Lyme borreliosis is known to be associated with several outdoor professions and activities, but the incidence of Lyme borreliosis in beekeepers has not previously been investigated. We distributed questionnaires based on various earlier investigations and 732 beekeepers responded. We found that in 31.1% of the responders, Lyme borreliosis had been diagnosed. Factors found to be associated with the likelihood of Lyme borreliosis infection were: the estimated percentage of tick bites received during beekeeping (number of tick bites during beekeeping compared to total number of tick bites); the total number of bites; the duration of beekeeping; migratory beekeeping and locating hives in forests. Stepwise discriminant analysis identified the estimated percentage of tick bites received during beekeeping as the most relevant factor. Protection against tick bites was used by only a minority of responders (11.4%). From our results, we must conclude that beekeepers should be considered as a high risk group for Lyme borreliosis when compared to the general population and even forest workers. Education to increase the use of protection must be encouraged in order to decrease the incidence of Lyme borreliosis.