To avoid bias, control and dogs in the 1× groups were handled similarly to the other two groups but were not dosed during the administration of the second fraction.
Food was offered to the dogs prior to and after each treatment. Dogs were observed at least hourly for 3 h after the first dose fraction was administered and hourly for 4 h after the second dose fraction was administered. Feces from all dogs were examined for the presence of whole undigested chews on the day after treatment. Personnel involved with recording of the in-life observations were blinded as to treatment. The pathologist performing the necropsy was unaware of dog’s group but the origin of the dogs was unblinded for the histologic evaluation. Physical examinations were performed weekly during the pre-test period and biweekly to Day 125, and included the evaluation of the general appearance, body weight, respiration rate, heart rate, selleck products and body temperature. Daily feed intake was monitored and recorded for analysis beginning on Day −1. In addition, blood hematology, plasma chemistry, and coagulation profiles were determined
twice during pre-test in conjunction with physical examinations on Days 14, 27, 42, 55, 70, 83, 97, 111 and 125. Standard laboratory techniques were used for collection and analysis of the samples. The Merial laboratory conducting the analysis provided reference ranges for the plasma chemistry, coagulation, and hematology profiles. The hematology profile included: red blood cell count (RBC), white blood cell count, white blood cell differentials (absolute count), platelet count, hemoglobin,
hematocrit, Selleckchem Epigenetic inhibitor Bay 11-7085 mean corpuscular volume, mean corpuscular hemoglobin, mean cell hemoglobin concentration, and RBC morphology. The plasma chemistry profile included: alanine aminotransferase, albumin, alkaline phosphatase, amylase, aspartate aminotransferase, calcium, chloride, cholesterol, creatinine, globulin, glucose, phosphorus, potassium, sodium, total bilirubin, total protein, triglycerides, and urea nitrogen. The coagulation profile included activated partial thromboplastin time, prothrombin time, and thrombin clotting time. Urine samples were obtained once during pretest and on Days 27 and 126 using either a metabolism pan or by cystocentesis (Day 126). Urinalysis included determination of urobilinogen, nitrite, glucose, bilirubin, ketones, blood, leukocytes, specific gravity, pH, and protein by use of MULTISTIX® SG Reagent Strips (Bayer Corporation). In addition to the reagent strips, a refractometer was used to determine urine specific gravity. Urine sediment was evaluated microscopically for at least the following: crystals, casts, red blood cells, white blood cells, and epithelial cells. Standard laboratory techniques were used. Reference ranges for the specific gravity was from Stockham and Scott (2002). Dogs were humanely euthanized on Day 126 and a complete post-mortem examination was conducted.
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