Acute exposure to the herbicide propanil is immunotoxic for selected immune functions, as well as causing changes in the weights of the thymus and spleen. Although spleen cellularity and weight increase with propanil exposure, the thymus: body weight ratio decreases with increasing doses of propanil. The present study analyzes the thymocyte subpopulations in the thymus, spleen, and mesenteric lymph nodes. C57BI/6 mice were treated with either 0, 100, 150, or 200 mg/kg propanil, and 7 d later thymocyte populations were analyzed by flow cytometry. In the thymus, propanil exposure resulted in a dose‐dependent decrease in total numbers of T cells, as would be expected with its reduced weight. Determination of the thymocyte subpopulation distribution in the thymus showed a significant reduction in the number of CD3+CD4+CD8−(CD3+4+8−), CD3+CD4−CD8+(CD3+4−8+), and CD3+CD4+CD8+(CD3+4+8+) cells. Percent distribution of these thymic cell subpopulations showed similar decreases only with the highest dose. Apparent dose‐related decreases in the numbers of CD3−CD4+CD8+(CD3−4+8+) cells were also noted and were attributed to the general decrease in total thymus cells. The percentage of CD3−subpopulations showed an increasing trend with dose, which suggests that at 7 d postpropanil exposure there may be a specific effect on this most immature population. Although the size and cellularity of the spleen were increased, no change in CD4+or CD8+cell distribution was observed. Similarly, mesenteric lymph nodes showed no changes in the cell subpopulation distribution between propanil‐treated and control animals.