Evaluation of the Hygienic Status in Food Serving Establishment

| This study was conducted to evaluate the bacteriological safety of raw and cooked meat products represented by Aerobic Plate Count, Enterobacteriaceae, Coliforms, E. coli and Staph. aureus counts as well as determination of pH, TVBN and TBA of a totally examined 110 samples (20 each of shish tawook, kofta, fillet and escalope pane, besides, 30 swabs (10 each of hands, food contact surfaces and cutting knifes). The obtained results in this study indicated that the Overall reduction the bacterial counts of the above-mentioned organisms according to their prevalence in the different products under study were significantly reduced by 2, <1, 1, and <2 log10cfu/g in raw examined samples of shish tawook, kofta, fillet and escalope pane treated with 5% vinegar as compared with raw untreated samples, respectively. In addition, cooked treated aforementioned sample types with vinegar 5% showed significant reduction represented by 2, 1, <2 and 1 log10cfu/g when compared with cooked untreated samples. So, using of vinegar either in raw or cooked products leads to a significant reduction in the number of bacteria contaminating products. Hand swab before cleaning and sanitizing recorded 3.31±0.52 (100%), 2.54±0.92 (100%), 2.47±0.74 (60%), <1 (0.0%) and 2.12±0.55 (80%), while after cleaning it was recorded 2.39±0.5 (100%), 1.59±0.25 (60%),1.36±0.24 (40%), <1 (0.0%) and 1.74±0.62 (60%). Surfaces swab recorded 3±1.04 (100%), 1.5±0.96 (50%), 2.06±1.49 (20%), <1 (0.0%) and 2.3±1.18 (30%), before cleaning and 1.73±1.02 (100%), <1, <1, <1 and (30%) 2±0.7 log10cfu/g after cleaning. Knife swabs recorded 3.35±0.76 (100%), 2.68±0.68 (50%), 2.5±0.91 (40%), <1and 2.25±1.0 (70%) before cleaning, while after cleaning they were recorded 1.77±1.19 (100%), 2.59±0.16 (20%), <1, <1and 1.97±0.58 for the aforementioned bacterial counts, respectively. Salmonella was not isolated from any of the examined samples or swabs. Meanwhile, chemical analysis of pH, TVB-N and TBA recorded 0.04±0.02, 11.86±1.24 and 5.86±0.18 for shish tawook; 0.05±0.02, 6.6±2.19 and 6.12±0.31 for kofta; 0.04±0.03, 7.22±2.49 and 5.62±0.13 for fillet and 0.41±0.16, 13.18±2.15 and 5.96±0.49 for escalope pane, respectively. Significance differences (P<0.05) were obvious between raw and cooked samples and for some extent between treated and non-treated one. meat,


INTRODUCTION
M eat, poultry and their products considered of high nutritive value for consumers because they are constituting an important source for animal protein, essential amino and fatty acids, minerals, vitamins and other beneficial nutrients. On the other hand, meat is considered as an ideal media for growth and multiplication of many types of food spoilage and food poisoning microorganisms (Moghnia et al., 2021) because of their high moisture Category of RTE foods as high risky foods because they do not require any heat treatment or any other process prior to consumption. In addition, food workers may transmit pathogens to food from a contaminated surface, from another food, or through contaminated hands with microorganisms from their gastrointestinal tract. Moreover, RTE food products provide a source of readily available and nutrition meals for the consumer. However, many questions have been raised about the safety and microbiological quality of these food products (Fang et al., 2003).
There are several factors that expose RTE foods to contamination with various microorganisms, as contaminated water sources, unhygienic toilets and inadequate washing facilities. The washing is normally carried out in large containers or buckets, that makes insects and rodents attracted to sites where there is no organized sewage disposal. In addition to, food which is not adequately protected from flies and not properly refrigerated (Mensah et al., 2002) Besides, there are many other factors that are exposed RTE foods to contamination by different microorganisms including food preparation using unhygienic traditional processing methods, inappropriate holding temperatures and poor personal hygiene of food handlers (Muinde and Kuria, 2005;Barro et al., 2006). Consumers who depend on such food are usually pay little attention to its safety, quality and hygiene. Many other factors affecting the quality and safety of RTE foods that explained by (Afolabi et al., 2012) represented in preparation of food and storage at ambient temperature for a long time before its consumption and inadequate cooling and reheating as well as improper cooking time and temperature of the food.
Sliced cooked meats are one of the most consumed readyto-eat food (RTE) products around the world. The major reasons that increase the demand for such products, they are meet the consumer requirements, its satisfaction, convenience and good acceptability. During preparation and sale, cooked meat products can be contaminated by spoilage and pathogenic bacteria causing hazard for consumers if sufficient doses to cause disease are consumed through contaminated food (EFSA, 2007;Norrung and Buncic, 2008). Cross-contamination arises from many sources as using of the same cutting board for chicken meat and salad without cleaning in between, for raw and cooked foods, spreading of pathogens via the kitchen contaminated environment, which seems to be of greater importance than the risk associated with undercooking of poultry meat (Petra, 2009).
Based on the (PHLS, 2000) microbiological Guidelines for some ready to eat foods sampled at the point of sale, 179 (40%) out of 449 chicken sandwich samples were satisfactory, 156 (35%) acceptable, and 114 (25%) samples were of unsatisfactory microbiological quality. None of the examined samples were exceeded the unacceptable microbiological quality limits. Enterobacteriaceae was the microbiological parameter most often associated with unsatisfactory results.
Poultry meat is considered as one of the main sources of spreading of Staph. aureus microorganism, proper handling of raw meat, while, adequate cleaning of hands, surfaces, equipment's, disinfection of poultry slaughter houses, vehicles and good personal hygiene can reduce its spreading (Karmi, 2013).
Lipid oxidation and auto-oxidation are major causes of deterioration and reduced shelf life of meat products. Lipids oxidation may produce changes in meat quality parameters such as color, flavor, odor, texture and even nutritional value (Fernandez et al., 1997). Meanwhile, TVB-N are considered as a reliable indicative measure for the quality of protein of various food articles specially meat, poultry and their products. Generally, microorganisms either flora or those induced by handling of food items grow at different levels and may resulted in protein deterioration (Warries, 2000).
Development of off-flavors was arise due to the effect of lipolytic bacteria resulted in lipid oxidation, which could be determined by sensory evaluation and measuring of thiobarbeturic acid. Fat content has a basic effect on

Advances in Animal and Veterinary Sciences
November 2021 | Volume 9 | Issue 11 | Page 1935 various physicochemical and sensory characteristics as flavor, juiciness, texture; handling, bite, heat transfer etc. and it cannot be modified and/or reduced with improper less fat or another type of fat (Pearson and Gillett, 1999).
So, the current work was planned to investigate the hygienic status in food serving establishment through determining the bacterial profile of served meat and poultry meals including raw and after cooking shish tawook, kofta, fillet and escalope pane, as well as determination of their physico-chemical criteria and examination of swab samples from different localities.

COlleCTiOn Of SAmpleS
A total of 110 different samples classified as follows: • Eighty samples of shish tawook, kofta, fillet and escalope pane (20 samples each); 10 of which were raw and 10 were cooked (five samples dipped in vinegar 5% while the other 5 samples remained as control). • Thirty swab samples of workers hands, food contact surfaces and cutting knifes (10 samples each). Were collected from food serving establishment, kept in its sailing bags and aseptically transferred without delay, in an insulated ice box to the laboratory without delay.

prepArATiOn Of SAmple hOmOgenATe
Twenty-five grams of the examined samples were aseptically transferred to a sterile stomacher bag and homogenized with 225 ml sterile buffered peptone water (0.1%) for 30-60 seconds to give an initial dilution of 1/10. Serial dilutions up to 10 6 was carried out according to (ISO 6887-2, 2003). Prepared homogenate was used to carry out the following examinations.

ChemiCAl AnAlySiS
Determination of pH, TVB-N and TBA according to AOAC (1995).

STATiSTiCAl AnAlySiS
Was carried out using Statistical Package for the Social Sciences (SPSS). One Way ANOVA, Ver. 20

RESULTS AND DISCUSSION
Contamination with foodborne pathogens particularly, E. coli, Salmonellae and Staph. aureus can occur across the multiple steps along the food chain, including production, processing, storage, distribution, retail marketing, exhibition and handling or preparation either by cooking, frying or grilling, E. coli as indicator bacteria are used to evaluate the hygienic status of the food establishment (Sofos et al., 1999;Ghafir et al., 2008). Worldwide, foodborne illness is often associated with consumption of meats and poultry products sold at unhygienic retail markets or poor sanitized restaurants. Moreover, chicken meat is considered an important source for transmission of foodborne pathogens to the consumers resulting in health risk (Matias et al., 2010). Aerobic plate count, Enterobacteriaceae, coliforms and E. coli can reflect the level of hygiene during food production and subsequently its shelf life and thus, the potential problems arises during the stages of food production and processing. Must focus on the potential problems that may arise due to poor handling or lack of general and personal hygiene, storage, unsafe handling, or preparation through the use of insufficient heat treatments to cook food and eliminate these microbes or reduce them to internationally accepted levels (HPA, 2009).

Advances in Animal and Veterinary Sciences
November 2021 | Volume 9 | Issue 11 | Page 1936 Higher APC in chicken filet than in chicken thigh (shish) was attributed to the processing stages of breast and divide into parts with removal of the skin in addition to soaking chicken fillet in unclean water to increase their weight, or using unclean knives and chopping tables which manufactured from wood. All these factors lead to further spread of contamination to the fleshy parts. Obtained results in Figure 2 illustrated that the mean counts (log 10 cfu/g±SD) of APC  Mousa et al. ( 2019) failed to detect Salmonella in grilled kofta. The variation in the results between different authors may be due to the differences in the effectiveness of hygienic measures applied during cooking, handling and serving of food High Staph. aureus counts are indicators of poor personal hygiene, poor handling and temperature control failure. The high count of Staph. aureus could be due to the neglected hygienic practices of the workers. Besides, the different pre and post slaughtering sources of contamination with Staph. aureus including feed, feces, feather, scald water, defeathering machines due to the presence of cracks of the rubber fingers, water chiller and the employees (Al-Dughaym and Altabari, 2010). Moreover, burning surfaces of grilled kofta are significantly impaired heat transfer in the food and eventually prevent further cooking of the inner portions of meat pieces being cooked which does not sufficiently affect the elimination of microbial contamination ( Jay, 1996).
In Egypt, the most ready to eat sandwiches and meals sold by street vendors and restaurants are chicken kofta, pane, fillet, shawerma and el-hawawshi. added that, E. coli in recent years has become recognized as a serious foodborne pathogen and has been associated with numerous outbreaks in the UK, Japan and USA Scotter et al. (2000). Moreover, Hassanin et al. (2014) concluded that the presence of Salmonella, E. coli and Staph. aureus in RTE foods, not only renders these sandwiches of inferior quality and unfit for human consumption, but also as an indication for fecal and environmental contamination and bad personnel hygiene which affect the food quality and the possibility the occurrence of consumers health risk. Figure 3 revealed
The results more or less agreed with (Álvarez-Astorga et al., 2002) (6.29 ± 0.64 to 7.28 ± 0.51), (Abd-El Aziz et al., 2012) examined three batches of raw meat delivered to three restaurants for processing of escalope pane and found that APC recorded 5.86 log 10 cfu/g of meat batch no III delivered to restaurant no."A", (4.6) for Enterobacteriaceae from batch II of restaurant "A", (4.6, 4.54 and 4.72) for Enterobacteriaceae collected from batch no III of restaurant a, B and C, respectively. The author also recorded similar results for E. coli for all three meat batches from the three restaurants with average of 2.6 log 10 cfu/g, while lower Staph. aureus counts which ranged from (0.52 t0 1.95 log 10 cfu/g), in escalope pane sandwiches, similar results for E. coli and Staph. aureus (<1log 10 cfu/g) while APC and Enterobacteriaceae of processed escalope pane recorded higher counts than that in the present study (5.34 and 3.99). Lerasle et al. (2014) (6.05 ± 0.18) for APC In escalope pane. Coliforms are used as indicator of water pollution or as a general indicator of sanitary condition in in the food processing environment. Also, high coliforms count indicates poor hygienic quality of meat and considered as a significant indicator of fecal contamination. Coliforms also have the ability to grow well over wide range of temperature below 10 o C up to 46 o C (Feng et al., 2002). Moreover, the major foodborne pathogens represented by Escherichia coli, Staphylococcus aureus, and Salmonella (Gaia et al., 2017).
The primary action of hand washing is the mechanical removal of viable transient microorganisms, whereas the primary action of antimicrobial soap includes both mechanical removal and killing or inhibition of both transient and resident flora (Larson, 1989). This is an indication of insufficient hand washing and sanitation, as one would expect a significantly reduced bacterial count on the workers' hands after they have cleaned and sanitized them. (Raspor, 2008) reported the importance of management training of all employees in the use of effective hand washing procedures, and that the safety of food chain supply can easily be broken proper enforcement these procedures. Sixty percent of the TPC samples analyzed exceeded the legal limit (<100 cfu.cm -2 ) stipulated by the Foodstuffs, Cosmetics and Disinfectants Act for food contact surfaces (Republic of South Africa. Regulation 962, 2012).    (8.18,33.85 and 35.35) for pane, shish tawook and grilled kofta, respectively.
In this regard, (Hassanin-Faten et al., 2017) recorded mean pH value of chicken breast and thigh (5.81 ± 0.05 and 5.73 ± 0.03), TVB-N (5.56 ± 0.24 and 7.12 ± 0.29) and TBA (0.09 ± 0.01and 0.14 ± 0.01), respectively. Several investigators recorded pH of chicken thigh and fillet including; (Shedeed, 1999)  The susceptibility of RTE cooked meat to lipid oxidation are closely related to levels of lipids, unsaturated fatty acids and iron present in the different species. Lipid oxidation had extensive economic importance for the meat industry as it results in development of rancidity and formation of potentially toxic reaction products as well as chemical spoilage in food (Murphy et al., 2001;Tang et al., (2001). Moreover, the significant increase of Thiobarbeturic acid TBA) value was due to prolonged storage time which will result in the presence of unpleasant detectable flavor and lower degree of acceptability of processed products due to fat oxidation which resulted from prolonged storage or due to the use of low-quality meat in production of such products (Mulla, 2002). In addition, pH value is an indicator of the keeping quality of meat where the pH measurement of meat and its products is used to assess the shelf life and quality of the products, the decrease in pH value may be due to the breakdown of glycogen with the formation of lactic acid and the increase of pH may be due to the partial proteolysis (Shaltout et al., 2014).

CONCLUSIONS AND RECOMMENDATIONS
This study provided useful information about the hygienic status in food serving establishment in Egypt. In general, findings have shown that treated cooked RTE meat and chicken products contained APC (< 5 log10cfu/g), Enterobacteriaceae (< 2), coliforms (< 3), Staph aureus (< 3) while, Escherichia coli and Salmonella failed to be detected in all the examined heat-treated samples. These limits are considered satisfactory according to the safety parameters submitted by general international organizations. While, for swab samples, the obtained results in the present study considered unsatisfactory as APC should not exceed 2 log10cfu/cm2 as well as coliforms and staphylococci should be absent /cm2. Therefore, training courses about the safe dealing with food and the importance to follow up general and personnel cleaning and application of good hygienic and manufacturing (GHP and GMP) practices should be submitted for workers in the preparation and provision of food in order to safeguard consumer health from food-borne pathogens Which is also contribute to improving the quality of products offered to the pioneers of those places.

ACKNOWLEDGMENTS
The authors are thankful to General Organization for Veterinary Services, Egypt for providing necessary facilities for this study. The authors did not receive any funds for this study.

NOVELTy STATEMENT
The current work was planned to investigate the hygienic status in food serving establishment through determining the bacterial profile of served meat and poultry meals including raw and after cooking shish tawook, kofta, fillet and escalope pane, as well as determination of their physico-chemical criteria and examination of swab samples from different localities which reflect the level of hygiene during food production and subsequently its shelf life and thus, the potential problems arises during the stages of food production and processing.

AUTHOR'S CONTRIbUTION
ASO collected the samples, and performed the laboratory work and data analysis. KST, AA and MEE planned the research and experimental design, supervised the work, and checked the data analysis. SHAH prepare the manuscript.

eThiCAl STATemenT
Ethical approval was not needed in this study.