The chemical composition of cactus forage becomes a favorable culture medium for accelerated microbial activity when exposed to air, as it contains high content of non-fiber carbohydrates and water. Thus, the aim of this study was to evaluate the bacterial community dynamics of different mixtures, using fresh forage of cactus and buffel grass hay as a function of the period of exposure to air. The experimental design used was a 5 × 5 factorial completely randomized (five levels of cactus forage × five times of exposure to air), with five replications. The peak of Escherichia coli population growth was after 16.06 h of exposure to air, observed in treatments of 90% and 100% cactus forage. There was an increase in microbial richness and uniformity of all treatments after six hours. The most abundant genera were Weissella, Lactobacillus, Bacteroides, Pseudomonas, Sphingobacterium, and Sphingomonas. The diet with 100% cactus forage showed a predominance of Weissella, Lactobacillus, and Leuconostoc. With 20% cactus forage, there was a greater apparent abundance of Pseudomonas, Sphingomonas, and Sphingobacterium. Aerobic exposure of mixtures of cactus forage with buffel grass hay increases the proliferation of microorganisms with pathogenic potential in the diet. Aerobic exposure of mixtures of cactus forage with buffel grass hay increases the proliferation of microorganisms with pathogenic potential in the diet. Therefore, an exposure period of fewer than six hours with 20% cactus forage is recommended to minimize levels of E. coli. Avoiding negative effects of the multiplication of pathogenic microorganisms on animal and human health.