Increased bioaerosol loadings in downwind plumes from concentrated animal feeding operations (CAFOs) may increase the risk for allergy and infection in humans. In this study, we monitored airborne concentrations of culturable bacteria and fungi at upwind (background) and downwind sites at a 10,000 milking cow dairy over the course of a year. The average bacterial concentrations at the upwind site were 8.4 9 10 3 colony forming units (CFU) m -3and increased to 9.9 9 10 5 CFU m -3 at the downwind edge of the cattle lots, decreasing to 6.3 9 10 4 CFU m -3 200 m farther downwind. At the same sites, the average fungal concentrations were 515, 945, and 1,010 CFU m -3 , respectively. Significant correlations between the ambient weather conditions and airborne fungal and bacterial concentrations were identified. Sequence analysis of PCR-amplified DNA from bacterial clones and fungal isolates revealed genus and species level differences between upwind and downwind sites. Although we could not cultivate gram-negative bacteria, bacterial clones at downwind sites identified as being gram-negative matched with the following genera: Acinetobacter, Bradyrhizobium, Escherichia, Idiomarina, Methylobacterium, Ralstonia, and Novosphingobium. Fungal isolates from downwind matched with the following genera: