284 ^^^ Philippine Journal of Science wn 



contained slightly turbid yellowish brown urine. No signs of 

 blood or haemoglobin were in evidence. The general appearance 

 was that of marked anaemia. 



Tissues were taken from the heart, spleen, lungs, liver, kidney, 

 fourth stomach, and duodenum. These were fixed in sublimate 

 acetic acid and sectioned. 



Five cubic centimeters of heart's blood were immediately taken 

 from cow 3929 and injected subcutaneously into bull 3926. The 

 latter animal was kept under observation until February 29, 

 1916, and never showed any ill effects from the injection, nor 

 were anaplasmalike bodies ever found in its blood. 



Bulls 3932 and 3939 showed a few anaplasmalike bodies in 

 their blood, ranging from 2 to 3 per cent, but both contracted 

 rinderpest after their removal from the quarantine shed and died 

 of that disease. 



The blood-smear preparations and sections of tissue were 

 stained with Giemsa's, Wright's, Jenner's, and Hastings's stains. 

 The best pictures were procured with Giemsa's following the 

 technic used by Sieber. (5) 



Cover glasses have to be cleaned well in alcohol, then a thin film of blood 

 is spread on, and, with the smeared side downwards, they are thrown into 

 hot sublimate alcohol (concentrated watery sublimate lotion two parts, 

 alcohol one part). The preparations remain in this solution for two to 

 twenty-four hours. Then they are taken out with small horn forceps, well 

 rinsed, and, in order to remove the sublimate, thrown into a solution of 2 

 per cent iodide of potash (100 parts) and Lugol's solution (3 parts) ; 

 rinsed again after ten to fifteen minutes, and in order to remove the iodine 

 put into a watery solution of sodium hyposulphite (0.5 per cent) . The pre- 

 parations having become colorless by the solution of iodine (after five to ten 

 minutes) are now carefully rinsed in water and fit for other staining 

 manipulations. * * * 



For Giemsa staining of the above-mentioned smears, diluted Giemsa 

 lotion is used (J to 1 drop in 1 ccm. aq. dist.). This liquid has to be 

 changed several times during the first two hours, then the preparations are 

 left in the solution for four to twenty-four hours. Then they are well 

 rinsed and brought through the following: 



Xylol 5, acetone 95. 



Xylol 30, acetone 70. 



Xylol 70, acetone 30. 



Xylol pure. 

 According to the degree of differentiation, the preparations are left for 

 a longer or shorter time in the acetone liquids. The preparations are taken 

 out of the pure xylol and placed at once in oil of cedarwood (not Canada 

 balsam). 



This method was slightly modified by using 1 to 1,000 potas- 

 sium carbonate solution instead of the distilled water in making 

 up the stain. This causes the preparations to stain deeper. 



