Detection of Ascosphaera apis, causing chalkbrood disease in the colonies of European honey bee, Apis mellifera in West Bengal, India

The decline of honey bee (Apis mellifera L.) populations is of great concern around the world. Among the several key drivers, dissemination of pests and pathogens is potential one. Chalkbrood is very common fungal disease of honey bee, caused due to Ascosphaera apis. In the present study, survey was conducted regarding the prevalence of diseases in A. mellifera beekeeping in Gangetic plains of West Bengal, India. Results confirmed the occurrence of chalkbrood disease in different apiaries, where dead and mummified bee larvae with cotton like chalky white or greyish-black covering were found as physical symptoms of the disease. From three surveyed apiaries, a total of 16 hives out of 113 hives were found to be infected with the diseases; and 46 frames out of 132 frames from the infected 16 hives were found to be affected by the pathogen. Microscopic examination reflected that nearly 87% of the samples collected from the infected frame were found to be positive for the spore of Ascosphaera. The fungus associated with the disease was isolated on Potato Dextrose Agar medium, pure cultured and its genomic DNA was isolated to perform PCR and based on 18s rDNA sequencing by using specific primer pair of ITS-1 and ITS-4, the fungus was identified as Ascosphaera apis. Keywords: 18s rDNA, Bees, Necrotrophs, Phylogen