One of the major limitations of current methods of biological detection of exposure to hazardous environmental agents is their inability to detect long-term exposures. In the current study we examined the potential of a new bioassay based on the hypothesis that serum of exposed individuals contains a toxic factor(s) produced by an affected cell/tissue. The procedure included exposure of neuronal PC12 cell cultures to sera of rats treated once with the organophosphate chlorpyrifos. Samples taken 4 weeks after chlorpyrifos exposure reduced nerve growth factor (NGF)-induced neurite outgrowth by 40%. This effect lasted 6 weeks after treatment, whereas motor activity and cholinesterase activity returned to normal levels within 1 week. These results demonstrate the potential of the proposed method to detect environmental exposures long after they have occurred.