MIL-PRF-12070E
APPENDIX
(Application for copies should be addressed to the Central Data Management National
Toxicological Program, P.O Box 12233, Research Triangle Park, NC 27709-2233.)
3. Procedure. Specific details concerning the design of the mouse skin painting cancer
bioassays are as follows.
3.1 Test mice. Male Swiss or C3H mice (other strains routinely used in mouse skin cancer
bioassays are acceptable) will be used. Mice should be observed for 14 days before the studies
begin and should be 6 to 8 weeks old when the first dose is applied.
3.2 Control groups. A negative untreated control group and a positive control group (the
positive control will be treated with either a known carcinogenic mineral oil or with benzo (a)
pyrene) will be included. The dose group and controls will contain 50 mice each.
3.3 Dosing. Undiluted test oil and positive controls will be applied directly to the
interscapular skin in either 37.5 µL aliquots twice weekly or 25 µL aliquots 3 times weekly to
yield a total weekly dose of 75 µL. Fur in the area of the skin receiving the dose application will
be clipped once every week. Animals are to be sacrificed after two years of treatment unless they
become moribund or a squamous cell carcinoma is grossly diagnosed before that time.
3.4 Examination. Mice will be housed individually throughout the studies and will receive
food and water ad libitum. Animals will be examined twice weekly for the appearance of dermal
tumors. Mice will be examined twice daily for mortality, changes in appearance or behavior, and
signs of toxicologic or pharmacologic effects.
3.5 Clinical findings. Clinical findings will be recorded weekly for the first 13 weeks, then at
4-week intervals thereafter until the end of the study. Body weights will be recorded weekly for
the first 12 weeks, then every 4 weeks until the end of the study.
3.6 Necropsy. Complete necropsies will be performed on all mice. During necropsy all
organs and tissues will be examined for gross lesions. Histopathologic examination will be
performed on skin from all mice and on any gross lesions observed during necropsy.
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