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PUBLISHED MONTHLY.-SUBSCRIPTION $1.00 A YEAR. VOL. XIX. AUSTIN, AUGUST, 1903. Original Contributions. No. 2. Experiments to Determine the Disinfecting Value of the Morris Formaldehyde Generator.* BY ALLEN J. SMITH, M. D., Professor of Pathology; Bacteriologist of the Medical Department, University of Texas, Galveston, Texas. The room used for all these investigations is a small one, containing 850 feet of air space, situated on the second floor of the Medical College building. It has one window, 3x4 feet, about 8 feet from the floor, and one large door. All the noticeable cracks and crevices were pasted over with strips of paper, other places were stuffed with old rags; the cracks about the door made as air tight as possible with cotton. Even after stopping the room in this manner, it presented the following difficulties in securing the best results. 1. It is small. and so a small charge of alcohol is to be used. It requires the same amount of alcohol to heat the asbestos disc, when a small amount of alcohol is used as when a large quantity is taken, so, in a small room, a comparatively greater amount of the alcohol is used up before the generation of formaldehyde begins. 2. A number of large water and steam pipes pass through the floor and ceiling of the room and it is difficult to pack the openings about the pipes airtight. 3. No way of withdrawing the infected material was had save by opening the door, and as this had to be done seven times in the twenty-four hours, it will be seen that a great diminution of the gas must have occurred. That the room leaked, was evidenced by the strong odor of formal [*Now called the Germicidal Gas Generator.-EDITOR JOURNAL.] dehyde which prevailed in the corridors and rooms during the experiments. Four pathogenic organisms were used, the streptococcus pyogenes, bacillus typhosis, bacterium anthracis, and mycobacterium diphtheria. These organisms represent a fair range in their resistance to destructive agents, the anthrax being most resistive. In each series, three conditions of exposure were prepared. The first was made by dipping bits of sterile glass in a bouillon culture of the organism, placing it on a bit of sterile cotton and covering it with a very loose bit of cotton. This gave a practically free exposure, with great danger of contamination subsequent to exposure to the gas. In the second, we dipped bits of sterile filter paper in the bouillon cultures and placed them between two layers of a felt (billiard cloth), weighing 367 grams per square meter. Thisfelt is quite compact and is more difficult of penetration than the ordinary blankets. The third condition was like the second, save the paper was protected by two thicknesses of felt. Also in each series (with the exceptions shown in the tables), inoculations were made at stated intervals into sterile peptone and bouillon tubes. These intervals are, at the close of the generation of the gas (usually one hour after starting), 2, 3, 6, 12, 18 and 24 hours. These inoculated tubes were incubated at 37 degrees C. (body temperature), and records were made at the end of 24, 48: and 72 hours. The fluid in the generator was methyl alcohol, to which 1 per cent carbolic acid had been added. Series five was done first, and must be regarded in the light of a preliminary investigation to determine whether the disc was packed properly. The results of this experiment were not satisfactory, so it was repeated in series one, after having properly adjusted the packing of asbestos. In series five, on April 10th, the generator was charged with 1500 cc. of the alcohol mixture, 750 cc. of water, with a temperature of 50 degrees C., were placed in the water pan and the apparatus started in the usual manner. From the first, it was noticed that the volume of gas usually obtained was not being given off and the results bore this out. Twenty-four bouillon cultures were used to infect the glass and filter paper. The anthrax culture apparently contained no spores, as least none were demonstrated by staining. Inoculations were made at the intervals given above, and the tubes were grown three days. The results are shown in tables A and B. Here it will be seen that the streptococcus was killed from the first; the bacillus typhosus was not destroyed entirely until after six hours; clean tubes not being obtained until twelve hours from the start. The anthrax was readily killed, the tube containing the bit of glass exposed twelve hours being infected, but by a coccus which must have gotten in subsequent to disinfection. At eighteen hours, the tube containing the filter paper, covered by two thicknesses of felt, was infected, but by a rod-shaped organism, too small and thin for the anthrax. All the tubes supposed to contain the mycob. diphtheria were contaminated, even after twenty-four hours exposure to the gas. This, however, was not the diphtheria germ, but has been identified by us as one form of the B. subtilis. (For further discussion, see on series four.) In series one, the disc had been properly packed and the amount of gas given off was satisfactory. Fifteen hundred cc. of alcohol were again used, 750 cc. of water, at 50 degrees C., placed in the evaporating pan. That the air was pretty thoroughly saturated was evidenced by the deposition of moisture on the window pane. Exposures were made as before, the material being taken from 24-hour bouillon cultures. By staining, no spores were demonstrated in the anthrax. By reference to tables C and D, it will be seen that complete destruction of the streptococcus pyogenes was effected at the end of two hours; with the B. typhosus this was done in three hours; with the Bact. anthracis, at the end of three hours; the tube containing the bit of glass was contaminated at the end of forty-eight hours. It is to be noted, however, that all the 2-hour tubes were clean. The Mycob. diphtheria was destroyed all the way through after two hours. In series two, 1000 cc. of fluid were used, and 750 cc. of water at 50 degrees C. At the end of twenty-four hours, 250 cc. of this water was still unevaporated. Spores were present in the culture of anthrax; 48-hour cultures being used. Inoculations, as before, were incubated. Tables E and F will give the results. All the streptococci tubes were clean at three hours; one of the 3-hour typhoid tubes was infected in seventy-two hours, but by a nonmotile, rod-shaped organism too thick to be typhoid, all the typhoid bacilli being killed in three hours. The anthrax was killed in six hours this time, while the diphtheria was killed all the way through. In series three, exposures were made under similar conditions, save here 750 cc. of alcohol and 750 cc. of water, at 50 degrees C., were used. Of this water, there remained at the end of twentyfour hours 400 cc. Again, inoculations were made at the stated intervals. Here (tables G and H), two hours were sufficient to destroy all the streptococci. The tube containing the bit of glass exposed for six hours is excluded, since it contained no cocci, but a rod-shaped organism, undoubtedly due to external contamination. |