Touch or external stimuli Moribundity.SMER28 supplier Clinical PathologyFor hematology analysis on the ADVIAH 120 (Siemens, Tarrytown, NY), blood was collected into EDTA tubes according to Table 1. For clinical chemistry analysis conducted on the ADVIAH 1200 (Siemens, Tarrytown, NY), blood was collected into SST tubes according to Table 1. The hematology parameters analyzed included: Red Blood Cell Count (RBC, 106 cells/mL), Hemoglobin (HGB, g/dL), Hematocrit (HCT, ), Mean Corpuscular Volume (MCV, fL), Mean Corpuscular Hemoglobin (MCH, pg), Mean Corpuscular Hemoglobin Concentration (MCHC, g/dL), Cell Hemoglobin Concentration Mean (CHCM, g/dL), Red Cell Distribution Width (RDW, ), Platelet Count (PLT, 103 cells/mL), Mean Platelet Volume (MPV, fL), White Blood Cell Count (WBC, 103 cells/mL), Neutrophils (103 cells/mL), Lymphocytes (103 cells/mL), Neutrophils/Lymphocytes Ratio (N/L Ratio), Monocytes (103 cells/ mL), Eosinophils (103 cells/mL), Basophils (103 cells/mL) and Large Unstained Cells (LUC, 103 cells/mL). The clinical chemistry parameters analyzed included: Total Bilirubin (mg/ dL), Aspartate Aminotransferase (AST, U/L), Alanine Aminotransferase (ALT, U/L), Sorbitol Dehydrogenase (SDH, U/L), Glucose (mg/dL), Alkaline Phosphatase (ALP, U/L), Gamma Glutamyl Transferase (GGT, U/L), Total Protein (g/dL), Albumin (g/dL), Globulin (g/dL), Albumin/Globulin Ratio (A/G Ratio), Blood Urea Nitrogen (BUN, mg/dL), Creatinine (mg/dL), BUN/Creatinine Ratio, Sodium (mEq/L), Potassium (mEq/L), Chloride (mEq/L), Calcium (mg/dL), and Phosphorus (mg/dL).Animals were sedated prior to administration of an overdose of euthanasia agent containing pentobarbital. Discomfort and distress were limited to that which was unavoidable in the conductance of the study. All study procedures were approved in 
accordance to the guidelines set by the Institutional 3-Amino-1-propanesulfonic acid biological activity animal Care and Use Committee. All work involving infected animals or virus was performed in the Biosafety level (BSL)-3 laboratory.Intranasal InoculationOn the day of challenge (Study Day 0), animals were anesthetized with TelazolH (16?2 mg/kg, intramuscular) and the challenge material was instilled slowly into the nares of the ferrets using a micropipettor, alternating sides during the instillation. Influenza challenge stocks were prepared in specific pathogen free embryonated chicken eggs and tested for sterility, Mycoplasma, and hemagglutinin sequence confirmation. Challenge material was diluted in calcium- and magnesium-free phosphate buffered saline (CMF-PBS) to a target dose delivered in 0.6 mL. A portion of the diluted challenge material was analyzed by Median Tissue Culture Infectious Dose assay (TCID50) to confirm viral dose.Temperature MeasurementsAnimals were implanted with programmable temperature transponder chips for monitoring body temperature prior to challenge. Temperature transponder chips (2 per animal) were implanted in the shoulder and rump area (1 per region). Two transponders were implanted in the event that one of the transponders 
was to fail during the course of the studies. Temperature 1317923 readings were taken from both the rump and the shoulder in the morning and the evening. These temperature values were then averaged for all statistical analyses for each animal at each time point. Similarly, baseline temperatures for each ferret were calculated by averaging morning and afternoon temperatures for seven days prior to challenge.Hemagglutination Inhibition Assay (HAI)Serum samples were initially treat.Touch or external stimuli Moribundity.Clinical PathologyFor hematology analysis on the ADVIAH 120 (Siemens, Tarrytown, NY), blood was collected into EDTA tubes according to Table 1. For clinical chemistry analysis conducted on the ADVIAH 1200 (Siemens, Tarrytown, NY), blood was collected into SST tubes according to Table 1. The hematology parameters analyzed included: Red Blood Cell Count (RBC, 106 cells/mL), Hemoglobin (HGB, g/dL), Hematocrit (HCT, ), Mean Corpuscular Volume (MCV, fL), Mean Corpuscular Hemoglobin (MCH, pg), Mean Corpuscular Hemoglobin Concentration (MCHC, g/dL), Cell Hemoglobin Concentration Mean (CHCM, g/dL), Red Cell Distribution Width (RDW, ), Platelet Count (PLT, 103 cells/mL), Mean Platelet Volume (MPV, fL), White Blood Cell Count (WBC, 103 cells/mL), Neutrophils (103 cells/mL), Lymphocytes (103 cells/mL), Neutrophils/Lymphocytes Ratio (N/L Ratio), Monocytes (103 cells/ mL), Eosinophils (103 cells/mL), Basophils (103 cells/mL) and Large Unstained Cells (LUC, 103 cells/mL). The clinical chemistry parameters analyzed included: Total Bilirubin (mg/ dL), Aspartate Aminotransferase (AST, U/L), Alanine Aminotransferase (ALT, U/L), Sorbitol Dehydrogenase (SDH, U/L), Glucose (mg/dL), Alkaline Phosphatase (ALP, U/L), Gamma Glutamyl Transferase (GGT, U/L), Total Protein (g/dL), Albumin (g/dL), Globulin (g/dL), Albumin/Globulin Ratio (A/G Ratio), Blood Urea Nitrogen (BUN, mg/dL), Creatinine (mg/dL), BUN/Creatinine Ratio, Sodium (mEq/L), Potassium (mEq/L), Chloride (mEq/L), Calcium (mg/dL), and Phosphorus (mg/dL).Animals were sedated prior to administration of an overdose of euthanasia agent containing pentobarbital. Discomfort and distress were limited to that which was unavoidable in the conductance of the study. All study procedures were approved in accordance to the guidelines set by the Institutional Animal Care and Use Committee. All work involving infected animals or virus was performed in the Biosafety level (BSL)-3 laboratory.Intranasal InoculationOn the day of challenge (Study Day 0), animals were anesthetized with TelazolH (16?2 mg/kg, intramuscular) and the challenge material was instilled slowly into the nares of the ferrets using a micropipettor, alternating sides during the instillation. Influenza challenge stocks were prepared in specific pathogen free embryonated chicken eggs and tested for sterility, Mycoplasma, and hemagglutinin sequence confirmation. Challenge material was diluted in calcium- and magnesium-free phosphate buffered saline (CMF-PBS) to a target dose delivered in 0.6 mL. A portion of the diluted challenge material was analyzed by Median Tissue Culture Infectious Dose assay (TCID50) to confirm viral dose.Temperature MeasurementsAnimals were implanted with programmable temperature transponder chips for monitoring body temperature prior to challenge. Temperature transponder chips (2 per animal) were implanted in the shoulder and rump area (1 per region). Two transponders were implanted in the event that one of the transponders was to fail during the course of the studies. Temperature 1317923 readings were taken from both the rump and the shoulder in the morning and the evening. These temperature values were then averaged for all statistical analyses for each animal at each time point. Similarly, baseline temperatures for each ferret were calculated by averaging morning and afternoon temperatures for seven days prior to challenge.Hemagglutination Inhibition Assay (HAI)Serum samples were initially treat.
