Arab Journal of Nutrition and Exercise                                                                                                       ISSN:2518-6590          Published on 31 st August, 2017 Abstract Background: Cross-contamination of foodborne pathogens from undercooked poultry meat to ready-to-eat food has been shown to be responsible for a number of foodborne disease outbreaks. Various studies have indicated that bacterial cross-contamination occurs during food preparation where bacteria present on food contact other surfaces and cause illness. Objectives: This study evaluated the ability of bacteria to survive and cross-contaminate other foods during the preparation of fresh chicken. Salmonella spp. cross-contamination from chicken to cucumber and utensils under various food handling scenarios was determined. Methods: Two scenarios were tested: in scenario 1, cutting board and knife used for cutting chicken without washing step were sampled.  In scenario 2, cutting board and knife was washed with tap water separately after cutting chicken, and subsequently used for cutting cucumber. In scenario 1, chicken, cutting board, knife, and hands were sampled, and in scenario 2, cucumber was tested. Fluorescent in situ hybridization (FISH) method, using published Salmonella specific gene probes was used for Salmonella detection in samples taken from cross-contamination scenarios. A culture-based detection by Hektoen enteric agar was used for the confirmation of Salmonella species . Results: All the samples analyzed were found to be positive for Salmonella spp. with different contamination levels. These results were further confirmed by culture based method. In scenario 1, Salmonella spp. was detected by Sal-1 and Salm-63 oligonucleotide probes in all four samples (chicken, cutting board, knife and hands). A high contamination level was observed in chicken samples in comparison to samples collected from cutting board, knife and hands. In scenario 2, Salmonella spp. was detected by Sal-1 and Salm-63 oligonucleotide probes in the cucumber with very low contamination level. Salmonella Enterica was also detected by Sal-3 and Sapath-3 in both scenarios but the contamination level was not high as compared to Salmonella spp. Conclusion: In conclusion, Salmonella spp. cross-contamination during fresh chicken preparation to read-to-eat-food (cucumber) was confirmed by this study. The experimental data obtained in this study clearly suggest that it is extremely difficult to prevent the spread of Salmonella spp. during the preparation of raw poultry-based meals. Therefore, extreme precautions such as proper cleaning and sanitization of utensils, equipment and surfaces should be carefully followed during the preparation of fresh poultry meat-based food items.