Aims: The contamination of groundnut by aflatoxins (AF) results in financial losses to farmers’ as well as severe food safety and public health challenges globally. This study was carried out to; (i) assess the levels of AFB1 in husked groundnut seeds in Makurdi (ii) determine the relationship between moisture content and AF levels, and (iii) investigate vendors’ knowledge, attitudes and practices (KAP) of AF and their approach towards groundnut storage. Study Design: Quantitative research method was employed in this study. Place and Duration of Study: This study was conducted at the Department of Microbiology, Benue State University, Makurdi from May – June 2019. Methodology: Duplicate groundnut samples were collected from ten market locations in Makurdi and analyzed using Enzyme Linked Immunosorbent Assay (ELISA) quantification method. Results: The moisture content of the groundnuts was determined, and data on Knowledge Attitude and Practice (KAP) relating to groundnut storage were obtained using mini-questionnaires. The results obtained showed that all the sampled groundnuts were contaminated with AFB1 levels ranging from 17.3 - 35.9 parts per billion (ppb). Furthermore, we found a correlation between high moisture content and high AFB1 levels and vice-versa. The knowledge of AF among the groundnut retailers was low (<40%), and 40.91% of the sellers confirmed that groundnuts were stored for ≤one month before sale. Conclusion: The levels of AFB1 levels in stored groundnuts are above the permissible limit of 20 ppb for stored groundnut in Nigeria. The data obtained raises concerns for food safety considering that groundnuts are widely consumed in Makurdi. Regular evaluation of AFB1 levels in food should be conducted in Nigeria.