586 
Journal of Agricultural Research 
Vol. XXIX, No. 12 
fever. The animals were described as 
fine Nubian milk goats costing from 
$200 to $600 each and the flock was 
mentioned as consisting of about 100 
head. Sterile containers were shortly 
afterward forwarded to Doctor Heck 
with the request that he obtain from 
some of the higher reacting goats speci¬ 
mens of blood, milk, and urine for cul¬ 
tural and inoculation work. The re¬ 
actors had been destroyed before the 
request was received; consequently no 
further action was taken at that time. 
About two months later, W. E. 
Whigham, of Donna, Tex., wrote ask¬ 
ing permission to forward for the appli¬ 
cation of the agglutination test for 
Malta fever blood-serum samples from 
two flocks of goats, approximately 100 
head each, which had been exposed to 
the flock previously tested. Doctor 
Whigham stated that he had had three 
or four cases of Malta fever in human 
beings contracted from drinking the 
milk of these goats, one of his patients 
being the owner of one of the exposed 
flocks. 
ORIGIN AND METHOD OF ISOLATING 
BACTERIUM MELITENSIS 
On November 23 and 25, 212 more 
serum samples from the two flocks 
mentioned were received. Of this 
number three reacted to the agglutina¬ 
tion test for Malta fever in a dilution 
of 1 to 1,000, one 1 to 500, nine 1 to 
200, eleven 1 to 100, seven 1 to 50, and 
eight 1 to 25. The remaining samples 
were negative in dilutions of 1 to 25 or 
higher. 
On November 6 a sample of human 
blood was received from Doctor Whig¬ 
ham. It was described as being ob¬ 
tained from one of his patients, a Mexi¬ 
can goat herder, 14 years of age, who 
had reacted to the Malta-fever test. 
The boy had been suffering with fever 
about 9 months, more or less contin¬ 
uously, his temperature reaching 102 to 
103° F. in the afternoon. The specimen 
was somewhat decomposed upon arrival, 
but was sown on serum agar, subjected 
to the agglutination test both with a 
Bacterium melitensis and Bad. abortus 
antigen,and used for animal-inoculation 
purposes. The sample reacted in a di¬ 
lution of 1 to 1,000 with both antigens. 
In making the inoculations 11 
guinea pigs were used; 6 were injected 
intra-abdominally with a physiological 
salt solution suspension of the blood and 
5 both intra-abdominally and intrates- 
ticularly. Intratesticular inoculations 
had been described by Meyer and his 
coworkers (21) as an effective way of 
transmitting the disease through injec¬ 
tions of Bacterium melitensis cultures. 
On December 22, between five and six 
weeks from the date of inoculation, five 
guinea pigs were destroyed. Two had 
received the material into both the ab¬ 
domen and one testicle, and three had 
received it free in the abdominal cavity. 
Blood serum from these animals gave 
negative results to the agglutination 
test for Malta fever. Serum-agar slants 
sown with the spleen tissue remained 
sterile. 
On December 28 the six remaining 
guinea pigs were destroyed. The 
autopsy findings and the cultural and 
serological results were as follows: 
Guinea pig No. 77012. Inoculated 
intra-abdominally and intratesticular- 
ly. Spleen was somewhat thickened 
and surface irregular. Mesenteric 
lymph glands were congested. The in¬ 
oculated testicles showed a constriction 
about midway between its extremities. 
On one side of the constriction was a 
small abscess containing semifluid pus. 
Other organs appeared normal. Cul¬ 
tures were made from the heart, liver, 
spleen, testicles, kidneys, and mesen¬ 
teric lymph glands. Serum-agar slants 
alone were used. Blood serum reacted 
to the agglutination test with both 
Bacterium abortus and Bad. melitensis 
antigens in a dilution of 1 to 1,000. 
Its end point was not determined. 
A portion of the cultures from each 
organ was incubated in closed jars in 
which 10 per cent of the air had been 
displaced by C0 2 gas and in which a 
ball of moistened cotton had been 
placed, providing conditions highly 
favorable for the growth of Bacterium 
abortus. Another lot was incubated 
under normal atmospheric conditions. 
The tubes in all cases were merely 
closed with cotton plugs, and the in¬ 
cubator was maintained at a tempera¬ 
ture of 37° C. On December 30 the 
cultures were examined with no evi¬ 
dence of growth. On January 2, 
two cultures from spleen incubated 
under C0 2 conditions showed from 100 
to 200 small opalescent colonies, two 
from affected testicle 15 to 20 colonies 
so small as to be recognized with 
difficulty, two from mesenteric lymph 
glands 10 to 15 colonies, one from 
kidney sterile, one from liver 3 colonies, 
one from heart sterile. Two cultures 
from spleen incubated under normal 
atmospheric conditions showed 200 to 
300 colonies somewhat larger than 
those developing in a 10 per cent C0 2 
atmosphere, one tube from testicle 50 
to 75 colonies, one from liver 40 to 50 
