Tissue tropism, pathology, and pathogenesis of West Nile virus infection in saltwater crocodile (Crocodylus porosus)

West Nile virus (WNV) is one of the leading causes of economic losses to the saltwater crocodile farming industry due to skin lesions, known as "pix", induced by the infection. Our previous study suggested a possible immunopathological pathway causing these lesions. We therefore resolved to investigate the kinetics of WNV-infection and the elicited immune responses in experimentally challenged saltwater crocodile hatchlings. Employing virus isolation, quantitation of viral genome loads in tissues by RT-qPCR and immunohistochemistry, we demonstrated that upon infection, the virus replicates in the spleen, liver and later in the pancreas. Transcriptomic analysis, based on RNA sequencing and RT-qPCR of kidney and liver tissues, revealed that the early host response is primarily via alteration of cellular structure and metabolism. As the infection progresses, the response becomes predominantly inflammatory and antiviral. The results suggest that the kidney and gastrointestinal tract are primary nidi of viral replication leading to cloacal shedding, but a link to skin lesion development remains to be fully clarified.