Ball Python Nidovirus: a Candidate Etiologic Agent for Severe Respiratory Disease in Python regius

Stenglein, Mark D.; Jacobson, Elliott R.; Wozniak, Edward J.; Wellehan, James F. X.; Kincaid, Anne; Gordon, Marcus; Porter, Brian F.; Baumgartner, Wes; Stahl, Scott; Kelley, Karen; Towner, Jonathan S.; DeRisi, Joseph L.

(ii) Collection 3. Three ball pythons (5, 6, and 7) from a private collection in Wisconsin were submitted to a veterinarian due to signs of respiratory disease. Ball python 5 was found dead and submitted for necropsy. The following tissues were collected, fixed in NBF, and processed for light microscopy: lungs, kidney, liver, stomach, spleen, and ovary with oviduct. Ball pythons 6 and 7 had acute onset of respiratory signs over a 2to 3-day period characterized by an increase in respiratory rate and effort. There also was elevation of the cranial third of the snake's body (from the ground), ultimately resulting in opisthotonos (stargazing). A decision was made to euthanatize and necropsy BP6 and -7. The following tissues were collected from each snake, fixed in NBF, and subsequently embedded in paraffin: trachea, lungs, heart, esophagus, stomach, intestines, liver, pancreas, kidneys, oviduct, brain, spinal cord, nasal sinuses, vomeronasal organ, and skin. The paraffin-embedded tissues were sectioned at 6 m, processed routinely for light microscopy, and stained with hematoxylin and eosin. Sections of the vomeronasal organ of one case were stained with Ziehl-Neelsen acid-fast stain, which was negative for acid-fast bacteria; methenamine silver staining was negative for yeast and hyphae. A portion of lung from BP6 was fixed in 4% paraformaldehyde-2.5% glutaraldehyde in 0.1 M sodium cacodylate buffer, pH 7.24, and processed for TEM (see below).