7
(vii) Plains and hill buffaloes injected subcutaneously with 3 c. c. of
serum and 48 hours later tested with virulent culture, were found at a later
date to possess an active immunity.
(viii) A dose of 35 c. c. of serum injected intravenously in hill cattle has
been found necessary to overcome the effects of a lethal dose of virulent
culture injected subcutaneously 15 hours previously ; the animals thus treated
presented no symptoms of the disease. Consequently, this mode of treatment,
on account of the large quantity of serum necessary, cannot be recommended
except in the case of valuable cattle being attacked, and even then if any
symptoms are exhibited, probably a much larger dose would be required than
the one above mentioned.
29. When bovines were hyper-immunized with the toxins produced by
the causal factor of Hæmorrhagic Septiæmia.—A plains bull, aged four and a
half years, body weight 421 lbs., susceptible to this disease, was hyperimmunized
with toxin prepared from the organism Hæmorrhagic Septicæmia allowed to
remain in an incubator at 37° C. from 11th October 1904 to 15th May 1905.
The doses of toxin injected subcutaneously varied from 1 c. c. to 110 c. c.
during a period of four months, in all a total of 626 c. c. were injected. The
body weight of the animal increased during this interval from 421 to 490 lbs.
On 18th July 1905, the animal was bled for the first time and serum
prepared in order to test its immunizing power after the 8th injection, nine
days after a dose of 80 c. c. had been injected. A dose of 5 c. c. serum and 48
hours later 0.1 c. c. virulent culture, failed to protect the animal, but 10 c. c.,
under similar conditions, only allowed of a slight thermal reaction, and a dose
of 7.5 c. c. proved sufficient to protect. The animal was bled 2,200 c. c. on
the 30th August and 3,000 c. c. on the 8th September, and later on the latter
date was inoculated with 0.1 c. c. of a culture of Hæmorrhagic Septicæmia,
to test the degree of susceptibility. A slight thermal reaction followed and
concurrently a small swelling occurred at the seat of inoculation 2 ×1 × ¼,
which was quickly absorbed.
30. Sera produced by hyper-immunizing bovines with certain organic
substances, other than the causal agent of Hæmorrhagic Septicæmia.—In
a previous communication I pointed out that serum prepared by hyper-immuniz-
ing plains cattle with phytoglobulin or some nearly allied substance obtained
from pumpkin seeds grown in the vicinity of Muktesar, on subcutaneous or
intravenous injection into susceptible animals, was capable of conferring a
resistance to the organism of Hæmorrhagic Septicæmia, which resistance
could be changed into an active protection, should the animal be inoculated
with a small quantity of culture of the " causal agent." Further, that this
active resistance was found to be maintained two years after the primary
protection had been conferred.
During the years 1902-1903, sera were also prepared by hyper-immunizing
cattle with certain substances other than the causal agent of Hæmorrhagic
Septicæmia, obtained from the testes of plains and hill cattle, and of
buffaloes, and also from the seeds of plants. Additional sera were made from
other organic substances such as milk, yolk of egg, fish sperm and spawn, cell-
protoplasm, mucin and yeast. During the year under review, certain of the
above sera, to which 0.5 per cent. carbolic acid was added in the first instance,
have been retested as to their protective effects on the " contagium " of
Hæmorrhagic Septicæmia, after they have been allowed to stand in the cold
room at Muktesar for periods of from 2½ to 3½ years.
The following interesting and important results were obtained :—
The minimal lethal dose of Hæmorrhagic Septicæmia was found to be
0.1 c. c. of bouillon culture 48 hours at 37° C. This killed an animal of 240 lbs.
body weight in 31 hours. Hill animals were respectively subcutaneously
injected with the following sera prepared from buffalo testes, yolks of eggs,
casinogen, Para-nucleo-proteid and pumpkin seeds. Each animal received
1 c. c. per lb. body weight of one of the respective sera, and 48 hours later
was subcutaneously inoculated with 0.1 c. c. of virulent Hæmorrhagic
Septicæmia culture. Some of the animals showed thermal reactions and
recovered, with one exception, namely, the one subjected to para-nucleo-proteid
serum. This animal succumbed in 35 hours 10 minutes, a little later than