The risk of Bovine Leukemia Virus (BLV) transmission by rectal examination was determined over 22 months in a commercial dairy herd. All 167 BLV seronegative cattle, of breeding age or greater, were divided randomly into two groups and identified by neck-chain color. In the treatment group, routine rectal palpation occurred after a BLV infected animal and without a change of sleeve, while in the other group, palpation occurred in a similar manner with the exception that sleeves were changed between animals. When BLV seronegative cattle in either group were palpated after BLV infected cattle, the event and identification of the cattle involved were recorded. Serologic testing was performed eight times during the 22 month study to determine the number of animals that became infected following a palpation (an event), Thirty-one animals seroconverted during the study; 24 in the treatment (no sleeve change) group and seven in the sleeve change group. Sixteen of the animals in the treatment group that seroconverted had been palpated prior to their seroconversion. A hazard ratio (relative risk) for BLV seroconversion was determined between the two groups. Cows palpated with no sleeve change had a 2.8-fold increase in risk (confidence interval 1.1-6.8) of BLV infection. The increased risk of BLV infection associated with rectal palpation may have been affected by the presence of some highly infectious cows in the herd. This study confirms that rectal palpation without a change of sleeve may be a significant risk factor in some herds, and if efforts are made to decrease the spread of BLV in a herd, the potential for rectal sleeve transmission must be considered.