2. MATERIAL AND METHODS
An adult female hairy dwarf porcupine (Sphiggurus spinosus , F. Cuvier, 1823) was admitted to a wildlife rescue center with a history of trauma. It was found in a periurban area of the metropolitan region of São Paulo, between the Atlantic rainforest and densely urbanized area (Figure 1). The animal was kept under medical care for almost five months, and, during the rehabilitation, the animal presented sporadic episodes of bilateral hind limb paresis (Supplementary material 1). Days before death, a purulent discharge in the right nostril was observed. Necropsy was performed, and tissue fragments (brain, heart, lungs, liver, spleen, kidney, adrenal glands, small intestine, uterus, and urinary bladder) were collected and fixed in 10% neutral buffered formalin, and fresh brain and liver fragments were frozen at −20°C. The samples were referred to Adolfo Lutz Institute within the framework of the wildlife epidemiological and laboratorial surveillance project: potential for the emergence and re-emergence of infectious diseases (SES-PRC-2020/32339 - PROC. 28/2020) in order to detect potential zoonotic diseases in wildlife.
For histopathology, the tissues were processed routinely, embedded in paraffin-wax, sectioned at 4 μm-thick and stained with hematoxylin and eosin (H&E) and Periodic Acid of Schiff (PAS), for routine microscopic analysis. Immunohistochemical and molecular methods are fully described in the Technical appendix 1.
The address where the porcupine was found was recorded. We geocoded the address using Geographic Information System ArcGIS 10.2.2. and its online imagery basemaps only for location purposes.