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Reference :--
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B. Serum taken from the blood drawn from Horse D that had been treated by a course of methylene blue followed by a course of perchloride of mercury was injected into the following animals:-
(a) Horse I
Calf M
received 50 c.c. serum from Horse D.
"
30 c.c.
"}
"
+
"
10 c.c.
D. D.
}
"
(d) Monkey B
10 c.c.
D.
11
"
(c) Dog S
Details of Cases under A which were Injected with Blood.
(a) Horse H (see Chart 30) was from 13th May to 17th May injected subcu taneously with blood every day from Horse C., the total volume of blood injected being 81 c.c.
Seven days after the last injection a slight rise of temperature occurred; try- panosomes appeared in the blood; by seven days later the horse had grown very weak, steadily grew worse, and seven days later died, on the 7th June.
On Post Mortem.--Horse much reduced in condition. Mucous membranes very pale. Two filariæ equinæ (as found in Horse B) in the peritoneal space in great colon; the gastrodiscus sonsinœ, or collinsi, very numerous, oxyuris in the rectum. Liver not enlarged, but uniformly studded both on the surface and within its sub- stance with calcareous nodules varying in size from a pin head to a millet seed. Owing to the hard calcareous structure of these nodules it was impossible to make microscopical sections.
Remarks. In this case, although the temperature did not show any great rise (as will be noted from the chart), and the blood showed very few trypanosomes, yet the animal succumbed very rapidly. This shows that the blood of Horse F was not free from trypanosomes; (2) that so large a volume of blood as 81 c.c. must have contained a great deal of toxic material which probably was the main cause of a rapid fatal termination combined with the fact that the general health of the horse could not have been very good, owing to the enormous number of entozoa in the alimentary canal.
(b) Calf K (see Charts 31a to 31g) was, from the 13th to 18th May, injected subcutaneously with blood from Horse F. The total volume of blood injected was 88 c.c.
The day after the first injection of 16 c.c. of blood there was a temperature reaction (probably toxic in nature), but no trypanosomes appeared in the blood; however, that day no injection of blood was made, owing to the temperature of 106° F. being reached. On the 20th and 21st May, the 2nd and 3rd day after the last injection of blood, there was a rise of temperature, but no trypanosomes were found in the blood. A few days later, although the temperature was down, trypano- somes were found in the blood for a few days, and then disappeared. Trypanosomes were never found in the blood again, and the temperature subsequently ran a normal course up to the 31st October, when temperature observations ceased. At first the calf fell off in condition, but subsequently improved, and grew very strong. Early in November, 1908, when left in charge of Dr. Alexander for general observation, he was in very good health and appeared to have recovered. News, dated January, 1909, from Dr. Alexander, said that the calf was quite well and strong.
Remarks. From this case it is evident that the blood of Horse F was not free from trypanosomes, for the disease was reproduced in the calf by inoculation, but its virulence had evidently been lowered, as shown by the following points :-
(1) A mild attack of the disease resulted.
(2) A slight rise of temperature occurred, but subsequently a normal
course ran.
(3) Trypanosomes appeared in the blood temporarily for one period of a few
days, and never again.
(4) The calf fell off in condition at first but improved again and became
strong.
(5) The animal was alive eight months after inoculation and the outlook
for the future favourable.
If this calf now survives he will be used for further experiment. He will be subjected to natural infection by being taken through a fly-belt or else he will be inoculated with virulent blood from an animal (horse or ox) that has acquired the disease naturally.
28 c.c.
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o) Bog Q (see Charts 32a to 32e) was injected with blood subcutaneously, from 18th to 20th April, from Horse F, the total amount of blood injected being From the chart it will be seen that a few days later the dog had a rise of temperature for a few days but no trypanosomes in his blood, and subsequently, although there were rises of temperature, no trypanosomes were ever found. Fle finally wasted away and died on the 12th August.
Remarks. It is evident that in this case, as in the other cases described under Section. II., that the virulence of the strain of the trypanosomes in Horse F had been lowered towards the dog, but that the blood contained some toxic material which proved fatal eventually without reproducing trypanosomes in the dog's blood.
(d) Dog R (see Charts 33a to 33d) was, on the 10th May, injected with 8 c.c. of blood from Horse C. The chart will show that during the third week the temperature rose but the blood failed to show any trypanosomes on examination, but at this time the dog fell off in condition very rapidly. Later the dog had a good deal of fever but no trypanosomes in his blood. He developed a corneal opacity and iritis. By August he became a skeleton, and on the 15th August he died.
The remarks made on the foregoing case of Dog Q apply to this case also.
(e) Monkey A (see Charts 34a to 34e) was, on the 10th and 11th May, inocu- lated subcutaneously with 16 c.c. of blood in all from Horse C. He had rises of temperature subsequently, but never showed any trypanosomes in his blood, and never appeared ill in the least. Later the temperature ran normal, and on the 30th September the temperature observations ceased. Now, the monkey was in good health. Early in November I took the monkey with me to Yola with the object of subjecting him to natural infection in the fly-belt, at the River Loko, but this failed, as no tsetse flies existed here at this time, so I handed over the .monkey to Dr. Dalziel, who promised to inoculate him with trypanosomes; but the
monkey got loose and ran away, so that the experiment could not be completed.
Remarks. This monkey was inoculated with blood from Horse C about the same time that Horse H was inoculated with blood from the same horse, and the fact that Horse H developed trypanosomes in his blood and died shows that in all probability the monkey was also inoculated with trypanosomes but did not get infected, proving that the virulence of the strain of trypanosomes in Horse C had been lowered towards the monkey by the treatment of mercury. Moreover, the monkey did not waste and die as all the dogs have done, showing that the monkey does not get a chronic infection by the inoculation of such blood as the dog does.
Detail of Cases under B. which were infected with Serum.
(a) Horse I (see Charts 35a and 35b) was, on the 16th May, injected subcuta- neously with 50 c.c. of serum obtained from the blood of Horse D. The tempera- ture rose a week later, and then trypanosomes also appeared in the blood. The temperature showed another rise later, and trypanosomes continued in the blood up to the time of death on the 1st July.
Remarks. This case proves that the blood of Horse D at this period, after the treatment by methylene blue and then by perchloride of mercury, was not free from trypanosomes, in that the disease developed in Horse I and proved rapidly fatal; the rapidly fatal course run by the disease was probably due to the large quantity of serum injected, which must have contained a good deal of toxic material.
(b) Calf M (see Charts 36a to 36f) was, on the 16th May, injected subcutaneously with 30 c.c. of serum obtained from the blood of Horse D. The chart will show that the temperature ran very nearly a normal course throughout and showed no trypanosomes in his blood all along. About the end of the period of incubation after inoculation, the calf fell off a little in condition, but soon improved again, and became healthy and strong. On the 31st October the temperature observations ceased, and the calf was left with Dr. Alexander at Maifoni to be kept under general observations, as in the case of Calf K. The last news of the calf, dated January, 1909, was that it was well and strong.
Remarks. It is evident from the above that although the blood serum of Horse D contained trypanosomes, and proved infective to Horse I, yet it failed to infect and reproduce the disease with trypanosomes in the blood in the case of Calf M, so that the virulence of the strain of trypanosomes in the blood of Horse D was lowered towards the calf, but not towards the horse, by the treatment first by