10 MASS. EXPERIMENT STATION BULLETIN 337 



of successful inoculations in the sciatic nerves may be due to the fact that this 

 nerve is larger than the wing nerve and retains larger doses of the pathological 

 cells. Further details of this experiment are given in Table 6. 



Next, a group of 15 chickens, one month of age, was inoculated intravenously 

 with four doses of 0.5 cc. suspensions of trypan-blue-resistant neurolympho- 

 matous cells at intervals of two days, and four of them came down with neuro- 

 lymphomatosis in the periods shown in Table 6. 



Also, 15 chickens from the same flock as the last were inoculated intraperi- 

 toneally with the same doses of the same suspension, and two developed 

 paralysis of the coeliac plexus. The results of this inoculation are reported in 

 Table 6, with the preceding tests, so that the results of the four routes of inoc- 

 ulation may be compared. 



This work indicates that neurolymphomatosis may be transmitted to chickens 

 by inoculation of trypan-blue-resistant cells intraneurally, intravenously, and 

 intraperitoneally. The sciatic nerve was preferable to the wing nerve because 

 it is larger and the suspension can be injected into it with a greater degree of 

 certainty. The intravenous route appeared to be more certain of positive 

 results than the intraperitoneal, but the suspension must be injected slowly 

 and in small quantities in order to prevent the formation of an embolus which 

 may cause the sudden death of the chicken. Regardless of whether the chickens 

 were two or four weeks of age, they showed symptoms from seven to ten weeks 

 after inoculation, which is about the age at which they begin to come down 

 with the disease in field Outbreaks. 



The Transmission of Neurolymphomatosis Through the Egg 



Previous to the beginning of this study the writer had observed cells histo- 

 logically indistinguishable from the pathological cells in neurolymphomatosis 

 in the venous and arterial blood, as well as infiltrations into tissues adjacent 

 to neurolymphomas. Aside from nervous tissue, ovarian and testicular tissue 

 appeared to be the most susceptible to such infiltrations from the coeliac plexus, 

 although other tissues were observed to be affected at times. In some infiltrated 

 ovaries the cells penetrated into the ovules (Figure 2), and in affected testicles 

 they passed into the tubules (Figure 3) and appeared to be free in the seminal 

 fluid along with the spermatozoa. An attempt to follow the passage of these 

 cells into the egg at the time of fertilization by histological methods had to be 

 abandoned, because of the lack of trained technical assistance for such work. 



Finally the problem was approached in a different manner. In this experi- 

 ment eggs secured from 20 hens and 6 roosters showing symptoms of neuro- 

 lymphomatosis were hatched and the chickens kept under observation for evi- 

 dence of paralysis. Eefore presenting the data it should be pointed out that 

 some of the hens were not in good condition and production was low. Also 

 fertility was inferior due to the fact that the roosters were not always able to 

 breed. These roosters were used because they were the only ones available 

 and because it was impossible to determine beforehand the course of the disease. 

 These handicaps necessarily limit the scope of this experiment; nevertheless, 

 the results indicate that neurolymphomatosis is transmitted through the egg, 

 as suggested by Doyle (1928, 1929), and in the 1932 and 1933 flocks the hen 

 appeared to be entirely responsible. The results of these observations are 

 reported in Tables 7, 8, and 9. 



