Technical Abstract: Aflatoxins present important food safely problems in both developed and developing countries. Contamination is monitored in developed countries using ELISA- and HPLC-based assays, both of which may be too expensive for routine use in many developing countries. There is a need for inexpensive alternative approaches to predict the likelihood of contamination in lots of foods and feeds. Reviewed here are culture-based methods, which determine if a sample is contaminated with aflatoxigenic fungi. These approaches include (i) blue fluorescence of aflatoxin B1, particularly when enhanced by including B-cyclodextrin in the culture medium, (ii) yellow pigment production and (iii) collor change on exposure to ammonium hydroxide vapor. Blue fluorescence detects the presence of aflatoxin B1, which is enhanced when the toxin complexes with the hydrophobic pocket of B-cyclodextrin. However, the yellow pigment and ammonium hydroxide vapor tests were found to be based on the production of yellow anthraquinone biosynthetic intermediates on the pathway to aflatoxins, which act as pH indicator dyes more visible in the red form at higher pH. Because the tests are based on two different mechanisms, it has been possible to combine them into a single test. In a study of 517 A. flavus isolates from the Mississippi Delta, the combined assay reduced false positives for aflatoxigenicity to 0% and false negatives to 7%. The increased predictive power of the combined cultural assay may enable its use for inexpensively identifying potential aflatoxin contamination in feeds and foods.