Enumeration of Fungal Population Isolated from Dried-Stored Spices
at Retailers in Traditional Markets in Medan, North Sumatera
Betriana Novi Lenta Gultom
1
, Kiki Nurtjahja
1*
, Aditya Bungsu
1
,
Jesica Simanullang
1
, Juwita Esterina Silalahi
1
and Riyanto
2
1
Department of Biology, Faculty of Mathematics and Natural Sciences, Sumatera Utara University, Jln. Bioteknologi no. 1,
Medan, Sumatera Utara, Indonesia 20155
2
Faculty of Biology, Medan Area University, Jln. Kolam no. 1 Medan, North Sumatera, Indonesia 20223
Keywords: Filamentous Fungi, Isolation, Spices, Traditional Market
Abstract : Fungal population on dried spices sold by retailers at traditional market in Medan was studied, the aim was to
investigate the number of postharvest fungi on each spice. A total of ten kinds of dried spices i.e. white and
black pepper, coriander, candle nut, cloves, cinnamon, star anise, nutmeg, cardamom and cumin were
obtained from retailers at five traditional markets in Medan. As much as 250 g of the each spice was taken
from five retailers at each traditional market. Fungal population was determined using a dilution method and
followed by a pour plate method on dichloran 18% glycerol agar medium. Each fungal species was isolated
and identified using czapex yeast agar and malt extract agar. Results showed that dried spices sold by
retailers in traditional markets were infected by storage fungi. Spice infection in traditional markets such as
Pringgan, Setiabudi and Sikambing was more predominant than that in Padang bulan and Simalingkar.
Among the fungi aspergilli was the most common found and Aspergillus chevalieri has the highest
population and almost found in all kinds of spice samples.
1 INTRODUCTION
Spices are one of Indonesia's main export
commodities. This commodity plays an important
role because it is used daily as a mixture of
traditional food, drinks and medicines. Most tropical
spices was harvested conventionally, the spices were
dried using sunlight and stored in gunny or
polypropylene sacks that permeable to water favor.
Among microorganisms, fungi are the main cause of
spice damage (Kneifel and Berger, 1994; Dimic et
al, 2000; Romagnoli et al, 2007). The increase of
water content during storage can cause fungal
infection. Postharvest fungi such as Aspergillus and
Penicillium are able to grow in water activities 0.78
(Koci-Tanackov et al., 2007). Fungal infection on
spices causes a decrease in quality such as physical
damage, loss of aroma and contain mycotoxins
(McKee, 1995; Koci-Tanackov et al., 2007). Dried
stored spices in traditional markets are generally
sold by retailers in open air. High humidity can
cause an equilibrium relative humidity between
dried spice and the surrounding air, as a result in
increase in spice water content. The purpose of this
study was to investigate the number of fungal
population in spices sold by retailers at traditional
markets in Medan.
2 MATERIALS AND METHODS
2.1 Sample Collection
A total of 250 g samples from ten kinds of dried
spices such as white and black pepper, coriander,
candle nut, cloves, cinnamon, star anise, nutmeg,
cardamom and cumin were obtained from 5 retailers
at five traditional markets, namely Simalingkar, Sei
Sikambing, Setiabudi, Pringgan and Padang bulan.
Each sample was put into a sterile polyethylene bag
and stored in a refrigerator at ± 12°C for further use.
Gultom, B., Nurtjahja, K., Bungsu, A., Simanullang, J., Silalahi, J. and Riyanto, .
Enumeration of Fungal Population Isolated from Dried-stored Spices at Retailers in Traditional Markets in Medan, North Sumatera.
DOI: 10.5220/0010199600002775
In Proceedings of the 1st International MIPAnet Conference on Science and Mathematics (IMC-SciMath 2019), pages 407-412
ISBN: 978-989-758-556-2
Copyright
c
 2022 by SCITEPRESS – Science and Technology Publications, Lda. All rights reserved
407

2.2 Determination Fungal Population
The population of each fungus was determined using
a dilution method followed by pour plate method on
the DG18 medium. Each 250 g sample was ground
using a Mill Powder RT-04 No. blender Series
980923 (Mill Powder Tech. Co. LTD, Taiwan) with
a speed of 25000 rpm for 30 seconds. As much as 25
g were put into a 500 ml erlenmeyer and suspended
with 250 ml of sterile distilled water and then
homogenized to obtain a 10
-1
suspension.
Furthermore, dilution is carried out on 10
-2
, 10
-3
and
10
-4
. Every 1 ml of dilution suspension was cultured
by pouring method using DG18 medium. Each
dilution was repeated 3 times. All plates were
incubated for 5 days (29
o
C). Fungal population in
each gram of spice (cfu/g) was determined using the
formula:
Fungal population
=
1
X.Y
. Z (cfu/g)
X = volume of suspension transferred to each petri
dish (1 ml)
Y = dilution which gives the fungus colonies
separately
Z = average number of colonies of each fungal
species from 3 petri dishes
2.3 Fungal Identification
Each fungus was cultured on the medium of malt
extract agar (MEA) and czapex yeasr agar (CYA)
and then identified according to Pitt and Hocking
(2009).
3 RESULTS AND DISCUSSION
All the dried spices sold by retailers in traditional
markets were infected by various species of fungi
and Aspergilli was the most common found. The
fungal infection on each spice was varies depending
on the kind of spice. Black pepper is the most
infected, whereas, star anise was the lowest. Fungal
population (cfu/g) in each spice sold by retailers in 5
traditional markets was shown as follows:
White Pepper. Fungal infection on white pepper
sold in the Sikambing market has the highest
population. Eight species of of the fungi were found
in white pepper and Aspergillus sydowi was the
highest population followed by A. candidus (Table
1).
Table 1. Fungal population (cfu/g) isolated from white pepper sold by retailers at traditional markets in Medan, North
Sumatera
Fungal species
Traditional markets / fungal population (cfu/g)
12345
A
s
p
er
g
illlus
f
lavus 0.3×10
3
00.3×10
3
3.3×10
3
0.6×10
3
A
. nige
r
01×10
3
1.6×10
3
2×10
3
0.3×10
3
A
.
sy
dowii 100×10
3
32.6×10
3
66.6×10
3
79.3×10
3
79.6×10
3
A
. tereus 0 0 0.3×10
3
00
A
. tamarii 0001.3×10
3
0
A
. candidus 12×10
3
17×10
3
0 22.6×10
3
2×10
3
A
. chevalieri 12.6×10
3
7×10
3
13.3×10
3
1×10
3
17×10
3
Penicillium sp. 6.3×10
3
1×10
3
1.3×10
3
0.6×10
3
15.6×10
3
Simalingkar (1), Padang bulan (2), Pringgan (3), Setiabudi (4), Sikambing (5).
cfu = colony forming unit
Black Pepper. Thirteen species of fungi were
successfully isolated by black pepper, A. niger had
the highest population in black pepper particularly in
the Padang Bulan market, followed by A. ustus in
Sikambing (Table 2). The infection of A. fumigatus,
A. ustus and A. chevalieri are found in all markets.
IMC-SciMath 2019 - The International MIPAnet Conference on Science and Mathematics (IMC-SciMath)
408

Table 2. Fungal population (cfu/g) isolated from black pepper sold by retailer at traditional markets in Medan, North
Sumatera
Fungal species
Traditional markets / fun
g
al population (cfu/
g
)
123 4 5
A
spergillus flavus 02×10
3
0.3×10
3
0.3×10
3
0.3×10
3
A
. ni
g
e
r
0 22×10
3
2.3×10
3
5×10
3
1.6×10
3
A
. oryzae 00.3×10
3
00 0
A
.
sy
dowii 000.6×10
3
00
A
.
f
umigatus 2.3×10
3
3×10
3
0.6×10
3
19×10
3
2×10
3
A
. tereus 0 0.3×10
3
0.3×10
3
1×10
3
0
A. ustus 5.6×10
3
1×10
3
1.6×10
3
2×10
3
16.6×10
3
A
. tamarii 0 0 0 0 0.3×10
3
A
. candidus 1×10
3
05×10
3
01.3×10
3
A
. chevalieri 3×10
3
1.3×10
3
6×10
3
2.6×10
3
6×10
3
Fusarium s
p
.000.6×10
3
00
M
ucor sp. 0 0 0 1.6×10
3
0.3×10
3
Penicillium s
p
. 0.6×10
3
00 0 0
Simalingkar (1), Padang bulan (2), Pringgan (3), Setiabudi (4), Sikambing (5).
cfu = colony forming unit
Coriander. Seven species of postharvest fungi
isolated from coriander were found, Aspergillus
chevalieri has the highest population and is found in
all markets followed by Penicillium sp. and
Cladosporium cladosporioides (Table 3).
Table 3. Fungal population (cfu/g) isolated from coriander sold by retailer at traditional markets in Medan, North Sumatera
Fungal species
Traditional markets / fun
g
al population (cfu/
g
)
123 4 5
A
s
p
er
g
illus
f
lavus 0 0.3×10
3
00 0
A
. candidus 04.3×10
3
00 0
A
.
f
umi
g
atus 0 0 1×10
3
00
Cladosporium
clados
p
oroides
0,3×10
3
0 0,3×10
3
0 0
A
. chevalieri 22,3×10
3
109×10
3
19.6×10
3
39.3×10
3
0.6×10
3
Penicillium s
p
. 1.3×10
3
2×10
3
00.6×10
3
12×10
3
Simalingkar (1), Padang bulan (2), Pringgan (3), Setiabudi (4), Sikambing (5).
cfu = colony forming unit
Candle nut. Eight species of postharvest fungi
isolated from candlenut were found and Aspergillus
chevalieri was the most predominant fungi (Table
4). Among the traditional markets candle nut in
Pringgan was the most infected (six species) by the
filamentous fungi.
Table 4. Fungal population (cfu/g) isolated from candle nut sold by retailer at traditional markets in Medan, North Sumatera
Fungal species
Traditional markets / fungal population (cfu/g)
12 3 4 5
A
spergillus flavus 0 1 ×10
3
00 0
A
. ni
g
e
r
0 0 0 0.6×10
3
0
A
.
s
ydowii 01×10
3
00.3×10
3
0
A
. wentii 01.3×10
3
00 0
A
.
f
umigatus 00 00.3×10
3
0
A
. tamarii 00.3×10
3
00 0
A
. chevalieri 0 4.6×10
3
8.6×10
3
00.6×10
3
Penicillium s
p
. 0 0.6×10
3
0.3×10
3
00
Simalingkar (1), Padang bulan (2), Pringgan (3), Setiabudi (4), Sikambing (5).
cfu = colony forming unit
Enumeration of Fungal Population Isolated from Dried-stored Spices at Retailers in Traditional Markets in Medan, North Sumatera
409

Cloves. Only 4 species of filamentous fungi isolated
from cloves and Cladosporium cladosporioides is
the highest population in cloves (0.6 × 104 cfu / g)
(Table 5).
Table 5. Fungal population (cfu/g) isolated from cloves sold by retailer at traditional markets in Medan, North Sumatera
Fungal species
Traditional markets / fun
g
al
p
o
p
ulation
(
cfu/
g)
123 4 5
A
s
p
er
g
illus
f
lavus 000.3×10
4
00
A
. chevalieri 000.3×10
4
00
Cladosporium
cladosporoides
0.6×10
4
0 0 0 0
Penicillium s
p
. 0 0.3×10
4
0 0 0.3×10
4
Simalingkar (1), Padang bulan (2), Pringgan (3), Setiabudi (4), Sikambing (5).
cfu = colony forming unit
Cinnamon. A total of 4 species of fungi that attack
cinnamon from all markets. Aspergillus chevalieri
has the highest population, found in Simalingkar
market and Setiabudi market (Table 6).
Table 6. Fungal population (cfu/g) isolated from cinnamon sold by retailer at traditional markets in Medan, North Sumatera
Fungal species
Traditional markets / fun
g
al
p
o
p
ulation
(
cfu/
g)
1234 5
A
spergillus flavus 0.3×10
3
000 0
A
. carboni
f
erous 00.3×10
3
00 0
A
.
f
ischerianus 00.3×10
3
00 0
A
. chevalieri 0.6×10
3
0 0 0.4×10
3
0
Simalingkar (1), Padang bulan (2), Pringgan (3), Setiabudi (4), Sikambing (5).
cfu = colony forming unit
Star anise. Among the spices, star anise is the least
infected by fungi (Table 7). Three species of storage
fungi were isolated from star anise. No fungal
infection on star anise sold in Pringgan and
Sikambing.
Table 7. Fungal population (cfu/g) isolated from star anise sold by retailer at traditional markets in Medan, North Sumatera
Fungal species
Traditional markets / fun
g
al
p
o
p
ulation
(
cfu/
g)
1234 5
A
spergillus fumigatus 0000.3×10
3
0
A
. chevalieri 0.3×10
3
00 0 0
Penicillium sp. 0.3×10
3
0.3×10
3
00 0
Simalingkar (1), Padang bulan (2), Pringgan (3), Setiabudi (4), Sikambing (5).
cfu = colony forming unit
Nutmeg kernels. A total of 10 species of fungi were
isolated from nutmeg. The infection was dominated
by Aspergillus chevalieri followed by Aspergillus
flavus. Nutmeg kernels in Padang bulan was the
highest infected by fungi followed by Setiabudi
market (Table 8).
Table 8. Fungal population (cfu/g) isolated from nutmeg kernels sold by retailer at traditional markets in Medan, North
Sumatera
Fungal species
Traditional markets / fun
g
al
p
o
p
ulation
(
cfu/
g)
12 3 4 5
A
spergillus flavus 0 0.3×10
3
80×10
3
2.6×10
3
30×10
3
A
. ni
g
e
r
0.3×10
3
00 0 0
A
. tamarii 0 0 0 0 11.6×10
3
A
. candidus 2.6×10
3
2×10
3
000
A
. ustus 24.6×10
3
4×10
3
000
A
. wentii 0 0 2.6×10
3
00
A
. chevalieri 7×10
3
67×10
3
0 56.6×10
3
0
Fusarium s
p
. 0 1.6×10
3
00.3×10
3
0
Penicillium citrinum 02.3×10
3
6×10
3
0.3×10
3
6.6×10
3
R
hizo
p
us stoloni
f
e
r
0.3×10
3
00 0 0
Simalingkar (1), Padang bulan (2), Pringgan (3), Setiabudi (4), Sikambing (5).
cfu = colony forming unit
IMC-SciMath 2019 - The International MIPAnet Conference on Science and Mathematics (IMC-SciMath)
410

Cardamom. All cardamom sold by retailers in
traditional markets have infected by fungi. A total of
12 species of fungi isolated and A. westerdijkiae, A.
niger and A. chevalieri were the most perdominant
found followed by A. restrictus and A. tamarii.
(Table 9).
Table 9. Fungal population (cfu/g) isolated from cardamom sold by retailer at traditional markets in Medan, North Sumatera
Fungal species
Traditional markets / fun
g
al
p
o
p
ulation
(
cfu/
g)
1 234 5
A
s
p
er
g
illus
f
lavus 1.6×10
3
000 0
A
. nige
r
17.3×10
3
1.3×10
3
00 0
A
. restrictus 0 0 0.3×10
3
0 16×10
3
A
. tereus 0 0 0 0.3×10
3
0
A
. tamarii 4.3×10
3
13.3×10
3
00 0
A
. ustus 2.6×10
3
0.6×10
3
3×10
3
00
A
. westerdijkiae 38×10
3
0.3×10
3
00 0
A
.
f
umi
g
atus 0 11×10
3
00 0
A
.
j
aponicus 0 0 0.6×10
3
00
A
. chevalieri 0.3×10
3
1.6×10
3
0.3×10
3
17.3×10
3
3.6×10
3
A
. repens 0003×10
3
0
Penicillium s
p
. 0 0 0 0 0.3×10
3
Simalingkar (1), Padang bulan (2), Pringgan (3), Setiabudi (4), Sikambing (5).
cfu = colony forming unit
Cumin. All cumin sold by retailers in traditional
markets was infected with postharvest fungi.
Setiabudi market has the highest infection.
Aspergillus chevalieri was the most common found
on Setiabudi followed by A. ustus on the
Simalingkar market (Table 10).
Table 10. Fungal population (cfu/g) isolated from cumin sold by retailer at traditional markets in Medan, North Sumatera
Fungal species
Traditional markets / fun
g
al
p
o
p
ulation
(
cfu/
g)
12345
A
spergillus fumigatus 00.3×10
3
000
A
. niveus 1×10
3
00.3×10
3
00
A
. ustus 2.3×10
3
0000
A
. chevalieri 00.3×10
3
1.6×10
3
2.6×10
3
1×10
3
A
. repens 0 0 1×10
3
0.3×10
3
1×10
3
Penicillium s
p
. 1×10
3
00.3×10
3
00
Simalingkar (1), Padang bulan (2), Pringgan (3), Setiabudi (4), Sikambing (5).
cfu = colony forming unit
Stored-dried spices sold by retailers in traditional
markets in Medan were commonly infected by
storage fungi. We assumed that the infection
occurred during pre and post-harvest handling.
Stored in open air and high relative humidity at
traditional markets lead to stored-dried spices
absorbed water vapor from the surrounding air as a
result spice water content increase during storage.
Nurtjahja et al. (2017) reported that dried nutmeg
kernels stored in various water activity (a
w
) showed
at a
w
0.75-0.97 were dominated by storage fungi
such as Aspergillus chevalieri and Penicillium
citrinum. However, some spices such as cinnamon,
star anise and cloves less infected than the other.
The presence of antifungal compounds in the spices
might inhibit fungal growth. Research conducted by
Freire et al. (2011) showed star anise (Illicium
verum) contain chemical compounds such as
limonene, terpineol, α-pinene, methyl-chavicol and
trans-anetol. The activity of antifungal of the
compounds at a dose of 1 µL / mL inhibit A. flavus
mycelia, whereas A. parasiticus is inhibited at 2.0
µL / mL. The ability of postharvest fungi to live at
a
w
(<0.85) lead to increase their population. Dried
spices sold by relatilers at traditional markets were
packed in gunny sack or polypropylene that
permeable to water vapor. Balesevic-Tubic et al.
2005 reported that fungal infection on agricultural
product including spices can not be avoided.
Preventing fungal growth on dried spices during
storage is required.
4 CONCLUSIONS
Dried spices are infected by fungi. The pre and
posthandling spices were required more optimally to
Enumeration of Fungal Population Isolated from Dried-stored Spices at Retailers in Traditional Markets in Medan, North Sumatera
411

get the quality of spices that are in line with food
safety standards.
ACKNOWLEDGEMENTS
The research was supported by Directorate General
Higher Education, Ministry of Research and
Technology Republic of Indonesia contract DRPM
Reseach no. 67/UN5.2.3.1/PPM/KP-DRPM/2018.
REFERENCES
Balesevic-Tubic, S., Malenèiæ D., Tatiæ, M., Miladinovic,
J., 2005. Influencing of aging process on biochemical
changes in sunflower seed. Helia 28(42):107-114.
Dimic, G., S. Krinjar, M.V., Dosen-Bogic, E., Dance,,
2000a potential producer and sterigmatocystin in
spices (Croatian). Tecnol. Mes., , 41 (4-6): 131-137.
Freire, JM., Cardoso, MG., Batista, LR., Andrade MA.,
2011. Essential oil of Origanum majorana L., Illicium
verum Hook.f. and Cinnamomum zeylanicum Blume:
chemical and antimicrobial characterization. Rev Bras
Pl Med., Botucatu 13(2): 209-214.
Koci-Tanackov, SD., Dimi, GR., Karali D., 2007.
Contamination of spices with moulds potential
producers of sterigmatocystine. APTEFF. 38: 29-35.
Kneifel, W., Berger, E., 1994. Microbial criteria of
random samples of spices and herbs retailed on the
Austrian market. J. Food. Prot. 57: 893-901.
Nurtjahja, K., O.S,. Dharmaputra, WP., Rahayu, R.,
Syarief, 2017. Fungal population of nutmeg (Myristica
fragrans) kernels affected by water activity during
storage. Agritech 17(3): 288-294.
McKee, LH., 1995. Microbial contamination of spices and
herbs. A review. Lebensm. Wiss.Technol. 28: 1-11.
Pitt JI, Hocking AD., 2009. Fungi and Food Spoilage.
New York (US): Springer.
Romagnoli, B., Menna, V., Gruppioni N, Bergamini C.,
2007. Aflatoxins in spices, aromatic herbs, herbs-teas
and medicinal plants marketed in Italy. Food Control.
18: 697-701.
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