الفهرس 
INTRODUCTION
RISK FACTORS from INFECTED FOOD HANDLERSOnly 14 pages are availabe for public view
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Abstract
Contamination of salads may occur at any point along the farm-to-fork continuum. during growing, harvesting, sorting, preparation, packaging, and storage of fresh fruits and vegetables. Most human pathogens associated with consumption of fresh fruits and vegetables are transmitted via the faecal-oral pathway, in which fecal material from an infected individual contaminates food that is then ingested by a healthy person. Therefore, set of guidelines to prevent microbial contamination should be considerably taken.
This study was conducted in the kitchen of 2 restaurants in the Eastern zone and in 2 restaurants in the middle zone of Alexandria. It aimed to assess the microbiological and the parasitic quality during green salads preparation, to assess the hygienic conditions under which green salads are prepared, to carry out an intervention program to correct problems encountered during salad handling, to reassess the microbiological and the parasitic quality of green salads after the intervention program and to reassess the hygienic conditions under which green salads are prepared.
To fulfil this aim the following method was adopted:
1. Assessment/Reassessment of the hygienic conditions under which
green salads are prepared.
Hygienic conditions under which green salads were prepared were assessed before intervention, this was done using a pre- deigned checklist containing the following parameters:
1. RESTAURANT’S ENVIRONMENT
1.1. Grounds
1.2. Kitchen design
2. SANITARY OPERATION
2.1. Pest control
2.2. Cleaning &sanitizing
3. SANITARY FACILITY AND CONTROL
3.1. Water supply
3.2. Toilet facilities
4. KITCHEN EQUIPMENT AND UTENSILS
4.1. Kitchen utensils
4.2. Cutting board
5. FOOD HANDLER
5.1. Disease control
5.2. Responsibility
5.3. Cleanliness
6. HANDLING OF SALAD
6.1. Receiving of salad (cucumber, tomato, lettuce)
6.2. Sources of salad
6.3. Preparation of salad
6.4. Storage of salad
2. Intervention stage
The intervention stage was carried out in only the 2 restaurants of the middle zone of Alexandria as they agreed to participate. In this stage handlers were first asked to fill the predesigned knowledge questionnaire to test their knowledge about the safe preparation of green salads. After that, they were invited to attend a training session to provide them with the basic knowledge needed to prepare safe salads. Slide show presentations were designed and used for that purpose. Immediately at the end of the training session, handlers were asked to answer a short knowledge quiz similar to the knowledge questionnaire to test their prompt acquired knowledge.
After approximately one week handlers were asked to refill the knowledge questionnaire to obtain a feedback on their retained knowledge.
The knowledge quiz had the following Parameters and items:
The knowledge questionnaire contained the following parameters:
1. SOURCES OF SALAD CONTAMINATION
1.1. Insects and rodents
1.2. Water
1.3. Soil
1.4. Air
1.5. Sewage
1.6. Equipment and utensils
1.7. Floors, walls, and ceilings
2. FOOD HANDLER AS A SOURCE OF FOOD CONTAMINATION
2.1. Handler’s nails
2.2. Jewellery and accessories
2.3. Hair and beard
2.4. Face, nose, mouth
2.5. Cuts and wounds
2.6. Salad handling practices
2.7. Hand washing
3. SAFE HANDLING OF SALAD
3.1. Receiving of salad
3.2. Hygienic preparation of salad
3.3. Storing of salad
3.4. Serving of salad
3.5. Cleaning and sanitizing
3. Assessment /Reassessment of the microbiological quality during green
salads preparation.
Samples of green salad and swabs from utensils and food preparation surfaces were collected from the kitchen of the visited restaurants and were subjected to microbiological examination.
A total of 79 green salad samples were collected. These were distributed as 19 samples from the East zone of Alexandria (9 from E1 and 10 from E2 restaurants) and 60 samples from the Middle zone of Alexandria (15 from M1 and 15 from M2 restaurants; before intervention +15 from M1 and 15 from M2 restaurants; after intervention ).
Samples from E1 restaurant were taken from the different steps of salad prepared, 3 from raw vegetables, 3 from prepared salads and 3 from stored salads. Samples of the other restaurants (E2, M1 and M2) were ready to serve only.
These samples were subjected to the following microbiological tests:
1. Total aerobic mesophilic plate count
2. Enumeration of coliforms [Most Probable Number, (MPN)]
3. Detection of fecal coliforms
4. Isolation of Coagulase and DNase positive Staphylococci
5. Mold and yeast count
6. Detection of parasites (Giardia spp,and Entamoeba histolytica)
Additionally, a total of 9 swab samples were collected from E1 restaurant only. These were as follows:
• 3 swabs from cutting boards.
• 3 swabs from surfaces.
• 4 swabs from knives.
Also, 2 hand washes were collected from the same restaurant.
The obtained results were as follows:
• This study was carried out in 2 zones of Alexandria Governorate; the Eastern Zone (E1 and E2 restaurants) and the Middle Zone (M1 and M2 restaurants).
1. Checklist of the visited restaurants
• Salads were received from unsafe sources in E1 restaurant (score percentage=0.00 %) and were more or less properly prepared and handled (score percentages =63.64 %,). As a total, the handling of salad in E1 restaurant was given a moderate score percentage of 65.38 %. As a total the kitchen was scored only 52.34 % which indicates poor hygienic environment and poor hygienic handling of salads.
• M1 restaurant’s environment was very poor as it was given a total score percentage of 33.33%.While that of M2 was moderate (total score percentage =71.43%).
Equipment and utensils in both restaurants were poor, they were given a very low total score percentages of 16.67 %.
• Moreover, salads in M1 restaurant received poor preparation (score percentage 36.36%) and more or less properly storage of salad (score percentage 62.50 %). As a total, the handling of salad in M1 restaurant was given a low score percentage of 46.15 %.
• The overall mean score percentages of the checklists were 52.34%, 35.16 %, 54.90 % in E1, M1, and M2 in which M2 restaurant had the highest overall score percentage.
• The highest parameter was sanitary operations in E1 and M1 restaurants (85.71 % and 57.14 %) And sanitary facilities & control in M2 restaurant 71.43 %
2. Knowledge of Food handlers
2.1 Before intervention:
• Food handlers in E2 restaurant had higher mean score percentage (85.26 %) concerning their knowledge about sources of contamination than E1 restaurant (63.85%).
• For the knowledge about serving of salad as well as cleaning and sanitizing; these were excellent for workers of E2 (mean score=100.00% in each item), while a mean score percentage of 60.00 % was given to E1 workers who had low perception about cleaning and sanitizing as being sources of contamination.
• Generally, knowledge of food handlers in E2 restaurant was higher (85.98 %) than that in E1 restaurant (73.02 %).
• Food handlers in M1 restaurant had higher mean score percentage (80.77%) concerning their knowledge about sources of contamination than in M2 restaurant (56.92%).
• Knowledge about hygienic storage of salad (95.00 %) was excellent in workers of M1 restaurant while the lowest mean score percentage (75.00 %) was given for their low perception about receiving of salad and serving of salad.
• Concerning M2 restaurant, knowledge of workers about storing of salad, cleaning and sanitizing (80.00 %) was good while the lowest mean score percentage was given to workers who had low perception about hygienic preparation of salad (57.00 %).
• Generally, knowledge of food handlers in M1 restaurant was higher (86.51 %) than in M2 restaurant (66.53 %).
• Comparison between Eastern and Middle zones, regarding the knowledge of their food handlers before intervention showed that E2 workers had the highest significant mean score (22.17 ± 1.33) for their knowledge about the different sources of salad contamination (P<0.05). However, their scores differed insignificantly for the other knowledge parameters. As a total, handlers in M1 restaurant had the highest significant mean score (54.50±6.61, P<0.05)
2.2 During slide show training
• Generally, food handler’s performance in M2 restaurant was higher (mean score percentage 87.33 %) than those in M1 restaurant (mean score percentage 80.83 %) during slide show training.
2.3 After intervention
• Food handlers in M2 restaurant had higher mean score percentage (92.31%) concerning their Knowledge about sources of contamination than there in M1 restaurant (78.85 %).
• Concerning M2 restaurant, knowledge of workers about serving of salad was excellent (100.00 %) while the lowest mean score percentage was given to workers who had low perception about receiving of salad and cleaning and sanitizing (0.00%).
• Generally, Knowledge of food handlers in M2 restaurant was higher (83.49%) than in M1 restaurant (79.76 %).
2.4 Comparison Before and After intervention
• Food handlers in M1 restaurant had higher mean score percentage (80.77 %) concerning their knowledge about sources of salad contamination before intervention than after intervention (78.85 %), while in M2 restaurant food handlers had higher mean score percentage (92.31 %) after intervention than before intervention (56.92 %).
• Concerning M1 restaurant, knowledge of workers about food handlers was higher (92.50 % mean score percentage) than after intervention (70.00 %), while in M2 restaurant they had higher mean score percentage after intervention (85.00 % ) than before intervention( 78.00 %)
• For knowledge about handling of salad in M1 restaurant was higher before intervention (88.40 %) than after (77.94 %), while in M2 restaurant knowledge was higher after intervention (68.24 %) than before intervention (67.06 %).
• Generally, It can be seen from the results that the mean score knowledge of M1 handlers decreased insignificantly, after applying the intervention as a whole in each of the questionnaire’s parameters. On the other hand, the mean score knowledge of M2 handlers increased significantly (P<0.05) for their knowledge about different sources of salad contamination before and after intervention (from 14.80 ± 5.430 to 24.00±0.000, respectively).
• Similar significant increase was obtained in their scores of the overall questionnaire, from
41.800 ±5.762 to 52.60 ± 0.548, before and after intervention, respectively.
3. The Microbiological and parasitic profile of green salads.
3.1 Green salad prepared and served at E1 restaurant
3.1.1 Microbiological and parasitic content of the different steps of green salad
preparation
• The aerobic plate count of examined samples was high. It was 1.00×106 ± 0.00 cfu/g for raw salad, decreased to 6.81×105 ± 5.52×105 cfu/g for prepared salad, then increased to 7.00×106±5.19×106 cfu/g for stored salad samples.
• The coliform counts of examined samples was 7.65×102 ± 1.08×103 MPN/g for raw salad, increased to 4.41×103 ± 5.78×103 MPN/g in prepared salad, then to 6.00×103 ± 4.58×103 MPN/g in stored salad.
• Staphylococci were not detected in raw salad samples, however, were found in the following salad preparation steps; prepared salad (3.57×104 ± 5.58×104 cfu/g) and stored salad (3.33×104 ± 5.77×104 cfu/g). Fortunately, coagulase and DNase staphylococci were not detected in raw, prepared or stored salad samples.
• Moreover, mold and yeast were found to be higher in the salad samples, they were up to 7.26×106 ± 4.73×106 cfu/gm for stored salad samples.
• Results also showed the absence of a significant difference in the APC, coliform, staphylococci and mould and yeast counts of either raw, prepared or stored salad.
• Additionally all examined samples were negative for fecal coliform and for parasites.
3.1.2 Microbiological profile of swabs collected from surfaces and utensils
• Both cutting boards and surfaces were also heavily contaminated with aerobic microorganisms. The APC was 1.00×103±0.00 and 1.72×103±2.51×103 cfu/cm2, respectively.
• The highest coliform count detected in E1 restaurant’s kitchen was 4.57×10 ± 5.61×10 MPN/g for the swabs taken from the cutting board.
• Staphylococci were found to range from a minimum of 1.50 ±1.00 cfu/cm2 for the knife surface to a maximum of 6.67×10 ±1.15×102 cfu/cm2 for the surfaces. Fortunately, none of the samples were positive for coagulase and DNase staphylococci.
• Mold and yeast were as high as 2.20×103±3.81×103 cfu/cm2 in the swab taken from the cutting board.
3.2 Green salad samples served at E2, M1, and M2 restaurants
3.2.1 Before intervention
Results in E2 restaurant showed that :
• Thirty% (30%) of the samples had APC ≥107 cfu/g and 70% had counts ranging from 1.00×106-9.20×106.
• The mould and yeast count was from 1.00×105-8.4×105 in 80.00% of the samples, and was 1.00×107in 20.00% cfu/g of them.
• For staphylococci, 60.00% of the examined samples had counts ranging from 4.20×102-6.80×104 and none of the examined samples were coagulase and DNase positive staphylococci.
• All examined samples had coliform counts ranging from 2.00×103-1.10×104MPN/g.
• Only one sample was positive for Giardia spp.
Results in M1 restaurant showed that:
• All examined samples in M1 restaurant had coliforms count 1.10×104 MPN/g. Additionally, all examined samples were negative for fecal coliform
• For staphylococci, 33.33% of the examined samples had counts ranging from 1.20×105-1.00×106, 46.67% had counts from 1.03×104-2.80×104, and 20.00% of them were <10 cfu/g.
• There were 2 positive samples of Coagulase and DNase positive Staphylococci .
• There were 3 samples positive for Entamoeba histolytica.
Results in M2 restaurant showed that :
• All of examined samples had coliform counts ranging from 2.00×103-1.10×104 MPN/g.
• Only one sample was positive for fecal coliform.
• For total staphylococci, 40% of the examined samples had counts ranging from 2.00×103-7.20×104, 47% of them had counts ranging from 1.00×105-1.00×106 and 13% were ≥107.
• For mold and yeast, 13% of the examined samples had count ≥107, 74 % of the samples had counts ranging from 3.20×105-8.40×106 and 13% had counts ranging from 2.00×103-3.00×104.
Results of the comparison between Eastern and Middle zones restaurants showed that:
• There were significant differences in the APC and coliform count of the examined served green salad samples collected from different restaurants at P<0.05.
• The highest mean APC was 5.52 ×106 ± 3.90 × 106 cfu/g for samples from E2 restaurant, followed by 5.46 ×106 ± 3.78 × 106 cfu/g for samples from M1 restaurant.
• Samples from E1 restaurant showed the least mean APC count of 6.81×105 ±5.52×105 cfu/g.
• The highest coliform count was1.10 ×104 ± 0.00 MPN/g which was detected in M1 restaurants followed by 9.20 ×103 ± 3.79× 103 MPN/g for samples collected from E2 restaurant, that of M2 was as high as 8.40 × 103± 3.91 × 106 MPN/g while the lowest was 4.41 × 103± 5.78 × 103 MPN/g for samples from E1 restaurant.
• Staphylococci were found to range from a minimum of 3.57×104± 5.58× 104 in E1 restaurant to a maximum of 5.51× 106± 1.80× 107inM2 restaurant.
• For E2 and M1 restaurants , the mean staphylococci were of 3.21× 105± 9.42× 105and 1.46× 105± 2.83× 105.
• Two samples of M1 restaurant were positive for coagulase and DNase staphylococci.
• Mould and yeast were found to be higher in M1 (5.56× 106± 4.28× 106,followed by E2 (4.05× 106± 4.05× 106) , then E1 (3.46× 106± 5.67× 106) and finally M2 (2.99× 106± 3.52× 106).
3.2.2 After intervention
3.2.2.1 Microbiological and parasitic profile of the green salad samples served at M1 and M2 restaurants
• The APC of the examined samples was higher in the M1 restaurant (6.51×106±4.09×106 cfu/g) than in M2 restaurant (4.57×106 ± 4.62×106 cfu/g).
• The highest Coliform count detected in M1 restaurant’s kitchen was (6.82×103 ± 4.30×103 MPN/g). That of M2 was as high as (5.64 ×103 ± 5.22 ×103 MPN/ml).
• Staphylococci were higher in M2 (7.17×105 ± 2.57×106) than in M1 restaurant (7.51×104 ±6.97×104).
• The results also shows that 6 samples of M1 and 2 samples of M2 restaurant were positive for coagulase and DNase staphylococci.
• Moreover, mold and yeast were found to be higher in M1 (5.51×106 ± 4.24×106) than in M2 restaurant (2.53×106±3.94×106).
3.2.2.2 Effect of intervention on the microbiological profile of green salads served at M1 and M2 restaurants
• Results showed the presence of a significant decrease (P<0.05) in coliform count of M1 salad samples from 1.10×104 ± 0.00 before intervention to 6.82×103±4.30×103 MPN/g after intervention. However, no significant change was found for the other microbial tests.
• Similar significant decrease was found in coliforms of M2 samples before and after intervention (8.40×103±3.91×103 and 5.64×103±5.22×103, respectively) but not in the other tested microorganisms.
• Both staphylococci and mold and yeast counts in M1 and M2 samples samples decreased after intervention but insignificantly.
• Neither fecal coliforms nor parasites were detected after intervention.
Recommendations
The following points are recommended:
1. Managerial support, the availability of equipment and tools, training and pre-
training motivation are urgently needed as they can all influence the extent to which individuals react to the training experience.
2. Training of managers is recommended as it is a necessary precursor to the
implementation of realistic food safety practices within the workplace.
3. Managers should also understand that the effectiveness of training is very
dependent on both managers attitude and their willingness to provide the resources and systems for food handlers to implement good practice.