Advia 120

A full CBC on blood preserved in EDTA was performed on FCGS (n = 23) and healthy (n = 8) cats (Bayer ADVIA 120; Bayer Diagnostics, Tarrytown, NY, http://healthcare.bayer.com, Veterinary Medical Teaching Hospital, School of Veterinary Medicine, UC Davis). Routine serum biochemistry was performed for all FCGS cats included in the study. Inclusion into this study was conditioned on no evidence of systemic illness and normal biochemistry panel with exception of hyperproteinemia and hyperglobulinemia (previously reported as common in cats with in FCGS) (10 (link)). All cats in this study were confirmed negative for FIV and FeLV. The cats were not tested for feline calicivirus. The mucosal tissues from FCGS cats were collected under general anesthesia from the area lateral to the palatoglossal folds using iris scissors. Full-thickness 6–8 mm biopsies were obtained from grossly affected mucosa and adjacent normal mucosa.

The total WBC count and WBC differential count provide classical clinicopathologic information including the total number and types of peripheral blood granulocytic and mononuclear leukocytes that are important in both the innate and adaptive immune responses (30 ). Serum protein electrophoresis is used to identify changes in alpha, beta, and gamma globulins that are produced in various inflammatory and infectious disease states in dolphins (31 (link)). Specifically, elevated alpha/beta globulins and elevated gamma globulins typically reflect an acute phase protein response and antigenic stimulation of humoral immunity, respectively (3 (link), 31 (link)).
Total WBCs were determined by an automated analyzer (Bayer ADVIA 120, Bayer Diagnostics) and the relative number of WBC types were determined by microscopic examination of modified Wright-Giemsa stained blood smears (18 (link)). Serum protein electrophoresis was evaluated on an automated analyzer (Rapid Electrophoresis, Helena Laboratories) as previously described (31 (link)).

At 4 h and 1, 3, 4 and 7 days after WBI, mice were humanely euthanized for whole blood, serum and tissue collection as described in previous reports [18 (link)]. The mice were deeply anesthetized prior to collecting whole blood through a cardiac blood draw and confirmatory cervical dislocation was performed while the animal was still anesthetized in accordance with the approved IACUC protocol. The blood was immediately divided into two tubes. The samples in EDTA tubes were used for peripheral blood cell counts by a clinical hematology analyzer (Bayer Advia 120, Bayer, Tarrytown, NY) at the AFRRI Veterinary Sciences Department facility, and samples in BD Microtainer Gold tubes were left unmoved on racks. Following 30 min coagulation at room temperature, sera were well separated from the gel by 10 min-centrifugation at 10,000×g, collected and stored at −80 °C for later study. After blood collection and euthanasia were completed for an individual mouse, tissues were collected. BM cells were collected from mouse femurs and humeri. After erythrocytes were lysed with lysis buffer (Qiagen GmbH, Hilden, Germany), total BM myeloid cells were collected for further use. Mouse spleens were excised, rinsed with phosphate buffered saline (PBS), and snap-frozen in liquid nitrogen, then stored at −80 °C for further use.

Hematological parameters, such as total white blood cell count, red blood cell count, packed cell volume, hemoglobin, and platelets (PLT), were determined using a fully automated hematology analyzer (Simens, Bayer ADVIA120, Germany).