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DL-LACTONE CAS N°:79-50-5 - page 85 / 113





85 / 113



ID: 79-50-5 DATE: 18.01.2006

calculated according to the Observations F0 animals Mortality/Viability: Twice distress or discomfort, whi nature or was likely to bec humane reasons. The time of precisely as possible. Clinical signs: Once daily made in all animals. Once p a week thereafter, this was cage in a standard arena du period. The time of onset, signs were recorded. Gradin according to fixed scales. these scales was as follows grade 0 = absent, grade 1 = grade 3 or 4 : grade 1 = sl

latest body weight.

daily. Animals showing pain, ch was considered not transient in ome more severe, were killed for

death was recorded as

detailed clinical observations were rior to start of treatment and once

also performed outside the home ring the pre-mating degree and duration of clinical g of the symptoms took place The definition of gradings within : Fixed scale with maximum grade 1:

present. Fixed scale with max. ight, grade 2 = moderate, grade 3 =

severe, grade 4 = very severe.

Cage debris of pregnant females was examined to detect potential abortions or premature births. Signs of difficult or prolonged parturition were recorded. Functional Observations: The following tests were performed in 5 males and 5 females, randomly selected from each group: hearing ability; pupillary reflex; static righting reflex; motor activity test (recording period: 12 hours during overnight for individual animals, using a computerised monitoring system, Pearson Technical Services, Debenham, Stowmarket, England), during the motor activity test, males were caged individually and females were caged with their offspring. The assigned males were tested during week 4 of treatment and the assigned females were tested during lactation (all before blood sampling). In order to avoid hypothermia of pups, dams were removed from the pups for not more than 30-40 minutes. Body weights: Males and females were weighed on the first day of exposure and weekly thereafter. Mated females were weighed on days 0, 7, 14 and 21 of gestation and during lactation on days 1 and 4. Food consumption: Weekly, for males and females. During the mating period analysis of food consumption was suspended. Food consumption of mated females was measured on gestation days 0, 7, 14 and 21 and during lactation on days 1 and 4. Water consumption: Subjective appraisal was maintained during the study, but no quantitative investigation introduced as no effect was suspected. Reproduction processes: Male number paired with, mating date, confirmation of pregnancy and delivery day were recorded.

Observations offspring Each litter was examined to determine the following if practically possible: - the numbers of live and dead pups at the First Litter Check (= check at day 1 of lactation) and daily thereafter (if possible, defects or cause of death were evaluated); - the individual weight of all live pups on days 1 and 4 of lactation; - sex of all pups (by assessment of the ano-genital distance); - the number of pups with physical or behavioural abnormalities, daily.

Clinical labooratory investigations F0 animals Blood samples were collected from 5 males and 5 females



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