Beekeeping is a breeding activity in which various products such as honey, pollen, royal jelly and propolis can be obtained by using honey bees (Apis mellifera L.) and plant sources. Bees have vital importance due to their role in plant pollination in the nature and the positive effects of bee products on human health. Bee diseases and pests are among the most important problems in beekeeping. One of these diseases, American foulbrood (AFB), is a very dangerous and highly contagious disease that causes the death of honey bee larvae. The causative agent of the disease, Paenibacillus larvae, is a Gram-positive bacterium that has two forms: vegetative and spore form. In this study, the presence of AFB was investigated in 207 samples taken from beekeeping enterprises in the Central and Eastern Black Sea regions between 2020-2023. Isolation, identification, and PCR analysis of P. larvae bacteria, the causative agent of AFB performed on the samples. As a result of the study, it was determined that 10 (4.83%) of 207 honey bee enterprises were infected with P. larvae. Effective monitoring of AFB disease with rapid detection methods can help beekeepers to take early and effective control measures and prevent the further spread of the disease.