In response to reported increased cancer risks among farmers, the Agricultural Health Study (AHS) was designed to examine health outcomes and environmental exposures among farm families in the United States. In the pilot phase of the AHS, food, beverage, air, dermal, dust, surface wipe, and biological specimens (blood and urine) were collected and analyzed for six farm families in two states (IA and NC). In addition, questionnaires were administered to examine previous pesticide use. This paper reports the organochlorine pesticide results of the serum and dietary analyses as well as questionnaire results from the pilot exposure study of farmers and their families. Note, no organochlorine pesticides were reported as currently being applied to the study farms. In all human serum samples examined, typical U.S. population levels were found for the majority of the pesticides. In addition, human serum levels of organochlorine pesticides showed no significant daily or seasonal variation. However, serum trans-nonachlor levels were found to be higher in people living on the two farms in North Carolina than in people living on the four farms in Iowa (p < 0.05). Further, unusually high dieldrin levels were found in serum samples from a farmer and spouse living on an Iowa farm, and these levels were significantly higher than those of people living on the other farms (p < 0.05). Dieldrin was persistent in the foods consumed on the same Iowa farm where family members showed elevated serum levels. In addition, dietary samples from the North Carolina farms exhibited high levels of chlordane. No organochlorine pesticides were found in any of the drinking water samples. Dietary dieldrin levels on the same Iowa farm exceeded the oral reference dose (RfD) eight- to eleven-fold (50 ng/kg-day). No other pesticide exceeded the RfD. However, dietary chlordane levels at a North Carolina farm reached 17% of the RfD. Previous use of aldrin on an Iowa farm corresponded to dieldrin found in the diet and in the serum of the farmer and spouse. Previous reported use of chlordane on the North Carolina farms corresponded with measurable dietary levels of chlordance and higher serum trans-nonachlor levels than the levels in Iowa farm families.