High pathogenicity avian influenza A (H5N1) clade 2.3.4.4b virus infection in a captive Tibetan black bear (Ursus thibetanus): investigations based on paraffin-embedded tissues, France, 2022.

High pathogenicity avian influenza viruses (HPAIVs) H5Nx of clade 2.3.4.4b have been circulating increasingly in both wild and domestic birds in recent years. In turn, this has led to an increase in the number of spillover events affecting mammals. In November 2022, an HPAIV H5N1 caused an outbreak in a zoological park in the south of France, resulting in the death of a Tibetan black bear (Ursus thibetanus) and several captive and wild bird species. We detected the virus in various tissues of the bear and a wild black-headed gull (Chroicocephalus ridibundus) found dead in its enclosure using histopathology, two different in situ detection techniques, and next-generation sequencing, all performed on formalin-fixed paraffin-embedded tissues. Phylogenetic analysis performed on the hemagglutinin gene segment showed that bear and gull strains shared 99.998% genetic identity, making the bird strain the closest related strain. We detected the PB2 E627K mutation in minute quantities in the gull, whereas it predominated in the bear, which suggests that this mammalian adaptation marker was selected during the bear infection. Our results provide the first molecular and histopathological characterization of an H5N1 virus infection in this bear species. IMPORTANCE: Avian influenza viruses are able to cross the species barrier between birds and mammals because of their high genetic diversity and mutation rate. Using formalin-fixed paraffin-embedded tissues, we were able to investigate a Tibetan black bear's infection by a high pathogenicity H5N1 avian influenza virus at the molecular, phylogenetic, and histological levels. Our results highlight the importance of virological surveillance programs in mammals and the importance of raising awareness among veterinarians and zookeepers of the clinical presentations associated with H5Nx virus infection in mammals.

Immunohistochemical evaluation of hepatic progenitor cells in different types of feline liver diseases.

Hepatic progenitor cells are periportally resident cells capable of differentiating into mature hepatocytes or cholangiocytes to ensure hepatic regeneration. This reaction is termed a ductular reaction. In the present study, regenerative response of the feline liver to different hepatic diseases was investigated immunohistochemically. Regeneration of the liver through hepatocellular replication and proliferation of progenitor cell compartment were comparatively evaluated. Histological and immunohistochemical stainings were conducted on feline liver samples (n=40) representing various hepatobiliary diseases. Cytokeratin (CK) 7, CK19, Proliferating cell nuclear antigen (PCNA), Ki67, and Human hepatocyte marker 1 (Hep Par-1) were used. The presence of progenitor cells within feline livers was proved, both as passive cells in normal liver and as active cells (ductular reaction) in hepatic lesions. CK7 was found to be a suitable antibody for immunohistochemically detecting feline progenitor cells. In acute events, regeneration was predominantly shaped by the division of hepatocytes. In chronic events and severe acute events, hepatocytes lost their ability to divide and regeneration mainly occurred through progenitor cells. Location of the ductular reaction varied between different hepatic diseases. Parenchymal ductular reaction was detected in fulminant hepatitis, chronic hepatitis, hepatocellular lipidosis and metastatic lymphoma, whereas septal ductular reaction was detected in chronic hepatitis and metastatic lymphoma. Ductular reaction exhibited positive staining for Hep Par-1 in chronic and severe acute events. This study indicates the major role played by hepatic progenitor cells in regeneration of the feline liver. Moreover, it shows how the activation pattern of ductular reaction varies according to the hepatobiliary disease type.