Molecular Characterization of Herpesviral Encephalitis in Cetaceans: Correlation with Histopathological and Immunohistochemical Findings

SIMPLE SUMMARY: In this study we describe the molecular and pathological characteristics of alpha- and gamma-herpesvirus infection of the central nervous system of stranded cetaceans and correlate them with viral load, immunohistochemical findings and biological data such as age, sex, and the presence of co-infections. The viruses (alpha- and gamma-herpesvirus) were detected in twelve out of 103 analysed stranded cetaceans and were associated with a wide range of histopathological lesions, as previously described for these and other species. In five out the twelve animals, lesions were severe enough (malacia, neuronal necrosis and neuronophagia) to cause death. Intranuclear inclusions bodies were present in brain tissue samples from half of the HV-positive animals, indicating that the injury was due to an infective agent belonging to a group of filterable viruses. These results are in accordance with immunohistochemical findings, as all the brain tissue samples with INIBs were immunolabeled with Anti-HSV1. Males, juveniles, and calves were predominantly infected among the analysed cetaceans and a 41.6% (5/12) incidence of co-infections in the brain was detected, with three animals co-infected with Dolphin Morbillivirus (DMV). In this study, we present, to the best of our knowledge, the first histopathological evidence of superinfection between HV and DMV pathogens in brain tissue. ABSTRACT: Herpesviruses are causative agents of meningitis and encephalitis in cetaceans, which are among the main leading known natural causes of death in these species. Brain samples from 103 stranded cetaceans were retrospectively screened for the presence of herpesvirus DNA in the brain. Molecular detection of Cetacean Morbillivirus was performed in HV positive brain cases. Histopathologic evaluation of brain samples included the presence or absence of the following findings (n = 7): meningitis, perivascular cuffings, microgliosis, intranuclear inclusion bodies, malacia, neuronal necrosis and neurophagic nodules, and haemorrhages. Histological evidence of the involvement of other etiological agents led to complementary analysis. We detected the presence of alpha and gamma-HVs in 12 out of 103 (11.6%) brain samples from stranded cetaceans of five different species: one bottlenose dolphin, six striped dolphins, three Atlantic spotted dolphins, one Cuvier’s beaked whale, and one common dolphin. Pathogenic factors such as viral strain, age, sex, and the presence of co-infections were analysed and correlated with the brain histopathological findings in each case. Herpesvirus was more prevalent in males, juveniles, and calves and a 41.6% incidence of co-infections in the brain was detected in our study: three with Dolphin Morbillivirus, one with Staphilococcus aureus septicaemia and one with Brucella spp.