European Journal of Medical Genetics and Clinical Biology
Volume 1, Issue 8 | 2024
FUNGAL CONTAMINATION IN FRESH RAISIN
JUICE IN MOSUL CITY
https://doi.org/10.61796/jmgcb.v1i8.887
May Akram Krmo1
nineveh Education Directorate,Mosul-Iraq
1mayakram9@gmail.com
Maha Akram Mohammad Ali Al_Rejaboo2
Department of Biology, College of Science, University of Mosul,
Mosul- Iraq
mahaalrejaboo2@uomosul.edu.iq
2
Received: Jun 22, 2024; Accepted: Jul 29, 2024; Published: Aug 18, 2024;
Abstract: Background: Fungal contamination in food and beverages, including fruit juices, poses significant
health risks. Specific Background: This study investigates the extent of fungal contamination in fresh raisin
juice collected from Mosul City and its suburbs, including Bashiqa, Al-Hamdaniya, Bartella, Al-Shekhan,
Aqra, and Dohuk, over a period from January 2023 to April 2024. Knowledge Gap: Despite the potential for
fungal contamination in fruit juices, detailed analyses of specific contamination rates and fungal species in
this region have been lacking. Aims: The study aimed to identify and quantify fungal contamination in fresh
raisin juice samples using Potato Dextrose Agar and Sabouraud Sucrose Agar, and to characterize the fungal
isolates morphologically, biochemically, and molecularly. Results: Of the 40 samples tested, 100% were
contaminated with fungi, yielding 121 isolates of various species. The highest contamination rates were
observed for Candida spp. and Aspergillus spp., each constituting 27.3% of isolates. Other notable
contaminants included Wickerhamomyces anomalus (12.39%), Saccharomyces cerevisiae (11.57%), and
Rhodotorula spp. (8.26%). Additionally, lower rates of contamination were found for Penicillium spp.,
Exophiala dermatitidis, and several other species. Novelty: The study revealed a high prevalence of fungal
contamination, including the identification of a novel Candida oleophila strain (GenBank accession
PP961930.1). Implications: These findings highlight the significant fungal contamination in fresh raisin
juice, which could have implications for food safety and quality control. The identification of specific fungal
species and the novel strain underscores the need for stringent monitoring and improved processing practices
to mitigate contamination risks.
Keywords: Fresh Raisin Juice, Fungal contamination, Mosul City, Its Suburbs
This is an open-acces article under the CC-BY 4.0 license
Introduction
Raisins are dried grapes Grape is a berry fruit that generally grows in a group of deciduous
woody vines belonging to the Vitaceae family. (Venkitasamy et al., 2019). Grapes are an important
and economically well-known fruit all over the world (Kui et al., 2020). Grape production is estimated
at more than 74.5 million tons per year, based on a cultivation area of 7.12 million hectares. China,
the United States, Italy, Spain, France, Turkey, Argentina, India and Iran are the largest producers of
grapes (Anjum et al., 2020). It is mainly consumed as food such as raisins, juice and wine and as fresh
fruit or used in various products such as vinegar. It is considered a rich source of vitamins, minerals,
carbohydrates, dietary fiber and substances chemical vegetarian like polyphenol and promotes general
health (2023,.Jaiswal et al). It is also considered antibacterial, antioxidant and anti-inflammatory due
to its content of Polyphenol (Imran et al., 2017). Black grapes have large amounts of antioxidants and
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grapes are considered a seasonal fruit that grows abundantly and thus they were dried and converted
into raisins for storage and use throughout the year, either by the old method of drying under the sun
or in the air (2015,.Wang et al). As for modern drying using microwaves, ovens and drying under
infrared rays (Wang et al.,2016 and Khiari et al., 2018a). Old and new drying techniques greatly affect
changes in the volatile organic compounds that contribute to the aroma of raisins.((Torres et al., 2015;
Wang et al., 2017). Raisins have benefits for blood pressure and cholesterol low-density lipoproteins,
triglycerides, and fatty acids oxidizing in the blood and cytokines that cause inflammation, this
indicates the possibility that raisins greatly reduce the risk of developing type 2 diabetes.) (T2DM,
cardiovascular disease, cancer, and gut health, indicating that raisins consumed as part of a healthy
diet are rich in nutrients (Das, 2008 and Restani et al., 2016).
Raisin juice is considered one of the popular juices it is used as a healthy alternative to
caffeinated drinks such as coffee, tea and soft drinks. It has many benefits such as antioxidants, anticancer and anti-diabetic properties, and vitamins and minerals play an effective role in preventing a
cancer, heart disease, diabetes and high cholesterol (Madappa et al., 2018). It contains 70% of its
weight sugar and 2% of its weight protein. It is also rich in elements, especially K, Na, P, Mg, and
Ca+ and vitamins A, B3, C. there are many contaminated according to the type of storage until
consumption and thus affects the production of juice and wine (Welke, 2019) . This results in
contamination of raisin juice with mycotoxins (Pena et al.,2010).Studies have shown that fungal
contamination attributed to climate conditions and physiochemical and for grapes and wine and its
effects on production (Freire et al., 2017).
Methods
(40) samples of fresh raisin juice were collected from Mosul city to the right and left and its
suburbs, including the areas of Bashiqa, Al-Hamdaniya, Bartella, Al-Shekhan, Aqra and Douw, with
50 ml of each sample, and placed in special sterile boxes to avoid contamination and take information
about each sample in terms of the source of the raisins, the date of making the juice, the method of
storing it and the components of the juice. The sample collection period was from January 2023 to
April 2024, and the samples were cultured using the dilution method on the media (PDA) Potato
Dextrose Agar and Sabouraud Sucrose Agar (SSA) (Ronald et al., 1959 and Marth, 1978). To
determine the extent of fungal contamination, it was diagnosed morphologically by relying on the
color of the colony in the dish from both sides, the diameter of the colony, the presence and color of
secretions, and the appearance of the fungal hyphae. Microscopically, a small piece of the colony was
taken with a small amount of the medium on a glass slide, then mixed with a drop of lactophenol dye
or crystal violet dye, then the cover glass was placed on it and examined under a microscope at a
magnification of 40x (de-hoog and Gurro,1997 ; Pitt and Hocking ,1997 ; Samson and Frisvad
,2004).Yeasts were diagnosed using differential media based on the taxonomic key (Pitt and Hocking,
1997), where 7 differential media were used, including MEA25, MEA37, MEA28, Malt Extract Agar,
Czapek Agar (CA), Malt Acetice Agar (MAA), Malt Extract yeast 50% Glucose Agar (MY 50G),
and Extract Yeast Malt 5% or 10% salt 12% Glucose Agar (MY10-12). The isolates were cultured
after purification on SSA medium for two days until diagnostic tests were performed, and the general
characteristics were recorded in terms of colony color, texture, and size. The diagnosis of the fungal
isolate was confirmed using molecular methods, where DNA was isolated from two-day-old yeast,
its purity was checked, PCR was performed, and DNA was isolated by electrophoresis and
sequencing. The result was then sent to the fungal isolate for the purpose of conducting a nitrogenous
base sequence determination test after the sample was diagnosed using ITS primers in the United
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Volume 1, Issue 8 | 2024
States of America.
Results and Discussion
Isolated and identified the contaminating fungi for (40) sample of natural raisin juice sold in
the markets of Mosul and its suburbs from December 2023 to April 2024. Elected(28)sample from
shops randomly on the side in Mosul right and left it included all of the shops in Al-Muthanna district,
Al-Zahour district, Al-Majmoua Al-Thaqafiya district, Mosul University, Al-Darkazliyya, AlMasarif district, three samples from Bashiqa district, two samples from Sheikhan district, two samples
from Bartella, and one sample from Al-Hamdaniya and two samples from Aqra district and four
samples from Dohuk governorate, where the location of the shop and the source of the raisins were
taken, as well as the date of making the raisin juice from all the selected shops. The fresh juice was
composed of (dried raisins, sugar, and a dried mint leaves and a Synthetic dye). The results of the
study showed pollution for large samples-Mold and Yeast ,there were fungi growing on both media
SSA, PDA and fungi grown on the only one medium. Sample R1, R2 and R5 isolated from Bashiqa
stores contained on a mold and yeast in both media SSA, SDP which was the source of raisins from
the perfumers of Mosul from it C.oleophila one types of Candida which was registered as a new
genotype in NCBI named Candida strain MAYKAIY and we have shown Aspergillus niger and
Saccaromyces cerevisiae and P.atramentosum was found in the plate containing SSA medium and
not in SDP medium. The fungus C.tropicalis was one of the most contaminated fungi in these
samples.
Figure (3): C.tropical yeast
As for the samples taken from Aqrah district, it was found that:R27 contaminated only With
A.nigerand R28 contaminated with A.niger, Rhodotorula and Wickerhamomyces anomula and
C.parapsilosis, where we grew upRhodotorula On each ofPDA did not grow on SSA, but the most
common species that appeared between the two samples was A.niger, while the Samples R3 and R13
TakenFrom the judiciary the Two old men grew mushrooms A.niger, C. oleophila, Saccharomyces
cerevisiae in both SDP The most frequent fungus in both samples A.niger
Figure (4): A. niger on the SSA medium
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European Journal of Medical Genetics and Clinical Biology
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the two samples R10, R11 from Bartella area were sourced in Mosul grew both Cladosporium
cladosporioides, A. niger, Saccaromyces cerevisiae, C. oleophila, C. tropicalis, Rhodoroula,
although both samples were taken that from two different places and also it was A.niger more visible
in both samples and the sample R12 from area the Hamdaniya was the source of raisins from northern
Iraq contaminated with- Penicillium, C. parapsilosis , A.niger , Saccaromycetes services and one
sample of Rhodotorula , the most common fungi are: A.niger collect four samples of R18,R23,R24,R7
from Dohuk was the source of raisins taken from scattered areas in northern Iraq contaminated withC.tropicalise and C. oleaphila, Wickerhamomyces anomula, Rhodotorula, and a species of
Aspergillus spp. While C. tropicalis was abundant, its proportion was large. As for the samples of
Mosul city on the left side, they were:R14, ,8R4,R6,R9,R19,R20,R25,R26,R27,R30,31R,
R32,R33,R34 R35, R36, R37,R38, R39,R40 most of all the raisin samples were from Aqra district
and northern Iraq. All kinds of fungi grew A. niger, C.oleaphila, Cladosporium cladosporioides,
Rhodotorula, C.krusei, C.albican, C.parapsilosis, C.tropicalis and species of Penicillium. The
contamination rate was high in some samples and low in others . As for the right side of the city of
Mosul, the samples wereR15, R16, R17, R18 are contaminated with Aspergillus sp and C.tropicalis
and Saccaromyces cereivsiae and Wickerhamomyces anomalus and C. parapsilosis and Rhodotorula
and C.krusei and C.albican , AThe more contaminated the samples were Rhotorula it was found that
all samples of raisin juice were contaminated to varying degrees with fungi, with the contamination
rate being- Aspergillus sp. 27.3% While she was Candida sp Contaminated by 27.3% and12.39%
Contaminated with- Wickerhamomyces anomalus followed by other fungi at a rate of Say and it was
kindAspergillus niger andCandida spFrom moreTypesContamination and spread in samples.
The shape (5): Shows the percentage of contamination of the study samples with fungal and
yeast species.
11.57%
8.26%
3.3%
3.3%
12.39%
2.47%
2.47%
0.82%
0.82%
0.82%
27.3%
27.3%
Aspergillus spp.
Candida spp.
Wickerhamomyces anomalus
Saccharomyces cerevisiae
Rhotodorula spp
Exophiala dermatitidis
Penicillium spp.
Cladosporium spp
Trichophyton mentagrophytes
Geotrichum spp
Epidermophyton floccosum
Fusarium spp
It was found that the most highly contaminated samples with fungi were:R1, R3, R4, R5, R7,
R9, R10, R11, R12, R14, R15, R16, R17, R19, R20, R21, R22, R26, R28, R29, R30, R35, R36, R38,
R39, R40 either samples are less contaminated it is:R2, R6, R8, R13, R18, R18, R23, R24, R25, R27,
R31, R32, R34, R40. In general, all samples contained yeasts and mold. The results that appeared to
us were consistent with what the scientist mentioned (Alomari et al., 2014) found that Aspergillus
niger was dominant in some dried fruits during isolation and also observed different species of
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Penicillum spp. It was agreed results the current study with (Iamanaka et al., 2005), Aspergillus
niger is the most prevalent in raisin samples in the Brazil , (Allam et al., 2020) it was found that the
genus Aspergillus spp is dominant by 27.3% over the rest of the other genera yeasts and other
filamentous fungi, this study agreed with our study.
Figure(6):New mushroom pattern registration Candida oleophila strain MAYKAIY and give
it an ID number PP961930 in NCBL.
Conclusion
The investigation of fungal contamination in fresh raisin juice from Mosul City and its suburbs
revealed a high prevalence of various fungal species. The study identified 121 fungal isolates from
40 samples, with Candida spp. and Aspergillus spp. being the most frequent contaminants, each
representing 27.3% of the isolates. Other notable contaminants included Wickerhamomyces
anomalus, Saccharomyces cerevisiae, and Rhodotorula spp., highlighting significant microbial
diversity. The results underscore the widespread issue of fungal contamination in raisin juice, which
could impact consumer health and product safety. This study's findings emphasize the need for
improved quality control measures during juice production and storage. Further research should focus
on assessing the health impacts of these contaminants and exploring effective methods for reducing
fungal contamination in raisin juice and similar products.
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