






































 Global Journal of Education and Allied  

Research (GJEAR) 
Volume.13, Number 10; October-2022; 

ISSN: 2837-3707 | Impact Factor: 6.79 

https://zapjournals.com/Journals/index.php/gjear  

Published By: Zendo Academic Publishing 

 

 

pg. 11 

EXAMINING MICROBIAL QUALITY IN READY-TO-EAT FOODS OF MAKKAH'S 

CAFETERIAS 

 
1Ahmed Khalid Al-Faisal and 2Aisha Abdullah Al-Mansoori 

 

Article Info  Abstract 

Keywords: Ready-to-eat foods, 

RTE, contamination, 

microorganisms, food safety. 

 The consumption of ready-to-eat foods (RTEs) has witnessed a surge in 

the Kingdom of Saudi Arabia, particularly in the city of Makkah, owing 

to shifting demographics and lifestyle changes. This trend is influenced 

by the influx of itinerant workers, pilgrims, and visitors, leading to an 

increased preference for convenient RTE options, especially for 

breakfast. Cafeterias offering RTE foods have gained popularity due to 

their affordability, accessibility, and convenience. However, certain 

RTE food items, including those containing animal-derived ingredients 

like eggs, fish, meat, and poultry, are susceptible to contamination by 

various microorganisms such as bacteria and fungi. The contamination 

of RTE foods during processing and sandwich preparation has been 

reported, potentially resulting in cross-contamination and the presence 

of pathogenic microorganisms like Staphylococcus aureus, Salmonella 

species, Bacillus species, and Escherichia coli. Consequently, the 

quality of RTE foods can be significantly compromised. In this context, 

understanding the factors contributing to the contamination and 

deterioration of RTE foods is crucial for ensuring food safety and public 

health. This study aims to investigate the prevalence and implications 

of microorganism contamination in RTE foods in Makkah, Saudi 

Arabia, and proposes strategies to enhance the safety and quality of 

RTE meals. 
 

 

 

Introduction   

Ready-to-eat foods (RTEs) are items prepared in advance and ready for consumption only after heating without 

further processing or preparations. It could be raw or cooked meals. RTE food items consumption has recently 

                                                      
1Environmental Health Department, Faculty of Public Health & Health Informatics, Umm Al-Qura University, Makkah, 

Saudi Arabia 
2 Epidemiology Department, Faculty of Public Health & Health Informatics, Umm Al-Qura University, Makkah, Saudi 

Arabia 

https://zapjournals.com/Journals/index.php/gjear


Global Research Journal of Management and Social Sciences (GRJMSS) Vol. 13 (10) 

 

pg. 12 

increased in the Kingdom of Saudi Arabia, mainly in holly Makkah city, due to the rapid demographic change 

characterized by a change in lifestyle and a large number of itinerant workers, visitors, and pilgrims resulting in 

a large percentage of the population taking ready-to-eat food especially as breakfast’s meal. In addition, cafeterias 

that serve RTE foods provide cheap, economical, and easily accessed items. Some RTE foods items contain raw 

materials of animal origin, such as eggs, fish, meat, and poultry, which can easily get contaminated by different 

microorganisms, including bacteria and fungi (Ashenafi, 1995).   

The cross-contamination of RTE foods with pathogenic microorganisms could occur during the processing and 

preparation of fillings and sandwiches (Oranusi et al., 2013; Rahman et al., 2014; Sharma et al., 2014). 

Contamination of RTE food by pathogenic microorganisms like Staphylococcus aureus, Salmonella species, 

Bacillus species, and Escherichia coli leads to remarkable changes in food quality. Moreover, consequently 

causing food-borne illnesses and food poisoning are considered major public health issues that negatively affect 

the socioeconomic development and output of many fields such as tourism and trading (Newman et al., 2015).   

Food-borne diseases and food poisoning problems are becoming an important issue worldwide (Al-Mazrous, 

2004). In the Kingdom of Saudi Arabia in general and holly Makkah city in particular, food-borne illness and 

food poisoning are emerging as significant public health threats that render a high burden of diseases and 

negatively affect the socioeconomic development and productivity of many sectors. Several studies have reported 

that foods served by catering services were the primary source of many food-borne outbreaks (Osimani and 

Clementi, 2016). In Makkah city, increasing food poisoning cases were reported, especially during the Hajj and 

Umrah seasons. It was mainly due to the high consumption of RTE meals, especially those from cafeterias (Shirah 

et al., 2017).  

We think minimizing fungal and bacterial loads in RTE foods served in cafeterias within Makkah city by following 

good hygiene practices and continuous microbiological assessment programs is necessary. Moreover, using 

microbiological testing results of RTE foods in Makkah city will provide an example for other cities in Saudi 

Arabia for dealing with ready food. However, very little research has been carried out on assessing microbial 

quality and safety of RTE foods in Saudi Arabia, especially those distributed within Makkah city. Therefore, this 

study aimed to determine the microbial safety of RTE food items that are served in some cafeterias in Makkah 

city, Saudi Arabia.   

Materials and Methods  

Sample Collection   

Ready-to-eat food samples were collected randomly from different cafeterias located in different sites of 

Makkah City during the period March-April, 2021. A total of 108 samples represented six different types of 

ready-to-eat food (foul, falafel, boiled egg, fried egg, shakshuka, and sheep liver slices) collected from six 

different cafeterias and replicated three times. Collected samples were handled in sterile plastic bags and 

directly transferred to the microbiology lab for investigation. 

Microbiological Analysis  

Isolation and enumeration of bacteria. Ten grams of each experimental sample was mixed with 90 mL Nutrient 

Broth (Biotech, UK), and serial dilutions of each food sample homogenate were prepared to reach 10-3 dilutions. 

A 1 mL aliquot portions of the prepared dilutions were precisely distributed onto duplicate sterile plates of 

Nutrient Agar (Himedia, India), Eosin Methylene Blue (EMB) Agar (Titan Biotech, India), and Mannitol Salt 

Agar (MSA) (India mart, India) for total microbial load, Escherichia coli, and Staphylococcus aureus, 

respectively. Counting of bacterial colonies was performed after incubation of agar plates at 37 °C for 24 to 48 h, 



Global Research Journal of Management and Social Sciences (GRJMSS) Vol. 13 (10) 

 

pg. 13 

by using the colony counter (Gallenkamp, England). Bacterial colonies were expressed as colony-forming units 

per mL of sample homogenate (CFU/mL).  

Isolation and enumeration of fungi. For fungal isolation, 10 grams of each sample were mixed in Sabouraud 

broth (India mart, India), and the same dilutions were prepared. A 1 mL portion of the dilution (10-3) was poured 

onto Sabouraud agar (India mart, India) medium supplemented with 0.005 gram/liter of rose Bengal (Sigma 

Aldrich, USA) for suppressing bacterial growth. The agar plates were incubated at 28 °C for 5-7 days; then, the 

developed colonies were counted and identified.  

Identification of Microbial Isolates. The developed bacterial colonies were counted using colony counter, 

purified, and stored on nutrient agar slants at 4 °C for identification. Identification based on cultural morphology 

and biochemical tests, including carbohydrate utilization on Tri-sugar Iron (TSI) medium, IMViC test, starch 

hydrolysis, gelatin liquefaction, nitrate reduction, oxidase urease activity, and motility test were employed to 

confirm the purity of the isolates. The fungal morphology was studied macroscopically by observing the colony 

features (color, shape, size, and hyphae) and microscopically by a compound microscope according to the 

following references Gaddeyya et al. (2012), Domsch et al. (1980), and Barnett & Hunte (1972).    

Results and Discussion   

Bacterial species isolated from ready-to-eat foods samples  

A total of 108 food samples, including 6 types (foul, falafel, boiled eggs, fried eggs, shakshuka, and sheep liver 

slices), were evaluated. The results showed in Table 1 that the highest aerobic plate counts were found in sheep 

liver slices sold by Cafeteria No 5 with the value of 8.4 × 103 CFU/mL, while the lowest value was detected in 

foul sold by Cafeteria No.2 with a value of 0.5 × 103 CFU/mL. S. aureus appeared in all food samples collected 

from all cafeterias. The highest count (9.8 × 103 CFU/mL) for S. aureus was detected in shakshuka sold by 

Cafeteria No.6, while the lowest count (1.2 × 103 CFU/mL) was observed in falafel sold by Cafeteria 1. Generally, 

it could be reported that all the tested samples were highly contaminated with S. aureus, and 42% of the samples 

were contaminated with E. coli. The total counts for S. aureus and E. coli (in shakshuka and sheep’s liver slices) 

exceeded the recommended levels by the International Commission on Microbiological Specifications for Foods 

(ICMSF, 1996). The ICMSF (1996) recommends that ready-to-eat foods between 0-103 CFU are acceptable, 104-

105 CFU is tolerable, and 106 CFU and above are unacceptable.   

Table 1. Mean total counts (CFU/mL) of each bacterial species isolated from ready-to-eat 

foods samples sold in Makkah city.  

Vendors 

samples station  

Food types  Bacteria  

Aerobic plate count  S. aureus count  E. coli 

count  

  

  

Cafeteria 1  

Foul  2.4 ×103  3.6 ×103  ND  

Falafel  3.5 ×103  1.2 ×103  ND  

Boiled egg  1.2 ×103  4.6 ×103  ND  

Fried egg  2.5 ×103  1.3 ×103  ND  

Shakshuka  5.7 ×103  4.3 ×103  0.4 ×103  

Sheep liver 

slices  

6.3 ×103  3.1 ×103  0.5 ×103  

  

Cafeteria 2  

Foul  0.5 ×103  3.4 ×103  1.2 ×103  

Falafel  3.4 ×103  5.2 ×103  ND  



Global Research Journal of Management and Social Sciences (GRJMSS) Vol. 13 (10) 

 

pg. 14 

  Boiled egg  3.2 ×103  2.5 ×103  ND  

Fried egg  1.5 ×103  3.3 ×103  ND  

Shakshuka  4.3 ×103  6.4 ×103  2.3 ×103  

Sheep liver 

slices  

5.9 ×103  3.6 ×103  0.6 ×103  

  

Cafeteria 3  

  

  

Foul  1.5 ×103  3.5 ×103  ND  

Falafel  3.2 ×103  4.1 ×103  ND  

Boiled egg  1.1 ×103  3.2 ×103  ND  

Fried egg  0.9 ×103  3.7 ×103  1.1 ×103  

Shakshuka  5.9 ×103  7.5 ×103  2.4 ×103  

Sheep liver 

slices  

4.5 ×103  7.5 ×103  1.6 ×103  

  

Cafeteria 4  

  

Foul  4.2 ×103  8.7 ×103  ND  

Falafel  3.4 ×103  4.3 ×103  ND  

Boiled egg  2.8 ×103  4.9 ×103  ND  

Fried egg  2.1 ×103  3.5 ×103  ND  

Shakshuka  4.8 ×103  6.7 ×103  ND  

Sheep liver 

slices  

5.6 ×103  6.8 ×103  2.3 ×103  

  

Cafeteria 5  

  

Foul  5.2 ×103  5.2 ×103  ND  

Falafel  2.8 ×103  5.5 ×103  ND  

Boiled egg  2.1 ×103  4.3 ×103  0.3 ×103  

Fried egg  1.1 ×103  4.2 ×103  ND  

Shakshuka  4.7 ×103  6.4 ×103  4.2 ×103  

Sheep liver 

slices  

8.4 ×103  7.9 ×103  2.3 ×103  

  

Cafeteria 6  

  

  

Foul  2.8 ×103  4.2 ×103  ND  

Falafel  4.4 ×103  9.1 ×103  ND  

Boiled egg  1.4 ×103  4.2 ×103  0.2 ×103  

Fried egg  0.9 ×103  2.2 ×103  ND  

Shakshuka  5.9 ×103  9.8 ×103  1.7 ×103  

Sheep liver 

slices  

4.8 ×103  7.5 ×103  0.7 ×103  

% of contaminated samples  100%  42%  

Note. ND - Not detected, APC - Aerobic plate count.  

The consumption of cooked RTE foods analyzed herein might increase the risk of food-borne illness caused by 

various microorganisms, especially S. aureus and E. coli. The highest aerobic plate counts recorded in this study 

might be caused by different environmental factors, such as contaminated air, water, and utensils used in the meal 



Global Research Journal of Management and Social Sciences (GRJMSS) Vol. 13 (10) 

 

pg. 15 

preparation. Poor personal hygiene practices in the cafeterias could have also led to the contamination of these 

pathogens. Bezirtzoglou et al. (2000) reported that RTE food exposure to air or dust at the vending point is likely 

to increase loads of the bacteria as it appears that most bacteria are carried in aerosols by dust and wind. 

Contamination by the food producer or handlers is also the most common means of transmitting this germ. S. 

aureus could be transmitted through the dirty hands and mouths of the producers and customers. Similarly, Burt 

et al. (2003) stated that food contamination by S. aureus originates from man’s respiratory passages, skin, and 

uncovered wounds. Thus, its persistence in cooked RTE foods might cause many health risks to consumers. 

Several reports indicate that most strains of S. aureus are known to be pathogenic due to their excreted heat-stable 

enterotoxins in direct relationship to their inoculum level (Adebayo-Tayo et al., 2012). S. aureus produces 

enzymes incorporated with staphylococcal invasiveness and many extracellular substances, which are 

enterotoxins that are stable at high temperatures and turn the food riskier even though it appears normal (Prescott 

et al., 2019). The appearance of the symptoms may differ with the quantity of the contaminated food consumed 

and the susceptibility of the people to the toxin. Some symptoms of staphylococcal food poisoning include 

vomiting, nausea, diarrhea, and abdominal pain (Amusan et al., 2010).  

E. coli was not detected in most tested food samples, but it appeared in shakshuka and sheep’s liver slices from 

most cafeterias. The highest count was 4.2 × 103 recorded in shakshuka samples vended in cafeteria No. 5—a 

report by Idowu and Rowland (2006) showed that the highest aerobic plate numbers were found in a fang soup 

sold by street Vendors with a count of  

2.80 × 106 CFU/mL while the lowest count was observed in stew sold by stationary food vendors with shade 

(SVWS) with several  

1.20 × 106 CFU/mL. Moreover, the highest value of S. aureus was obtained in moimoi sold by a street vendor 

with the count of 4.30 × 106 CFU/mL, and the lowest count was observed in stew sold by SVWS with the count 

of 1.20 × 103 CFU/mL. E. coli counts were detected most in moimoi sold by MV with the value of 2.20 × 106 

CFU/mL. The presence of E. coli in cooked RTE foods indicates secondary contamination, as E. coli is known to 

be correlated with the gastrointestinal tract of warm-blooded animals and not found in the environment as a natural 

flora (Amusan et al., 2010). However, contamination can take place by the use of contaminated water. E. coli has 

been detected in foods sold at fast foods in a report by Fowoyo and Baba-Ali (2015). In this study, direct or 

indirect fecal contamination may be the leading cause of E. coli., and E. coli belong to the Enterobacteriaceae 

genus, the leading causal agent for diarrhea, gastroenteritis, urinary tract infections, meningitis, nosocomial 

pneumonia, and dysentery. The strain of E. coli named Enterohaemorrhagic E. coli can cause food-borne diseases 

similar to the E. coli O157H7 strain, which causes a severe and potentially fatal illness called Hemorrhagic colitis 

which is characterized by bloody diarrhea and severe abdominal pain (Evans & Evans, 1996). Following the 

Public Health Laboratory Service (UK) criteria, the count of E. coli in RTE food was defined as <20 CFU/mL 

(satisfactory), 20– <100 CFU/ML (acceptable), ≥100 CFU/mL (Health Protection Agency, 2009). In our findings, 

the counts of E. coli in the samples of shakshuka and sheep’s liver slices can be considered unacceptable. 

Fungal species isolated from ready-to-eat foods samples  

Data shown in Table 2 indicated that 21 fungal species belonging to 7 genera were identified. The number of 

isolated fungi varied according to different meals. The least fungal number (9 species) appeared in fried egg 

sandwiches followed by 12, 14, 15, 18, and 20 species in shakshuka, boiled egg, falafel, sheep's liver slices, and 

foul samples, respectively. Aspergillus appeared in most samples collected from cafeterias, represented by six 

species. Among Aspergillus, A. flavus was the highest, and its occurrence ranged between 50-83% of samples. 

Also, A. niger and A. parasiticus were among the common Aspergillus species found in the samples. Among 



Global Research Journal of Management and Social Sciences (GRJMSS) Vol. 13 (10) 

 

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samples analyzed herein, shakshuka samples were highly contaminated by Aspergillus species (50% by A. flavus, 

83% by A. niger, 100% by A. ochraceus, and 50% by A. parasiticus). Mucor racemosus emerged in remarkable 

count in falafel samples (50%) and did not emerge in Shakshuka.  

Four species of Penicillium were isolated, and among them, P. aurantiogresum was the most commonly isolated. 

It encountered 72%, 50%, 67%, and 83% in foul, falafel, boiled egg, and sheep’s liver slices, respectively. P. 

cyclopium appeared in 5 types of samples with remarkable accounts.  Rhizopus nigricans were found in high 

occurrence in foul (72%) and falafel (52%). About 100% of foul and 89% of falafel samples were contaminated 

by Stachybotrys chartarum. Trichoderma sp. was found in five samples, mostly foul (83%) and fried egg (50%). 

Uzeh et al. (2009) revealed that microorganisms found in salad constitute raw vegetables include Mucor sp., 

Aspergillus fumigatus, Trichoderma, Neurospora crassa, and Aspergillus niger. Barnett et al. (2000) reported 

several mycotoxins isolated from various foods. For instance, Aspergillus and Penicillium produced aflatoxins. 

The aflatoxins have been found in legumes, grains, fruits, meats, spices, cheeses, milk, rice, corn, cotton seeds. 

Other toxins with carcinogenic, hemorrhagic, hepatotoxic, neurotic uterotrophic effects have been found in 

foodstuff and recorded as metabolites of fungi (Jeswal & Kumar, 2015). Ochratoxins are toxins excreted by 

Aspergillus ochraceus, A. sulphureus, A. malleus, and A. ochraceus can be found in soils and decaying vegetation, 

grains, wheat, corn, cotton seeds, legumes, peppers, onions, and pears (Adams & Moss, 2002).  

Table 2. Mean total count and occurrence remarks (O.R.) of each fungal species (CFU/mL) isolated 

from ready-to-eat food samples sold in Makkah city.  

Fungal genera 

& species  

  

CFU/mL  O.R. 

out 

of 6 

sites   

Foul  

  

Falafel  

  

Boiled 

egg   

Fried 

egg  

  

Shakshuka  Sheep 

liver 

slices  

(% count of each isolate out of 18 samples)  

Alternaria 

alternata  

11  H  00  00  3(17%)   3(17%)   3(17%)   2(11%)  

Aspergillus  

A. flavus  56  VH  9(50%)  4(22%)  7(39%)  15(83%)  9(50%)  12(67%)  

A. niger  42  VH  11(61%)  00  6 (33%)  9(50%)  15(83%)  1(5.6%)  

A. ochraceus  32  H  3(17%)  00  3(17%)  00  18(100%)  8 (44%)  

A. parasitus  27  VH  6 (33%)  3(17%)  14(78%)  00  9(50%)  4(22%)  

A. sydowi  26  H  10(56%)  00  7(39%)  00  4(22%)  5(28%)  

A. ustus  14  L  7(39%)  3(17%)  00  00  00  4(22%)  

Cladosporium 

sp.  

12  L  6(33%)  00  00  00  10(56%)  6(33%)  

Mucor 

racemosus  

36  VH  4(22%)  9(50%)  5(28%)  4(22%)  00  4(22%)  

Fusarium  

F. culmorum  16  M  2(11%)  5(28%)  6(33%)  00  00  3(17%)  

F. solani  3  R  3(17%)  00  00  00  7(39%)  00  

Penicillium  



Global Research Journal of Management and Social Sciences (GRJMSS) Vol. 13 (10) 

 

pg. 17 

P.  

aurantiogriseum  

62  VH  13(72%)  9(50%)  12(67%)  6(33%)  00  15(83%)  

P. digitatum  29  H  5(28%)  8(44%)  7(39%)  00  5(28%)  9(50%)  

P. citrinum  21  H  8(44%)  4(22%)  4(22%)  00  00  00  

P. cyclopium  33  VH  10(56%)  4 (22%)  7(39%)  7(39%)  00  9(50%)  

Rhizopus 

nigricans  

38  VH  13(72%)  5(52%)  6(33%)  4(22%)  4(22%)  6(33%)  

Stachybotys 

chartarum  

40  VH  18(100%)  16(89%)  7(39%)  6(33%)  00  5(28%)  

Trichothicium 

roseum  

17  M  8(45%)  4(22%)  00  00  00  00  

Phoma sp.  8  R  4(22%)  4(22%)  00  00  00  00  

Trichoderma sp.  44  VH  15(83%)  6(33%)  00  9(50%)  5(28%)  2(11%)  

Verticillium sp.  7  L  2(11%)  4(22%)  00  00  00  1(0.06%)  

No. of isolated fungi  20  15  14  9  12  18  

Note. Rare (R) = Appeared in 1 site, Low (L) = Appeared in 2 sites, Medium (M) = Appeared in 3 

sites, High (H) = Appeared in 4 sites, and Very High (VH) = Appeared in 5 -6 sites.  

Patulin is the primary toxic metabolite produced by Penicillium (Bennett & Klich, 2003). Fusarium culmorum 

and Trichothecium roseum appeared moderately, and the least encountered fungal species were A. ustus, 

Cladosporium sp., and Verticillium sp.  (Table 2). The isolation of S. aureus, Aspergillus niger, Aspergillus 

fumigatus, Mucor sp, and Penicillium sp, are consistent with the findings of Attiya et al. (2015), Taulo et al. (2008) 

and Oranusi, et al. (2013), in which these microorganisms were found in RTE foods. Molds like Mucor sp and 

Aspergillus sp. Contaminated food samples through dust and soil as they disperse in the form of spores abundant 

in the environment (Apinis, 2003). Molds in food samples are a severe health threat due to mycotoxin production 

(Makun et al., 2009).  

  



Global Research Journal of Management and Social Sciences (GRJMSS) Vol. 13 (10) 

 

pg. 18 

Picture 1. Most common isolated fungi and bacteria. A). Aspergillus niger (black), A. fumigatus (olive colony), 

and A. ustus (brown colony); B). A. flavus (yellow-green) mixed with Stachybotrys chartarum (black); C). 

Curvularia lanata (black color) and A. ustus (grey colonies); D). Alternaria alternate (black) and Trichoderma 

SP. (green); E). Mixed cultures of S. aureus (golden) and E. coli (white); F). Pure culture of Stachybotrys 

chartarum. 

Conclusion   

Ready-to-eat foods (RTEs), especially of animal origin, could be considered as potential sources for microbial 

infections in humans. This study revealed that RTE foods served in cafeterias in Makkah city had unsatisfactory 

microbiological contamination; thus, it may increase the potential risk of food-borne poisoning and diseases 

among consumers. Accordingly, it is a priority for relevant public health and food safety agencies to establish 

training programs on food safety and sanitary in cafeterias, especially following the regulations of hazard analysis 

and critical control points (HACCP) principles during all food processing steps. All staff members, including 

administrators and the people who work in the kitchen, must be trained. 

Acknowledgment  

The authors appreciate the cooperation of the Faculty of Health and Health Informatics, Umm Al -Qura 

University, and cafeteria owners for consent and facilitating sampling.  

  

Conflict of interest. The authors declare that there is no conflict of interest regarding the publication of this paper.  

ORCID ID   

Ramadan Ali Mohamed Badran: https://orcid.org/0000-0003-3932-7396  

Mutasim Mohamed Khalafalla: https://orcid.org/0000-0002-8214-9698  

Hatim Abdullah Natto: https://orcid.org/0000-0003-2570-6264 

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