Microbiological Contamination and Food Hygiene Practices In old Restaurants In Nablus Governorate, Palestine


 Introduction: Personal hygiene is important in maintaining food quality and health safety, so the low level of personal hygiene causes contamination leading to food spoilage and the occurrence of sick cases for consumers.Objective: This study was conducted under the Corona pandemic to verify food safety in Nablus old restaurants, Palestine, in the year 2021.Research methodology: This study covered 32 restaurants and 160 employees randomly selected. Data collection began between April and June in 2021 using 160 questionnaires that included 20 questions were answered by workers in these restaurants.Results: Contamination with coliform bacteria was found in the restaurant samples with a percentage (42.5%), where the samples have taken from the tables having the highest contamination of this bacterium (50%). Furthermore, the data revealed that there was a relationship between coliform bacteria contamination and age, educational level, and food safety training, as the contamination being seen in the group that got the training, In contrast to those who didn't at the P-value <0.20.Conclusion: There was an association of bacterial contamination with age, especially among the younger group who were less concerned with hygiene and food safety practices compared to the elderly group. And it turns out that restaurant workers must be properly trained in food safety and monitored regularly.


Introduction
Because food is one of the indispensable basics of life, the health and hygiene of food have a major role in the spread of diseases as a result of the presence of microbes [1]. Bacterial contamination occurs when food is contaminated by microorganisms such as bacteria, viruses, molds, fungus, and toxins [2]. This can occur in a variety of ways, including species that generate poisons that are hazardous to humans. Despite the Ministry of Health's efforts to prevent the growth of these microbes, the food remains exposed to spoilage or contamination with them [2,3].
Coliform bacteria are Gram-negative, non-spore-forming, facultative anaerobic rods that ferment lactose rapidly into acid and gas at 35 2°C within 24 or 48 hours, Some types of coliform bacteria are characterized by their ability to cause disease, showing symptoms similar to those of the u to a large extent, such as stomach upset and diarrhea, but these symptoms may be mild for healthy adults, while those with weak immunity and the elderly may suffer from severe symptoms [4,5].
Therefore, this study was conducted in the city of Nablus, a city north of Jerusalem that stands between two mountains with a variety of natural springs, and is one of the largest Palestinian cities in terms of population, with hundreds of restaurants [6].
Despite the fact that Nablus is one of the most attractive tourist destinations in Palestine, this study was restricted to the old restaurants in the city. Tourists from all over the world visit it, especially the old town, which has restaurants that are tens of years old, such as Al-Aqsa restaurant for kunafa and others with people constantly visiting them. These visitors vary in age, nationality, and race [7].
Food workers (restaurants) have a signi cant role in reducing the transmission of foodborne diseases[8], so this study took into account the detection of the presence of coliform bacteria -total coliform bacteria -using violet-red media, From food (direct) and non-food (indirect) samples such as a

Materials And Methods
This descriptive and observational study comprised of randomly selected 32 old restaurants that have been open for at least 20 years. In addition, 160 employees working in those restaurants gave their consent to ll out a questionnaire about food hygiene practices after our explanations about the aim and method of the study.
Samples were collected between April and June in 2021, ve samples were taken from the same restaurant (table, plate, spoon, disc, and knife) using transport media, and then kept in portable refrigerators until it was transferred to the incubator in a lab, taking into account public safety precautions during the COVID 19 era such as wearing masks, gloves, continuous sterilization of hands after each collection.

Study Tool:
An interviewer-administered questionnaire was performed to obtain data on restaurant food hygiene procedures while keeping absolute privacy by not providing the restaurant names or any other information about them.
The questionnaire is divided into two sections: the rst section includes 6 questions about the general characteristics of the participants in the study.
The second contained 14 questions about the participants' food health practices. Food hygiene practices were measured using a ve-point Likert scale that ranges from always to never.

Microbiological Sample Analysis:
After ensuring that the samples were correctly organized and preserved, the culture and media preparation processes were carried out in the research and microbiological laboratories of the Faculty of Medicine at An-Najah National University, with the following numbers: (17G0020, 17B1140) respectively.
Violet Red Bile Agar Media (VRBA) was prepared for the cultivation of coliform bacteria by mixing 38.5g of powder with 1 liter of distilled water, boiling them using a magnetic eld to speed up the process, then we waited until the media had cooled down a little to facilitate the process of emptying it into Petri dishes under sterile conditions by a Bunsen ame [9], VRBA is recommended by American Public Health Association (APHA) and International Organization for Standardization (ISO) [10,11].
Due to the addition of bile salt, crystal violet, which suppresses the growth of no target Gram-positive bacteria. Coliforms rapidly metabolize the lactose in VRBA Agar and decrease the pH of the medium which detect by a neutral red indicator, resulting in redpurple colonies with red-purple halos [12]. See picture 1 <<Insert pictures 1 here.>> Data analysis: Descriptive statistics were used and expressed using frequencies, percentages, mean, standard deviation (SD), median and interquartile range (IQR).
Mann Whitney U test, which is known as a nonparametric test that focuses on frequencies was used to connect the relationship between gender and safety training with contamination of coliform bacteria in samples at P-value < 0.20. Furthermore, the Analysis of variance (ANOVA) test was conducted for normally distributed data with more than three groups within the independent variables which link between education levels and bacterial contamination at the P-value < 0.20.

Study Ethics:
To start this study, approval was obtained from Institutional Review Board (IRB) at An-Najah National University. In addition, the consent of the restaurant owners and employees was also obtained to collect samples and ll out the questionnaire. The results also showed that the participants who attended university were slightly more than half (57.5%) while the participants who attended high school were (2.5%). The work experience of our study participants range was from one year to 45 years with a median of 7 years and IQR of 44 years. (45%) of the food handler were servers. And nally, more than half (61.3%) had never received training for food safety.

Coliform Contamination in Samples
Based on the coliform contamination results in the samples given in Table 2. The largest proportion of contamination was found in the samples obtained from the tables (50%), which was due to the tables' proximity to food, which contributes to the spread of foodborne illnesses. these results were agreed by Galie    While the results showed that the percentage of contamination from spoons was the lowest (34.37%) because most restaurants during the study period (Corona Virus) relied on the use of single-use plastic spoons.

Food Hygiene Practices
The food hygiene practices of food handlers were shown in Table 3. (100%) of the participants in this research washing their hands after using the toilet, and 91.9% washing their hands after cooking, which were both considered healthy behaviors. Furthermore, more than half of them (60.6%) never keep their ngernails long as we have noticed.

Age and Coliform Contamination
In this study, age was related to the contamination of coliform bacteria through the Pearson correlation coe cient. As the bacterial contamination of dishes and disk samples had a correlation with age on its value (0.163, 0.091) respectively, at the Pvalue <0.20, this correlation was in the favore of the younger group, which was less concerned with hygiene and food safety than the older group according to the results that were appeared, and that was opposite of previous research, which found that age had an impact on food safety from contamination, particularly in the elderly [16,17]. Moreover, there is no correlation between bacterial contamination and work experience as shown in Table 4. In Table 5, we used the Mann Whitney U test (P-value <0.20), which connects the relationship between gender and contamination in samples, showing that there was no relationship between them. It also studied the relationship between safety training and contamination in samples, where there was a correlation between bacterial contamination in samples of the spoon (0.002) and knife (0.007) with safety-trained workers.  The correlation was higher in the category of employees who got safety training as mentioned above, which suggests that the approach used to train them was ine cient and that we need to properly train and certify them in this area. At the same time, awareness programs should be launched for restaurant employees to increase knowledge of food safety and this is consistent with a study conducted in Bangladesh that talked about this[18].

Educational Level And Coliform Contamination
Regarding educational level, which is one of the most important factors that contribute to high food safety [16], The relationship between bacterial contamination and education levels was performed by ANOVA test as shown in Table 6. The results showed that there was a high correlation between participants who attended secondary school and contamination samples in dishes and knives separately compared to the rest of the educational levels.
This indicates that those who attended high levels of education had low levels of contamination with bacteria. Therefore, we must train secondary groups with more education on food safety and health issues.

Conclusion
There was an association of bacterial contamination with age, especially among the younger group who were less concerned with hygiene and food safety practices compared to the elderly group. And it turns out that workers must be properly trained on food safety and monitored regularly.

Limitations
This study was conducted during the Coronavirus period, as most restaurants adopted the single-use plastic cutlery system, and this affected the results. So we recommended conducting it in another period to compare it with the current study results.