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C O N F E R E N C E 16 24 January 2018 Joint Pathology Center Veterinary Pathology Services WEDNESDAY SLIDE CONFERENCE 2017-2018 C o n f e r e n c e 16 24 January 2018 CASE I: F16-0079.3 (JPC 4101317). proliferative activity was noted in all epithelial cell layers of the diseased lung via Signalment: 5-year-old, male, Green tree a PCNA marker. Additionally, the python, Morelia viridis, reptile. immunohistochemical examination for the detection of active-caspase 3 revealed an History: The animal showed acute increased number of apoptotic epithelial cells respiratory distress and discharge of mucus in the lung of the infected animals. from the oral and nasal cavity. Two days after Microscopic Description: The two the onset of symptoms the animal was found histologic hallmarks are the epithelial dead. thickening in the entire lung and excess mucus in the lumen of lungs and airway. The Gross Pathology: A marked thickening of multilayered respiratory epithelium covering the lung parenchyma and a severe the smooth muscle at the luminal end of the accumulation of clear mucus in the upper and trabeculae displays increased cellularity with distal airways (larynx, trachea, lung, air sacs) an increase in cell layers and irregular cell were noted at gross examination. The animal arrangement (hyperplasia). In the faveolar was in moderate body condition and the space, the epithelium is characterized by carcass was slightly dehydrated. All other patchy to diffuse proliferations of cuboidal to internal organs showed no macroscopic columnar cells carrying short microvilli (type changes. II pneumocytes) and rare flat cells with short Laboratory results: The presence of nidoviral RNA (Morelia viridis Nidovirus, “MVNV”) was confirmed through RT-PCR of the affected lung tissue and intracellular nidoviral protein was detected via immunohistochemistry in respiratory and faveolar epithelial cells of the lungs. Infected epithelial cells were also found in the trachea and the nasal cavity. A markedly increased 1 Lung, green tree python. Subgross image of the lung demonstrates marked edema within the airway and faveolae. Faveolar septa are markedly expanded and the respiratory epithelium is markedly hyperplastic. (HE, 400X). cytoplasmic projections covering the and mucus accumulation in the faveolar capillaries (type I pneumocytes). The space. inflammatory component is represented by moderate, multifocal to diffuse interstitial Contributor’s Comment: The Morelia infiltration of lymphocytes, plasma cells and viridis nidovirus belongs to the subfamily of heterophilic granulocytes. Multifocal the Torovirinae (Family Coronaviridae), heterophilic granulocytes and lymphocytes which have so far mainly been associated also infiltrate the respiratory epithelium. The with enteric diseases (humans, horses, cattle, lymphoid aggregates (equivalent to the swine).4,11,13,16 However, recent studies on mammalian bronchiolar-associated tissue) toroviruses have shown that they can be both are activated. Abundant homogenous entero- and pneumotropic.14 In the last two eosinophilic fluid (mucus) is filling the years snake nidoviruses have been identified faveolar space, admixed with numerous as the causative agent of a severe respiratory heterophilic granulocytes, erythrocytes and disease in ball pythons, green tree pythons3 cell debris. and Indian pythons.2,8,9,12 Nidoviruses were also identified in the lungs of cattle and wild Contributor’s Morphologic Diagnosis: shingleback lizards with pneumonia, though Lung: Proliferative pneumonia, severe, their direct association with disease has so far diffuse with type II pneumocyte hyperplasia not been examined.9,14 2 Lung, green tree python. Histological appearance of a healthy snake lung (HE, 400X). (Photo courtesy of: Institute of Veterinary Pathology Vetsuisse-Faculty (University of Zurich), Winterthurerstrasse 268, CH-8057 Zurich, Fax number +41 44 635 89 34, www.vetpathology.uzh.ch) In the python lung, nidovirus infection is electron microscopy in type II pneumocytes. associated with a distinct proliferative Following the nidoviral infection, activity and a degree of apoptotic cell death “transformed” type II pneumocytes of both type I and type II pneumocytes, containing intracytoplasmic serous/mucous leading to an increased epithelial turnover granules are noted ultrastructurally in the and hyperplasia.3 faveolar space.3 This epithelial cell type has been previously described in various reptile In the faveolar space of the healthy snake species and is likely to explain the lung, thin cytoplasmic extensions of the type pathognomonic excess in mucus production I pneumocytes cover the capillary walls, in nidoviral pneumonia7,10 and indicate an forming the gas-blood barrier.10 This case decrease in surfactant production.3 In the nicely demonstrates a marked replacement present case viral protein could be detected in of type I pneumocytes by type II the epithelium of the entire respiratory tract pneumocytes in the faveolar space, known as (nasal cavity, larynx, trachea, lung and air an unspecific regenerative response of snakes sacs) and indicates a respiratory transmission to infectious agents, similar to response to (droplet) as the most likely infection route. lung injury in mammals.5 In the healthy Nidoviruses cause sudden outbreaks with reptilian lung, lamellated bodies containing high mortality in breeding collections, surfactant can be demonstrated through symptoms prior to deceasing/death are rare. 3 have not been formally classified to date.1 JPC Diagnosis: Lung: Pneumonia, Regardless of the phylogenetic differences, bronchointerstitial, necrotizing, diffuse, mild toroviruses all share similar tissue tropism for with marked respiratory and type II the gastrointestinal and respiratory tracts. pneumocyte hyperplasia and faveolar edema, Green tree python (Morelia viridis), reptile. Snake-associated nidoviruses were first identified in ball pythons (Python regius) and Conference Comment: Nidoviruses are a Indian rock pythons (P. molurus) in diverse group, affecting humans and a 20142,12,15 and green tree pythons (Morelia growing population of veterinary species M. viridis) in 2017.3 Gross findings in these including: snakes, wild shingleback lizards, snakes include: stomatitis, sinusitis, cattle, and nematodes. Nidoviruses that affect pharyngitis, tracheitis, esophagitis, and animal species are from the Coronaviridae proliferative pneumonia with abundant family and Torovirinae subfamily. mucus secretion. Specifically, the Torovirus genus affects mammals and the little known Bafinivirus More and more over the past several years, genus affects ray-finned fish. There have nidovirus infection has been associated with been disagreements regarding the fatal respiratory disease in several species of classification of reptile nidoviruses which python, but Koch’s postulates had not Lung, green tree python. Marked thickening of the entire epithelium (hyperplasia) with a moderate mixed-cellular interstitial inflammation and accumulation of mucus, cell debris and heterophils in the lung lumen (HE, 400X) (Photo courtesy of: Institute of Veterinary Pathology Vetsuisse-Faculty (University of Zurich), Winterthurerstrasse 268, CH-8057 Zurich, Fax number +41 44 635 89 34, www.vetpathology.uzh.ch) 4 definitively proven that it was the cause. In a recent study,6 it was demonstrated through experimental infection of three ball pythons (Python regius) that nidovirus infection results in mucinous chronic inflammation and proliferative interstitial pneumonia. Possible transmission routes were suggested by identifying infectious virus in oral secretions and fecal material. Additionally, it was suggested that choanal and oral swabs were the best sample locations for antemortem diagnosis. Lung, green tree python. Multifocal infected type I and type II pneumocytes (Immunohistochemistry for Conference participants thought there was nidoviral-antigen, 400X). (Photo courtesy of: Institute of Veterinary Pathology Vetsuisse-Faculty (University of enough necrosis within the submitted Zurich), Winterthurerstrasse 268, CH-8057 Zurich, Fax sections to include it as a modifier in the number +41 44 635 89 34, www.vetpathology.uzh.ch) morphologic diagnosis. Morphologically, the participants suggested that the material Contributing Institution: present within the faveolar spaces suggested Institute if Veterinary Pathology proteinaceous edema fluid rather than mucus. Vetsuisse-Faculty (University of Zurich) The location descriptor of Winterthurerstrasse 268, CH-8057 Zurich bronchopneumonia as used in the JPC Fax number +41 44 635 89 34 morphologic is based on the presence of www.vetpathology.uzh.ch hyperplastic and inflammatory changes in the respiratory epithelium at the tips of the References: faveolar septa as well as the type II 1. Adams MJ, Lefkowitz EJ, King AMQ, et pneumoncyte hyperplasia deep in the al. Changes to taxonomy and the faveolae. international code of virus classification and nomenclature ratified by the international committee on taxonomy of viruses. Arch. Virol. 2017;162:2505- 2538. 2. Bodewes R, Lempp C, Schurch AC, et al. Novel divergent nidovirus in a python with pneumonia. The Journal of General Virology. 2014;95(Pt 11):2480-2485. 3. Dervas E, Hepojoki J, Laimbacher A, Romero-Palomo F, Jelinek C, Keller S, Smura T, Hepojoki S, Kipar A, Hetzel U. Nidovirus-associated proliferative Lung, green tree python. Evidence of proliferative activity pneumonia in the green tree python in all epithelial cell layers of the multilayered respiratory epithelium (PCNA, 200X). (Photo courtesy
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