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DL-LACTONE CAS N°:79-50-5 - page 100 / 113





100 / 113




ID: 79-50-5 DATE: 18.01.2006

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; Jejunum; Testes (fixed in Bouin’s); Kidneys; Thymus; Larynx; Thyroid including parathyroid; Lachrymal gland, exorbital; Tongue; Liver; Trachea; Lung (infused with formalin); Urinary bladder; Lymph nodes (mandibular, mesenteric); Uterus; Nasopharynx; Vagina; Oesophagus; All gross lesions. From all adult animals: Cervix; Clitoral gland; Coagulation gland; Epididymides (fixed in Bouin’s); Ovaries; Preputial gland; Prostate gland; Seminal vesicles; Testes (fixed in Bouin’s); Uterus; Vagina; All gross lesions. Organ weights: Terminal body weight was recorded for all parental animals. The following organ weights were recorded. From 5 surviving animals/sex/group: Adrenal glands; Brain; Epididymides (total weight for both); Heart; Kidneys; Liver;

Spleen; Testes; Thymus. From all adult males: (total weight for both); Testes. Histotechnology: All organ and tissue samples


, as defined

under Histopathology (following), were processed, embedded and cut at a thickness of 2-4 µm and stained with haematoxylin and eosin. Of the selected 5 males/group of the control and high dose group, additional slides of the testes were prepared to examine staging of spermatogenesis. The testes was processed, sectioned at 3-4 µm, and stained with PAS/haematoxylin. Histopathology The following slides were examined by a pathologist: The preserved organs and tissues of the selected animals of groups 1 and 4. - The additional slides of the testes of the selected


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