Neuropathological Characterization of Dolphin Morbillivirus Infection in Cetaceans Stranded in Italy

SIMPLE SUMMARY: There is abundant literature reporting demyelination in dogs and pinnipeds affected by morbillivirus infection, but myelinopathy is poorly investigated in stranded cetaceans affected with the virus. Also, the neuropathogenesis of cetacean morbillivirus infection has not been fully clarified, leaving questions on cell tropism unanswered. A novel dolphin morbillivirus lineage of Atlantic origin circulating in Italian waters replaced the previous Mediterranean strain in late 2015; however, differences in virulence and pathogenesis between the two strains have not yet been documented. The aims of the present study were to: describe histopathological changes and immunohistochemical findings in the central nervous system of 31 cetaceans which tested positive on molecular investigations for the two dolphin morbillivirus strains; characterize by double indirect immunofluorescence staining the areas of myelin damage. The most frequently observed morbillivirus-associated lesions were astro-microgliosis, neuronal necrosis, spongiosis, malacia, and non-suppurative meningoencephalitis. Demyelination was detected by means of a specific myelin biomarker. Inside and around the demyelinated areas there were morbillivirus antigen-bearing cells of mainly neuronal and microglial origin, associated with marked astro and microglia reactivity. Molecular and immunohistochemical analysis suggested a higher neurotropic affinity of the novel circulating strain. ABSTRACT: Cetacean morbillivirus (CeMV) is responsible for epidemic and endemic fatalities in free-ranging cetaceans. Neuro-inflammation sustained by CeMV is a leading cause of death in stranded cetaceans. A novel dolphin morbillivirus (DMV) strain of Atlantic origin circulating in Italian waters since early 2016 has caused acute/subacute lesions associated with positive immunolabelling of the virus. To date, myelin damage has not been fully documented and investigated in cetaceans. This study describes neuropathological findings in the brain tissue of 31 cetaceans found stranded along the Italian coastline and positive for DMV infection on molecular testing. Cell changes in the areas of myelinopathy were revealed by double indirect immunofluorescence. The most frequent DMV-associated lesions were astro-microgliosis, neuronal necrosis, spongiosis, malacia, and non-suppurative meningoencephalitis. Myelin reduction and areas of demyelination were revealed by means of a specific myelin biomarker. Morbilliviral antigen immunolabelling was mainly observed in neurons and microglial cells, in association with a marked activation of microglia and astrocytes. These findings extend our knowledge of DMV-associated brain lesions and shed light on their pathogenesis.