PATHOGENICITY OF THE OONOCOCCUS 549 



accompanied by hypopyon ; in the pus cells gonococci could not be detected microscopic- 

 ally, but could be cultured for the first two days. 



Nicolaysen (1897) injected an aqueous suspension of gonococci into the knee joint of 

 a rabbit. Arthritis followed with abundant purulent exudate, which persisted for a week • 

 no organisms could be demonstrated in it, however. The same result occurred when a 

 culture killed by heat at 70° C. for 1 hour was injected. 



From these results it is seen that the gonococcus does not live for long in the 

 animal body ; cultures are rarely positive after 2 days, and then only when the 

 injections are made directly into serous cavities or the blood stream. In fact 

 it is doubtful whether true proliferation occurs at all. De Christmas (1897), for 

 instance, found that if the cocci were introduced, enclosed in collodion sacs, into 

 the peritoneal cavity of laboratory animals, they failed to grow. The reaction 

 following the injection of gonococci is due therefore, not to a true infection, but 

 to a toxic action of the organisms. That this is true is abundantly clear from the 

 numerous experiments on gonotoxin. 



Toxin Production. — De Christmas (1897, 1900) recorded experiments from which he 

 concluded that the gonococcus forms a true exotoxin. His results have not, however, been 

 confirmed by other workers. Wassermann (1898), for instance, found that heat-killed 

 cultures were as fatal to mice, injected intraperitoneally, as living cultures ; on the other 

 hand, filtered cultures proved innocuous, unless a filtrate of a 2 to 3 weeks' culture was used ; 

 even then, the filtrate was never as toxic as the whole culture. He grew the gonococci in 

 33 per cent, ascitic broth in a thin layer of fluid in flat-bottomed flasks. After a week, the 

 cocci were collected, dried, and ground in an agate mortar ; weighed quantities of the 

 powder were suspended in water, and sterilized by steaming or in the autoclave. Injected 

 intraperitoneally into mice this powder was fatal in a dose of 0-01 gm., death occurring in 

 24 hours or later. Injected into rabbits or guinea-pigs subcutaneously, it caused wide- 

 spread doughy infiltration, often passing on to necrosis. Injected into the anterior chamber 

 of the eye of rabbits, it caused corneal turbidity, hypopyon, and sometimes complete 

 destruction of the eye. He was unable to extract the toxin by heat, by distilled water, 

 or by N/10 NaOH ; the cocci still remained toxic. The toxin was not destroyed in the 

 bacterial bodies by drying or by heat at 120° C. ; it was not destroyed by absolute alcohol, 

 or by prolonged boiling ; nor was he able to immunize rabbits or mice against it. These 

 results point strongly to the conclusion that the toxin is an endotoxin — a body contained 

 in the cell substance, and adhering strongly to it. 



SchafFer (1897) found that filtered ascitic broth cultures, 2 to 6 days old, were without 

 effect when injected in large doses — 3-10 ml. — into guinea-pigs and rabbits. Maslovski 

 (1900) likewise found that the filtrate of a 9-days' serum broth culture, injected subcutane- 

 ously into rabbits, gave rise to nothing but a slight rise of temperature and loss of weight. 

 He concluded too that the toxic substance was an endotoxin. Scholtz ( 1900) came to the 

 same conclusion. On the other hand, Vannod ( 1907) working with filtrates of 17-20-days' 

 cultures in de Christmas's medium — concentrated veal broth containing 75 per cent, of 

 ascitic fluid — found that they were fatal to rabbits injected intraperitoneally. Post mor- 

 tem, there was a purulent effusion into the peritoneal cavity ; the serosa was congested and 

 covered with purulent deposits ; the suprarenals were enlarged and hj^persemic. Vannod, 

 however, used large and repeated doses — a total of 18 to 24 ml. in 10 days to 3 weeks. 



We may conclude that the gonococcus contains a toxin which can be extracted 

 by grinding the dried organisms and suspending the resultant powder in water. 

 In cultures in fluid media the toxin may be liberated from the cocci by autolysis ; 

 after 2 or 3 weeks' incubation, the organisms may have autolysed to such an extent 

 that a certain amount of toxin may be present in filtrates of the cultures. The 

 available evidence leaves little doubt that the toxin belongs to the class of so-called 

 endotoxins. 



