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DL-LACTONE CAS N°:79-50-5 - page 86 / 113





86 / 113




ID: 79-50-5 DATE: 18.01.2006

randomly selected from each group under isoflurane anaesthesia immediately prior to scheduled post mortem examination, between 07:30 and 09:30 am. The animals were fasted overnight (with a maximum of 20 hours) before blood sampling, but water was provided. Blood samples were drawn from the retro-orbital sinus of all rats/sex/group and collected into tubes prepared with EDTA for haematological parameters (0.25 ml), with citrate for clotting tests (1.0 ml) and Li-heparin-treated tubes for clinical biochemistry parameters (1.0 ml). The following parameters were determined. Haematology: Erythrocytes count (RBC); Haemoglobin (HB); Haematocrit (HCT); Mean corpuscular volume (MCV); Mean corpuscular haemoglobin (MCH); Mean corpuscular haemoglobin concentration (MCHC); Platelet count; Red cell distribution width; Total leucocytes count (WBC); Differential leucocyte count; Clotting Potential; Prothrombin time (PT); Partial thromboplastin time (APTT). Clinical Biochemistry: Alanine aminotransferase (ALAT); Alkaline phosphatase (ALP); Aspartate aminotransferase (ASAT); Bilirubin, total; Chloride; Cholesterol, total; Creatinine; Glucose; Phosphorus (inorganic); Protein, total; Protein, albumin; Urea; Calcium; Potassium; Sodium.

Pathology, F0 animals Termination: All animals surviving to the end of the observation period and all moribund animals were anaesthetised using iso-flurane and subsequently exsanguinated. All animals were fasted overnight (with a maximum of 20 hours) prior to necropsy, but water was provided. Males were killed after the mating period when the minimum total dosing period of 28 days had been completed. Females with litter were killed at day 4 post partum or shortly thereafter. Females without litter were killed around the same time as the females with litter. In case a female was not pregnant, the uterus was stained using the Salewski technique in order to determine any very early post-implantation losses (=implantation site scars). Based on macroscopic findings (uterus enlarged and greenish contents), no Saleweski staining was performed on the uterus of female 63. Macroscopic examination: After sacrifice or death all parental animals were subjected to macroscopic examination of the cranial, thoracic and abdominal tissues and organs, with special attention being paid to the reproductive organs. Descriptions of all macroscopic abnormalities were recorded. Samples of the following tissues and organs were collected and fixed in neutral phosphate buffered 4% formaldehyde solution (except the epididymides and testes): From 5 surviving animals/sex/group and from all animals that died spontaneously or were killed in extremis: Identification marks; not processed Ovaries; Adrenal glands; Pancreas; Aorta; Peyer's patches (jejunum, ileum) if detectable; Brain (cerebellum, mid-brain, cortex); Pituitary gland; Caecum; Preputial gland; Cervix; Prostate gland; Clitoral gland; Rectum; Colon; Salivary glands (mandibular, sublingual); Coagulation gland; Sciatic nerve; Duodenum; Seminal vesicles; Epididymides (fixed in Bouin’s); Skeletal muscle; Eyes with optic nerve and Harderian gland; Skin; Female mammary gland area; Spinal cord (cervical, midthoracic, lumbar); Femur including joint; Spleen; Heart; Sternum with bone marrow; Ileum; Stomach;


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