ZOOLOGY AND BOTANY, MICROSCOPY, ETC. 
703 
Subcultures from the fluffy glassy growth at the bottom of the broth 
cultivations were made on agar-gelatin and on broth. On agar at 37° 
developed circular small translucent droplets, having no tendency to 
become confluent. In the broth subcultures at 37° glassy fluffy masses 
developed at the bottom of the tubes, the supernatant fluid remaining 
quite clear. The gelatin tubes incubated at 20° failed. Cover-glass 
specimens of the broth cultures stained with rubin (2 per cent, watery 
solution), methyl-blue-anilin water, and in gentian-violet-anilin water, 
showed the growth to be made up of strings and filaments ; the filaments 
were composed of short, thin rods, most of which had a granule at each 
end. The growth in agar also showed filaments, but also many groups 
of minute bacilli resembling those found in the sputum. 
Method of Examining Saliva for Pathogenic Organisms."' — Dr. 
W. D. Miller finds that, owing to the large number and different sorts 
of micro-organisms infesting the mouth, it is almost impossible to arrive 
at any conclusion regarding the presence or absence of any particular 
kind by a simple microscopical examination. Agar-cultures also often 
fail, first, because many pathogenic mouth bacteria do not grow on this 
culture medium ; or, secondly, they grow so slowdy that they are soon 
overgrown and hidden by the more proliferous oral saprophytes. Gelatin 
is still less adapted to the purpose. Pathogenic organisms must there- 
fore be isolated through the medium of an animal body. 
The person whose saliva is to be examined should be instructed to 
rub the tongue against the cheeks and gums, so as to make the saliva 
mix with the dead epithelium and other deposits. One or two drops thus 
obtained were injected into the abdominal cavity of a white mouse. 
When the mouse died within five days the cause of death was found to 
be acute peritonitis, or blood poisoning, or a combination of both; if 
later, death was nearly always due to the local suppuration processes. 
Thus, from these experiments the author (111 mice were injected) was 
able to divide the pathogenic mouth bacteria into two classes, and he 
also found that injections made with the blood or peritoneal exudations 
of the dead mice produced the same results as injections with saliva. 
Preparing the Antitoxic Serum of Tetanus.f — MM. E. Boux and 
L. Vaillard, in a contribution to the study of tetanus, in which they 
deal with the prevention and treatment of this disease, state tbeir method 
of obtaining an antitoxic serum. 
They use tetanus cultures in peptonized bouillon about four or five 
weeks old ; these cultures, filtered through unglazed porcelain, furnish 
a clear liquid. This liquid is the author’s tetanotoxin in a condition of 
extreme activity, as 1/4000 ccm. kills a mouse. This toxin mixed with 
a solution of iodine loses in great measure its harmful properties, and 
forms the vaccinal fluid, which is in no way caustic. 
The serum is obtained from a rabbit (say of 2J kilo, weight). On 
the first day the animal is injected subcutaneously with a mixture of 
3 ccm. of toxin and 1 ccm. of Gram’s iodine solution. On the fifth day, 
5 ccm. of toxin and 2 cc. of Gram’s solution. On the ninth day, 12 ccm. 
of toxin and 3 ccm. of the iodide solution. Eight days after the third 
* Trans. Seventh Iniernat. Congress Hygiene, ii. (1892) p. 46. 
t Ann. Inst. Pasteur, vii. (1893) pp. 72-7. 
