J. H. KELLOGG, M. D., Supt.

PAUL PAQUIN, M. D., Director.

RESEARCHES RESPECTING THE PURITY OF VACCINE.

THE director of this laboratory began investigations and experiments in 1887, on the relative purity and activity of the

vaccine of commerce. Owing to multiplied duties, he was unable to pursue his work steadily to completion. He carried on various series of investigations at long intervals, in 1887, 1888, 1889, and 1890, at the University of Missouri, and in 1891 at the institution now under his direction. To this date, one hundred and ten specimens of commercial vaccines from twenty different sources have been studied bacteriologically. During these studies, which are still in progress and will be prosecuted more closely than heretofore, there were isolated by culture eight forms of bacteria, which produced, after inoculation, lesions or symptoms of disease more or less pronounced in rabbits, guinea-pigs, rats, mice, cattle, and man, or in several of the species, and seven which produced no lesion or symptom. Among the pathogenic forms

were

Staphylococcus pyogenes,

Staphylococcus pyogenes aureus,
Staphylococcus pyogenes citreus,
Bacillus pyogenes fœtidus,
Bacillus septicus,

a bacillus which was connected with, and probably the cause of, gangrene; a micrococcus in connection with another case of gangrene; and a saprogenous bacillus in connection with septic lesions.

The last three not identified with known forms, were isolated in 1889, from two different crusts purchased on the market. All of these forms were pathogenic to rabbits, and some of them to certain of the other animals mentioned,

The quantity inoculated varied from 1 to 3 drops of a liquid culture diluted

1This report of progress gives more details and more experiments than that made to the American Health Association at its last meeting in Kansas City.

BATTLE CREEK, MICH., DECEMBER, 1891.

in from two to six or seven drops of distilled sterilized water. Forty rabbits inoculated with these bacteria (7 animals with each kind of the pathogenic germs enumerated above) resulted in 28 local lesions, of which five were nodules more or less pronounced; thirteen were abscesses which healed; ten were abscesses or infectious wounds followed by general infection and death. Nine cases died from septicaemia without local lesions. Only three remained apparently free from symptoms of disease. Among the seven kinds of bacteria which were found in these vaccines, and which proved to be harmless by inoculation, one was the bacterium termo, which may, under certain circumstances complicate a wound, though it is not generally considered a parasitic microbe.

cial product supposed to be safe, had local A mulatto, vaccinated with a commersepticemia of a very severe and painful type. Constant local cold-packing reduced it after two days' close attention. A coccus and bacillus existed in the sanius secretion of the angry, deep ulcer that took place at the seat of the pustule. Both these microbes were accidentally lost before any inoculation.

Twenty-six specimens of virus on ivory points, from twelve different sources, purchased in the market, were tested by inoculation to rabbits. Three points of each specimen were wiped with a sterile cloth, soaked from one to three hours in two drams of distilled sterilized water, and ten drops were inoculated hypodermically. The result was, nineteen local swellings, diffused or circumscribed, and four abscesses, one of which caused general infection and death. Three samples proved harmless.

Microscopic analysis of the material inoculated, revealed a degree of purity or impurity in ratio with the symptoms observed. The points offering the smallest number of septic germs produced the slightest lesions, and rice versa. Those

that produced no lesions at all appeared about free from any foreign microbes.

Four rabbits were tested with twelve half quills. Of these, two produced local swellings, and one an abscess which healed.

Fourteen inoculations made with vaccine from cones or crusts, produced results as follows: Every rabbit had marked swellings, and ten of them had abscesses; of the whole lot, one died within 48 hours, from general infection, and two died after long suppuration.

On the other hand, thirty rabbits were inoculated with vaccine virus, diluted in sterilized water, in which (vaccine) the microscope and cultures indicated the greatest relative degree of purity. The result was a slight enlargement at the point of inoculation in fifty per cent, but there was not a single diffuse swelling, not one abscess, not a death. Possibly the vaccine virus itself produced the slight symptoms observed.

At this stage of our experiments, taking into account extensive observations in several hundred cases of vaccination in man and beast, none of which are recorded here, and from which the writer gathered statistics, we believe that:

a. Most of commercial vaccines are more or less impure, bacteriologically speaking.

b. Much of it is dangerously contaminated with pathogenic germs.

c. Crusts, scabs, and cones are very impure and wholly unfit for vaccination of human beings.

d. That the extensive inflammatory symptoms, such as diffused reddish or violet swellings, painful or painless œdema; angry or sanious ulcers; lesions with tough, purple, adhering scabs, surrounded by painful enlargement; swellings extending several inches, or even a foot or more away from the point of inoculation, and involving the lymphatics at considerable distance; little abscesses here and there, remote from the spot vaccinated, are so many lesions or symptoms of complication of vaccinia.

e. That such complications are secondary, the vaccinia cocci preparing the field for the septic forms to grow.

f. That frequently the foreign germs destroy the vaccinia growth in the inoculated spot before the latter has produced its effects, and the patient is left unprotected,' and may be vaccinated again successfully or take smallpox.

3 The writer has seen several cases in which second vaccination took well, after such complications, which doctors had thought typical vaccinia.

g. That the foreign germs found associated with vaccinia cocci are not necessary to produce the true, simple vaccinia pustule.

h. That many doctors mistake marked complications for good vaccinia pustule.

i. That it is possible to produce absolutely safe' vaccine by antiseptic and aseptic methods, though extremely difficult to produce vaccine absolutely free from certain air germs.

j. That vaccine from a second dipping in the same pustule a day or two after the first one, as is done in many commercial institutions, is a bad and dangerous practice, as such vaccine is almost always very impure. (See frontispiece plate.)

HOW TO STERILIZE MILK.

SINCE the subject of the sterilization of milk has been agitated, the question is often asked, What is the best method of sterilizing milk? that is, destroying the germs of disease or fermentation which it may contain. We have made many experiments, and from our experience can confirm the results of many observers who have given attention to this subject, that the complete sterilization of milk is a matter of no small difficulty. Milk which has been boiled for half an hour once or twice each day for several days in succession, will keep for a number of days, but ultimately sours. This is due to the fact that some of the germs contained in milk are particularly hard to kill, and require a temperature above that of boiling

milk.

We have recently been conducting some experiments upon this subject, with results so satisfactory that we are glad to be able to communicate them to the readers of the BACTERIOLOGICAL WORLD AND MODERN MEDICINE. The first experiments were made with a tin receptacle capable of resisting a pressure of twentyfive pounds. This was partly filled with water and placed in boiling water, to the action of which it was exposed for half an hour. The pressure indicator showed no very considerable increase in pressure within the closed receptacle. We then tried boiling the tin vessel in a saturated solution of salt in water, when the pressure, as indicated by the pressure gauge, rose to four pounds. This was the result which we expected. We accordingly proceeded to a further experiment, which consisted in boiling milk tightly sealed in 1 This is done at this Laboratory by scientific methods.

strong bottles, in a saturated solution of salt. Milk sterilized in this way, by boiling in the salt solution for half an hour, will keep perfectly for an indefinite length of time. We opened, a few days ago, a bottle of milk which had thus been sterilized last June (1891), and found it to be as fresh as when placed in the bottle. It is only necessary to take the precaution to allow the solution of salt, in which the bottles are boiled, to cool before removing the bottles. If the bottles are removed from the solution while hot, they will almost instantly burst. The vessel containing the bottles of boiling milk should be set aside and allowed to cool gradually, when the bottles should be removed and placed in an ice chest or an ordinary refrigerator. Ordinary soda-water or beer bottles are excellent for the purpose; or beer bottles may be used. Ordinary corks may be used for the purpose, but they should be previously boiled for half an hour. They should be pressed in tightly, and fastened with wire or with a patent fastener. After the bottles have been cooled and removed from the boiling kettle, the tops should be carefully dried, and if corks are used, covered with sealing wax, such as is ordinarily used for canning purposes.

The efficiency of the salt solution is due to the fact that its boiling point is 227°F., while that of boiling milk is less than 200° F. By using different salts, a still higher temperature may be attained. For example, a saturated solution of carbonate of potash, or saleratus, boils at a temperature of 275° F., while a saturated solution of chloride of calcium boils at 355° F. These high temperatures are, however, unnecessary.

THE BACILLI OF TUBERCULOSIS.

THESE little rod-shaped parasites vary between 2 and 6 in length according to special conditions of their life, and o # 3 to o 5 in width. Their average length is about 3, and the average width about oμ 4. They may be straight, or slightly curved, or irregular in their aspect. They appear either as homogeneous little rods, or as finely dotted bacilli; these dots are

1 The μ (mikron), a unit in certain microscopic meas. urements, means micromillimeter, and is equal to one thousandth of a millimeter. A millimeter is, as the word implies, the one thousandth of a meter, and a meter is equal to 39.37 inches. A millimeter, then, is nearly 1-25 part of an inch, and a μ (mikron) is the 1-1,000 part

of 1-25 of an inch.

due to the presence of fine ovoid or spherical bodies placed end to end like a very minute string of beads. These germs can scarcely be seen without coloring reagents except with high-power lens, and then the dots or spherical bodies, which are looked upon as spores, do not appear. The organisms unstained, particularly if treated with potassa, have the appearance of hyaline, or motionless rods.

The stained bacilli of tuberculosis can be appreciated when magnified 350 to 400 diameters, but it is better to examine them under greater increase, say 480, and it is still better to see them at 700 or even 800 if possible. This high increase. is not necessary for diagnosis.

The bacilli may appear shorter or longer in the same specimen, according to their age. For instance, fresh sputum may at first show very fine, short rods. Let it stand a day or two, and if it does not desiccate, the forms will be found more clearly dotted and longer. After standing for days or weeks, many fine, loose dots - spores doubtless may be observed, and long bacilli formed by them, disposed end to end. In patients having large lung cavities, the bacilli are sometimes longer, and the loose spores more numerous, than in milder cases.

The quantity of bacilli in sputum, and in all fluids, varies greatly; it depends on the extent and age of the lesions. They are usually much more numerous in extensive cavities than in secretions from small lesions resulting from slight tubercles. A given mount may contain only a few-half a dozen or less- and the next, hundreds.

Sometimes we find bacilli in a large cell, a giant cell, indicating an attempt, perhaps, on the part of nature to destroy the germs by cellular digestion — phagocytosis.

Usually specimens to be analyzed, particularly sputum, contain numerous other germs which have no direct relation to the disease so far as the cause is concerned, but are doubtless very harmful complications; for they must be, as in wounds, dangerous and most potent factors in the destruction of tissue, in producing pus cavities, and in creating very damaging ptomaines.

The good influence of high altitudes on consumptives is due not a little to the fact, I imagine, that pus or decomposition germs are comparatively rare in high altitudes, and complications such as almost always exist in other atmospheric condi

tions in tubercular lesions opened to the air, find there no destructive agent of the class of ptomaine makers and the like. These foreign germs, in making analysis, can be made to appear unstained at the side of stained tubercle bacilli, or may be stained as the background, and may therefore be readily distinguished.

Tubercle bacilli may be found in sputum, milk, urine, tubercular abscess of the skin, joints, etc., and also in intestinal discharges. In cases of these discharges, one may discover the bacilli in analyzing the coating of the feces, the liquid of an evacuation obtained by a clyster, given a few minutes after a thorough washing of the intestines by careful but extensive enema. At other times it is necessary to analyze much of the fecal matters, and frequently to repeat the operation. Hundreds of other germs may be found in these matters.

The Durability of Apochromatic Lens. As is well known by microscopists, whatever advantage an apochromatic lens has over the ordinary kind, its durability seemed inferior to the latter. At least it has been the experience of several workers to find themselves with a depreciated, costly lens, which previously did splendid work. The director of this laboratory had this experience with one of Zeiss's apochromatic lenses. Prof. Burrill, of Illinois, in a paper read before the American Society of Microscopists, gives the following experience on the subject:

Last

"I have now to add a word in regard to the durability of the apochromatic, the want of which has been frequently questioned. After about two years' use it became evident that this lens was in some way impaired, and by looking through it from the back with a magnifier, a hazy-granular appearance was noticeable, not due to dust on the back lens. March the objective was sent to the makers for examination and repair. It reached me again in July, as good as new, with the statement that the front lens had been slightly decentered, and that the repair had been easily made, and was without charge. I have no other information upon this point, neither do I know what interpretation to place upon the granular appearance noted. There is certainly nothing of the kind visible now."

TECHNIQUE.

To Extract Ptomaines from Urine. -The necessity of more frequent analysis of urine from a diagnostic standpoint, is made more and more apparent every day. The following is an additional argument in that line:

In a note to the Academy of Science, Paris, Mr. A. B. Griffiths presents the following method of extracting ptomaines from urine in certain infectious maladies: "A considerable quantity of urine is alkalinized by the addition of a little carbonate of soda, and mixed afterward with half its volume of ether. After deposit and filtration, the ether is shaken with a solution of tartaric acid, which fixes on the ptomaines to form soluble tartrates. After evaporation of the dissolved ether, the acid tartaric solution is again alkalinized by carbonate of soda and shaken with half its volume of ether. This ether solution is allowed to evaporate spontaneously. The ptomaines remain as residue.

It

"a. Scarlet Fever: The ptomaine thus extracted from urine in case of scarlet fever is a white crystalline substance soluble in water, slightly alkaline. forms a crystallized chlorhydrate and a chloraurate. Phosphomolybdique acid produces a white yellowish precipitate; phosphotungstic acid gives a white precipitate; picric acid, a yellow precipitate. It is also precipitated by Nessler's solution. The chemical formula of this ptomaine is as follows: CH12AZO1.

"Pure cultures of micrococcus scarlatina, gave the same ptomaine by Gauthier's method.

"b. Diphtheria: The urine of diphtheritic cases is also a white crystalline substance. It gives a chlorhydrate and a chloraurate. Tannic acid precipitates it yellow; phosphomolybdic, white; picric acid, yellow; and Nessler's solution, brown. Formula: CHAZO". The bacillus diphtheria No. 2, Klebs and Loeffler, gives the same ptomaine in pure cultures.

"c. In a case of congestion of the kidneys, the parotid glands and the sub-maxillary glands, a ptomaine, crystallizing in prismatic white needles, was formed. Formula: CH1Az O2. It is very poi

As doubtless most of you are aware, the Sanitarium, although opened in 1866 and incorporated in 1867, began its real work at its re-organization in 1876. At this time, the institution was placed under a new management; new plans and methods were adopted, a broader and more liberal policy was introduced, and the medical management was placed upon a thoroughly rational and scientific basis.

At the time of its re-organization, the assets of the institution were about $45,000, most of which had been contributed by the friends of the institution and subsequent to its incorporation, as nondividend paying stock. The stock was issued in twenty-five dollar shares, and was purchased in amounts ranging from one to forty shares, by the friends of the enterprise. The total number of stockholders at the present date is 642.

The purpose of the promoters of this enterprise, as defined in the By-laws, was the treatment of the sick by rational methods and the promotion of sanitary reforms.

At its first organization, it was expected that small dividends would be rendered, but within a year after the incorporation of the institution, the stockholders voted to devote their dividends to charitable and benevolent purposes, the increase of the facilities of the institution, the treatment of the sick poor, the diffusion of sanitary knowledge, etc.

All stock issued since the time referred to has contained a clause equivalent to the above, and all dividends have been used as indicated. It will thus appear that the institution is simply a self-supporting charitable enterprise. All officers, managers, and other persons connected with the institution, are employed upon moderate salaries. Not a single dollar of the earnings of the institution has ever been distributed to stockholders, or to any other persons, except to objects of charity. Each year a portion of the earnings of the institution has been devoted to improvements, repairs, etc., and as large a sum as possible has been appropriated annually to the charitable treatment of the sick poor. This feature of the institution has steadily increased from year to year within the last fifteen years, until, in the last year, from Oct. 1, 1890, to Oct. 1, 1891, a total of more than $32,000 was reached. This sum includes not only those cases treated in the Sanitarium proper, but also expenses incurred in the support of free

The amount ex

beds in the Sanitarium hospital. pended during the last year in this way was somewhat injudiciously large, as it left nothing from the net earnings for repairs and improvements; nevertheless, it is hoped that by the exercise of greater economy in some departments, the same or nearly as large an amount may be expended during the coming year. In fact, the plans which the managers have made with reference to charity treatment for the year to come are considerably more liberal than for the year past. The conditions upon which charity patients are received, are chiefly as follows:

1. Only worthy, indigent, and friendless persons will be received as free patients.

2. No one will be admitted without proper credentials and recommendations.

3. No one will be admitted for more than three months at a time, and each month the Medical Superintendent in charge shall examine the patients with reference to the propriety or necessity of their prolonged stay, and shall sign a statement of the patients' condition, with a recommendation.

4. Each patient must deposit, on entering the hospital, a sum sufficient to defray his expenses home, and also sign an agreement to leave the institution when requested to do so by the Board of Managers. 5. Incurable persons will not be received, or if received, will not be retained.

6. Insane persons, epileptics, and patients with offensive, loathsome, or contagious diseases, will not be received.

7. Persons wishing to enter the hospital must make arrangement by correspondence beforehand.

The total number of patients who have been made the recipients of charitable treatment during the last two years, has been 837. This is, of course, a much smaller number than is treated gratuitously at many hospitals, but it should be remembered that the institution has no source of income whatever except from its own earnings, and consequently its charitable work must be, to a degree, limited. It should also be taken into consideration that the expense of caring for patients in a Sanitarium is very much greater than for hospital cases. A few items of expense will make this point clear: The number of employees in the institution, during the last two years, has varied from 325 to 365. As stated above, the salaries are small, and yet the total sum is by no means inconsiderable, amounting, for the last two years, to $115,657.62. During the greater part of the year, the milk supply of the institution amounts to from ten to twelve hundred quarts daily. The expense for fuel during the cold season, is from forty