Varroa destructor is considered the most dangerous external parasite affecting honey bee (Apis mellifera) colonies, and the principal factor associated with colony collapse, whether through its direct effects or through its role as a vector for several viruses, most notably Deformed Wing Virus (DWV). Although the parasite entered Libya in 1976, with its first appearance recorded in the Jabal Al-Akhdar region, recent epidemiological information on infestation levels in this region remains limited. This study aimed to provide preliminary epidemiological data on the prevalence of Varroa mite infestation in honey bee colonies in the Jabal Al-Akhdar region of Libya, and to describe the associated clinical manifestations, by combining beekeeper-reported information with direct laboratory examination. The study comprised two parts: an electronic survey of 46 beekeepers covering twelve items on apiary characteristics and monitoring/treatment practices, and field visits to ten apiaries during March–June 2026, in which 4 hives were randomly examined per apiary (40 hives total) and fully opened manually to extract reproductive mites, alongside phoretic mites collected from dead bee samples; mites were morphologically identified, and a composite infestation severity index was calculated as the mean number of Varroa individuals per examined hive. 84.8% of beekeepers reported personally observing Varroa in their apiaries, while 80.4% relied on direct visual inspection as their sole monitoring method. Laboratory examination confirmed the parasite's presence at all ten sites without exception (100% prevalence), including both phoretic and reproductive stages, with an overall infestation severity index averaging 4.98 ± 1.81 mites/hive (95% CI: 3.68–6.27; range: 2.50–7.25). A chi-square goodness-of-fit test showed a statistically significant difference in infestation density among the ten apiaries (χ² = 23.66, df = 9, p = 0.0049). Varroa destructor was present in every apiary subjected to field examination, with variation in the infestation severity index among sites, while beekeepers showed limited reliance on quantitative monitoring methods. These findings indicate a need to strengthen epidemiological surveillance and awareness programs on appropriate detection and control methods across the Jabal Al-Akhdar region................... Keywords:.................. Varroa destructor mite; Jabal Al-Akhdar; Libya; honey bees; infestation severity index; Deformed Wing Virus (DWV).