MICRO-ORGANISMS BELO\<,I\<; TO THE HIGHER BACTERIA 421 



Isolation of Actinomyces. Wright 1 recommends that granules, 

 preferably obtained from closed lesions, are first thoroughly washed in 

 sterile water or bouillon and then crushed between two sterile glass 

 slides. In bovine cases make sure the granule has filamentous masses, 

 for if not no culture will grow. The crushed granule is transferred 

 to a tube of melted 1 per cent, glucose agar at 40 C. The material 

 is thoroughly distributed by shaking and the tube placed in the incu- 

 bator. A number of granules after washing should be placed on the 

 inside of a sterile test-tube and allowed to dry. In this way, should the 

 material be contaminated, the drying of the granules for several weeks 

 may kill off the other bacteria. The tube should be examined daily. 

 If a number of living filaments were added to the agar a large number 

 of colonies will develop. These will be most numerous in the depth in 

 a zone five to twelve millimetres below the surface. 



From this primary culture a colony is cut out and the bit of agar 

 washed in bouillon and then inserted in a tube of melted agar. The 

 growth in this will give material for transplants. 



Is actinomycosis due to a single micro-organism or to a group of 

 organisms having widely different characteristics? 



Wright 2 has recently made an important research study on this 

 question. His conclusions were as follows : From thirteen human cases 

 and from two in cattle the organisms seem to be all of one species, for 

 the differences among the various strains are no greater than among 

 various strains of tubercle or diphtheria bacilli. 



This micro-organism grows well only in agar and bouillon cultures 

 and in the incubator; in the other usual culture media and at room 

 temperature it grows only very little or not at all. It is essentially an 

 anaerobe. It does not form spore-like reproductive elements. In 

 cultures its colonies are similar in character to colonies of the micro- 

 organism in the lesions of actinomycosis. If colonies of the micro- 

 organism are immersed in animal fluids, such as blood serum and serous 

 pleuritic fluid, the filaments of the colonies in immediate contact with 

 the fluid may, under certain unknown conditions, become invested with 

 a layer of hyaline eosin-staining material of varying thickness, and the 

 filament may then disappear. Thus structures are produced that seem 

 to be identical with the characteristic "clubs" of actinomyces colonies 

 in the lesions. 



Inoculation experiments on animals were made with the cultures of 

 the micro-organism from thirteen cases, including the two bovine cases. 

 All of these strains were found to be capable of forming the characteristic 

 "club"-bearing colonies in the tissues of the experimental animals. 

 These colonies were either enclosed in small nodules of connective tissue 

 or were contained in suppurative foci within nodular tumors made up 

 of connective tissue in varying stages of development. With the cultures 

 from most of the cases nodular lesions identical in histological character 

 with those of actinomycosis were produced in inoculated animals and 



i Journal of Medical Research, May, 1905. 2 Loc. cit. 



