238 MICROBIOLOGY 



make animal inoculations with the suspected material. For 

 this purpose guinea pigs are to be used. After death, the 

 organism can be obtained in pure culture from the liver or 

 spleen. If the suspected material is from the nasal ulcers or 

 skin abscesses the inoculation should be made subcutaneously, 

 to reduce the danger of death from sepsis. If the material is 

 uncontaminated, more rapid results will follow an intraperi- 

 toneal inoculation. 



Strauss pointed out that male guinea pigs should be em- 

 ployed, as this organism within a few days produces orchitis, 

 when the guinea pig may be chloroformed and cultures made 

 directly from the inflamed testicles. This is known as the 

 Strauss method. 



Bacteriological examination. (1) Cultures. By inocu- 

 lating suitable media with the discharge from the nasal sep- 

 tum ulcers, contents of the skin abscesses or from the lesions 

 found on post mortem in the lungs or other organs. The mi- 

 croscopic examination is helpful but can not be considered 

 entirely positive in making the diagnosis, as Bacterium mallei 

 does not possess differential morphological or staining proper- 

 ties. (2) Animal inoculation. By inoculating guinea pigs 

 with the suspected material (Strauss method). In addition to 

 these glanders is diagnosed in the living animal by several 

 methods of which mallein w r as the first. 



Preparation of mallein. Mallein is prepared by culti- 

 vating virulent Bact. mallei in acid glycerin bouillon, in flasks 

 containing 250 cc. The culture is allowed to grow in an incu- 

 bator at a temperature of 35 to 37 C. for from four to six 

 weeks. At the end of this time, the cultures are sterilized at 

 100 C. for 30 minutes by boiling in a water bath or by steam- 

 ing. They are then passed through a Berkefeld filter to remove 

 the organisms. The filtrate is then evaporated one fifth (the 

 original method was to evaporate the filtrate to one tenth its 

 original volume and to give 0.25 cc. diluted to 2 cc. in a 0.5% 

 carbolic acid solution as a dose) and 0.5% carbolic acid is 

 added to preserve it. The dose is 2 cc. for the average size 

 horse. 



