During the spring-summer 2019, the presence and prevalence of the Black Queen Cell Virus (BQCV), Chronic Bee Paralysis Virus (CBPV), Deformed Wing Virus (DWV), Kashmir Bee Virus (KBV), Israeli Acute Paralysis Virus (IAPV), Lake Sinai Virus 1 (LSV1), Lake Sinai Virus 2 (LSV2) and Sacbrood Virus (SBV) were investigated in 125 Apulian honey bee colonies. A total of 10 honey bees were sampled from each colony and were pooled as 1 sample, then analyzed by using Reverse Transcription Polymerase Chain Reaction (RT-PCR). The obtained RT-PCR results showed that the most prevalent virus was DWV, present in 88% of the tested samples, followed by CBPV, LSV1, LSV2, BQCV, and SBV (33%, 27%, 26%, 23%, and 10% respectively). Whereas, KBV and IAPV were not detected in any of the surveyed apiaries. Interestingly, most of the detected viruses were reported in multiple infections. The phylogenetic analyses of
the detected viruses confirmed the high sequence identity at the nucleotide level with other isolates distributed worldwide. In addition, this study reports the first detection of LSV1 and LSV2 in Apulia region, and the first molecular characterization of CBPV, LSV1, LSV2, BQCV and SBV in Apis mellifera colonies of this area.