International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-4, July-Aug- 2017

http://dx.doi.org/10.22161/ijeab/2.4.10 ISSN: 2456-1878

Isolation and Identification of Fungi from fast

food restaurants in Langa Bazar
Sewgil Saaduldeen Anwer1, Gazang A. Ali2, Chra Z.Hamadamin3, Hanan Y. Jaafar4
1
Department of Clinical Biochemistry, College of Health Science, Erbil, Kurdistan Region, Iraq.
2,3,4
Department of Medical Microbiology, College of Health Science, Erbil, Kurdistan Region, Iraq.

Abstract A total of (218) samples from Eleven different Microbial spoilage is a major concern for so-called
foods were processed between October 2016 and February perishable foods such as fresh fruits, vegetables, meats,
2017 which include (Tomato, Chicken meat, red meat, poultry, fish, bakery products, milk, and juices. Meat and
falafel, potato, bread, eggplant, cabbage, celery, cucumber dairy products, with their high nutritional value and the
and onion). Samples were collected from 4 different fast presence of easily metabolized carbohydrates, fats, and
food restaurants inoculated on Potato dextrose agar and proteins provide ideal environments for microbial spoilage
Sabouraud Dextrose Agar. Isolated fungus identified (3).
morphologically and microscopically in accordance with A wide variety of fungi, including species of Rhizopus,
standard procedures. Results showed that six fungal genera Alternaria, Penicillium, Aspergillus, and Botrytis spoil
were associated with the selected fast food restaurants. The foods. Since fungi grow readily in acidic as well as low-
isolated fungal genera were Aspergillus sp., Alternaria sp., moisture environments, fruits and breads are more likely to
Mucor sp., Rhizopus sp., Saccharomyces sp., Brettanomyces be spoiled by fungi than by bacteria. AspergillusFlavus
sp. The number of total colonies in October were 236 and in infects peanuts and other grains, producing aflatoxin, a
February were 119 and the number of colonies were higher potent carcinogen monitored by the food and drug
when cultured on Potato dextrose agar than Sabouraud Administration (4).
Dextrose Agar. There was variation in the pattern of The factors that affect microbial growth in food and
occurrence of the fungus in fast foods Aspergillus sp. constantly the association that develop also determine the
appears to be the most pathogenic fungi that present in the nature of spoilage and any health risks posed for
food samples. convenience they can be divided into four groups Intrinsic
Keywords Fungus, Fast food restaurants, Food factors Intrinsic factors of a food include nutrients, growth
spoilage. factors, and inhibitors or (antimicrobials), water activity,
pH, and oxidationreduction potential. The inuence of
I. INTRODUCTION each factor on growth in a food system the factors are
Food spoilage is any change in the appearance, smell, or present together and exert effects on microbial growth in
taste of a food product that makes it unacceptable to the combination, either favorably or adversely (5,6). , Extrinsic
consumer. Spoiled food may still be safe to eat, but is factors Which include environmental factors like
generally regarded as unpalatable and will not be purchased temperature, Relative humidity and gases, Implicate factors
or readily consumed. Food spoilage causes losses to Include Specific growth, Synergism, Antagonism and
producers, distributors, and consumers in the form of commensalism competition among the microorganism that
reduced quality and quantity and higher prices (1). Another cause changes of physical and chemical structure of the
case of spoilage could be that the nutrients (e.g. vitamin food and spoiled the food (7,8,9).and Processing factors
content) in the food have deteriorated to the point that the Include the tools with slaughtering and cutting the water
food product no longer meets its declared nutritional value. which use in washing, vehicles which are used in
The time it takes for a food product to reach one of these transporting, and retail market (8).
spoilage conditions is generally termed the products shelf-
life potential food spoilage microorganisms include Food-borne illness by fungi:
bacteria, fungi (mold and yeast), viruses, and parasites.(2). A food-borne illness occurs when a person becomes ill after
Action by microorganisms is a common means of food eating or drinking contaminated foods or beverages.
spoilage and the most common cause of foodborne illness. Nothing that we eat or drink is completely pure, and many

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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-4, July-Aug- 2017
http://dx.doi.org/10.22161/ijeab/2.4.10 ISSN: 2456-1878
microorganisms live in the foods and beverages that we Identification:
consume. Microorganisms or microbes are tiny organisms Fungal isolates were transferred to sterilized plates for
such as bacteria, Fungi, viruses, and parasites that exist in purification and identification. The grown fungi were placed
our bodies as well as in plants, animals, food, water, air, and on a slide, stained with gram stain for yeast identification
soil. Many of these microbes do us no harm, and some and lacto phenol cotton blue to detect fungal structures
"friendly" ones are necessary for a healthy body, but other covered with a cover slip, examined under microscope and
microbes can cause infections that lead to serious, or even identified on the basis of their colony morphology and spore
fatal, illness. Additionally, there are harmful chemicals and characteristics (12).
other substances in food that can cause a food-borne illness Macroscopic and microscopic observations were carried out
(10). on the cultures. The physical characteristics of the mycelia
Fungi include mold and yeasts which are more adaptable to such as the colour and structure were noted as well as the
various conditions than other microorganisms. Have high microscopic characteristics (13,14).
tolerance for acidic condition and more often responsible
for food spoilage than for food-borne illness (11). III. RESULTS
3.1.Variation of colony numbers in Fungus during
II. MATERIAL AND METHODS October:
Media preparation: After collecting the samples and culturing we obtained the
Two media can be used, Potato Dextrose Agar (PDA) and colonies and after examining the colony under microscope
Sabourated Dextrose Agar (SDA). They were prepared we obtained some fungal genera that associated with food
according to the manufacturers instructions 39gm (PDA) spoilage. Figure 3.1.showspercentage of colonies isolated
powder in 1 L of distilled water for PDA medium. And from fast food restaurants during October ( 22/10/2016).
62gm (SDA) powder in 1L distilled water for SDA medium
, then sterilized and used for fungal cultivation.

Collection of samples and Isolation:

A total of 218 sample from 11 different foods were
obtained from 4 restaurant three times a day respectively (
9:30 AM, 12:00 PM, 4:30 PM) during the months of
October 2016- February 2017. The food samples were
taken from restaurant in sterile plastic bags according to
Cheesbrough 1984 Direct plate used for culturing the
Individual yeast and mold species (figure 2.1) The samples
transferred with sterile forceps into Petri dish contain
sterilized SDA Saboured dextrose agar and PDA Potato
Dextrose Agar and the plates were incubated at 25C for 5- Fig.3.1: Percentage of total colony isolated in food during
7 days(12) October in (PDA, SDA).

3.1.2. Variation of colony of Fungus number during

February:
The number of colony decreased when sample collected and
examined by cultivation to obtain colony.figure 3.2 shows
the percentage of colonies isolated from fast food restaurant
during February (14/2/2017).

Fig. Sampling and Culturing of fungi on SDA and PDA.

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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-4, July-Aug- 2017
http://dx.doi.org/10.22161/ijeab/2.4.10 ISSN: 2456-1878
activities researchers (15,16,17) .Yassin and his coworkers
isolated same fungal strains from restaurant and meat shops
in al Samawa (18).
3.3. Cultural and Microscopic properties of Fungal
isolates:
Microscopically examination for isolates from Cucumber,
Red meat, Chicken meat and onion showed that
Conidiophores and septate, unbranched hyphae. With a
swollen apex (vesicle) .Phialides born on the metulae
(biseriate). Conidia in dry chains forming compact columns
(columnar) that can be identified as Aspergillus sp.
Fig: Percentage of total colony isolated in food during Colonies on PDA agar at 25oC within 7 days, and consisting
February in (PDA, SDA) medium of a dense felt of green conidiophores. (Figure 3.3)

3.2. Detection of Fungus In different foods:

From a total of (218) collected samples and 11 different
foods from different fast food restaurants a total of 6 genera
(mold and yeast)
were isolated as shown in the table 3.1

Table.3.1: Occurrence of fungal genera in different food

sample.
No. Food type Fungal genera
1 Tomato Alternaria sp.
2 Eggplant Mucorsp.
3 Falafel Saccharomyces sp.
4 Cucumber Aspergillus sp.
5 Potato Mucor sp.
6 Chicken meat Aspergillus sp. Fig.3.3::Microscopic examination of Aspergillus sp.
7 Red meat Aspergillus sp.
8 Onion Aspergillussp. Microscopically examination for isolates from falafel
9 Cabbage Brettanomycessp. .showed that cells quite large, globules to sub globes,
10 Bread Rhizopus sp. reproduce by budding as showed in figure 3.4.and the
colonies on PDA agar at 25 oC within 7 days, and whitish
11 Celery Alternariasp.
mucous texture that can identified as Saccharomyces sp.
The major problem associated with fast food is the frequent
incidence of contamination .Due to the nature of these foods
and their methods of preparation involving extensive
handling. The results of the present study revealed that
different molds were isolated from 11 different food
samples namely eggplant, falafel, chicken meat, red meat,
celery, cabbage, potato, tomato, bread and cucumber
collected from four locations fast food restaurant in Langa
Bazar. The isolated molds were identified as food-borne.
The molds include Aspergillus sp., Alternaria sp., Mucor
Fig.3.4: Microscopic examination of Saccharomyces sp.
sp., Rhizopus sp. and Saccharomyces sp. and Brettanomyces
sp. Several studies have reported similar molds from
Isolates from cabbage under microscope appears that the
different snacks and fast food contamination from water,
cells are olive to cylindrical shaped and Colonies on PDA
air, storage distribution facilities, environment and human
agar at 25 oC appear as white small soft cotton textures as

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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-4, July-Aug- 2017
http://dx.doi.org/10.22161/ijeab/2.4.10 ISSN: 2456-1878
showed in figure 3.5. and the isolates identified as
Brettanomyces sp.

Fig.3.7: Microscopic examination of Mucorsp

Microscopically appearance of isolates from tomato and

Fig.3.6: Shows microscopic examination of Brettanomyces celery showed that septate branched pale brown to olive
sp. brown with secondary conidiophores, conidia in long
branched chains. Colonies on PDA at 25oC for 7 days,
Microscopic features that was used to help in identification olivaceous-black or greenish black surrounded with white
of isolates from Bread showed that sporangiospores, sub cotton hyphae and is identified asAlternaria sp. ( figure
globes, and irregular in shape with Apophysis and having 3.8.).
stolone and Rhizoids and colonies on PDA, was whitish
becoming grayish-brown. Figure 3.6. And can be identified
as Rhizopus sp.

Fig.3.8: Microscopic examination of Alternaria sp.

As showed in figure 3.9: There was no growth of fungus on

plate contain PDA that incubate in 25 oC for 7 days (used as
Fig.3.5: Microscopic examination of Rhizopus sp. control).

When isolate from Eggplant and Potato examined under

microscope hayphae was non septate, sporangiophores
simple at first, later slightly sympodially branched, with
sporangium at terminal ends without apophysis and colonies
on SDA at 25oC, resemble white-to-gray cotton candy,
darkening with time. Fegure 3.7. The isolates can be
identified as Mucor sp.

Fig.3.9: PDA medium used as control.

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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-4, July-Aug- 2017
http://dx.doi.org/10.22161/ijeab/2.4.10 ISSN: 2456-1878
After collecting the samples and after subculturing them on the number of colonies on PDA was more than SDA.Our
PDA and SDA the number of colonies that obtained were suggestion for fast food restaurant isto keep the foods away
(96) colonies at 9:30 AM (81) colonies at 12:00 PM and at from contamination and spoilage and this will be by
4:30 PM (59) colonies as showed in table 3.1 the number of commitment with health rules that deal with restaurants
colonies at 9:30 AM were more than other times (12:00 PM cleanly and proceeding regular tests for restaurants workers
and at 4:30 PM) the reason for this result is using and to Study presence or absence t of mycotoxin in the
previously made foods that is not stored properly at 9:30 food products.
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