TECHNIC FOR PREPARING BACTERIAL VACCINES 211 



In the collection of sputum special care is required: the patient 

 should be instructed to brush the teeth with a sterilized brush, rinse the 

 mouth several times with boiled water, and after swallowing several 

 mouthfuls of water, to cough and expectorate into a wide-mouthed 

 sterilized bottle. The sputum may be plated at once, or washed sev- 

 eral times in sterile Petri dishes with sterile water and then cultured. 



Lung Puncture may occasionally be required in infective lung con- 

 ditions in which sputum is not obtainable or is too badly contaminated. 

 A 5 to 10 c.c. all-glass syringe with a strong needle is sterilized by boiling, 

 and 2 or 3 c.c. of peptone broth introduced. The skin of the chest-wall 

 over the site of infection, as shown by clinical evidence, is sterilized with 

 tincture of iodin, and puncture made into the pulmonary tissues. When 

 the desired depth has been reached, 1 c.c. of the broth is injected gently 

 into the tissues, and after the lapse of a few seconds reaspirated as far 

 as possible into the syringe. During the operation the patient should 

 refrain from respiratory movements, in order to minimize any risk of 

 lacerating the pulmonary tissues (Allen). 



Urine should always be withdrawn with a sterile catheter after 

 thoroughly cleansing the meatus. This last is especially important, for 

 the infection may be due to a certain strain of Bacillus coli, and unless 

 we are successful in obtaining a culture of this particular strain, the 

 vaccine will have little value. 



Blood specimens are taken with a sterile syringe from a prominent 

 vein at the elbow after the skin has been cleansed and sterilized with 

 tincture of iodin, and cultured in large amounts on proper culture-media. 



2. Preparing Pure Cultures. This is frequently the most difficult 

 step in the whole technic, for some microorganisms, as, for example, the 

 gonococcus and Bacillus influenzse, grow slowly, require special culture- 

 media, and their colonies may easily be overlooked. To secure pure 

 cultures, and especially to select one or at most two organisms that may 

 be the chief offenders, considerable bacteriologic knowledge is necessary, 

 and no simple rules or directions can be given in the limited space of this 

 volume. 



1. Stained smears of the secretions of a lesion may indicate the nature 

 of the infection and the best culture-medium and technic to use for pur- 

 poses of isolation. 



2. Cultures of the lesion may be made upon solid media, and isola- 

 tion carried out after a primary growth has been secured. With proper 

 care, primary cultures may be grown, such as staphylococci from the 

 pus of a freshly incised abscess, or the microorganism of a case of cystitis 



