Aim: To assess the prevalence of Non-Typhoidal Salmonella spp isolated from food and stool samples in Port Harcourt. Study Design: This was a cross-sectional study with simple randomized sampling. Methodology: In this study, 210 stool samples and 210 food samples collected from December 2022 to November 2023 were tested for Salmonella using standard bacteriological and biochemical tests. The Salmonella species were isolated from the samples using Salmonella-Shigella agar (SSA), and Bismuth Sulfite Agar (BSA) after pre-enrichment and enrichment methods had been carried out using peptone water and Selenite F Broth after which biochemical tests were carried out for further identification. Data collected was analyzed with Graph pad prism version 8. Results: The prevalence and distribution of Salmonella were presented in frequencies and percentages with all analysis done at a 95% confidence interval and P-values less than .05 were considered significant. From chi-square analysis there was a 3.3% prevalence rate in food samples in comparison to stool samples that had a 2.4% prevalence (P value = .56). A higher prevalence was also reported in the female subjects (3.7%) compared to the male subjects (1.0%) (P value = .19). There was a statistically significant difference in relation to the age groups with the ‘above 50’ and ‘0 – 10’ age groups having a higher prevalence (11.1% and 10% respectively) compared to other age groups (P value = .02). For the food samples, samples in the chicken category had the highest prevalence (8.7%) (P value = 0.11). Conclusion: This study reports a relatively lower prevalence of Non-Typhoidal Salmonella species at 2.9% with the age, education and occupation of the subjects being significantly associated (P value < .05) with the prevalence of the infection. Health promotion and appropriate surveillance system should be put in place to continually reduce the burden of this disease.