662 APPENDIX: LABORATORY METHODS 



VIII. Sterility Tests on Antipneumococcic Serum 

 Bulk Sterility Tests (National Institute of Health Regulations) 



For sterility tests, from each bulk container holding more than 1 liter 

 at least 10 cc. should be planted into broth fermentation tubes; for bulk 

 containers of less than 1 liter capacity at least 3 cc. should be planted 

 into enough fermentation tubes so that the preservative shall not exceed 

 0.01 per cent of the broth used. If the bulk containers are opened at 

 any subsequent manipulation, two 5 drop and two 20 drop fermenta- 

 tion tubes should be planted from each container. If contaminations ap- 

 pear in any of the bulk tests, the test may be repeated, but no lot shall 

 be passed until such tests show no growth, and if the same contaminat- 

 ing organism appears in more than one test, the material shall be dis- 

 carded or resterilized. 



Inoculated tubes are incubated at 37° for seven days and should be 

 examined on the second, fourth, and seventh days after planting, and 

 agitated only after 48 hours incubation to ensure initial anaerobiosis ; 

 the lowest part of the bend should be examined for slight growths, as 

 well as both arms. In case the material is turbid, transplants to fresh 

 fermentation tubes, as well as smears, should be made on the seventh 

 day. 



Sterility Tests on Final Containers (National Institute of Health 

 Regulations) 



Of the final containers selected at random from each lot, three to ten 

 shall be tested in accordance with the following schedule : 



At least one 5 drop and one 20 drop fermentation tube shall be 

 planted from each container. In the case of products to be administered 

 in doses larger than 1 cc, the entire dose up to 5 cc. from each con- 

 tainer tested shall be planted in fermentation tubes containing enough 



