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Malaysian Journal of Microbiology, Vol 14(2) 2018, pp. 691-698
DOI: http://dx.doi.org/10.21161/mjm.116117
Malaysian Journal of Microbiology
Published by Malaysian Society for Microbiology
(In since 2011)
691 ISSN (print): 1823-8262, ISSN (online): 2231-7538
*Corresponding author
Mold Contamination and aflatoxin B1 levels in salted fish commodities from
traditional market in Yogyakarta and Surabaya, Indonesia
Zulfa Kamil Rafli, Yulius Darma Putra Damara, Sidar Andika, Setyaningsih Widiastuti, Sri Anggrahini, Pamudji
Rahardjo Agustinus, Sutriswati Rahayu Endang*
Department of Food and Agricultural Product Technology, Faculty of Agricultural Technology, Universitas Gadjah Mada,
Jl. Flora No 1 Bulaksumur Yogyakarta 55281 Indonesia.
Email: endangsrahayu@ugm.ac.id
Received 11 April 2018; Received in revised form 14 August 2018; Accepted 16 August 2018
ABSTRACT
Aims: To identify mold contaminant on salted fish, from two different market locations (Kenjeran market, Surabaya and
Beringharjo market, Yogyakarta). Furthermore, levels of AFB1 (aflatoxin B1) in salted fish samples were assayed.
Methodology and results: The samples were cultivated on DRBC (Dichloran Rose Bengal Chloramphenicol Agar) and
DG-18 (Dichloran (18%) Glycerol Agar) medium for enumeration, then transferred on MEA (Malt Extract Agar) medium
for isolation and identification, followed by ELISA test to measure the AFB1 level. Meanwhile aflatoxin biosynthesis
correlated genes (i.e. aflR, nor-1 and omtB genes) were identified using Polymerase Chain Reaction (PCR) method. The
results showed that Aspergillus tamarii and A. flavus being contaminant on salted fish along with A. sydowii, A. niger, A.
versicolor, Penicillium citrinum, and P. chrysogenum. Rhizopus sp. contamination was also found. AFB 1 was positively
detected in all of samples with the highest concentration measured was 75.81 µg/kg which belong to Lidah salted fish
and the lowest concentration measured was 4.33 µg/kg which belong to Rese salted fish. The suspected A. flavus and
A. tamarii isolated from salted fish was positively detected in the presence of aflR, nor-1 and omtB genes.
Conclusion, significance and impact of study: Mold contamination was detected in salted fish from two different
markets and all of those samples were contaminated by AFB1. These can be important information related to food
safety aspect for salted fish.
Keywords: Salted fish, mold contamination, Aflatoxin B1
INTRODUCTION
Indonesia is an archipelago country with the abundant of
fish commodities, which are non-perishable due to high
moisture and nutrient content of fish. According to the
statistical data on Indonesia’s fisheries year of 2015, fish
production in 2014 reached 6.50 million tons.
Furthermore, in Indonesia, the most common method to
prolong the self-life of fish is fish salting, which are done
by adding salt and drying. In Indonesia, salted fish is the
second preference after fresh fish to be consumed. It is
also often used as an ingredient or condiment in several
Indonesian foods.
According to Sutarni (2013), the variation of salt
concentration used in the salting process affects the salt
penetration into the fish’s body. Although salting the fish
may extend the self-life of fish, due to moisture content
reduction, the product is often sold uncovered. This can
lead to contamination of the product either from dust or
insect, such as flies. Sources of contamination also come
from poor personal hygiene from both the sellers and the
buyers. Based on fact that mentioned before, mold
contamination may occur in salted fish product. Moreover,
Indonesia’s tropical climate which has a high relative
humidity also gives a contribution to mold contamination.
In addition, based on Indonesia National Standard of
salted fish (SNI 01-2721-1992), which mentioned that
mold must not be detected.
The mold can grow at temperatures of 20-30 °C, with
a relatively low aw at 0.85 (Rahayu et al., 2014) and
affects the appearance of salted fish e.g. color and smell.
Furthermore, the most negative effect from mold
contamination is mycotoxins, because it cannot be easily
detected by the eyes. However, the growths of mold and
mycotoxins production are dependent on several factors
such as the mold strain and competitor, substrates,
temperature and relative humidity.
The occurrence of mold contamination has been
reported in dried salted fish, dried fish and smoked fish.
Smoked fish from tropical area has the potential for
Malays. J. Microbiol. Vol 14(7) 2018, pp. 691-698
DOI: http://dx.doi.org/10.21161/mjm.116117
692 ISSN (print): 1823-8262, ISSN (online): 2231-7538
contamination by toxigenic molds, such as Aspergillus
flavus (Adebayo-Tayo et al., 2008). Aspergillus flavus is
also the dominant species identified on smoked fish from
Sierra Leone area, followed by contamination of
A.ochraceus, A. niger and A. tamarii (Jonsyn and Lahai,
1992). In addition, A. flavus is also isolated from dried fish
in Sri Lanka and Indonesia (Atapattu and Samarajeewa,
1990). Furthermore, A. tamarii is found to be the
predominant contaminant in Maldives fish (Mohamed,
2013). Aspergillus sydowii is also isolated from dried fish
sold in traditional markets in Jakarta (Santoso et al.,
1999).
Some mold can produce mycotoxins as their second
metabolite. Aflatoxin, which majority produced by A. flavus
and A. paraciticus, has hepatotoxic, mutagenic,
teratogenic, and carcinogenic effect for both humans and
animals (Alberts et al., 2006). An outbreak of aflatoxin was
first noticed in the early 1960s in the UK (Turkey X
disease) (Do and Choi, 2007). Furthermore, according to
The International Agency for Research on Cancer (IARC,
2002), AFB1 is classified as a group 1 carcinogen which
means carcinogenic to humans. The occurrence of
aflatoxin is very common in developing countries (Samuel
et al., 2009). Moreover, aflatoxin is very heat stable,
therefore, they are not easily removed from food, even
after processing by high temperature (Rahayu et al.,
2003). According to Firsvad et al. (2005), there are other
molds that have the ability to generate aflatoxin. There are
A. nomius, A. pseudotamarii, A. parvisclerotigenus, and A.
bombycis of section Flavi, A. ochraceoroseus and A.
rambellii from section Ochraceorosei and Emericella
astellata and E. venezuelensis from Nidulatans section.
Because of those reasons, detection and identification
of mold contamination on salted fish from traditional
market in Indonesia is required as food safety and good
manufacturing processes information. The aim of this
study was to identify the mold that can contaminate salted
fish, and to quantify the AFB1 level in salted fish.
MATERIALS AND METHOD
Sample collection and preparation
A total of 20 different salted fish samples were used in this
study and purchased from two different traditional markets
to illustrate mold domination from different supply chain.
Salted fish from Kenjeran market, Surabaya were sold in
the open-air stalls near the production site, while salted
fish from Beringharjo market, Yogyakarta were supplied
from another city with semi-permanent stalls. All salted
fish were produced traditionally by adding salt followed by
sun-drying. Each sample was stored in polyethylene
plastic with specific code. Samples were stored in a cool
room in the microbiology laboratory, Center Study of Food
and Nutrition, Universitas Gadjah Mada. Before
performing the tests, the salted fish was allowed to heat
up to room temperature (±25 °C).
Mold Enumeration
All salted fish samples were inoculated on DRBC media
(Oxoid) and DG-18 (Oxoid) directly, large-sized samples
were cut with sterile scissors before inoculated. Samples
were then incubated at room temperature (±25 °C) for 5
days. Colonies formed were differentiated by appearance
and color (i.e. bluish grey, green, brownish green, black,
yellow, bluish green and velvet blue colony) for
enumeration. Enumeration shows percentage of samples
contaminated. The total pieces of analyzed samples can
be seen in Table 1.
% of samples contaminated =
100%
Identification of mold contamination
Colonies that grew on the surface of the sample were then
isolated based on the colour differences on MEA media
(Oxoid). Then the incubation was done for 5 days at room
temperature (±25 °C). Isolates obtained were identified
using macromorphology and micromorphology according
to Rahayu et al. (2014).
Determination of Aflatoxin B1 contamination
Detection of AFB1 in salted fish was assayed using
ELISA. Twenty-five grams of crushed samples was
extracted using 75 mL of 70% methanol and shook for 3
min. There was no pre-treatment to reduce the salt
content in salted fish samples. The extract was then
separated from the cake using Whatmann paper No.1.
AFB1 content was tested in accordance to the manual on
RIDASCREEN Aflatoxin B1 ELISA kit (Bioo Scientific)
then absorbance at 450 nm wavelength was measured.
AFB1 standard solutions concentrations of 0, 2, 5, 20, and
50 ng/mL were used as the calibration curve. The
calculated AFB1 in extract was then converted to AFB1 in
weight of samples.
Detection of aflatoxigenic genes in Aspergillus sp
Molecular detection of aflatoxigenic genes were
performed based on PCR method for amplification of
several genes correlated to aflatoxin production, such as
aflR, nor-1, and omtB genes. Reaction of PCR
amplification was performed in 25 µL of mix PCR using
Ready To Go PCR kit mixed with 21 µL ddH2O, 1 µL of
each primer, and 1 µL of DNA template. The PCR
program of aflR gene based on the procedure operated by
Rahimi et al. (2008), amplification was started with 1 cycle
initial denaturation at 94 °C for 5 min, followed by 35
cycles of denaturation at 94 °C for 1 min, annealing at 62
°C for 1 min, and extension at 72 °C for 2 min, as well as
1 cycle of final elongation at 72 °C for 10 min. In addition,
nor-1 gene amplification was performed according to
Geisen (1996), which consisted of 1 cycle initial
Malays. J. Microbiol. Vol 14(7) 2018, pp. 691-698
DOI: http://dx.doi.org/10.21161/mjm.116117
693 ISSN (print): 1823-8262, ISSN (online): 2231-7538
denaturation at 94 °C for 5 min, 30 cycles of denaturation,
annealing, and extension at 95 °C, 1 min; 65 °C, at 2 min;
and 72 °C, at 4 min, respectively. While, The PCR
program for omtB gene was based on Rodrigues et al.,
(2007) comprising of 1 step initial denaturation at 94 °C for
3 min continued by 35 cycles denaturation, annealing,
extension for 1 min at 94 °C, 1 min at 55 °C, 1 min at 72
°C, respectively and 1 cycle final extension for 10 min at
72 °C.
RESULTS AND DISCUSSION
Mold contamination of salted fish
From observation of the incubated samples, it became
clear that all of the samples on DRBC and DG 18 media
are contaminated by mold as seen on Figure 1. Yeast
colonies were also found on the salted fish, but was not
identified. Contamination on examined salted fish was
dominated by Aspergillus, followed by Penicillium. Hence,
suspected aflatoxin producer was also successfully
isolated (Figure 1) and identified as A. flavus and A.
tamarii. This was in alignment with result from Hassan et
al., (2011) that the most dominant contamination of salted
fish which is randomly collected from different shops and
retail markets at Giza Governorate was by Aspergillus sp.
members, at about 83.3% with A. flavus contamination
being at 66.6%. According to Pitt and Hocking (2009),
Aspergillus and Eurotium were the most dominant
contamination species found on dried food in tropical and
subtropical regions. Aspergillus and Eurotium were also
dominant on smoked dried fish from warm water regions
(Adebayo-Tayo et al., 2008). This was caused by
Aspergillus which usually grows faster than Penicillium but
takes longer to sporulate (Rahayu et al., 2014).
The samples obtained from Kenjeran market Surabaya
had different dominant mold from those obtained in
Beringharjo market, Yogyakarta. This was due to
differences in the supply chain and the handling. Samples
from Kenjeran market, Surabaya were sold in coastal and
open roadside stalls, in addition the production sites and
the dried fish stalls were close to each other. While the
samples from Beringharjo market, Yogyakarta consisted
of salted fish supplied from Pati, Central Java and the
stalls were all in one room, with not only salted fish stalls
but also another staple food (i.e. vegetables, fruits, tubers,
and beef). According to Sharma (2012), A. niger is
commonly found in indoor environment, fruit and
vegetable. This explains that mold domination in
Beringharjo market comes from the environment of the
stalls. On the other hand, Aspergillus sp. are closely
associated with the soil (Owaga et al., 2009). Therefore, it
is possible that the sun-drying of salted fish on the
roadside of Kenjeran market could have exposed with
Aspergillus sp., particularly from A. flavus.
The frequency of mold contamination on salted fish is
shown in Table 2. Samples from Kenjeran market,
Surabaya showed that all samples were found to have
mold contamination. The dominant mold was 100% A.
tamarii (9/9 samples) and 89% A. flavus (8/9 samples).
Lidah salted fish had the highest frequency of A. flavus
contamination which was about 92%. Spores of A. flavus
were found in the tropical air. In Indonesia itself, which is a
tropical country, the drying of salted fish still takes the
advantage of sunshine, thus increasing the likelihood of
contamination by fungal spores. The samples from
Beringharjo market, Yogyakarta were also contaminated
by mold. The dominant molds were A. niger, A. flavus, A.
tamarii, and Rhizopus sp with the frequency of each
contamination approximately 45% (5/11 samples). The
highest A. flavus contamination was found on Pethek
salted fish at the level of 50%. By the presence of A.
flavus on salted fish, it is possible to find aflatoxin
contamination.
Figure 1: (a) Mold diversity contaminating on salted fish (upper-side is on DG-18 medium and down-side is on DRBC
medium). (b) Suspected aflatoxin producer identified as A. flavus (upper-side) and A. tamarii (down-side).
Malays. J. Microbiol. Vol 14(7) 2018, pp. 691-698
DOI: http://dx.doi.org/10.21161/mjm.116117
694 ISSN (print): 1823-8262, ISSN (online): 2231-7538
Table 1: Macro- and micro-morphology of identified mold.
Isolate code
Macro morphology
Micro morphology
Mold identified
HAF1
Green colony, ±50 mm colony diameter,
reverse colony creamish-yellow
Biseriate, spherical vesicle,
hyaline, long conidiophores
A. flavus
BAAS1
Bluish gray colony, ±15 mm colony
diameter, produce exudate, reverse
colony brown
Biseriate, circular vesicle, hyaline,
cylindric phialides, small conidia
A. sydowii
CAT2
Solid brownish green colony, ±50 mm
colony diameter, reverse colony
creamish-yellow
Biseriate, long conidia, spherical
vesicle, hyaline, cylindric phialides
A. tamarii
KAV2
White colony at the beginning and turn to
yellow, velvet-like appearance, ±10 mm
colony diameter, reverse colony cream
Biseriate, oval (spoon-shaped)
vesicle, spreading conidia, hyaline
A. versicolor
HAN1
Black colony, ±60 mm colony diameter,
reverse colony dark creamish-yellow
Biseriate, black spherical vesicle,
long conidiophores, circular
conidia, cylindric phialides
A. niger
HBPC1
Velvet blue colony, ±25 mm colony
diameter, reverse colony pale yellow,
produce exudate
Tervertisilata, hyaline, cylindric
phialides
P. chrysogenum
HAPC2
Bluish green colony, ±25 mm colony
diameter, produce exudate, slow growth,
reverse colony pale yellow
Bivertisilata, hyaline, cylindric
phialides
P. citrinum
Another important genus which was identified is
Penicillium, although it had a lower frequency than the
Aspergillus genus. Penicillium citrinum and P.
chrysogenum were also identified on some salted fish
samples. The highest P. citrinum contamination was found
on Ebi salted fish, at about 50% and the highest P.
chrysogenum contamination was found on Lidah salted
fish, at about 33%. P. citrinum produces citrinin toxin
which is nephrotic and carcinogenic for animals (Flajs and
Peraice, 2009).
Seven isolates identified had halotolerant and
xerophilic properties. Aspergillus sydowii is a mold mostly
found on marine organisms. According to Prakash et al.
(2011). Aspergillus flavus and A. niger, which originated
from dried seafood products in India showed salt
resistance of up to 18%. Aspergillus flavus was also found
to be the dominant contaminant on smoked dried fish
(Adebayo-Tayo et al., 2008). In the case of Maldives fish,
which is a salted fish from the Maldives region, A. tamarii
contamination was higher than A. flavus contamination,
because Maldives fish has a low salinity at an average
2.08% (Mohamed, 2013). It is similar to the total mold
contaminant in samples from Kenjeran market, Surabaya.
Aflatoxin B1 contamination in salted fish
The presence of A. flavus indicates the possibility of
contamination by AFB1 in salted fish. Out of the samples
analyzed, all were positively contaminated by AFB1. The
highest AFB1 contamination was on the Lidah salted fish
taken from Kenjeran market, Surabaya which had 75.81
µg/kg. And the lowest AFB1 contamination was on Rese
salted fish taken from Beringharjo market, Yogyakarta
which had 4.38 µg/kg. The AFB1 levels of the salted fish
are shown in Table 2.
The concentration of aflatoxin in samples was
determined from the standard calibration curve plotted in
the range of 0, 2, 5, 20, and 50 ng/mL using ELISA.
Linear regression analysis was further used for the
quantification of aflatoxin present within the samples. The
standard calibration graph showed excellent linearity with
R2 value of 0.9967. The limit of detection and
quantification of ELISA method were 0.34 and 1.02 ng/mL
respectively.
If the result of ELISA test is correlated with the
frequency of A. flavus contamination on the samples
(Figure 2), it can be fully understood that the amount of
AFB1 in the samples was not affected by the frequency of
A. flavus contamination. Hence, if the ELISA result
compared to mold species contaminated on samples, not
all samples were contaminated by A. flavus which is an
AFB1 producer. Meanwhile, all of samples were positively
contaminated by AFB1 in different amounts.
Figure 2. Corellation frequence of A. flavus and aflatoxin
B1 level.
Malays. J. Microbiol. Vol 14(7) 2018, pp. 691-698
DOI: http://dx.doi.org/10.21161/mjm.116117
695 ISSN (print): 1823-8262, ISSN (online): 2231-7538
Table 2: Frequence of mold contamination and Aflatoxin B1 levels in salted fish samples.
Salted
Fish
Species
Analyzed
sample
Source
Aflatoxin B1
(µg/kg)
Frequence of mold contamination (%)*
A. flavus
A. tamarii
A. niger
A. sydowii
A. versicolor
P. citrinum
P. chrysogenum
Rhizopus sp
Bulu
bebek
Thryssa mystax
8
Kenjeran
market,
Surabaya
(9
samples)
8.52
-
25
-
38
25
-
-
-
Campur
-
18
9.63
22
28
39
-
-
-
-
-
Glomo
Johnius coitor
12
19.86
33
50
-
33
-
25
-
-
Janjan
Anguilla rostrata
12
16.14
42
25
42
17
33
-
-
-
Keteng
Mystus planiceps
12
9.63
50
25
8
50
25
-
-
-
Kindo
Unknown
11
13.29
18
36
-
27
-
-
-
55
Lidah
Cynoglossus
lingua
12
75.81
92
25
8
25
25
-
33
-
Talang
Chorinemus tala
12
13.77
42
50
-
-
-
-
-
-
Udang
Litopenaeus
vannamei
12
27.42
25
25
8
-
17
-
-
-
Banyar
peda
Rastrelliger
kanagurta
12
Beringharjo
market,
Yogyakarta
(11
samples)
26.37
8
-
8
-
-
-
-
-
Belek
Unknown
12
7.59
-
-
-
-
-
-
8
-
Ebi
Litopenaeus
vannamei
12
6.24
-
-
-
-
-
50
-
17
Kacangan
Charangidae sp
12
9.39
-
-
17
-
-
-
25
-
Layur
Trichurus savala
12
33.72
-
8
-
8
-
17
-
8
Pedho alit
Rastrelliger
scombridae
12
7.98
33
8
-
-
-
-
-
17
Pethek
Leiognathus
equulus
8
5.61
50
-
-
-
-
63
-
100
Rese
Acetes indicus
30
4.38
33
7
7
-
-
-
30
-
Sero
Unknown
12
10.80
-
-
-
-
-
-
-
23
Teri
Stolephorus tri
10
6.48
10
-
20
-
10
20
-
-
Teri Nasi
Stolephorus
commersonii
22
11.19
-
-
9
-
-
-
14
-
*The number expressed as the percentage was calculated in relation to the amount of analyzed samples.
Aflatoxin contamination has no correlation to the
frequency of A. flavus on samples because it does not
grow well. The growth of A. flavus was resisted by other
more dominant molds, so there were some samples that
were not contaminated by A. flavus. In addition, according
to Abidin et al. (2010) A. tamarii also can produce AFB1
and AFB2, probably the presence of aflatoxin was
produced by A. tamarii instead of A. flavus. Moreover, the
ELISA method has low specificity to detect other
components which have a similar structure to AFB1, such
as AFB2, AFG1 and AFG2 (Leszczynska et al., 2001).
The salt content in the samples caused a matrix effect that
can affect the absorbance measurement. It is also
possible a cross-linking component to occur in the
sample, thus affecting the selectivity and specificity of
ELISA measurement (Rachmawati et al., 2004). Because
of those reasons, ELISA method has a limitation for
analyzing AFB 1 in salted fish samples, regarding there
was no pre-treatment to reduce the salt content in
samples.
Molecular detection of aflatoxin biosynthesis genes in
Aspergillus strain
Fungal group of Aspergillus (green Aspergilli and black
Aspergilli) found in salted fish were then selected for
aflatoxigenic detection based on PCR amplification
method. Three types of genes involved in aflatoxin
biosynthesis namely aflR, nor-1, and omtB were used in
detecting aflatoxin production ability. Based on
amplification result, primer aflR, nor-1, and omtB were
able to amplify an expected for approximately 600 bp, 400
bp, and 1000 bp fragment in aflatoxigenic isolates,
respectively (Figure 3). Aflatoxigenic and nonaflatoxigenic
Malays. J. Microbiol. Vol 14(7) 2018, pp. 691-698
DOI: http://dx.doi.org/10.21161/mjm.116117
696 ISSN (print): 1823-8262, ISSN (online): 2231-7538
strains could be determined on the basis of amplification
of these three target DNA fragments.
Based on Table 3, it demonstrated that almost all
isolates of green Aspergilli showed positive results in the
presence of aflR, nor-1, and omtB genes. The positive
results against those primer were A. flavus and A. tamari.
Meanwhile, none of black Aspergilli have those three
genes detected. Two isolates only have both regulatory
genes, aflR and nor-1, while the other two isolates have
none of three target genes.
The existing of three genes target involved in aflatoxin
biosynthesis such as aflR, nor-1, and omtB may indicate
that the isolate was able to produce aflatoxin. PCR
analysis was able to amplify aflatoxin biosynthetic genes,
i.e aflR, nor-1, and omtB in almost green Aspergilli group,
while none of omtB gene detected in black Aspergilli.
Aflatoxin, a secondary metabolites which is a polyketide-
derived, is produced via the following conversion path:
acetate polyketide anthraquinones xanthones aflatoxin
(Bhatnagar et al., 2003; Yu, 2012). However, aflatoxin
formation depends on the final step of aflatoxin
biosynthesis pathway. In this case, norsolorinic acid
(NOR) is the earliest step and the first stable aflatoxin
precursor in the aflatoxin biosynthetic pathway thus it play
a role in polyketide synthase (Yu et al., 2004; Yu, 2012).
In contrast, O-methyltransferase (omtB) involve in the
later step of aflatoxin formation. Meanwhile, aflR is a
positive regulatory gene which is required for
transcriptional activation of most of the structural genes
such as nor-1 and omtB (Yu et al., 2004). Thus, the
absence of omtB gene in such black Aspergilli isolate
might indicate that those are unable to produce aflatoxin,
while the presence of those three target genes in green
Aspergilli are possible to support the correlation of ability
in aflatoxin production.
Table 3: Detection of aflatoxigenic and non-aflatoxigenic
Aspergillus group based on PCR amplified product.
Code
Mold
species
Aspergillus
group
Primer AFB1
nor-1
aflR
omtB
IC1
A. tamarii
Green
Aspergilli
-
-
-
IC2
A. tamarii
+
+
+
IHK1
A. flavus
+
+
+
IHK2
A. flavus
+
+
+
IB
A. niger
Black
Aspergilli
-
-
-
IHTM
A. niger
+
+
-
IH
A. niger
+
+
-
IHIT
A. niger
-
-
-
Figure 3: Amplified product of five target genes in Aspergillus. (A) Amplified product of aflR gene in Aspergillus; (B)
Amplified product of nor-1 gene in Aspergillus; (C) Amplified product of omtB gene in Aspergillus.
Malays. J. Microbiol. Vol 14(7) 2018, pp. 691-698
DOI: http://dx.doi.org/10.21161/mjm.116117
697 ISSN (print): 1823-8262, ISSN (online): 2231-7538
CONCLUSIONS
Aspergillus tamarii, A. flavus, A. sydowii, A. niger, A.
versicolor, P. citrinum, P. chrysogenum and Rhizopus sp
are identified being contaminant on salted fish. Salted fish
samples from Kenjeran market were dominated by A.
tamarii and A. flavus, while salted fish samples from
Beringharjo market were dominated by A. flavus and A.
niger. The difference of supply chain from both sources of
samples affects the mold domination. In addition, AFB1
was detected positively in all of samples with highest level
belong to Lidah salted fish (75.81 µg/kg) and the lowest
AFB1 level belong to Rese salted fish (4.33 µg/kg).
Besides, ELISA test has a limitation for quantify AFB 1
level from salted fish, due to the high salt content that can
be interference for absorbance measurement. The
suspected A. flavus and A. tamarii isolated from salted
fish were positively detected in the presence of aflR, nor-1
and omtB genes.
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