90-Day Inhalation Study with Cigarettes containing G7-25 variation seen in previous studies (Coggins et of animals in the experiment. Surplus animals al., 1989a, b). thereafter will be removed from the study and Thererare currently no data available on made available for methods development the toxicology of the addition of G7-25 sheet work. to cigarettes: the present study thus is not a During the week after allocation into duplicate of any other work. groups, animals will be tail-tattooed (Animal Animal husbandry Identification & Marking Systems, A minimum of 520 (2608, 2607) Piscataway, NJ) with their permanent animals weighing 126-150 g will be ordered, identification number. The animals will be to provide the required number of animals for returned to cages with cards attached, testing. A reputable vendor (e.g. Charles recording the study number, animal number, River Raleigh, NC will be used). Animals will sex, pre-study number, and Study Director. be housed individually in transparent The following animal numbers will be polycarbonate cages and acclimated to used: sham O 101-132 OF 151-182, sentinel d' laboratory conditions for 2-3 weeks prior to 201-220 251-270, test low d' 301-332 OF the first exposure. 351-382, medium d' 401-432 451-482, high Two days after delivery, but before d' 501-532 551-482, reference low d' group allocation (see below), 5 animals per sex 601-632 651-682 medium O' 701-732 HO will be randomly chosen and killed (see 751-782 high d 801-832 of 851-832. "Euthanasia", below) for collection of sera. The animals will be housed and cared Sera will be tested for the following antibodies for in accordance with the Animal Welfare Act to disease: Reovirus Type 3, cilia associated of 1970 and amendments (Public Law 91-579), respiratory bacillus, Kilham's rat virus, as set forth in CFR Title 9, Part 3 Sub-part E, Toolan's H-1 virus, pneumonia virus of mice, Specifications for the humane handling, care, Sendai, rat coronavirus / sialodacryoadenitis treatment, and transportation of virus, lymphocytic choriomeningitis virus, warm-blooded animals other than dogs, cats, Mycoplasma pulmonis Antibody testing will rabbits, hamsters, guinea pigs and non-human again be made on sera obtained from 5 animals primates Reference will also be made to the per sex at the beginning, mid-point and end of DHHS document Guide for the Care and Use the inhalation part of the experiment, and of Laboratory Animals (NIH 86-23), and to again at reversibility (a total of 25 animals per the Society of Toxicology's Code of Ethics. sex for serology testing). The lungs from the The animals will be housed in a sentinel animals will be taken and examined vivarium and inhalation suite with controlled histopathologically, to ascertain health status. lighting (12 hours of darkness, from 6 p.m.), The start of the inhalation part of the temperature (20-24°C), and humidity (40-60% study is dependent upon negative serology relative humidity, RH). Seven-day continuous data being obtained on the pre-study samples, recordings will be kept of RH and temperature. and upon a histopathology statement on the When empty, the animal rooms will be animals killed at delivery, releasing the certified as being Class 100 (less than 100 animals from quarantine. particles per cubic meter). Any animals with questionable health Animals will be housed individually in (as assessed by a veterinarian or experienced polycarbonate cages with Alpha-Dri cellulose assistant) will be excluded from the following bedding (Shepherd Specialty Papers, procedure. One week after delivery the Kalamazoo, MI); cages will be cleaned thrice animals will be allocated by sex into 7 groups weekly and refilled with clean bedding. (32 animals per sex per group, giving a total of Animals will have unrestricted access to 224 animals per sex), such that the body certified feed (Purina Rodent Chow # 5002, weights in the groups are as homogeneous as presented as pellets) and distilled water. No possible. A record will be kept of quarantine feed or water will be available during number, weight at group allocation and inhalation exposures. Chemical analyses of permanent identification number. feed, water or bedding will not be performed, Of the animals discarded from the above because it is unlikely that contaminants would process, 20 per sex will be allocated to the adversely affect the experiment. sentinel group, thus minimizing the total usage Invasive Techniques DRAFT: DO NOT QUOTE. Page 2 Source: https://www.industrydocuments.ucsf.edu/docs/ffbb0103