Honey bees, Apis mellifera L. (Hymenoptera: Apidae), are the most important pollinators of agricultural products and plants in the natural environment. Honeybees are an important ecosystem component due to their role in nature and agricultural production. Bacterial, fungal, viral, and parasitic factors in bee farms are among the most important causes of honey bee colony losses. Honey bee diseases (bacterial, fungal and viral) and parasites are among the most important factors limiting beekeeping development and production efficiency in Türkiye. In addition to diseases caused by bacterial and fungal agents, diseases caused by viral agents are very diverse. Viruses, especially mixed infections, cause colony losses and are the most important factors in the decline of honey bee colonies. In this study the presence and prevalence of honey bee pathogens (Varroa destructor, Nosema ceranae, Paenibacillus larvae, and nine viruses) in suspicious samples with colony losses were investigated in Sivas province. For this purpose, microscopic, microbiological, and molecular methods were investigated on larvae and adult bee. The results showed that the most common viral pathogens in samples from Sivas province were Deformed Wing Virus (70%), Apis mellifera Filamentous Virus (60%), Black Queen Cell Virus (60%), Sacbrood Virus (55%) and Varroa destructor virus-1 (40%), respectively. In some samples, it was observed that there was a double (17.5%), triple (30%), quadruple (22.5%), or even quintuple (17.5%) association of viral agents. The viral infection/varroa coexistence rate was determined to be 50%. It was determined that 22.5% of the samples examined contained Nosema spores, while 12.5% were positive for P. larvae. Revealing the distribution of bee diseases will help beekeepers in disease-fighting and taking measures. This study showed the presence of the AmFV and the Varroa destructor virüs-1 in the Sivas province of Türkiye for the first time.