Viral Encephalopathy and Retinopathy in Dusky Groupers (Epinephelus marginatus, Lowe 1834) from Two Marine Protected Areas of the Northern Mediterranean Sea

: Betanodavirus infection poses a significant threat to marine fish species in the Mediterranean, affecting both aquaculture and wild populations. Despite increasing evidence of viral circulation in farmed and wild fish, data on natural outbreaks in wild groupers remain limited. This study investigated mortality episodes in wild dusky groupers (Epinephelus marginatus) within two marine protected areas (MPAs): Portofino MPA (Liguria, Italy) and Larvotto MPA (Principality of Monaco) during 2018-2019. Pathological examinations and virological diagnostics confirmed that the causative agents were betanodavirus strains belonging to the RGNNV genotype. Phylogenetic analyses revealed high genetic similarity among viral strains detected at geographically distant sites and across host species, suggesting potential regional connectivity mediated by mobile vectors or environmental transport. Seawater temperature analysis indicated that extreme and prolonged high-water temperatures were prodromal and coincided with observed outbreaks, supporting a role for thermal stress in triggering outbreak onsets. These findings highlight the susceptibility of wild dusky grouper populations to betanodavirus and underscore the interplay between host behavior, environmental conditions, and pathogen dynamics. The study emphasizes the importance of integrated health surveillance strategies within and around MPAs to monitor fish health and environmental parameters, thereby conserving wild fish populations and biodiversity.