15 



serum of immune fowls, which is not to 

 be wondered at. 



The preparation of anti-epithelioraa- 

 tous serum, as it is being carried out, 

 consists in triturating material from epi- 

 thelioma in a stone mortar, straining it 

 through muslin and filtering it through 

 paper, this being a technic similar to 

 the one we employed in preparing scrum 

 against foot-and-mouth disease and other 

 sera which are being worked upon. In 

 the case that was being studied I avoided, 

 in opposition to the one of foot-and- 

 mouth disease, intravenous injections, al- 

 though I consider them the best for su- 

 perimmunisation, because in the case of 

 fowls its difficulty renders it unpractica- 

 ble; I preferred, therefore, intraperito- 

 neal inoculations. 



To avoid virus-carriers, I resolved to 

 take only not very young fowls in big 

 lots and to carry out a first test-inocu- 

 lation on them. Although using birds 

 that by their ages should have had chan- 

 ces of infecting themselves before, I was 

 constantly obliged to set aside more or 

 less 5O0/0, because they would either ac- 

 quire a marked diphtheria, or else be- 

 cause they would manifest the chronic 

 coryza to which I have referred. 



The birds, which in the course of 

 a week show no symptoms of disease, 

 save a slight green diarrhoea, are isola- 

 ted for superimmunisation. This is done 

 according to the general technic which 

 we adopt in current practice for tlie pre- 

 paration of serum against the filterable 

 germs, with the alterations in dose and 

 mode of inoculation required by the 

 case, vir. ; 



Irst Inoculation (Test Inoculation'* 

 Hypodermic injection of 0,1 gr. 

 of the substance of the skin loca- 

 lisations (boubas) triturated and 



Irst 



Chicks one month of age inoculated 

 serum. 



dissolved in physiological saline 

 solution, filtered through muslin 

 and through filter-paper. 

 2nd inoculation; in the peritoneum» 

 of 0,5 gr, of the substance of the 

 skin localisations properly tritu- 

 rated, diluted and filtered. 

 3rd Inoculation; intra-peritoneal ino- 

 culation of 1 gr. of the material 

 referred to. 

 4th Inoculation; intra-periloneal ino- 

 culation of 2 grs. of the material 

 referred to. 

 5th Inoculation; intra-peritoneal ino- 

 culation of 4 grs. of the material 

 referred to. 

 Superimmunised fowls are killed on 

 the tenth day after the fifth inoculation. 

 The serum should be used the same day, 

 as the preservation of it by means of 

 Carbolic Acid (Phenol) produces in young 

 chicks convulsions sometimes causing 

 death and a great irritation at the site 

 of inoculation. The Assistant-Chemist of 

 the Station. BAETA VíANNA, purposes 

 to obviate this drawback by substituting 

 this chemical substance and besides by 

 concentrating the serum through reduc- 

 tion of volume. 



The chicks chosen should be very 

 young in order to avoid the ones that 

 should have become immunised by a 

 previous attack of the disease. 



Chicks of more than three months 

 of age are already more resistant to 

 infection and older ones and adults still 

 more so; the latter recover without treat- 

 ment. 



After various tentative experiments 

 I ascertained that a dose of 2,5 cm^ 

 already protects, when inoculated pre- 

 viously and, at times even when inocu- 

 lated shortly after the virus. 



The following tables are résimiés of 

 the more recent experiments: 

 Series, 

 with 2,5 cm' of antiepitheliom.itous 



