Nidovirus-Associated Proliferative
Pneumonia in the Green Tree Python
(Morelia viridis)
Eva Dervas,a Jussi Hepojoki,a,b Andrea Laimbacher,c Fernando Romero-Palomo,a
Christine Jelinek,a Saskia Keller,a Teemu Smura,b Satu Hepojoki,a Anja Kipar,a
Udo Hetzela
Institute of Veterinary Pathology, Vetsuisse Faculty, University of Zurich, Zurich, Switzerlanda
; University of Helsinki, Medicum, Department of Virology, Helsinki, Finlandb; Institute of Virology, Vetsuisse Faculty,
University of Zurich, Zurich, Switzerlandc
ABSTRACT
In 2014 we observed a noticeable increase in the number of sudden
deaths among green tree pythons (Morelia viridis). Pathological examination revealed
the accumulation of mucoid material within the airways and lungs in association
with enlargement of the entire lung. We performed a full necropsy and
histological examination on 12 affected green tree pythons from 7 different
breeders to characterize the pathogenesis of this mucinous pneumonia. By histology
we could show a marked hyperplasia of the airway epithelium and of
faveolar type II pneumocytes. Since routine microbiological tests failed to identify
a causative agent, we studied lung tissue samples from a few diseased
snakes by next-generation sequencing (NGS). From the NGS data we could assemble
a piece of RNA genome whose sequence was 85% identical to that of
nidoviruses previously identified in ball pythons and Indian pythons. We then
employed reverse transcription-PCR to demonstrate the presence of the novel nidovirus
in all diseased snakes. To attempt virus isolation, we established primary
cultures of Morelia viridis liver and brain cells, which we inoculated with homogenates
of lung tissue from infected individuals. Ultrastructural examination of
concentrated cell culture supernatants showed the presence of nidovirus particles,
and subsequent NGS analysis yielded the full genome of the novel virus
Morelia viridis nidovirus (MVNV). We then generated an antibody against MVNV
nucleoprotein, which we used alongside RNA in situ hybridization to demonstrate
viral antigen and RNA in the affected lungs. This suggests that in natural
infection MVNV damages the respiratory tract epithelium, which then results in
epithelial hyperplasia, most likely as an exaggerated regenerative attempt in association
with increased epithelial turnover.