by United States consuls and sanitary inspectors or officers of the Marine Hospital Service serving in foreign ports, and published by this division. Among such reports of sanitary importance, published during the fiscal year, attention may be called to the preliminary report of Dr. S. Kitasato on the "Bubonic plague;" the report on the use of "Antitoxic serum” in diphtheria by P. A. Surg. J. J. Kinyoun; reports on the progress of cholera in Europe by Surgeon Irwin during the summer and fall of 1894; a report on the water supply of the city of Habana by Sanitary Inspector D. M. Burgess; a report on the efficacy of vaccination as exemplified in the epidemic at Bradford, England; reports on the prevalence of plague in China, and on the prophylactic inoculations for hydrophobia as practiced at Odessa, Russia. During the fiscal year the following number of reports were received from foreign countries, all relating to matters of sanitary importance. These are exclusive of the regular weekly reports mentioned above: Brazil, 55; Cuba, 45; Germany, 39; Turkey, 35; Holland, 22; Belgium, 18; France, 17; China, 18; Russia, 16; Mexico, 15; West Indies, 15; Great Britain, 13; Canada, 10; Gibraltar, 8; Nova Scotia, 6; Japan, 6; Austria-Hungary, 4; Greece, 4; United States of Colombia, 2; Sweden, 2; Argentine Republic, 2; Italy, 2; Venezuela, 2; and one each from Denmark, Switzerland, Nicaragua, Panama, Chile, Norway, India, Honduras, Philippine Islands, Siam, Syria, and Sierra Leone. Translations are made from foreign sanitary journals and publications, and are published in the abstract of sanitary reports. These translations relate principally to statistical information concerning cholera and to quarantine measures against cholera enforced by foreign governments. In this division also have been prepared tables showing the preva lence of cholera and yellow fever in foreign countries, the prevalence of smallpox in the United States, and the table showing the years in which yellow fever has invaded seaboard cities of the United States, these tables being inserted in the portions of this report relating to the diseases mentioned. INVESTIGATION OF WATER SUPPLY, SEWAGE, AND DISPOSAL OF GARBAGE IN THE CITIES AND TOWNS OF THE UNITED STATES. In view of the wide-spread prevalence of typhoid fever during this fall-this being a water-borne disease-and in view of the general interest now manifested concerning the water supply of cities of the United States and the pollution of rivers which furnish the water supply of some cities by the sewage of others, a preliminary investigation has been begun by this Bureau with the object of collecting information. from every city of the United States, showing the source of its water supply, the method of its collection, storage, distribution, and disposal, whether it is under municipal, corporate, or individual control, and whether subject to sewage contamination. A circular is now being prepared, addressed to the mayors of the cities and towns of the United States, calling for the required information. Following is a copy of a letter convening a board to report upon the best method of obtaining and collating the information desired: TREASURY DEPARTMENT, OFFICE OF THE SUPERVISING Surgeon-GenERAL, M. H. S., SIRS: You are hereby appointed a board for the purpose of formulating a plan for obtaining the information hereinafter mentioned. Section 3 of the act of Congress approved February 15, 1893, entitled an act granting additional quarantinē powers and imposing additional duties upon the MarineHospital Service, provides as follows: "That the Supervising Surgeon-General of the Marine-Hospital Service shall cooperate with and aid State and municipal boards of health in the execution and enforcement of the rules and regulations of such boards and in the execution and enforcement of the rules and regulations made by the Secretary of the Treasury to prevent the introduction of contagious or infectious diseases into one State or Territory or the District of Columbia from another State or Territory or the District of Columbia, and the Secretary of the Treasury shall make such additional rules and regulations as are necessary for this purpose." Section 5 of the same act of Congress provides: "That it shall be the duty of the Supervising Surgeon-General of the MarineHospital Service, under the direction of the Secretary of the Treasury, to perform all the duties in respect of quarantine and quarantine regulations which are provided for by this act, and to obtain information of the sanitary condition of foreign ports and places," etc.; "and the Secretary of the Treasury shall also, as far as he may be able, by means of the voluntary cooperation of State and municipal authorities, of public associations and private persons, procure information relating to the climatic and other conditions affecting the public health," etc. # In pursuance of these sections of the law, and in view of the necessity, which is daily becoming more apparent, for an accurate knowledge concerning the water supply and disposal of same of the various cities of the United States, and because the water supply and disposal of cities in one State are affected by and affect cities in other States by transmission of the germs of contagious diseases, it is my desire to begin a preliminary investigation relating to this subject. It is necessary to have on file in this Bureau a consolidated report showing, with regard to every city of the United States, the source or sources of its water supply, the method of its collection, storage, distribution, and disposal; second, whether this water supply is under municipal, corporate, or individual control; and third, the cost to consumers. Inas much as there is so much contamination of water supply by sewage, the inquiry will also include the method of sewage disposal. It is suggested that the inquiries should be arranged under separate heads or divisions, and that a circular embodying the facts required be prepared by the board for distribution to the various health officers and others from whom the information is desired to be obtained; also that a card, with blank form thereon, be provided for each separate head or division, to be filed in the Bureau, after the manner of the usual card index. You will make report upon this matter as soon as practicable. Respectfully, yours, WALTER WYMAN, Supervising Surgeon-General, Marine-Hospital Service. P. A. Surg. C. E. BANKS, Chairman. P. A. Surg. J. J. KINYOUN. Asst. Surg. W. J. S. STEWART, Recorder HYGIENIC LABORATORY. REPORT OF THE MEDICAL OFFICER IN CHARGE. TREASURY DEPARTMENT, HYGIENIC LABORATORY, M. H. S., Washington, D. C., November 1, 1895. SUPERVISING SURGEON-GENERAL, United States Marine-Hospital Service. SIR: I have the honor to submit the following as the report of the Hygienic Laboratory of the Service for the past year: From August 15 to December 5, 1894, I was absent in Europe, where I was detailed to the International Congress of Hygiene and Demography, which held its session at Budapest, September 1 to 9. Afterwards I visited the bacteriological laboratories of Vienna, Prague, Munich, Paris, Berlin, Hamburg, and London. The greater part of the time was spent in Paris and Berlin, where I made a careful study of the new remedy for the treatment of diphtheria. Under the dates of October 20 and November 6 I transmitted reports thereon, as follows: The serum therapy of diphtheria. INSTITUTE PASTEUR, Paris, October 20, 1894. SIR: While attending the eighth session of the International Congress of Hygiene and Demography, held in Budapest in September last, Prof. E. M. Roux, of the Pasteur Institute, read a paper before the section of hygiene on the serum therapy of diphtheria, in which he gave to the world the results of his labors during the past three years. No subject at any congress has, I dare say, been the cause of so much discussion as this, and, on the whole, elicited nothing but praise. The results obtained by Professor Roux in the treatment of cases of diphtheria are so astounding that at first one is almost compelled to ask oneself, "Is this possible?" But when the methods are known and the array of statistics are given, there can hardly remain a trace of doubt. A greater part of what I am going to say has, I know, been published in the daily and medical press, and the only apology I offer for the repetition is that it is well worth reading twice. It appears that at last we have found a method which is not only good in one disease, but the principle of the method can be applied to many. It at last has opened up a new field for work in infectious diseases. Availing myself of the kind invitation of Professor Roux to come to the institute, and there learn by practical experience the exact methods employed in the preparation of the antidiphtheritic serum, and also to observe the effects of the new treatment at the hospital for diphtheria, I arrived in Paris on September 20, and immediately commenced my work. Every facility has been afforded me by the gentlemen connected with the institute to make my stay pleasant and profitable. My sincere thanks are due to all, and especially so to Professor Roux and Drs. Martin and Chaillu. I have been in no hurry to report on what I have seen, nor to form an opinion of the merits of the treatment. After spending a month at the institute and hospital, I have seen sufficient to enable me to form an intelligent estimate of its value. There is still more to be said in its favor than was claimed for it by Professor Roux at Budapest. It has passed the experimental stage, and will in the future be reckoned in value for the treatment of diphtheria as vaccine is for the prevention of smallpox. The steps necessary in the preparation of the serum antitoxine may be divided into three: First, the preparation of the toxines of diphtheria; second, the immunization of animals; third, preparation of and conservation of the serum. Preparation of the toxines.-The toxines are prepared from a bouillon culture of virulent bacilli of diphtheria. As the methods employed in the institute are somewhat different from those of other continental laboratories, and in many instances, as in this, are superior, I will give them in detail: A virulent culture of the bacillus diphtheria is selected-one which is fatal to a 500-gram guinea pig in from twentyfour to thirty hours. From this culture a flask of alkaline peptone bouillon is inoculated. After it has remained in the thermostat for twenty-four hours, at a temperature of 36 C., it will be found rich in bacilli. This culture may be termed the stock culture for others which are destined for the toxines. For this latter a special form of culture flask is employed, in order to permit of a special method of cultivation of the bacilli, whereby the toxines formed are much stronger and, it is claimed, somewhat different in their character than when the ordinary methods are employed. The flasks used are known as the Fernbach flasks, and are large, flat-bottomed Florentine flasks, provided with a tubulature on the side within about an inch of the bottom. The neck and tubulature are constricted near the mouths for the recep tion of the proper cotton plugs. These flasks are filled to a short distance below the tubulature with an alkaline peptone bouillon and then sterilized. Soon after this the flasks are inoculated with the bouillon culture of the bacillus diphtheria, about 40 c. c. to each flask. They are then placed in the thermostat for twenty-four hours in order to "start" them, when each flask is connected with an aspirator and a current of air is slowly kept moving through the flask in the direction from the mouth and to the tubulature. The air before entering the flask is passed through an ordinary wash bottle, in order that it may be moist, so as to prevent the evaporation which would occur as well as to maintain the best conditions for culture. This method, so far as I know, is practiced only in this institute. It is the discovery of Dr. Fernbach, who observed that so long as the bacterium remains in the active liv ing state-or, in other words, maintains its integrity-little or none of those substances known as toxines or ptomaines are set free; but as soon as you present the conditions most favorable for the development of bacteria the life of the individual bacterium is shortened and it completes its cycle, ending in proliferation and setting free the nucleins from it. When a bacillus such as that of diphtheria is grown under the same conditions as cited above there is a greater quantity of the nucleins formed than would occur under the ordinary conditions of culture. These flasks are kept at a temperatere of 37 C. for from three to four weeks. At this time the bouillon is rich in flaky masses of the bacilli. If examined microscopically the masses are found to be nearly, if not all, disintegrated bacilli. Sometimes a few bacilli are encountered, but they have in a great measure lost their characteristic form. Filtration of the cultures.-Without further preparation the cultures are filtered through a Chamberland filter tube into sterilized flasks and kept until required for use. Each lot of the toxines is tested for virulency by standardizing it by injections into guinea pigs. The usual strength, being c. c., will kill a 500-gram guinea pig within twenty-four hours. The filtrate will preserve its virulency for a considerable time, provided it is kept away from light and maintained at an equal temperature. Boiling the cultures of even heating them to a degree that will kill the bacilli is not practiced, for it has been found that either process impairs the strength as well as changes some of its properties. Large quantities of cultures are kept growing, a special room being employed for this purpose, as each horse will require a large amount of the toxine to immunize it and to maintain the antitoxine in the blood after immunization is completed. Immunization of animals.-The antitoxine for the treatment of diphtheria is in solution in the blood of an animal rendered refractory to the disease. The manner of producing immunity in an animal may be performed in one of two ways-by injections of the toxines or by inoculations of the bacilli. The former method has been found to be the best, and at present is the only one in use. In the experiments of Professor Roux and Dr. Martin animals of all kinds were used, but now they use the horse, as it has been found to be the most satisfactory. It stands the process of immunization better, and gives a serum stronger than other larger animals, i. e., in the same length of time, besides furnishing a larger amount of serum. The present method of immunizing the horse is somewhat different in its details from that given in Professor Roux's paper, being much simpler. A horse is selected which is sound, having been subjected to injections of tuberculin and mallein, the age not playing any particular part; usually it is from six to eight years-a cab horse, which has seen better days. At first a trial injection of the toxine is made, usually less than a cubic centimeter, carefully noting the general and local reaction. In some horses even the trial dose has a profound effect, but usually there is quite considerable local and general reaction. If the animal becomes quite ill, a small quantity of Gram's solution is added to the next dose, and even the next following if the reaction is too strong. After this the horse bears the increasing dosage with little or no discomfort. The general plan for the injections is as follows: First day, c. c. of pure toxines, of which c. c. fatal to 500 grams of guinea pig; eighth day, 1 c. c.; fourteenth day, 14 c. c.; twentieth day, 2 c. c.; twenty-eighth day, 3 c. c.; thirty-third day, 5 c. c.; thirty-eighth day, 8 c. c.; forty-third day, 10 c. c.; forty-seventh day, 20 c. c.; fifty-first day, 30 c. c.; fifty-sixth day, 50 c. c.; sixty-second day, 50 c. c.; sixtyeighth day, 60 c. c.; seventy-fourth day, 100 c. c.; eightieth day, 250 c. c.; eightyeighth day, 250 c. c. When the first injections are given there is quite a marked local and general reaction to the poison; there is an edema at the point of the injection, which is followed by a distinct inflammatory process, hard in the center and soft and ædematous at its periphery. The general reaction is manifested by a rise in the temperature 10-2o C., loss of appetite, and occasionally cramps. The reaction must be taken as the guide in the future dosage, and a sufficient time must be allowed to elapse between the injections for the complete recovery from the general and local effects. As the quantity of the toxines is increased, the general effects generally decrease, perhaps a rise of a degree for twenty-four hours. The local effect partakes more of an adema, and has the character of an inflammation. At a certain stage, usually after two months' treatment, when 50-60 c. c. can be injected without harm, there is no general reaction, but a large œdema at the site of the injection, which disappears within from twenty-four to forty-eight hours. Toward the last, even when 200-300 c. c. are given, there is only an enormous edema, which disappears within from twelve to eighteen hours. When these inordinately large quantities can be given with only a local reaction being manifest, the horse has come well under the influence, and the blood will be found to be rich in the antitoxine. There is a curious fact well worth noting: At the end of the second month of the treatment, when the horse can bear as much as 50-60 c. c. of the toxines without discomfort, the blood will be found to contain but little of the antitoxine. The antitoxine only appears after repeated stimulation of the cells (?) by the large and frequent doses of the toxines. The subcutaneous injections do not yield a serum as rich in the antitoxines as |