TETANUS 47 



Examination of Pus from Suspected Cases of Tetanus. 



Requisites. — i. Slides and cover-glasses. 



2. A stiff platinum loop. 



3. Bunsen burner or spirit-lamp. 



4. Loffler's blue or carbol thionin. 



5. Materials for Gram's staining. 



6. Balsam. 



If cultures are to be taken, add a pipette (see Fig. 146), a deep 

 tube of agar to which 2 per cent, of grape-sugar has been added 

 previous to sterilization, a flask of water, and a thermometer. 



Method. — Scrape the deeper portions of the wound with the 

 platinum loop, and spread out the secretion thus obtained on tin 

 surface of a slide. Prepare several of these slides, and fix the 

 film by heat. Stain some by the simple stain for two minutes 

 and others by Gram's method. 



The bacillus of tetanus is about as long as the tubercle bacillus 

 and is very slender. It stains by Gram's method. A very 

 characteristic feature is its method of spore-formation. The 

 spores are spherical bodies which are formed at the extremities oi 

 the bacilli, giving them the appearance of pins or drumsticks 

 The spores do not stain by the ordinary stains, and appear a; 

 colourless and highly refractile bodies (Plate II., Fig. i). 



The cultures are made in agar to which 2 per cent, of grape 

 sugar is added, and the needle or pipette used in making the 

 inoculation is plunged deep down into the medium. The bacillui 

 of tetanus is an anaerobe — i.e., it grows only in the absence o 

 oxygen. The stabs are made deep in order to inoculate th( 

 material far away from the air, and the glucose is added to absorl 

 any oxygen which may be in the medium. To increase ou: 

 chances of obtaining this bacillus in pure culture, the materia 

 to be examined is to be heated to a temperature which will kill al 

 developed bacteria, but which will not be injurious to spores ; thi 

 tetanus bacillus is the only anaerobic organism with a spherica 

 terminal spore which is at all likely to occur in a wound. 



Method. — The inoculations are to be made with a pipette. I 

 the pus which comes from the wound can be drawn up into thi 

 capillary tube of a glass pipette such as is described on p. 35 

 the material should be collected in this way. If this is not th 

 case, the wound must be scraped with a sterilized platinur 

 needle or other suitable instrument, and the material thus obtaine 



