650 APPENDIX: LABORATORY METHODS 



Date of test, including month, day, and hour 

 Operator's initials 



Date of last animal passage of the culture 

 Period of exposure of diluted culture to room temperature 

 Period of time necessary for injection of all mice 



Hours of survival of mice dying up to 96 hours, indicated by number 

 Survival of mice for 96 hours, indicated by S 



Contamination to be indicated by circle around the number of hours of sur- 

 vival 



Control serum 

 Test serum 

 Test serum 

 Test serum 

 Control serum 



Date: OcT 0,1^3 



Operator! ABC 



Culture ebetilca 

 Ulllionth. 10 millionth. 100 millionth. Billionth 



V<J +3 *1 (E> +v +3 4-1 £ 



Serum dose C"v cc 



Culture passed through mloe OfT. , " w »J^ v1i 

 Dilution of culture began II "48 A M. 

 Injection of serum 'began IV. ov P-M- 

 Injection of culture completed IV.61 PM. 



SAMPLE PROTOCOL OF MOUSE PROTECTION TESTS, NATIONAL 

 INSTITUTE OF HEALTH 



League of Nations (as taken from Parish 1051 ) 



The test is intended to ascertain the amount of serum that is equiva- 

 lent in protecting power to a certain dose of a standard serum, the se- 

 rum and culture being mixed for 5 minutes at room temperature and in- 

 jected intraperitoneally in 1.0 cc. amounts of mixture. The culture dose 

 suggested by the League of Nations' Subcommittee is 0.001 cc. In each 

 experiment, varying dilutions of serum in 0.9 per cent saline solution 

 are mixed with equal volumes of the culture dilution in broth. Decreas- 

 ing amounts of serum are used, such as 0.001, 0.00,05, and 0.00,025 cc. 



a. Animals. The mice selected for the tests should weigh 18 to 22 

 grams. After the injections of serum and culture, the dead and sick ani- 

 mals should be noted morning and evening. Smears are made from the 

 hearts' blood of the dead mice and only those animals showing the pres- 

 ence of large numbers of pneumococci are counted as fatalities for the 



