms from the division Basidiomycota, (0.046 mg/g). Phytochemical analysis of Termitomyces
2 Evidence-Based Complementary and Alternative Medicine

microcarpus revealed the presence of volatile oil, alkaloid, the filtrate was concentrated by a rotary evaporator sepa-
carotenoid, steroid, triterpenoids, fatty acid, emodins, fla- rately. The concentrate was then stored in air-tight con-
vonoid, coumarin, anthracene glycoside, anthocyanadine tainers and refrigerated.
glycoside, tannins, saponins, glycosides, polyurenoid, and
polyoses in the ethereal, methanolic, and aqueous extracts
[9–11]. 2.4. Qualitative Phytochemical Screening of Extracts. The
It has also been reported that there are antioxidant and methods used to test for the presence of alkaloids, flavo-
antimicrobial potentials of extracts obtained from four wild noids, sterols and steroids, saponins, and tannins were as
mushrooms: Termitomyces clypeatus, Termitomyces robus- described in [19, 20].
tus, Lentinus subnudus and Lenzites species collected in
Nigeria [12]. In their study [13], the authors observed that
the mushrooms T. clyeaptus and L. squarrosulus possess 2.5. Determination of Alkaloids. The mushroom sample was
considerable quantities of bioactive compounds. stirred with 1% hydrochloric acid (HCL) on a steam bath.
Termitomyces striatus is a mushroom from the genus of The solution obtained was filtered and 1 ml of the filtrate was
edible mushrooms which are commonly consumed in Africa treated with two drops of Mayer’s reagent. The two solutions
and Asia among the mushrooms collected from the wild. The were mixed and made up to 100 ml with distilled water.
Termitomyces mushrooms grow as symbionts in the termite Turbidity of the extract filtrate on addition of Mayer’s re-
molds [14]. This mushroom contains important phyto- agent was regarded as evidence for the presence of alkaloids
chemical compounds such as alkaloids [8] and flavonoids in the extract.
[15]. They also have phenols, saponins, and steroids that
have been recorded as possessing antifungal effects [16, 17].
The various extracts of this mushroom also possess anti- 2.6. Determination of Flavonoids. To 1 ml of the mushroom
oxidant and antimicrobial activity [18]. extract in a test tube, a small piece of magnesium ribbon was
The objective of this study was to determine the quali- added followed by drop-wise addition of concentrated
tative phytochemical composition of aqueous extracts of hydrochloric acid. Formation of pink or magenta red colours
T. striatus and the antimicrobial effects of the aqueous indicated the presence of flavonoids.
extract and methanolic and DCM extracts of T. striatus on
various strains of bacteria and fungi.
2.7. Determination of Sterols and Steroids. One milliliter of
the mushroom extract was put into a test tube in which
2. Materials and Methods 0.5 ml of sulphuric acid, acetic anhydride, and chloroform in
2.1. Collection of Termitomyces striatus. The mushroom was similar amount was added. A red coloration indicates
collected from western Kenya, washed, air-dried, and then presence of sterols while a green colour indicated the
ground into fine powder awaiting extraction. The mushroom presence of steroids.
was authenticated from the National Museum of Kenya
(REF: NMK/BOT of 31 MAR 2017) and coded voucher
2.8. Determination of Saponins. One milliliter of the
specimens were kept for future reference in the Botany
mushroom extract under test was put into a test tube and
Department of the National Museum of Kenya in Nairobi.
50 ml of distilled water was added. The mixture was then
shaken vigorously. Foaming which persists on warming was
2.2. Aqueous Extraction of Mushroom. Mushroom powder taken as an evidence for the presence of saponins. However,
weighing 300 grams was added to 800 ml of distilled water the results went through further test for confirmation, which
and boiled for twenty minutes in in a conical flask. It was involved dissolving of one milliliter of the extract in a
allowed to cool until it reached room temperature; thereafter mixture of carbon tetrachloride and 4 drops of concentrated
the supernatant was removed. It was centrifuged at 5400x sulphuric acid. A blue, green, or red colour accompanied by
gravity for 10 minutes after which it was filtered through a pink ring confirmed the presence of saponins.
®
Whatman GF/C glass microfiber filter paper; it was frozen
at −15°C and then dried in a freeze-drier. The extract was
kept desiccated at 4°C after being weighed. 2.9. Determination of Tannins. Extract of the mushroom
The percentage yield of the extract was determined sample was stirred with 10 ml of distilled water and then
according to the expression provided by [19] filtered. To the filtrate, two drops of 5% iron III chloride
(FeCl3) reagent was added. Blue-black or blue-green col-
weight of the extract
percentage yield � . (1) oration was an indication of the presence of tannins.
weight of the mushroom powder

2.10. Determination of Carbohydrates. This was done using

2.3. Methanol and Dichloromethane Extraction of Mushroom. Molisch’s test. To 2 ml of extract, 2 drops of alcoholic-
Each three hundred grams of mushroom powder was soaked naphthol solution was added in a test tube. Formation of
in one liter of methanol and dichloromethane separately and violet ring at the junction indicated the presence of
left standing for two days. The extracts were then filtered and carbohydrates.
Evidence-Based Complementary and Alternative Medicine 3

2.11. Determination of Proteins. To 2 ml of the extract, a few 2.15. Test of the Mushroom Extracts for Antifungal Activity.
drops of concentrated nitric acid were added. Formation of The experiments were done using the protocol described
yellow colour indicated the presence of proteins. in [26] with few modifications. The culture medium was
prepared using Sabouraud dextrose agar. The preparation
was done in accordance with the manufacturer’s direc-
2.12. Evaluation of Antimicrobial Activities of T. striatus tion. The medium was prepared in the conical flask,
Extracts on Selected Pathogens. The various extracts of the boiled, and then sterilized by autoclaving. The sterilized
mushroom T. striatus were subjected to tests to evaluate medium was cooled to around 50 and exactly 20 ml was
their antibacterial and antifungal activities. These were done dispensed into sterile Petri dishes and allowed to solidify.
by use of the disc-diffusion method. The bacterial strains Thereafter,. Candida albicans (ATCC 90028) and
which were selected were Pseudomonas aeruginosa, S. cerevisiae were aseptically inoculated on Petri dishes
Escherichia coli, Bacillus subtilis, and Staphylococcus aureus, which were then incubated at 31°C for 48 hours to give
while the fungi selected for the study were Candida albicans white round colonies against a yellowish background. The
and Saccharomyces cerevisiae. The organisms were obtained well isolated colonies of the fungal strains were scooped by
from a culture collection maintained in the Microbiology help of a sterile wire loop and suspended in sterilized 0.9%
Laboratory at Mount Kenya University. The purity of the sodium chloride solution (normal saline). The turbidity of
bacteria was tested by culturing on nutrient agar and being the inoculum was compared to McFarland solution. The
maintained on nutrient agar slants. microbial suspension (1 ml) in normal saline was added to
74 ml of sterile medium, kept at 45°C to give concentration
2.13. Preparation of Inocula. The stock cultures were kept on of 2 × 107 cells/ml. Sterilized Petri dishes (9 cm diameter)
slopes of nutrient agar. The cultures which were used for the were inoculated with 1 ml of fungal strains. Into the Petri
experiments were made by picking a loopful of cells out of dish containing 1 ml of the fungal strains, 15 ml of the
the stock cultures and putting in test tubes that contained sterile SDA media was added and swirled to mix the fungal
Mueller–Hinton broth (MHB) for bacteria. Meanwhile, strain and the media homogenously. Disks impregnated
Sabouraud dextrose broth (SDB) was used for fungi. They with extracts at concentrations of 6.25, 12.5, 25, 50, 100,
were reactivated by culturing them overnight at 37°C. and 200 mg/ml were laid on the solid Sabouraud dextrose
Cultures were diluted with fresh MHB and SDB and agar medium with the help of a sterile pair of forceps and
compared with McFarland standard to achieve values cor- gently pressed. The treated Petri dishes were placed at 4°C
responding to 2 × 106 colony-forming unit for bacteria [21] for 1-2 hours and then incubated for 48 hr at 37°C.
and 2 × 107 spores/ml for fungal strain [22]. Nystatin 0.2 mg/ml was used as the reference drug [27]. At
the end of the period, the inhibition zones formed on the
media were measured with a transparent ruler in
2.14. Evaluation of Antibacterial Activity of the Extracts. millimeters.
This was done by use of the disk diffusion method as de-
scribed in [23]. The selected strains of bacteria used were
Escherichia coli (ATCC 25922) and Pseudomonas aeruginosa 2.16. Statistical Data Analysis. Raw data was recorded in a
(ATCC 27853) representing the Gram-negative bacteria and data book, entered in Microsoft Excel spreadsheet, and then
Staphylococcus aureus (ATCC 25923) and Bacillus subtilis exported to STATA statistical software version 14.2 for
(ATCC 6633) representing the Gram-positive bacteria. analysis. Descriptive statistics were expressed as mean-
Sterile filter paper discs (Whatman No.1) with a diameter ± standard deviation. One-way analysis of variance was used
of 5 mm were soaked with mushroom extracts at the con- to determine the statistical difference among different
centrations of 6.25, 12.5, 25, 50, 100, and 200 mg/ml. The disks treatment groups followed by Bonferroni post hoc test for
which were soaked in dimethylsulfoxide represented the comparison of means of different treatment groups. The
negative controls [24]. All the bacteria were incubated at 30°C level of significance was set at 95% (p ≤ 0.05).
for 24 hours by inoculation into nutrient broth. Sterilized
Petri dishes were inoculated with 0.01 ml of one of the above
culture media (105-106 bacteria per ml). Mueller–Hinton agar 3. Results
sterilized in a flask and cooled to 45–50°C was distributed in
all the Petri dishes that had been inoculated; this was then 3.1. Yields of T. striatus Extracted. The aqueous extract of
swirled to enable the medium to be distributed homoge- T. striatus recorded higher yields of 16.0%, followed by
nously. The disks which had been injected with the mush- dichloromethane with a yield of 6.5%, while methanol ex-
room extracts were placed on the solid agar medium by tract had the least yield of 3.4% (Figure 1).
pressing gently. Ciprofloxacin 0.2 mg/ml [25] was used as the
standard drug for the test bacteria strains. The Petri dishes
which were treated were kept at 4°C for 1-2 hours and 3.2. Qualitative Phytochemical Analysis. The qualitative
thereafter they were incubated at 35°C for 18–24 hours. The screening of aqueous extract of T. striatus showed that the
experiments were done in triplicate. When the set time following compounds were present: alkaloids, flavonoids,
elapsed, the zones of inhibition which had been formed on the sterols and steroids, saponins, phenols, carbohydrates, and
media were measured with a transparent ruler in millimeters. proteins. However, the tannins were absent (Table 1).
4 Evidence-Based Complementary and Alternative Medicine

18
16

Extract percentage yield (%)

14
12
10
8
6
4
2
0
Aqueous Dichloromethane Methanol
Solvent
Figure 1: Percentage yields of T. striatus extracts.

Table 1: Qualitative phytochemical analysis of aqueous extract of P. aeruginosa, E. coli, B. subtilis, and S. aureus (Table 3;
T. striatus. p < 0.05). The negative exhibited no antimicrobial activity on
all bacteria strains tested (Table 3).
Substance Present/absent The zones of inhibition of MeOH extract at the con-
Alkaloids + centration of 200 ml/ml were statistically higher than those
Flavonoids + of 6.25, 12.5, 25, and 100 mg/ml against P. aeruginosa
Sterols and steroids + (Table 3; p < 0.05). Similarly, the antibacterial effect of the
Saponins +
MeOH extract at the concentrations of 100 and 200 mg/ml
Tannins −
Carbohydrates + was statistically higher than those of 6.25 and 12.5 mg/ml
Proteins + against E. coli (Table 3; p < 0.05). However, the zones of
Phenols + inhibition at all concentrations tested were not significantly
Note: +: present; −: absent. different against B. subtilis and S. aureus (Table 3; p > 0.05).
Further, the effect of the negative control was comparable to
3.3. Antibacterial Activities of Dichloromethane, Methanol, the effect of methanol extract at all tested concentration
and Aqueous Extracts on Selected Bacterial Strains. The DCM against S. aureus (Table 3; p > 0.05).
extract revealed antibacterial activity against P. aeruginosa, Also, the aqueous extract at higher concentrations
E. coli, B. subtilis, and S. aureus at concentrations of 6.25, revealed antibacterial activity against P. aeruginosa, E. coli,
12.5, 25, 50, 100, and 200 mg/ml. This was indicated by and S. aureus. However, the aqueous extract demonstrated
various zones of inhibition of greater than 5 mm in diameter antibacterial effect at all concentrations tested against
after impregnation of paper discs. The negative control B. subtilis (Table 4). The concentrations of aqueous extract
(DMSO) recorded no antimicrobial effect against all bacteria that never showed the antibacterial effect had zones with a
strains tested (Table 2). The antibacterial activity of the diameter of 5 mm (Table 4). The zones of inhibition of the
reference drug, ciprofloxacin, was significantly higher reference drug, ciprofloxacin, were significantly higher
compared to that of DCM extract at all concentrations tested compared to those of aqueous extract at all the tested
against all the bacterial strains used (Table 2; p < 0.05). concentrations against P. aeruginosa, E. coli, B. subtilis, and
The antibacterial activity of DCM extract against S. aureus (Table 4; p < 0.05).
P. aeruginosa was significantly higher at the concentrations The antibacterial effect of the aqueous extract at the
of 12.5, 25, 50, 100, and 200 mg/ml than that of 6.25 mg/ml concentrations of 50, 100, and 200 mg/ml was not signifi-
(Table 2; p < 0.05). Similarly, the antibacterial activity of the cantly different against P. aeruginosa (Table 4; p > 0.05).
DCM extract at concentrations of 50, 100, and 200 mg/ml However, the aqueous extract at concentrations of 6.25, 12.5,
was significantly higher than that of 6.25 mg/ml against and 25 mg/ml never showed antibacterial effect and was
E. coli and S. aureus (Table 2; p < 0.05). However, the an- comparable to the negative control (Table 4; p > 0.05).
tibacterial effect of DCM extract at all concentrations tested The zones of inhibition of aqueous extract at the con-
showed no significance on B. subtilis (Table 2; p > 0.05). centrations of 100 and 200 mg/ml were significantly higher
The methanolic extract revealed antibacterial activities at than those of 25 and 25 mg/ml against E. coli (Table 4;
different concentrations tested against P. aeruginosa, E. coli, p < 0.05). However, the aqueous extract at the concentra-
B. subtilis, and S. aureus. However, the MeOH extract never tions of 6.25 and 12.5 mg/ml never showed the antibacterial
revealed antibacterial activity against E. coli at the con- effect against E. coli (Table 4). The effect of aqueous extract at
centration of 6.25 mg/ml (Table 3). The antibacterial effect of the concentrations of 6.25, 12.5, and 25 mg/ml was not
the ciprofloxacin was significantly higher compared to that significant different against E. coli and was comparable to
of MeOH extract at all the concentrations tested against that of the negative control (Table 4; p > 0.05).
Evidence-Based Complementary and Alternative Medicine 5

Table 2: Antibacterial activity of DCM extract against P. aeruginosa, E. coli, B. subtilis, and S. aureus.
Zones of inhibition (mm)
Treatment
P. aeruginosa E. coli B. subtilis S. aureus
200 mg/ml 10.67 ± 0.58b 7.67 ± 0.58b 10.33 ± 1.15b 9.33 ± 0.58b
100 mg/ml 10.67 ± 0.58b 6.50 ± 0.50bc 9.33 ± 1.15b 9.33 ± 0.58b
50 mg/ml 10.33 ± 0.58b 6.50 ± 0.50bc 9.00 ± 1.00b 9.33 ± 0.58b
25 mg/ml 9.67 ± 0.58b 6.33 ± 0.58bcd 9.00 ± 1.00b 8.67 ± 0.58bc
12.5 mg/ml 9.33 ± 0.58b 6.33 ± 0.58 bcd
8.33 ± 0.58b 8.33 ± 0.58bc
6.25 mg/ml 7.33 ± 0.58c 5.83 ± 0.29cd 8.33 ± 0.58b 7.67 ± 0.58c
Negative control 5.00 ± 0.00d 5.00 ± 0.00 d
5.00 ± 0.00c 5.00 ± 0.00d
Ciprofloxacin 19.33 ± 0.58a 32.67 ± 0.58a 20.67 ± 0.58a 14.00 ± 0.00a
Descriptive statistics expressed as mean ± standard deviation for three replicates. Values with the same letter along the column are not significantly different
by one-way ANOVA followed by Bonferroni post hoc test (p > 0.05).

Table 3: Antibacterial effect of MeOH extract against P. aeruginosa, E. coli, B. subtilis, and S. aureus.
Zones of inhibition (mm)
Treatment
P. aeruginosa E. coli B. subtilis S. aureus
200 mg/ml 8.33 ± 0.58b 8.33 ± 0.58b 10.66 ± 0.58b 5.58 ± 0.38b
100 mg/ml 7.33 ± 0.58bc 7.33 ± 0.58b 10.00 ± 1.00b 5.42 ± 0.14b
50 mg/ml 6.67 ± 0.58c 6.67 ± 0.57bc 9.67 ± 0.58b 5.42 ± 0.14b
25 mg/ml 6.67 ± 0.58c 6.33 ± 0.58 bcd
9.33 ± 0.58b 5.33 ± 0.14b
12.5 mg/ml 6.33 ± 0.58cd 5.83 ± 0.29 cd
8.67 ± 0.58b 5.25 ± 0.00b
6.25 mg/ml 6.00 ± 0.00cd 5.00 ± 0.00d 8.33 ± 0.58b 5.25 ± 0.00b
Negative control 5.00 ± 0.00d 5.00 ± 0.00 cd
5.00 ± 0.00c 5.00 ± 0.00b
Ciprofloxacin 19.33 ± 58a 31.67 ± 0.58a 21.00 ± 1.00a 12.33 ± 0.58a
Descriptive statistics expressed as mean ± standard deviation for three replicates. Values with the same letter along the column are not significantly different
by one-way ANOVA followed by Bonferroni post hoc test (p > 0.05).

Table 4: Antibacterial effect of aqueous extract against P. aeruginosa, E. coli, B. subtilis, and S. aureus.
Zones of inhibition (mm)
Treatment
P. aeruginosa E. coli B. subtilis S. aureus
200 mg/ml 6.33 ± 0.29b 11.67 ± 0.58b 7.67 ± 0.58b 5.42 ± 0.14b
100 mg/ml 6.33 ± 0.29b 10.33 ± 0.58b 7.67 ± 0.58b 5.42 ± 0.14b
50 mg/ml 6.17 ± 0.29b 8.67 ± 0.58 c
7.33 ± 0.58b 5.33 ± 0.14b
25 mg/ml 5.00 ± 0.00c 6.33 ± 0.58d 7.17 ± 0.29b 5.25 ± 0.00b
12.5 mg/ml 5.00 ± 0.00c 5.00 ± 0.00 d
7.00 ± 0.00b 5.00 ± 0.00b
6.25 mg/ml 5.00 ± 0.00c 5.00 ± 0.00 d
7.00 ± 0.00b 5.00 ± 0.00b
Negative control 5.00 ± 0.00c 5.00 ± 0.00d 5.00 ± 0.00c 5.00 ± 0.00b
Ciprofloxacin 19.33 ± 58a 32.33 ± 0.58 a
22.67 ± 1.15a 11.67 ± 0.58a
Descriptive statistics expressed as mean ± standard deviation for three replicates. Values with the same letter along the column are not significantly different
by one-way ANOVA followed by Bonferroni post hoc test (p > 0.05).

The antibacterial activity of aqueous extract at all con- extract demonstrated antifungal activity against C. albicans
centrations tested was insignificant against B. subtilis (Ta- and S. cerevisiae at the concentrations of 6.25, 12.5, 25, 50,
ble 4; p > 0.05). Besides, the zones of inhibition of aqueous 100, and 200 mg/ml. Different zones of inhibition of greater
extract at the concentrations of 25, 50, 100, and 200 mg/ml than 5 mm were observed after C. albicans and S. cerevisiae
were not statistically different against S. aureus. However, were treated with DCM extract (Figures 2 and 3). The
the aqueous extract at the concentrations of 6.25 and negative control never revealed antifungal activity (Figures 2
12.5 mg/ml never revealed antibacterial activity against and 3).
S. aureus (Table 4). The effect of aqueous extract at all The antifungal activity of DCM extract at the concen-
concentrations tested against S. aureus was statistically trations of 100 and 200 mg/ml was statistically higher than
similar to that of the negative control (Table 4; p > 0.05). that of 6.25, 12.5, and 25 mg/ml against C. albicans (Figure 2;
p < 0.05). The effect of DCM extract at the concentrations of
6.25, 12.5, and 25 mg/ml against C. albicans was comparable
3.4. Antifungal Activities of Dichloromethane, Methanol, and to negative control (Figure 2; p > 0.05). The antifungal effect
Aqueous Extracts on C. albicans and S. cerevisiae. The DCM of the reference drug, Nystatin, was significantly higher
6 Evidence-Based Complementary and Alternative Medicine

14
a
12

Zones of inhibition (mm)

10
b b
8 bc
cd cd cd
6 d

0
200 mg/ml

100 mg/ml

50 mg/ml

25 mg/ml

12.5 mg/ml

6.25 mg/ml

Negative
control

Nystatin
Figure 2: Antifungal activity of DCM extract on C. albicans. Bars with the same letter are not significantly different by one-way ANOVA
followed by Bonferroni post hoc test (p > 0.05).

8
a
7 b
bc bc bc
Zones of inhibition (mm)

6 cd
d d
5

0
200 mg/ml

100 mg/ml

50 mg/ml

25 mg/ml

12.5 mg/ml

6.25 mg/ml

Negative
control

Nystatin

Figure 3: Antifungal activities of DCM extract on S. cereviceae. Bars with the same letter are not significantly different by one-way ANOVA
followed by Bonferroni post hoc test (p > 0.05).

compared to that of DCM extract at all the concentrations antifungal effect of Nystatin was significantly higher com-
tested against C. albicans (Figure 2; p < 0.05). pared to that of MeOH extract at all concentrations tested
The zones of inhibition of DCM extract at the con- (Figure 4; p < 0.05).
centrations of 200 mg/ml were statistically higher than those Further, the aqueous extract revealed the antifungal
of 6.25, 12.5, 25, and 50 mg/ml against S. cerevisiae (Figure 3; effect against C. albicans at all the concentrations tested.
p < 0.05). The zone of inhibition of DCM extract at the However, the aqueous extract showed no antifungal activity
concentration of 6.25 mg/ml against S. cerevisiae was not against S. cerevisiae at all the concentrations tested (Figures 6
significantly different from the negative control (Figure 3; and 7). The antifungal effect of the aqueous extract was
p > 0.05). The reference drug, Nystatin, never revealed the statistically insignificant against C. albicans and was com-
antifungal effect against S. cerevisiae and was comparable to parable to the negative control (Figure 6; p > 0.05). However,
negative control and extract at the concentration of 6.25 mg/ the antifungal activity of Nystatin was significantly higher
ml (Figure 3; p > 0.05). compared to that of aqueous extract at all concentrations
The MeOH extract also reported the antifungal effect tested (Figure 6; p < 0.05).
against C. albicans at all the concentrations tested. However,
the MeOH extract revealed no antifungal activity against 4. Discussion
S. cerevisiae at all the concentrations tested (Figures 4 and 5).
The antifungal activity of the MeOH extract was not sig- The extract of the test mushroom T. striatus possessed
nificantly different against C. albicans and was comparable important phytochemical compounds, alkaloids, flavonoids,
to the negative control (Figure 4; p > 0.05). However, the steroids, saponins, and phenols which are responsible for
Evidence-Based Complementary and Alternative Medicine 7

10
a
9
8

Zones of inhibition (mm)

7
6 b b b b b b
b
5
4
3
2
1
0
200 mg/ml

100 mg/ml

50 mg/ml

25 mg/ml

12.5 mg/ml

6.25 mg/ml

Negative
control

Nystatin
Figure 4: Antifungal activity of methanol extract on C. albicans. Bars with the same letter are not significantly different by one-way ANOVA
followed by Bonferroni post hoc test (p > 0.05).

5
Zones of inhibition (mm)

0
200 mg/ml

100 mg/ml

50 mg/ml

25 mg/ml

12.5 mg/ml

6.25 mg/ml

Negative
control

Nystatin

Figure 5: Antifungal activities of methanol extract on S. cereviceae. Bars with the same letter are not significantly different by one-way
ANOVA followed by Bonferroni post hoc test (p > 0.05).

12

a
10
Zones of inhibition (mm)

6 b b b b b b b

0
200 mg/ml

100 mg/ml

50 mg/ml

25 mg/ml

12.5 mg/ml

6.25 mg/ml

Negative
control

Nystatin

Figure 6: Antifungal activity of aqueous extract on C. albicans. Bars with the same letter are not significantly different by one-way ANOVA
followed by Bonferroni post hoc test (p > 0.05).
8 Evidence-Based Complementary and Alternative Medicine

Zones of inhibition (mm)

4

0
200 mg/ml

100 mg/ml

50 mg/ml

25 mg/ml

12.5 mg/ml

6.25 mg/ml

Negative
control

Nystatin
Figure 7: Antifungal activities of aqueous extract on S. cereviceae. Bars with the same letter are not significantly different by one-way
ANOVA followed by Bonferroni post hoc test (p > 0.05).

various biological activities important in health research broad spectrum antibiotic activity. However, aqueous ex-
[13]. Phytochemical compounds such as saponins, alkaloids, tract shows very weak antibacterial activity against S. aureus.
phenolics, steroids, and flavonoids have been reported to This differs from the study in [18] in which they found out
possess antibacterial activities [28–30]. Phenolics, flavo- that hot water extract of both Auricularia and Termitomyces
noids, steroids, and saponins have also been documented to mushrooms species showed strong antibacterial activity
possess antifungal effects [16, 17]. Also present were car- against S. aureus.
bohydrates and proteins. This was in conformity with studies Furthermore, in their study using Trametes spp and
by Due et al. [31] where they also found that wild edible Microporus spp mushrooms, [32]; found out that hot water
mushroom Termitomyces heimii Natarajan from Côte extracts had the strongest antimicrobial activity against all
d’Ivoire mainly contains proteins and carbohydrates. tested organisms as compared to chloroform and ethanol
T. heimii is a good source of nutrients and it could be utilized extracts, which suggested that hot water extraction could be
in human diet to alleviate undernourishment caused by capable of producing several antimicrobial compounds such
protein deficiency. These findings concur with the study in as flavonoids, tannins, and terpenoids. Like the methanolic
[9] and those in [11] whose authors found out that a sample extract, the aqueous extract shows antifungal activity against
of Termitomyces microcarpus when screened revealed the C. albicans but not S. cerevisiae.
presence of alkaloids, steroids, flavonoids, saponins, and Comparatively, the antibacterial activity of DCM extract
other compounds in the ethereal, methanolic, and aqueous was higher (6.25 mg/ml) than that of the other two extracts
extracts. The results also agreed with a study in [8] which that is methanol (12.5 mg/ml) and water (25 mg/ml). This
indicated the presence of alkaloids in the extracts of wild means that the DCM has a higher ability of extracting the
edible termitophilous mushrooms. Besides, in their study, secondary metabolites responsible for these activities better
Adejumo et al. [15] found out that the Termitomyces than the two.
mushrooms also contain flavonoids. Test of the extracts with the two fungi showed that only
In the current study, dichloromethane extract showed the extracts for dichloromethane had higher activity against
antimicrobial activity against all test microorganisms C. albicans and S. cerevisiae. The results for antifungal tests
(P. aeruginosa E. coli, B. subtilis, and S. aureus) at all con- in various mushroom extracts concur with the finding of
centrations. DCM demonstrates broad spectrum antibac- [33], in which the author found out that the various extracts
terial activity by affecting both Gram-positive and Gram- of mushroom Termitomyces exhibited poor activity against
negative bacteria. It was also observed that DCM affects both various fungal pathogens which he used in the study
fungi used in the test that is C. albicans and S. cerevisiae. (A. flavus, C. albicans, and M. racemosus). The results are
The methanolic extract exhibited antibacterial activities also in conformity with observations by [34] in which he
at different concentrations tested against P. aeruginosa, found that aqueous extract had little antifungal activity in
E. coli, B. subtilis, and S. aureus. However, there was no the plants (Asparagus setaceus Kunth and Caesalpinia
antibacterial activity against E. coli at the concentration of volkensii).
6.25 mg/ml. The methanolic extract showed antifungal ac-
tivity effect against C. albicans but not S. cerevisiae. 5. Conclusion
Furthermore the aqueous extract demonstrated anti-
bacterial activity against all the bacterial isolates tested. This study concluded that the dichloromethane, methanol,
However, there was higher activity in E. coli and B. subtilis. and aqueous extracts of Termitomyces striatus revealed
This shows that aqueous extract like DCM extract exhibits potent antibacterial effect against Gram-negative
Evidence-Based Complementary and Alternative Medicine 9

(Escherichia coli and Pseudomonas aeruginosa) and Gram- from Tanzania,” Food Science and Quality Management,
positive (Bacillus subtilis and Staphylococcus aureus) bac- vol. 7, pp. 13–23, 2012.
teria. Similarly, the three extracts demonstrated antifungal [10] H. P. Aryal and U. Budhathoki, “Phytochemical screening of
effect against C. albicans. However, the methanol and termite’s mushroom in Nepal,” Journal of Natural History
aqueous extracts never showed antifungal activities against Museum, vol. 27, pp. 107–119, 2013.
[11] P. Mitra, N. Mandal, A. Roy, and K. Acharya, “Phytochemical
S. cerevisiae. The extracts of T. striatus, therefore, may be
study and antioxidative property of ethanolic extract from
used as antibacterial and antifungal agent. termitomyces clypeatus,” Journal of Applied Pharmaceutical
Science, vol. 6, no. 7, pp. 120–124, 2016.
Data Availability [12] V. O. Oyetayo and C. O. Ogidi, “Phytochemical screening and
antibacterial properties of a wild macrofungus, coriolosis
The data used to support the findings of this study are polyzona against microbial isolates from wastewater and
available from the corresponding author upon request. leftover foods,” Asian Journal of Pharmaceutical and Bio-
logical Research, vol. 1, no. 4, pp. 486–492, 2011.
Conflicts of Interest [13] S. D. Ghate and K. R. Sridhar, “Bioactive potential of Lentinus
squarrosulus and Termitomyces clypeatus from the south-
The authors declare that they have no conflicts of interest. western region of India,” Indian Journal of Natural Products
and Resources (IJNPR), vol. 8, no. 2, pp. 120–131, 2017.
Acknowledgments [14] H.-M. Hsieh and Y.-M. Ju, “Medicinal components in ter-
mitomyces mushrooms,” Applied Microbiology and Biotech-
We owe our gratitude to the Technologists in Mount Kenya nology, vol. 102, no. 12, pp. 4987–4994, 2018.
University Microbiology Laboratory and those in the [15] T. O. Adejumo, M. E. Coker, and V. O. Akinmoladun,
Government of Kenya Biology Laboratory situated at Jomo “Identification and evaluation of nutritional status of some
Kenyatta University of Agriculture and Technology for their edible and medicinal mushrooms in Akoko area, Ondo State,
support in running the experiments. Nigeria,” International Journal of Current Microbiology and
Applied Sciences, vol. 4, no. 4, pp. 1011–1028, 2015.
[16] C. de Andrade Monteiro and J. Ribeiro Alves dos Santos,
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