Bc 2800vet

After 8 months, blood samples were collected from the tail vein under isoflurane anesthesia (n = 12/per group). For routine blood analysis, coccygeal vein blood (1 ml) was collected with a Mindray automatic hematology analyzer (BC-2800vet, Shenzhen, China).

Peripheral blood and plasma samples of experimental mice were collected following standard protocols, and the complete blood count was conducted with Mindray veterinary automatic blood cell analyzer BC-2800Vet.

Blood samples collected in test tubes with K₃EDTA were utilized for estimation of various hematological parameters by hematology auto analyzer (Mindray BC-2800 Vet, China). On the day of blood collection, red blood cells, hemoglobin (Hb), packed cell volume mean corpuscular volume (MCV), mean corpuscular Hb (MCH) and MCH concentration (MCHC), total leucocytes counts white blood cells and platelets were estimated. Serum biochemical parameters were estimated in clinical serum biochemistry analyzer (NOVA 2021 Biochemistry Analyzer, Analytical Technologies Limited, Gujarat, India) including creatinine, uric acid and blood urea nitrogen (BUN). Serum XO activity was detected by using Coupled enzyme assay kit, purchased from Sigma-Aldrich (Spruce Street, St. Louis, MO, USA) by following the manufacturer’s protocol. XO activity was determined by spectrophotometric multiwell plate reader (Infinite M200, NanoQuant, TECAN) having 570 nm wavelength. XO activity was reported as milliunit/mL, where one milliunit of XO is defined as the amount of enzyme that catalyzes the oxidation of xanthine, yielding 1.0 µmol of uric acid and hydrogen peroxide per minute at 25°C.

After waiting three days after receipt of the animals, each guinea pig was rendered neutropenic by intraperitoneal injection (IP) of 100 mg/kg of cyclophosphamide (TCI (Shanghai) Development Co., Ltd, Shanghai, China) at a frequency of one injection per day for three days. Blood was drawn from the heart and leukocytes were counted with an automatic blood cell analyzer (Mindray BC-2800Vet, Shenzhen, China). An H. parasuis standard strain of serotype 13 (13R) was used to infect the neutropenic guinea pigs via intraperitoneal injection, specifically, a single inoculum of a 0.2 mL aliquot solution containing approximately 10⁹ CFU of the H. parasuis strain, which was a 95% infective dose (ID₉₅) that was previously determined in pilot studies. Ten guinea pigs were used to detect blood and lung tissue bacterial loads in a 72 h-period, and 256 guinea pigs were used for PKs of tulathromycin in a 168 h-period. To obtain antimicrobial PK data, infected neutropenic guinea pigs were given single IM doses at two levels, 1 or 10 mg/kg, via tulathromycin Injection (Draxxin®) (Zoetis, New York, USA).

Fifty microliters of peripheral blood were collected from the retro-orbital complex of 10 mice per group derived from longitudinal assay➂ and transferred to tubes containing 10% EDTA (Sigma Chemical Co) as an anticoagulant. An Auto Hematology Analyzer apparatus (Mindray BC-2800Vet, Hamburg, Germany) was used to evaluate parameters related to white blood cells (total leukocytes, lymphocytes, monocytes and polymorphonuclear cells) and platelets. Data was calculated as the fold change relative to the control group and then displayed as a heatmap.

For blood glucose test, mice were fasted at 8 a.m. for 6 hours before measurements as suggested by Andrikopoulos 57 (link). For blood glucose tolerance test, mice were injected intraperitoneally with 2 mg/kg glucose and blood glucose were measured at 0 min, 15 min, 30 min, 60 min and 120 min after glucose administration. All experiments concerning blood glucose were performed via tail vein using glucometer ACCU-CHEK (Roche, Germany). For other bloods tests, blood samples were extracted from eyeballs of mice after anesthesia. 50-100 µl whole blood was stored in anticoagulant tube attached with EDTA for whole blood cell test, the rest of the blood was placed at room temperature for 20 min and then centrifugated at 3,000 ×g at 4 °C. After centrifugation, supernatants were harvested for further analyses. Indicators for serum testing include lipids contents such as total triglyceride (TG), total cholesterol (TC), liver function markers including alanine aminotransferase (ALT), aspartate aminotransferase (AST), cardiac function markers including creatine kinase (CK), L-lactic dehydrogenase (LDH-L) and creatine kinase- myocardial isoenzymes (CK-MB). Blood biochemical assays were done by corresponding assay kits (HUILI, China) and blood cell analyses were performed on automated hematology analyzer BC-2800vet (Mindray, China).