DIAGNOSIS OF GLANDERS 487 



2. Cultures. Pus, scrapings of organs and other material must be collected 

 with the necessary aseptic precautions and should invariably be sown on 

 potato. The appearance presented by the glanders bacillus on this medium 

 is characteristic (vide ante) and is an important factor in the diagnosis. 

 Several tubes of potato should be sown in order to isolate the organism 

 which may not always be present in pure culture in the material used. 



3. Inoculations. Before the discovery of mallein the inoculation of animals 

 with pus, nasal discharge and other material from suspected cases of glanders 

 was an experiment of first rate importance in the diagnosis of the disease. 

 It has however been pointed out above that enlarged glands taken from 

 animals suffering from the disease may fail to produce glanders on inoculation 

 into healthy animals, and the method is to this extent a less certain means 

 of diagnosis than the mallein test : it however affords valuable confirmatory 

 evidence. 



The suspected material should be inoculated into a guinea-pig, an ass or a dog. 



(a) Guinea-pigs. The inoculation of suspected material into the peri- 

 toneal cavity of a guinea-pig has been recommended by Strauss as at once 

 the simplest and most certain method of diagnosing a case of glanders. The 

 difficulty however is that the material used for inoculation must contain 

 no organisms capable of setting up peritonitis in the inoculated guinea-pig, 

 and in practice it is found that about one-half of the animals inoculated 

 with the discharge from the nose die of septic peritonitis in 2436 hours. 



If the material therefore contains organisms other than the glanders bacillus 

 it should be inoculated beneath the skin of a guinea-pig, and a second guinea- 

 pig should be inoculated intra-peritoneally with a portion of a lymphatic 

 gland from the first animal. (In these cases however it is often better to 

 inoculate an ass with the suspected material.) 



For purposes of inoculation rub up a little of the pus or nasal discharge 

 or other material in a mortar with a little sterile water [or normal saline 

 solution] and inject the emulsion into the peritoneal cavity of a male guinea- 

 pig. In 2-3 days the characteristic enlargement of the testicle will become 

 apparent and the animal will die in a week to a fortnight. 



For a long time the appearance of an enlargement of the testicle Straus' sign 

 following the inoculation of material from a suspected case of glanders into the 

 peritoneal cavity of a male guinea-pig was regarded as pathognomonic of glanders 

 and as absolute proof of the material having been derived from a case of the disease. 

 But Kutschen has isolated from the nasal discharge of a glandered horse an organism 

 which while differing from the glanders bacillus in- other respects, on inoculation 

 into the peritoneal cavity of a male guinea-pig produces an orchitis similar to the 

 orchitis produced by the glanders bacillus. Hallopeau and Bureau observed a 

 similar orchitis develop after inoculating pus from a case of human mycosis into the 

 peritoneal cavity of a guinea-pig. And Nocard has recorded nineteen cases of a 

 slightly contagious, farcy-like lymphangitis in horses due to a bacillus which though 

 it produced an orchitis on inoculation into guinea-pigs was absolutely different from 

 the glanders bacillus both in its cultural characteristics and in its reaction to Gram's 

 stain. The inoculation of a guinea-pig therefore can only be regarded as one factor 

 in the diagnosis of a doubtful case of glanders and must be supplemented in every 

 case by a microscopical examination of the pus in the testicle and by the mallein 

 test (Nocard). 



(b) Asses. The susceptibility of the ass to glanders renders inoculation 

 of that animal a valuable means of diagnosis : the animal should be inoculated 

 through superficial scratches on the skin. If the material used for inoculation 

 contain the specific bacillus the animal will almost invariably show the 

 characteristic symptoms of the disease before the end of the second week 

 (but see p. 480). 



