Molecular Detection and Characterization of Chelonid Alphaherpesvirus 5 (Scutavirus chelonidalpha5) Associated with Fibropapillomatosis in Sea Turtles Rescued in Santa Marta, Colombia: Implications for Disease Surveillance and Marine Turtle Conservat

Fibropapillomatosis, a disease associated with Scutavirus chelonidalpha5, commonly known as Chelonid alphaherpesvirus 5 (ChHV5), manifests as benign tumors that impair the motor, visual, and physiological functions of affected sea turtles. In this study, blood and tissue samples were collected from turtles exhibiting fibropapilloma-like lesions as well as from clinically healthy individuals. A nested PCR approach was employed to amplify the viral UL30 and UL28 genes for the detection and characterization of the virus variants. The mitochondrial control region was used to assess the relationship between the turtle population and the viral variant. Among the 19 turtles analyzed, six tested positive for ChHV5, including both symptomatic and asymptomatic turtles. Phylogenetic analysis revealed that three positive samples belonged to the Western Atlantic/Caribbean clade, whereas the other three grouped within the Atlantic clade. New oligonucleotides and probes were designed for ChHV5 qPCR detection, accounting for the globally accumulated genetic variability. The qPCR test parameters demonstrated an optimized assay with an efficiency of 101.4% and a detection limit of 2.4 genome copy equivalents (GCE)/μL. This study confirms the presence of two ChHV5 viral variants in rescued turtles from the Caribbean region of Colombia, including both clinically affected and asymptomatic individuals. Therefore, these results support the association between ChHV5 and fibropapillomatosis. Furthermore, analysis of the mitochondrial control region supports the hypothesis of horizontal transmission of the virus. A novel qPCR protocol with a synthetic control is proposed to improve early diagnosis and strengthen conservation and prevention strategies.