EXPERIMENTS ON TUBERCULOSIS.

By GEORGE DEAN, M.A., M.B., C.M., Bacteyiologist, Serum Department, Jenner Institide of Preventize Mediciihe ; and CHARLES TODD, M.D., D.P.H., Assistant Bactwiologist, Sel-urn Depavtment, Jenner Institute of Pyeventiw Medicine.

THE main object of these experiments, which were carried out a t Lord Lister's suggestion and under his direction, was to ascertain whether the tubercle bacillus of human origin undergoes any marked change in virulence for the bovine species by passage through certain other animals.

The animals employed for this passage were the pig, cat, rabbit, rat, and mouse.' The material empIoyed was the sputum from well- marked cases of human pulmonary tuberculosis. The sputum, after microscopical examination to determine the presence of the tubercle bacillus, was administered to the intermediate animals either by injection or by feeding.

These intermediate animals were at first tested with tuberculin, but in view of certain objections, e.g. the possible creation of immunity, in some of the smaller animals the method of controls was employed subaequen tly.

We may here note that during the course of the experiments no single control animal was found to be tuberculous.

The intermediate animals after contracting tuberculosis were either allowed to die, or were killed on their manifesting signs of a severe infection and approaching death.

A post-mortem examination was made, and the most marked foci of infection were used for the inoculation of the calves.

A considerable number of intermediate animals of each of the species was used (wide Table), and the tuberculous material for the inoculation of the calves was taken from those animals first suc- cumbing to the disease, in the hope that the early death of the animals was some index of the virulence of that particular race of the bacilIus for the species in question.

In thc case of the mouse we failed to cause tuberculosis by injection of sputum, as the animals almost invariably died from the adventitious organisms present in the rnaterinl. Twelve mice injected with a virulent pure culture of human tubercIc, eight months ago, are alive and apparently healthy.

Over 100 animals were injected.

EXPERIMENTS O N TUBERCULOSIS. 459

The calves which had been previously tested with tuberculin (see Charts and Tables in Appendix) were injected with an emulsion of the diseased portions of the most seriously infected organs of the intermediate animals ; this emulsion being always proved microscop- ically, and in some cases experimentally, to contain tubercle bacilli.

About four months after the injection the calves were tested with tuberculin, killed,'and submitted t o a rigorous post-mortem ex- amination.

E:xPERl~E,vis.-General precautions.-To eliminate the possibility of the animals having been previously infected with bovine tuberculosis, and to avoid the possibility of their being so infected during the course of the experiments, certain precautions were adopted.

During the whole course of this research no esperiments on bovine tuberculosis were carried out in the department, nor had the attendants anything to do with other cattle.

Buildings provided for the experinwnts.-For the pigs two styes were provided; one of these was quite new ; in the other only one pig had been housed during the past five years, and this animal at the post-mortem examina- tion was found to be free from tuberculosis. This sty was thoroughly disinfected with crude carbolic acid (1 in ZO), and was limewashed with a carbolised wash.

Three new wooden sheds which had never been used for the housing of animals were provided for the other experimental animals. One of these was arranged for the small animals to be employed in the first part of the experiment. A second shed was employed for receiving the calves on their arrival, and for carrying out the tuberculin tests, and a third for receiving the calves which had passed the tests, and had beeh 'adapted for the experiments.

All these sheds were at some distance from'other buildings, and the third was situated in the middle of a large meadow on which no cattle had grazed for a t least five years. Two acres of this meadow round the shed were fenced off to form a paddock for the experimental calves. No cattle could come near the paddock.

At one side, which was formed by a hedge, a double fence was made to prevent the horses which were in the neighbouring field from coming near the isolated area. The use of such an open-air method of treating the animals was no doubt in their favour in resisting infection.

The shed used for carrying out the tuberculin tests, after every test was thoroughly washed with crude carbolic (1 in ZO), then freshly limewashed, the wash containing about the same proportion of carbolic acid. The whole shed was moved on to a fresh foundation.

The calves after admission to the testing-house were tested in three separate groups. The temperatures were taken night and morning for three or four days, and were also taken immediately before the injection of the tuberculin {vide Charts and Tables).

0.5 C.C. of Koch's tuberculin (Dr. Libhertz) was injected, and the tempera- ture was then taken on the third, sixth, ninth, twelfth, and fifteenth hours after the injection. A temperature of 104", or above, was regarded as a reaction, and the animals giving this reaction were rejected.

Where any member of a group reacted, the animals which were retained for the experiment were regarded as '' contacts," and kept isolated, and were again tested after the lapse of about a month, the precautions mentioned in regard to the testing-house being rigidly enforced.

The first group consisted of five animals, and, as will be seen from the charts, two of these reacted,

3 1 - ~ ~ OF PATU -vor,. viii

460 GEORGE D E A N A N D CHARLES TODD.

The second group consisted of ten calves. One calf of 3 weeks old died from acute gastro-enteritis. The other calves were 3 to 4 months old, and of these five gave a reaction, a sixth was rejected on account of its unhealthy appearance.

Three were retained for the experiment and passed a second testing. The third group, consisting of six calves about 4 months old, was obtained from a dairy farm where all the cows are tested with tuberculin. As will be seen from the Charts and Tables in the Appendix, none of these six calves gave a reaction. Under these circunistances it was considered unnecessary to regard these animals as contacts, and in their case no second tuberculin test was carried out. Although we were aware of the possibility that some degree of immunity might be established by the tuberculin injected, yet in view of the fact that we had to deal with animals of uncertain breeding and history, we considered it essential to carry out the tuberculin tests as described. If the experiinents were to be of any value, criticism as to the possible existence of a naturally occurring bovine tuberculosis had to be carefully forestalled. The matter resolved itself into the choice of the lesser of two evils. The chief interest of the testing is the large number of animals in the first two groups giving a reaction, and the absence of any reaction in the third group.

I n the case of hand-fed animals, when the milk from a number of cows is mixed, the presence of one animal giving milk containing tubercle bacilli gives rise to the possibility of a widespread infection. The third group seems to point to the value of the tuberculin test from the breeder’s point of view.

Tuberculin tests after t?/e inoculation.-These were carried out about four months after the injection of the tuberculous material. The charts of all t he calvea, both those utilised for the experiments and those rejected, are appended.

Morphology of the bacilli-The microscopical examination of the sputa from different cases showed that, in certain of these, fairly definite special morphological characteristics of the bacilli could be. determined. I n one particular sputum the predominant form of the bacillus was very long and beaded, whereas in others a type of medium length seemed bo be most frequent.

With the view of ascertaining whether these types persisted in cultures, and in the tissues of the experimental animals, very numerous microscopical examinations were made, but neither in the culture nor in the animal body could this persistence be demonstrated. For example, in preparations from cultures of the very long beaded bacillus referred to, in one field of the micro- scope, short, medium, and long bacilli were present. Nor in the experimental animals could we find that there was a uniform tendency for the bacillus to approximate to any one particular type.

IXTERMEDIATE ANIMALS.

Apart from a large number of the intermediate animals which died from infections due to the presence of adventitious organisms in the sputum, e.g. the pneumococcus, Streptococcus pyogenes, etc., the following is a statement of the animals which died of tuberculosis, either local o r general :-

In the tables the lethal period, the source of the infective material, and the nature of the infection are shown.

The data in regard to pigs will be found in the table in the section on Pigs.

EXPERIMENTS O N TUBERCULOSIS.

I

461

I 1:

Animal. ‘ Lethal Period. I - _ - -~

Lesions. I i I Sputum Eniploj 4.

SUBCUTANEOUS INJECTION. I __ - .

General tuberculosis. , t

Loc2 General ,, Local ,* General ,,

.- -

Average lethal period . . . 95 days Maximiim ,, ,, 185 ,, Minimum ,, ,, . . 45 ),

___ - - _ _ -- - - _ _ -

INTRAVENOUS INJECTION.

Rats. - _ _ _ - _ _ _ _

Lethal Period Sputuni Einplo) ed. Lesions.

SUBCUTANEOUS.

1 i Animal. -

__ -

Patient E.

:: R.

General tuberculosis. 1 , ,,

, 9

,

Average . . . . . . 127 days Maximum . . . . . . 146 ,, Minimum . . . . . . 111 ,,

ISTRAPERITONEAL.

. - _ _ _ _ - _ _ - _ _

- -

General tuberculosis.

, I

Average . . . . . . 106 days Maximum . . . . . . 184 ,, Minimum . . . . . . 73 ,,

I

.

465

so. 1' 13 days. Patient S.

, I 2 129 ,> ,, R.

GEORGE B E A N A N D CHARLES TODD.

Tubercle bacilli in IocaI lesion, luru- bar glands, and in spleen pulp.

Local tuberculosis.

Cats. - -_ . - _ _

! Aniniai. Lethal Penotl. Syutuin EmpIoyed. L e 8 ion 8.

DETAILS OF EXPERIMENTS ON CALVES.

Expel-iment on Calf No. 2.

Sputum, from a case of advanced phthisis (Patient H.), mas found to contain numerous tubercle bacilli ; wphologically, the organism in the sputum showed long forms frequently beaded and segmented. This type was constantly observed in numerous examinations of the sputum.

CuLTURALLY.-After passage through the guinea - pig the bacillus grew very freely on glycerin-potato, and on coagulated horse and dog serum (with and without glycerin). The cultures on glycerin-potato showed rather more yellowish-orange pigment than most other races grown on the same batch of glycerin-potato tubes.

MoRPHoLoQIcALLY.-The bacillus from these cultures presented all the types usually found in the case of the tubercle bacillus, and no preponder- ance of the long beaded form, or indeed of any other, could be ascertained.

White female, of the same litter as the three feeding and the three control pigs. Weight, 35 lb. This animal was not tested with tuberculin, as it was one of a litter bred from a healthy sow, and as three pigs of the same litter were reserved as controls, and these when killed five months later were found to be free from tubercle.

The material used for inoculation was the same sputum (Patient H.) that was employed in the case of pigs No. 4 and No. 6. 2 C.C. of this sputum was injected into the left thigh on October 29, 1901. There was a slight swelling at the site of inoculation, and this was followed by the,formation of a nodule the size of a horse bean. The animal wasted, and, died on November 29, 1901, i.e. thirty-one days after the injection.

Average weight of three control pigs of the same litter was 53 Ib. At the site of inoculation in the left thigh was a nodule the size of a horse bean, in which a number of minute grey points the size of millet seed was visible.

Round the knee - joint the capsule was thickened, and contained small caseating areas. I n dissecting the thickened capsule of the joint a smaIl abscess cavity was found, containing about half a drschm of thin pus showing numerous organisms-bacilli and cocci. The abscess did not communicate. with the joint cavity. In the infiltration round the knee-joint tubercle bacilli were present in enormous numbers. Many of these were very long, but beading was not a marked feature. The inguinal glands on this side were greatly enlarged, forming a mass the size of a large walnut, with projections the size of horse beans. This mass was congested at parts, and on section

PASSAQE OF HUMAN BACILLUS (PATIENT H.) THROUQH THE PIQ (NO. 7).-

POST-MORTEM EXAMINATION (Pro 7).-Weight, 24 Ib.

EXPZRhJfZ+NTS ON TUBERCI/J;USIS. 463

showed numerous millet-seed-iike nodules, io which tubercle bacilli were demonstrated. The lumbar glands on this side presented a eirnilar appearance, and were about the same size as the inguiiial (glands. Wistologically, both inguinal and lumbar glands showed beautiful giant cells containing namerous tubercle bacilli.

The liver, spleen, and kidneys appeared on naked-eye examination t o be iiormal.

The lungs were congested and had pnevmonic patches scattered through them, but no definite tubercrtlolis nodides could be made out on naked-eye examination.

The bronchial glads appeared to be slightly enlarged, the tOn8iIS normal.

K~STOLO~~CAL ~ X A M I X A T I O N . - ~ i i e affected joint capsule was swarming with tubercle bacilli. The inguinal and lumbar glands showed giant cells which had less branching. a t the periphery than in the case of pig No. 6, and showed no acid-fast property as in the case of pig No. 6. Tubercle bacilli were frequently present within these.

Tubercles could be seen in the ludgs OD microscopical examination, and these showed numerous tubercle bacilli.

The liver, spleen, and kidneys showed no tubercle. Pure cultivations of the tubercle bacilfus were not obtained from this case. Inoculation of Caif.-Bed heifer, at. I I months at the time of injection.

This animal had been twice tested with tuberculin, and had faiied to react an both occasions (vide Tubercrilin Tests and Previous History).

Received on 29th November 1901, on the right side of the neck in front of the shoulder, deep in the muscle, 35 C.C. of an emuIsian of the inguinal gland of the above pig (No. 7). The microscopicdl examinatian of this cmrdsion showed numerous tubercle bacilli. Beyond the slight swelling resulting from the injection, this animal sho\sred no symptoms. Zt was tested svith tuberculin 131 days after the injectiau, the highest temperature reached

‘IXS calf (Xo. 2) was killed on 2 4 t h Ali~il 1902 (136 days after the iiijection). S o definite macroscopic lesion codd be found, though perhaps tbere was a slight cicatricial thickening of the interniuscuIar septa in the inoculated region. A careful microscopical examination of the Iyniphatic glands ant1 viscera gave RO evidence of tubererrlaus infection.

beiog 105‘.5.

Ezperiment CaviVo. 3. Sputum from a case of advanced phthisis (Patient ft.) containing numerous

tubercIe bacil ti. MORPEOLOGY OF BAcrLLoa IN SP(ITUM.-Of medium size with marked

beading--a few very long forms observed. CULTURALLY.-After passage through t h e guinea-pig it grew with great

freedom on glycerin-serum agar and on glycerin-potato. A considerable degree of orangeyellow pigmentation was present, though

this was not so marked as in some of the other races. PASSAGE OF THE HUMAN B~crctus (PATIENT R.) THROUQII THE Pra-Black

male ; weight, 19 Ib.; was tested nine days before inoculation with tuberculin, and gave no reaction, The material used far irracukAion was sputum from the same ease (Patient R.) from which the matetiat was obtained from pig No. 2. 5 C.C. of a thick emulsion of this wz33 injected into the abdominal wall on tb8 23rd August 1801. Slight swelling tOak place at the site of hocdation, which, within a month, gave place to a small naduIe the size of a large pea;. The animal wasted and died on the 9th October 1901-€orty-seven days after inoculation.

POST-MORTEM EXAMINATION (PIG Xo. Jj.-Weight, 165 lb.; body much emaciated.

464 GEORGE DEAN AND CHARLES TODD.

Local lesion.-At the site of inoculation beneath the skin a hard nodule the size of a large pea could be felt, and on cutting into this it was found to have undergone caseous degeneration. Behind it there was a plaque of caseou'S material the size of a shilling.

The lumbar glands on this side were enlarged to the size of horse beans, and several of these had caseous points the size of millet seed.

Lungs.-The upper and middle lobes were consolidated, and portions did not float in water. They were of a deep reddish-purple colour, with yellow patches scattered throughout the consolidated areas. Bronchial glands enlarged.

Abdomen.-A few strands of fibrinous lymph on the peritoneal surface of the gut.

Liver and spleen.--Both studded with pale grey nodules the size of millet seed.

Kidneys.-A few tubercles the size of millet seed. With the exception of the lumbar, bronchial, and perhaps the mesenteric,

HISTOLOGICAL ExaMINATIoN.-The nodules above described in the organs

CuLTrvATIoN.-Nodules from the liver placed on glycerin-potato tubes gave, These had

The bacilli, on subculture, gave a luxuriant

Guinea-pigs inoculated from these pure cultures died of general tuberculosis. Inoculation of Calf.-Small Alderney heifer, 12 months old a t the time of

inoculation. This animal had been twice tested with tuberculin, and on both occasions failed to react (wide Tuberculin Tests).

An emulsion from the first culture generation on glycerin-potato was used for the inoculation of this animal. The emulsion was opaque, and was found microscopically to contain very numerous tubercle bacilli. 34 C.C. of this emulsion was injected into the left external jugular vein on 2nd December 1901.

A guinea-pig, weighing 360 grms., which received subcutaneously 0.2 C.C. of this emulsion, died in sixty days of very extensive general tuberculosis. The calf showed no signs of illness, and was tested with tuberculin 131 days after the injection, the highest temperature recorded being 104" (vide Chart).

This calf (No. 3) was killed on 14th April 1902, 136 days after the injection.

POST-MORTEM OF CaLF.-The only lesions found were several small pale grey areas, the size of a millet seed, in the liver. Similar areas of doubtful nature were observed in certain of the glands.

MICROSCOPICAL EXAMINATION of the suspected tissues showed no tubercles or tubercle bacilli.

Guinea-pigs (4) inoculated from liver, lung, cervical, and omental glands remained healthy.

the glands did not appear to be markedly enlarged.

and lymphatic glands showed the presence of tubercles and tubercle bacilli.

in thirty-one days, very distinct colonies of the tubercle bacillus. a marked orange-yellow colour. growth on all the ordinary media used for the tubercle bacillus.

Experiment on Calf iVo. 4. I n this experiment the sputum used was the same as in the case of calf

No. 3 (Patient R.). PASSAQE OF THE HUMAN BACILLUS (PATIENT R.) THROUGH THE RABBIT

(No. 6).-A sandy and white male rabbit, weighing 1050 grms., which nine days previously had been tested with tuberculin and had not reacted, was injected subcutaneously (on 23rd August 1901) in the abdominal wall with 1 C.C. of a thick emulsion of the sputum. An abscess developed, which per- sisted until the animal was killed on 4th December 1901.

At the site of inoculation in the abdominal wall was a nodular swelling the size

POST-MORTEM EXAMINATION OF RABBIT So. 6.-Weight, 1400 grms.

EXPERILVENTS O N TUBERCULOSIS. 465

of a walnut, and from it a small sinus opened externally. Section of the mass showed that it contained curdy pus, in which tubercle bacilli were demonstrated. The inguinal glands on the same side as the local lesion were the size of a pea and contained tubercle bacilli. The lungs had numerous tubercles scattered through them, the size of a millet to a hemp seed; a t points these had run together, forming plaques 1 in. in diameter, and almost cartilaginous in appearance and texture.

Liver and spleen each had a few nodules the size of hemp seed. Axillary, cervical, lumbar, mediastinal, and bronchial glands all enlarged.

Inoculation of CaV-An emulsion of the affected areas of the lungs of the rabbit was prepared by grinding these in a mortar with normal saline solution and filtering through wool. 80 C.C. of this emulsion, which contained numerous tubercle bacilli, was injected into the muscles of the neck of calf No. 4, roan heifer, aet. 5 months, on 4th December 1901, which had previously given no reaction on being tested with tuberculin.

Beyond slight swelling at the site of injection, the animal showed no symptoms. It was tested with tuberculin 139 days after the injection and failed to react, the maximum temperature recorded being 103O.2 (wirle Chart).

This calf (No. 4) was killed on 24th April 1902, 141 days after the injection.

POST-MORTEM OF CALF No. 4.-All the organs were examined micro- scopically and macroscopically, and no lesions were found.

A guinea-pig inoculated with an emulsion of the lymphatic gland nearest the point of inoculation did not develop tuberculosis.

Experintent o n Calf ATo. 5 . This is practically a duplicate of the experiment on calf S o . 4, sputum

from the same case being used, and also being passed through a rabbit. PASSAQE OF THE HUMAN BACILLUS (PATIEST R.) THROUQH THE RABBIT

(No. 5).-A sandy male, weighing 950 grms., which six days previously had been tested with tuberculin and shown no reaction, received subcutaneously on 20th August 1901, in the abdominal wall, 1 C.C. of an emulsion of the sputum (Patient R.). A swelling, followed by ulceration, resulted at the site of inoculation. The animal was killed on 4th December 1901.

At site of inoculation a plaque the size of a penny was present. This contained tubercle bacilli. The appearances were identical with those of the rabbit in the previous experiment, except that the liver and spleen were free from visible nodules.

Inoculation of Calf.-An emulsion was made fmni the lungs of the infected rabbit, and 60 C.C. of this emulsion was injected deep into the neck muscles of calf No. 5, red and white male, aet. 5 months, on 4th December 1901. This animal had been previously tested with tuberculin and had failed to react.

Before injection the emulsion was examined microscopicdly and found to contain many tubercle bacilli.

The calf showed no symptoms. It was tested with tuberculin 139 days after the injection, with the result that the temperature reached 104O.4 (vide Chart).

This calf (No. 5 ) was killed on 24th April 1902, 141 days after the injection.

POST-MORTEM ON CALF No. 5.-At the site of inoculation was a nodule, which was about the size of a hazel nut, and was found on section to have undergone caseous degeneration, and to contain tubercle bacilli. All the organs were found on microscopical examination to be healthy.

A guinea-pig inoculated with material from the site of inoculation contracted a general tuberculosis.

POST-MORTEM EXAXINATION (RABBIT No. 5).-Weight, 1440 gms.

466 GEORGE D E A N AND CHARLES TODD.

Experiment OR Gulf Avo. 6.

In this experiment the sputum used was the same as in the caw of calves Nos. 3, 4, and 5.

PASSAGE OF THE HUMAN BACILLUS (Patient R.) THROUGH THE WHITE RAT ( X O . 32).-A white rat, weighing 93 grms., which had been tested with tuberculin thirteen days previously, and had given no reaction, was injected intraperitonealIy with I C.C. of an emulsion of the sputum on 27th Augrrst 1901. He died on 4th December 1901, i.e. ninety-nine days after the iiijection.

opening the abdominal cavity, the cd iac lymphatic glands were seen to be enlarged to the size of peas. Both lungs had scattered through them very numerous, minute, pale yellow tubercles, which were only just visibIe to the naked eye. Histological examination revealed the presence in great numbers of very slender long tubercle bacilli.

fnoculation of Ca@--Red male, st. 5 months, which had failed to react to tuberculin. Received, on 4th December 1901, 40 C.C. of an emulsion of the lungs of the above rat, deep in the pr~scapular muscles. Beyond slight sweIL ing, the animal showed nothing locally, nor could iiny glandular enlargement be made out.

This calf WGS tested with tuberculin on 22nd April 1902, i.e. 139 days after the injection, the maximum temperature recorded being 105"-8 (vide Chart).

The calf was killed on 24th April 1902, i.e. 141 days after the in- jection.

PosT'-&Tom&&f OF CALF No. 6.--Deep in the tnnscles of the neck there was 3 thin layer of pale jd low cnseous-Iooking material, containing semi- calcareous nodules the size of millet seed, exhibiting tubercle bacilli.

I n such a lesion as this the question naturally arises as to whether the tubercle bacilli found were not those originally put in, but this appears unlikely from the number of the bacilli found, and from the presence of the semicalcareous nodules.

No other lesion could be discovered, either on macroscopical or micro- scopical examination. A guinea-pig inoculated with a small quantity of an emulsion of the material from the site of inoculation contracted a general t ubercul osis.

Eqe r imen t on Calf No. 7.

pO3T-%rORTEM EXABIINATION OF R A T NO. 32.-Weight, 162 grms. 011

In this experiment sputum from a case of advanced phthisis (Patient S.) was used. The bacilli in the sputum were numerous, of medium length, and with very distinct beading.

CumURALLY.-After passage through the guinea-pig the bacillus grew freely on glycerin-potato, and the growth showed a considerable amount of pigmentation.

PASSAGE OF HUMAS BACILLUS (Patient S.) TBROUQX TBE Warm RAT (No. 30).-A white rat, weighing 100 grms., was injected intraperitoneally, on 37th August 1901, with 1 C.C. of an emulsion of the sputum (Patient S.). It died on 6th December 1901, i.e. 101 days after the injection.

POST-MORTEM EXAKISATKON (RAT No. SO).--Weight, 90 grms. The peritoneum appeared to be perfectly healthy. There were no tubercles to be made out in the liver, spleen, or kidneys. The lungs had scattered through them numerous grey tubercles, from the size of a millet to the size of a hemp seed. These contained numerow tubercle bacilli, long and slender, and some markedly beaded.

Had previously failed to Inoculation of CaV-Red heifer, at. 5 months.

E X P E R I M E N T S O N TUUEKCULOSIS . 467

react to tuberculin. Was injected on 6th December 1901 with 20 C.C. of an emulsion from the lungs of the above rat, The inqjection was made into the praescapular muscles. The emulsion contained numerous tubercle bacilli. The animal had a transitory local swelling. It was tested with tuberculin 137 days after the injection, the maximum temperature reached being 102”.8, and was killed on 24th April, i.e. 139 days after the injection.

POST-MORTEM EXAMINATION (CALF KO. 7).--Neither macroscopically nor iiiicroscopically could m y tuberculous lesions be found. A few suspicious pale yellow points were found in the liver.

Two guinea-pigs inoculated with an emulsion of the liver containing these suspected areas did not develop a tuberculosis.

Expc?ime?it o n Calf No. 8. I n this experiment the sputum employed was that of Patient R., and this

experiment is a duplicate of the experiment on calf 6, except that in the present experiment the injection of the rat was subcutaneous and not in traperitoneal.

PASSAGE OF HUMAN BACILLUS (PATIENT R.) TEROUQH THE RAT (WHITE RAT No. 9).-Weight, 180 grms., which had been previously tested with tuberculin, received, on 23rd August 1901, 2 C.C. of an emulsion of the sputum (Patient R.) subcutaneously in the abdominal wall. The animal died on 12th December 1901, i.e. 111 days after thc injection.

POST-MORTEM (RAT No. 9).-Weight, 125 grms. The site of the inocula- tion was only just visible as a slight thickening of the abdominal wall. Peritoneum normal. No visible tubercles in the liver, spleen, or kidneys. Inguinal, axillary, and mesenteric glands normal. A mass of glands the size of a horse bean under the jaw. The surface of each lung showed through the pleura about a dozen tubercles, each the size of a millet seed, and on section numerous similar tubercles were seen scattered through the substance. These contained numerous tubercle bacilli.

Inoculation of CuZf-Red heifer, s t . 5 months. Had been previously tested with tuberculin, and had failed to react. Was injected deep in the muscles in front of the shoulder with PO C.C. of an emulsion of the lungs of the above rat on 12th December 1901. This emulsion contained abundant tubercle bacilli. This animal showed a distinct swelling a t the site of inoculation. This swelling diminished in size, but was still palpable when the animal was killed.

The calf was tested with tuberculin 151 days after the inoculation, the maximum temperature reached being 105O.4, and was killed on 16th May 1902, i.e. 155 days after the injection.

posT-&foRTEhf (CALF KO. 8).-At the site of inoculation in the neck was a nodule the size of a hazel nut ; it had undergone caseous degeneration and appeared to be softening. One or two minute grey tubercles, showing slight caseation, were visible in the prsscapular glands. The organs appeared healthy, and no further lesions could be found.

MICROSCOPICAL EXAMINATION. - The local lesion contained tubercle bacilli.

A guinea-pig inoculated with material from one of the prsscapular glands on the inoculated side contracted a general tuberculosis.

Experiment on Calf No. 9. I n this case sputum from the PATIENT R. was used. PASSAGE OF HUMAN BACILLUS (PATIENT R.) THROUGH THE CAT.-A male

cat (black clip, No. 4), weighing 1800 grms., received, on 23rd August 1901, 3 C.C. of an emulsion of the sputum (Patient R.) subcutaneously in the abdominal wall. The injection WRS followed by an ulcer which persisted till 14th

468 GEORGE DEAN AND CH;QRLES TODD.

October, but at that date showed signs of healing. The animal died on 30th December 1901, i.e. 129 days after the injection.

Body considerably emaciated. At the site of inoculation there was an area of infiltration the size of a florin, with numerous nodules, the size of hemp seed, scattered through it. No other lesion could be found, either on macroscopic or microscopic examination.

Inoculation of Calf.-Red heifer, at. 5 months, which had failed to react to the tuberculin test, was injected in the left side of the neck and the left groin, on 30th December 1901, with 5 C.C. in each place of an emulsion of the local lesion in the cat. This emulsion contained a fair number of tubercle bacilli.

This animal showed a small Itodule at the site of injection in the neck, and this nodule persisted until the animal was killed. The calf was tested with tuberculin on 15th May 1902, i.e. 136 days after the injection, the maximum temperature being 104"*8 (vide Chart), and was killed on 18th May 1903, i.e. 139 days after the injection.

POST-MORTEM ON CALF KO. 9.-At the site of inoculation in the neck was a nodule the size of a hazel nut, which had undergone caseous degeneration, and resembled the lesion found in the previous calf. The organs appeared healthy, and no other tuberculous lesiom could be found.

POST-MORTEM ON CAT No. 4.-Weight, 1130 grms.

In these tubercle bacilli were demonstrated.

MrcRnscoPrcAL Exa~rrNaTroN.-~he local lesion contained tubercle bacilli.

Expcrimeiit O?L Calf A'o. 10.

In this experiment the sputum of the patient H. was employed (for description of bacillus, etc., vide Experiment on Calf 2).

INJECTXON OF Pm No. 6.--Blsck male; weight, 30 lb. Tested with tuberculin seventy-six days before injection, and gave no reaction. The material used for inoculation was the same sputum (Patient H.) 88 in pig No. 4 ; 2 C.C. of this undiluted was injected into the left groin OR 23rd August 1901. A swelling occurred, which went on increasing until the time of death. The animal wasted, and died on 16th January 1902, i.e. 146 days after the injection.

Body greatly emaciated. At site of inoculation, in the left groin, mas an abscess cavity the size of a cocoanut, containing a thin grumous pus, in which were present numerous tubercle bacilli. The lymphatic glands in the groin were enlarged to the size of a walnut, and were caseating. On opening the abdominal cavity there were seen, running in front of the lumbar vertebrae and from the affected inguinal region, a chain of enlarged lymphatic glands, several of which had reached a walnut size, and bad undergone extensive degeneration. The cervical and axillary glands on both sides were slightly enlarged. The mesenteric glands shoaed no marked enlargement. The lungs had scattered through them an enormous number of small grey tubercles, from the size of a millet to the size of a hemp seed. There were small pneumonic patches surrounding these. Liver and spleen had numerous similar grey nodules. In the kidneys the tubercles were lees evident, but were present as minute grey points.

HISTOLOGICAL EXAMrNATfoN.-The inguinal and lumbar glands showed magnificent giant-cell systems ; the giant cells appeared to have undergone a hyaline degeneration, the whole body of the cell having become slightly acid- fast (contrast condition of cells in pig KO. 7). The nodules in the lung con- tained tubercle bacilli, but these were not numerous. The liver and spleen showed the structure of tubercle.

CULTIVATION.-PUre cultures of the tubercle bacillus were obtained from the liver and spleen. These cultures on glycerin-potato had less of the pale orange-yellow pigment than those similarly obtained from pig No. 3.

POST-MORTEM EXAXINATION (Pro No. 6).--Weight, 24 Ib.

EXPERIMENTS OiV TUBERCULOSIS. 469

Iuoculation o j Calf.--Red and white bull, aet. 6 months ; liad been twice tested with tuberculin, and had failed to react. Was injected, on 16th January 1902, in the left side of neck and left thigh with an emulsion from the lungs and a caseous gland of the above pig, 50 C.C. of the emulsion being injected in each place. In addition to this he received 20 C.C. of pus from the abscess cavity of the pig on the inside of his right thigh. 150th the emulsion and the pus injected contained numerous tubercle bacilli. The injection of pus into the thigh was followed by a very large abscess, which was opened and allowed to drain. The pus of this abscess Contained numerous tubercle bacilli. The abscess healed rapidly, and when the animal was killed only a cicatrix could be found. This calf was tested with tuberculin 139 days after the injection and reacted strongly, the maximum temperature recorded being 105"*S (vide Chart), and was killed on 5th June 1902, i.e. 140 days after the inoculation.

POST-MORTEM ON CALF Xo. 10.--Beyond the cicatrix in the thigh already mentioned, nothing could be found at the sites of inoculation, but a t the root of the neck, near the anterior opening of the thorax, a gland, the size of a hazel nut, with numerous yellow caseous and partly calcareous areas, was found. I n this animal no other lesion whatever could be found.

MICROSCOPICAL ~~xAMINATION.-The above - mentioned gland contained tubercle bacilli.

A guinea-pig inoculated from the gland developed a general tuberculosis.

Experimeitt o n C a v No. 1 1.

All other organs healthy.

This animal was inoculated directly with human tuberculous material, with- out its having been previously passed through another animal. For this purpose the sputum of a phthisical patient (Carr) was employed.

MORPHOLOGY IN Spmuni.-The bacilli were somewhat scanty in the sputum. They were of medium size, and showed no marked tendency to beading.

CuLTunaL-The bacilli, obtained from an infected guinea-pig, grew freely on the ordinary media, for the tubercle bacillus and pigmentation were not a marked feature.

Had been twice prc- viously tested with tuberculin, and on both occasions had failed to react; was injected, on 25th February 1902, with 20 C.C. of a thick emulsion of the sputum (Patient C.) in the left thigh. A swelling resulted a t the site of inoculation ; this continued to increase until 17th April, when it had reached a great size and was fluctuating. It was then opened, and there were evacuated about 3 to 4 pints of thin grumous pus, containing a fair number of tubercle bacilli, and numerous other bacilli and cocci. On plating this pus, Bacillus coli communis, Staphylococcus pyogenes aureus, and Streptococcus pyogenes, were identified. A guinea-pig of 400 grms., inoculated with the same sputum, died in seventy-seven days, with very advanced general tuberculosis. The calf was tested with tuberculin ninety-nine clays after the injection, giving a marked reaction, the highest temperature recorded being 106" (vide Chart), and was killed on 5th June 1902, i.e. 100 days after the injection.

POST-MORTEM ON CALF No. 11.-At the site of inoculation in the thigh, a nodule, the size of a hazel nut, could be felt through the skin, which was freely movable over it. On opening this, it was found to contain caseo-purulent materiaL This nodule proved to be the centre of an extensive tuberculous infection, which infiltrated the muscular septa over an area of about 3 to 4 in. On section, the infiltrated area showed irregular patches of caseation. The inguinal glands showed caseous points. There was a chain of enlarged lumbar glands, the largest the size of a walnut, on the same side. These, on section, were typically tuberculous in the stage of caseation. The glands on the opposite

Inoculation of Calf.--White male, aet. 7 months.

The abscess healed rapidly.

470 GEORGE DEAN AND CHARLES TODD.

side showed no evidence of tuberculous infection ; a few fibrous adhesions in the pleural cavities.

TWO guinea-pigs, inoculated with material from the lumbar glands of this calf, contracted a general tuberculosis.

GENERAL REMARKS OX THE EXPERIMENTS.

As regards the main object of this research, the experiments cited appear to demonstrate that the human tubercle bacillus is not markedly exalted in its virulence for the calf by a single passage through the pig, cat, rabbit, or rat.

A t the same time, tbe experiments show that the human tubercle bacillus is by no means innocuous to the calf, as the control animal injected directly with sputum contracted an extensive glandular tuber- culosis.

At first sight, the experiments might appear to show that passage through the intermediate animals had caused a diminution in the virulence of the bacillus for the calf, but i t must be remembered that in the one case the associated organisms had been eliminated by passage through the intermediate animals, whereas in the other case they may have played an important rSIe in aiding the attack of the tubercle bacillus.

The important r61e that these associated organisms are capable of playing in certain diseases is a matter of general knowledge ; one need only refer to the case of tetanus, when large quantities of living tetanus spores can he injected without causing the disease, but where the addition of other organisms to the spores enables these to develop and give rise to the disease. A p~opos of this, i t is interesting to note that the only pig in the series which failed to show tuberculous infec- tion of any so1 t was tbe anixnaI inoculated with tuberculous material obtained from another pig, and in this material the tubercle bacillus was unaccompanied by otber organisms.

The problem is also complicated by the possibility of the juices of the emulsified organs having played some part in the process.

As will be discussed in the special section on the infection of the pig, an important result wits obtained, as the experiments conclusiveIy prove that this animal is capable of contrncting a rapidly fatal general tuberculosis as the result of infection with the human bacillus.

OX THE RESULTS OBTAINED I N THE PIG.

In the course of these experiments on tuberculosis we obtained certain results in the case of the pig, which seem sufficiently important to merit a most detailed description.

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472 GEORGE DEAN AND CHARLES TODD.

Professor Koch, in the paper in which he formulated his recently expressed views as to the essential difference between the bacillus of human and of bovine tuberculosis, and as to the consequent slight danger to the human subject from ingestion of tuberculous material of bovine origin, based his views largely on certain experiments on various domestic animals.

As these experiments are fundamental and form part of the chain of evidence on which Professor Koch bases his views, it is important to ascertain whether different results might follow under slight modi- ficcztion of the experimental conditions.

Professor Koch, a t a later period, published the details of the experiments on which he based his views. We have recently come into possession of these detailed reports, and find that the results obtained by us in the case of the pig, especially by subcutaneous injection, differ materially from those obtained by him. As will be seen from the extract from his report, Professor Koch-using two pigs-was unable to produce infection with the human bacillus in either, by subcutaneous inoculation.

In our experiments, on the other hand, the six animals employed mere all infected with the human bacillus, and the character of the infection was a t least as severe as that described by Professor Koch as the result of inoculation of the pig with the bovine bacillus.

Professor Koch employed pure cultures of the bacillus, while in our experiments tuberculous sputum was used.

The difference in the results obtained is probably to be attributed to this fact, in which case the greater infective power of the tuber- culous sputum may be due to various factors, eg.-

1. In the sputum the bacillus probably retains its original virulence and has undergone no modification, as may be the case on cultivation.

2. The associated organisms or their products in the sputum may assist in the process of infection.

We are aware that Professor Koch’s experiments are comparative, but when the numbers used are so small-two animals for subcutaneous inoculation with human tubercle bacilli, and two with bovine tubercle bacilli-and when only one race of each type of bacillus is used, grave risks of error are incurred. One of these may be indicated, namely, that the race of human tubercle bacillus employed may have been one of comparatively low virulence.

With reference to the experiments on feeding with tuberculous sputum, Professor Koch obtained some form of infection in the three of the six animals employed by him.

In our experiments only three pigs were used, and all contracted a local but very definite infection, with great wasting followed by death. The tuberculous lesions appeared hardly sufficient to account for death or for the emaciation, but i t is to be noted that the animals

EXPERIMENTS ON TUBERCULOSIS. 4 i 3

were somewhat younger than those employed by Professor KO&, and that the feeding was commenced at the period of weaning when young animals are liable to catarrhal intestinal disturbances. Moreover, the sputum employed, having been sent by post, wa13 usuaIly tweaty-four hours old, and contained numerous putrefactive organisma and their products.

The following is a transbtioo of yrofessor Koch'e description of the results obtained by the subcutaiieous injection into pigs of tubercle Bacilli of human origin :-

PIO NO. 1.-A single injection of 2 C.C. of a suspension of bacilli of human tulterculosis-which was obtained by rubbing up I grm. of pure culture of the bacilli with 100 parts of water-under the skin of tha inner side of the left thigh.

Duration o j the exye~znzent.-l(illed after 135 days. Increase in 2oeigh,t.--53 kilos. CZiniea1.-At the site of injection there formed during the course of the

observation a nodule the size of a cherry stone; but neither rise of temperature nor any other change in the genera) condition was observed.

Pathological resubts.-A focus of calcification, the size of a bean, at the site of injection in the left thigh-otherwise all organs normal.

Micro.Tcopical.--In the calcified masses of the nodule, tubercle bacilli could be demonstrated.

INFECTION OF GIJINEA-PI~S.- Guinea-pigs injected with parts of the calcified site of injection contracted general tuberculosis.

PIQ KO. %--A single injection of 2 C.C. of a suspension of bacilli of human tuberculosis (dilution, 1 : 100) under the skin of the left thigh.

Duration of the ezperiment.--Killed after 100 days. Increase i ? h z~eigltt.-Z 1 kilos. ClinicaZ.-On the inner surface of the left thigh, at the point of inoculation,

there formed a nodule about the size of a hazel nut. From eight to fourteen days after the injection the temperature rose to 40"%-40"~3 C. During the whole of the rest of the time it remained normal.

PQthoZogieal.-Caseous tuberculous focus at the site of injection in the left thigh. Tubercle in the left external inguinal gland. PartiaI slight catarrhal pneumonia.

Mic~oscqpicd-At the site of injection, and in the left external inguinal gland, tubercle bacilli were demonstrated. The other organs had no tubercle bacilli.

INFECTION OF GuINEA-Pras.--Guinea-pigs inoculated under the akin with parts of the site of inoculation contracted general tuberculosis, while those inoculated with portions of the diseased lnng remained healthy.

THE AUTWORS' EXPERIMENTS. PIG KO. 1.-Elack Male. Weight, '294 Ib. Tested six days previously

with Koch's tuberculin (0.2 c.c., Dr. Libbertz),z gave no reaction. The materia1 used for inoculation was the sputum a€ an advanced case of phthisis (Patient 6.) (vide Reports on Calves).

Arch. f. ztiissenseh. u. prakt. Thierh., Berlin, 1902, Bd. xrviii. Hefte I und 2. In aII the pigs tested with tuberculin this dose was employed.

47-1- GEORGE DEAN AND CHARLES TODD.

5 C.C. of a thick emulsion of this was, injected subcutaneously iuto the abdoniinal wall on the 25th August 1901. The injection was followed by swelling, which subsided, but left a nodule which at the end of a month was the size of a walnut. The animal did not become emaciated, as was the case with several of the other pigs, but there was a certain arrest of development. The animal was killed on the 8th April 1902, 327 days after the injection.

Local lesion.-At the site of inoculation a nodule rather larger than a walnut, surrounded by smaller noduIes the size of millet seed to peas. Some of these were fibrous, others caseous or cretaceous. The lumbar glands on the same side were enlarged to the size of hazel nuts, and were also caseous or cretaceous.

Lzmgs.-Scattered through thesewere numerous tuberculous nodules, the size of millet seed, glass-like a t the periphery, grey opaque and cicatricial looking at the centre.

POST-MORTEM ExaMrr?aTioN.-Weight, 56 Ib.

Liver.-Showed a number of nodules similar to those in the lungs. Spleen.-Rather fibrous looking, but showed no naked-eye nodules. Kidneys.--Naked eye showed iiothing abnormal. Lympimtic glands.-The bronchial, prevertebral, and anterior mediastinal

were markedly tuberculous, the latter, the size of horse beans, were in a caseous state with a few calcareous points. The mouth, tonsils, and cervical glands showed nothing abnormal.

~1s~oLoarca1, E x a a r i ~ a ~ ~ o ~ . - T h e only point worthy of note is that the nodules in the lungs were sharply circumscribed and spherical with a dis- tinctly formed fibrous capsule.

EXPERIMENT ON GU~NEA-PZGS..--TWO guinea-pigs inoculated with 1 C.C. of an emulsion of the lumbar glands of this pig both died of general tuberculosis.

PIG So. 2.-Black Male. Tested six days before inocula- tion with tuberculin, gave no reaction. The material used for inoculation was sputum from an advanced case of phthisis (Patient R.). 5 C.C. of this, in the form of a thick emulsion in normal saline solution, was injected sub- cutaneously into the abdominal wall on the 20th August.lSO1. There resulted, at the site of inoculation, swelling, going on to the formation of a hard nodular mass, which before death underwent softening and spontaneous opening, giving rise to the formation of a small sinus. I n this case there was not only arrest of development but great emaciation. The animal died on 4th January 1902, 137 days after the injection.

At the site of inoculation in the left lumbar region there was a swelling the size of a small orange, from which the above-mentioned sinus. opened.' On section of the mass a small cavity was found in the centre, the.walls of the mass were very dense and fibrous, the centre necrotic pale yellow and apparently partially caseated. The inguinal and lumbar glands on that side were enlarged to the size of hazel nuts, and in them were caseous areas. The corresponding glands on the other side were apparently healthy.

Lungs.-Left lung had a depressed area on the pleural surface of about 2 in. diameter, corresponding to a partially consolidated dark red portion of the lung. A number of pale grey nodules, the size of hemp seed, projected through the pleura in this region. On section numerous similar nodules were found scattered throughout both lungs.

These were grey, cartilaginous-looking, with. a 'pale yellow centre.

They contained very few tubercle bacilli.

Weight, 30 Ib.

POST-MORTEM EXAMINATION.-~'eight, 24 lb.

Liver.-Had a few scattered nodules the size of millet seed.

Xpleen.-Had in it similar nodules. MICROSCOPICAL ExaarrNaTroN.-Revealed the presence of tubercle bacilli in

the local lesion, lung, liver, inguinal, lumbar, and mesenteric glands.

PIG No. 3.-Black Male. Weight, 19 Ib. Was tested nine days before inoculation with tuberculin, gave no reaction. The material used for inocula-

EXPERIMENTS O N TUBRRCULOSIS. 475

tion was sputum from the same case (Patient R.) from which the material was obtained for pig No. 2. 5 C.C. of a thick emulsion of this was injected into the abdominal wall on the 23rd August 1901. Slight swelling took place at the site of inoculation, which within a month gave place to a nodule the size of a large pea. The animal wasted and died on the 9th October 1901, forty-seven days after inoculation.

POST-MORTEM EXAMINATION.-Weight, 16g Ib. Local lesion.-At the site of inoculation beneath the skin a hard nodule,

the size of a large pea, could be felt, and on cutting into this it was found to have undergone caseous degeneration. Behind it there was a plaque of caseous material the size of a shilling.

The lumbar glands on this side were enlarged to the size of horse beans, and several of these had caseous points the size of niillet seed.

Lungs.-The upper and middle lobes were consolidated, and portions did not float in water. They were of a deep reddish-purple colour with yellow patches scattered throughout the consolidated areas.

Abdomen.-A few strands of fibrinous lymph on the peritoneal surface of the gut.

Liver and spleen.-Both studded with pale greynodules the size of millet seed. Kidneys.-A few tubercles the size of millet seed. With the exception of

the lumbar, bronchial, and perhaps the mesenteric, the lymphatic glands did not appear to be markedly enlarged.

HXSTOLOQICAL ExaMINATioN.-The nodules above described in the organs and lymphatic glands showed the presence of tubercle and tubercle bacilli.

CULTIVATION.-Nodu~es from the liver placed on glycerin-potato tubes gave in thirty-one days very distinct colonies of the tubercle bacillus ; these had a marked orange-yellow colour. The bacillus, on subculture, gave a luxuriant powth on all the ordinary media used for the tubercle bacillus.

Guinea-pigs inoculated from these pure cultures died of general tuberculosis.

Body much emaciated.

Bronchial glands enlarged.

PIG No. 4.-BIack Male. Weight, 58 lb., at time of inoculation. Tested with tuberculin seventy-six days before inoculation, gave no reaction. The material used for inoculation was the sputum from an advanced case of phthisis (Patient H.). I n this case the inoculations were made into the thorax as well as subcutaneously, and the undiluted, rather thin, sputum was used ; 2 C.C. of this was injected into the right pleural cavity, and 2 C.C. into the left inguinal region.

A swelling occurred a t the site of inoculation in the inguinal region, which was succeeded by a nodule and enlargement of the inguinal glands on that side.

The injections were made on 29th October 1901, and the animal died on 8th February 1902, 102 days after the injection.

POYT-MORTEH ExaMINATIoN.-weight, 70 Ib. A chain of enlarged glands ran along the inside of the left thigh. The inguinal glands were enlarged to the size of hazel nuts, and contained spherical nodules, containing semi-purulent caseous material, in which were found microscopically cocci and tubercle bacilli. After hardening, the central portion of these nodules had shrunken and could be shelled out, leaving n smooth spherical cavity-an appearance identical with that which is found in the naturally occurring infection of the pig. Cultivation gave colonies of Stapliylococcus pyoyenes aureus and albus and Streptowccus pyogenes.

The lumbar glands were enlarged to the size of small hazel nuts, and had in them caseous areas.

There mere a number of fibrous adhesions of the pleura on the right side. On this side a number of tubercles, the size of hemp seed, could be seen pro- jecting through the visceral pleura, and at one point, corresponding to the site of inoculation, was a plaque the size of a threepenny-piece, which was caseous, and covered a nodule the size of a pea in the lung substance. Both lungs had

32-JL. OF PATH.-VOL. VIII.

476 GEORGE DEAN AND CHARLES TODD.

pneumonic patches, probably of tuberculous nature. The liver, spleen, and kidneys showed no tubercles, and no other definite lesions could be found.

HISTOLOGICAL ExAMIr;ATroN.-The lungs, the inguinal and lumbar glands, presented the structure of old fibrous tubercle with giant cells and fibrous net- work. Bacilli mere not numerous, but in some parts a fair number was found.

Tested with tuberculin fifty-six days before injection, gave no reaction. The material used for inoculation was an emulsion from the nodules of the liver of pig No. 3. The only organisms contained in this emulsion were tubercle bacilli. 4 C.C. of this emulsion was injected into the left groin on 9th October 1901. Beyond a slight transient swelling a t the site of inoculation, the animal showed nothing abnormal, and mas killed on 16th April 1902, i.e. 189 days after the injection.

PosT->foRTEnf ExAMINATIoN.-The animal mas well-nourished, and the most careful examination revealed nothing abnormal.

PIG No. 6-Black Male. IVeight, 20 lb.’ Tested with tuberculin seventy- six days before injection, gave no reaction. The material used for inoculation was the same sputum (Patient H.) as in pig No. 4. 2 C.C. of this, undiluted, was injected into the left groin on 23rd August 1901. A swelling occurred, which went 011 increasing until the time of death. The animal wasted, and died on 16th January 1902, i.e. 146 days after the injection.

POST-MORTEM ExAaiINATIoPI’.--Wei~ht, 24 lb. At site of inoculation in the left groin was an abscess cavity, the size of a

cocoanut, containing a thin grumous pus, in which were present numerous tubercle bacilli, along with other bacteria. The lymphatic glands in the groin were enlarged to the size of a walnut, and were caseating. On opening the abdominnl cavity there was seen, running in front of the lumbar vertebm and from the affected inguinal region, a chain of enlarged lymphatic glands, several of which had reached a walnut size and had undergone extensive degeneration. The cervical and axillary glands on both sides were slightly enlarged. The mesenteric glands showed no marked enlargement. The lungs had scattered through them an enormous number of small grey tubercles, from the size of a millet seed to the size of a hemp seed. There were small pneumonic patches surrounding these.

Liver and spleen had numerous similar grey nodules. I n the kidneys the tubercles were less evident, but were present as minute grey points.

HISTOLOGICAL ExAMINATION.-The inguinal and lumbar glands showed niagnificent giant cell systems ; the giant cells appeared to have undergone a hyaline degeneration, the whole body of the cell having become slightly acid- fast (contrast condition of cells in pig No. 7). The nodules in the lung con- tained tubercle bacilli, but these were not numerous. The nodules in the liver and spleen showed the structure of tubercle.

CuLTIvaTrox.-Pure cultures of the tubercle bacillus were obtained from the liver and spleen. These cultures on glycerin-potato had less of the pale orange-yellow pigment than those similarly obtained from Pig KO. 3.

PIG So. 7.-White Female, of the same litter as the three feeding a d the three control pigs. This animal was not tested with tuberculin, as it was one of a litter bred from a healthy sow, and as three pigs of the same litter were reserved as controls. These, when killed five months later, were found to be free from tubercle.

The material used for inoculation was the same sputum (Patient H.) which was employed in the case of pigs No. 4 and No. 6. 2 C.C. of this sputum was injected into the left thigh on 29th October 1901. There was slight swelling at the site of inoculation, and this was followed by the formation of a nodule the size of a horse bean. The animal wasted and died on 29th November 1901, i.e. thirty-one days after the injection.

Average weight of

Pro KO. 5.--131ack Male. Weight, 36 Ib.

Body greatly emaciated.

Weight, 35 lb.

POST-~ZORTEJI EXAMIXATION (Pro 7).-Weight, 24 lb.

EXPERIMENTS O N TUBERCULOSIS. 477

three control pigs of the same litter was 53 lb. At the site of inoculation in the left thigh was a nodule the size of a horse bean, in which a number of minute grey points, the size of millet seed, were visible. Round the knee-joint the capsule was thickened, and contained small caseating areas. I n dissecting the thickened capsule of the joint, a small abscess cavity was found containing about half a drachm of thin pus showing numerous organisms-bacilli and cocci.

In the infiltration round the knee-joint tubercle bacilli were present in enormous numbers. Xany of these were very long, but beading was not a marked feature. The inguinal glands on this side were greatly enlarged, forming a mass the size of a large walnut, with projections the size of horse beans. This mass was congested at parts, and on section showed numerous millet-seed-like nodules, in which tubercle bacilli were demonstrated. The lumbar glands on this side presented a similar appearance, and were about the same size as the inguinal glands.

The liver, spleen, and kidneys appeared on naked-eye examination to be normal.

The lungs were congested, and had pneumonic patches scattered through them, but no definite tuberculous nodules could be made out on naked-eye examination. The bronchial glands appeared to be slightly enlarged. The tonsils normal.

HISTOLOGICAL ExalrIxATioN.-The affected joint capsule was swarming with tubercle bacilli. The inguinal and lumbar glands showed giant cells which had less branching at the periphery than in the case of pig No. 6, and showed no acid-fast property as in that case. Tubercle bacilli were frequently present in large numbers within these. Tubercles could be seen in the lung, on micro- scopical examination, and these showed numerous tubercle bacilli. The liver, spleen, and kidneys showed no tnbercle.

The abscess did not communicate with the joint cavity.

FEEDING EXPERIMENTS ON PIGS.

The following is a translation of the detailed description of Pro- fessor Koch’s experiments (Zoc. ci t . ) :-

Six pigs together received daily about 100 grins. of the sputum of a tuberculous patient ; the sputum being examined at stated intervals with reference to the number of tubercle bacilli. The sputum was rubbed up in a mortar and carefully mixed with 2 litres of milk, whicll had been sterilised in the steam steriliser.

The pigs took this iiiixture with the greatest eagerness, and showed no change in their general condition. The temperature always remained within the normal limits.

PIG No. 1.-Received daily about 15 grms. of the sputum of a tuberculous patient.

Duration of the jeeding.-l04 days. Dusation of fire eq)eriment.-Killed after 138 days. Increase in zoeighf.-40& kilos. PATHOLOGICAL RESULTS.-AI~ organs normal. PIO No. 2.-Receivetl daily about 15 grms. of the sputum of a tuberculous

Duration of the feeding.-l04 days. Duration of the everiment.-Killed after 138 days. Ineyeme in zcei&’.-50 kilos. PATHOLOGICAL RESULTS. -Tuberculosis with partial calcification of the

mesenteric glands, and of the lymphatic gland beneath the right parotid. Several miliary tubercles in the lungs.

patient.

478 GEORGE DEAN AND CHARLES TODD.

MIcRoscoPrcaL.-Tubercle bacilli could be demonstrated in the caseous and calcareous masses in the mesenteric glands, and in the gland beneath the right parotid.

INOCULATION OF GuINEa-PIGs.-The guinea-pigs inoculated with portions of the affected lymphatic glands and lungs contracted general tuberculosis.

PIG KO. 3.-Received daily about 15 grms. of the sputum of a tuberculous patient.

Duration of the feeding.-l04 days. Duration of the experiment.-Killed after 138 days. Increase in weight.-49 kilos. PATHOLOGICAL REsULTs.-several caseous and calcareous nodules the size

Beyond this all the

MrcRoscopIcaL.-Tubercle bacilli mere found in the nodules of the above-

INOCULATION OF GuINEa-PIGs.-Guinea-pigs inoculated with portions of

PIQ No. 4.-Received daily about 15 grmn. of the sputum of a tuberculous

Duration of the feeding.-l04 days. Duration of the experiment.-Killed after 104 days. Increase in weight.--27 kilos. PATHOLOGICAL RESULTS.-AII the organs normal. PIG KO. 5.-Received daily about 15 grms. of the sputum of a tuberculous

Duration of the feeding.--I04 days. Duration of the experiment.- 104 days. Increase in zaeight.--25 kilos. PATHOLOQICAL K.EsuLTs.-Tiiberculosis of the pharyngeal lymphatic glands.

MrcRoscoPIcaL.-Tubercle bacilli could bc deinonstrated in the lymphatic

INOCULATIOX OF GUINEA-PIGs.-Guinea-pigs inoculated with portions of

PIG No. 6.-Received daily about 15 grms. of the sputum of a tuberculous

Duration of the feeding.--lO4 days. Duration of the experiment.-Killed after 104 days. Increase in zueight.-20 kilos. PATHOLOGICAL RESULTS.-A~~ the organs normal.

of pin-heads, in the post-pharyngeal lymphatic glands. other organs normal.

mentioned lymphatic glands.

nodules contracted general tuberculosis.

patient.

patient.

Chronic inflammation of the liver.

glands which had undergone caseous degeneration.

the lymphatic glands contracted general tuberculosis.

patient.

THE AUTHORS' FEEDING EXPERIMENTS ON PIGS.

The three animals used for these experiments were from a litter of seven, bred from a healthy sow.

Three were kept as controls, and one (No. 7) was used for another experiment (subcutaneous injection).

When the animals were two months old they were removed from the sow, and were fed on coarse wheat meal, '' middlings," mixed with water, to which tuberculous sputum was added.

The animaIs received as a rule each about 15 C.C. a t one feeding, but on two occasions very much larger quantities were given, about

EXPEA'JMENTS O N TUBERCULOSIS. 479

100 C.C. to each animal. Owing to the supply of suitable sputum being somewhat irregular, the feedings of sputum could no t be given every day, but the number of feeds in the w e of each animal ie separately stated.

It wae impossibIe to feed each animal with a particular sputum, owing to the death of patients supplying the material during the course of the experiments.

PIG So. 8.--Femaie. Weight, 35 ib. The feeding with tuberculous sputum was commenced on the 3rd October 2902. The animal received twenty-four feeds of sputum (Patient H.). It rapidly emaciated, and died on the 24th December 1901 (eighty-two days).

Body g d i y emaciated. The tonsillar lymphatic glands mere enlarged to the size of a walnut, and on section presented a number of caseous areas the size of split peas. The deeper cervical glantls wePe enlarged to the size of peas, and also showed some caseous areas.

The mucous membrane of the mouth and pharynx appeared to be normal. The other lymphatic glands were not markedly enlarged, and showed no definite lesion.

POST-hIORTEM EXAM1NhTIoN.-\?'eight, 26 Ib.

N o other naked-eye lesion could be found. HISTOLOGICAL ExaMrsATtox-In the tonsillar and cervical glands tubercle

bncilli and the typical structure of tubercle, including giant-cells, mere found. The lungs showed small areas of catarrhal pneumonis, but no tubercles or titbercle bacilli could be demonstrated. The exaniination failed to reveal the presence of tubercles in siiy of the other organs.

PIG So. 9.-Male. \\'eight, 30 11). The feeding with tuberculous sputum was commenced on the 3rd October 1901. The animal received twenty feeds of sputum (Patient H.). It rapidly wasted, and died on the 14th December 1901 (seventy-two days).

Body greatly emaciated. The tonsillar lymphatic glands were of the size of hazel nuts, and on section presented numerous caseating areas. The mucous membrane of the mouth and pharynx normal.

HISTOLOGXCAJ, s x A w i v A ! f I o x , - h tho tonsiIIar glands the structures of tubercle including giant cells, as well as tubercle bacilli, were found. KO lesions could be found in any of the other organs.

POST-MORTEM ~XAMINATION.-weight, 23 Ib.

A few fibrinous adhesions between the intestinal coils.

PIG No. 20.-White male. Weight, 36 lb. This animal-for reasoils previously stated-was fed with sputum from three different patients, namely, sixteen feeds of sputum (Patient H.), one feed (Patient J.), eight feeds (Patient S.), i.e. twenty-five feeds in all.

The pig wasted rapidIy, and died on 20th December 1901, i.e. seventy-eight days after the commence- nient of the feeding.

POST-MORTEM ~xAbiIwATroN.-Weight, 26 Ib. The tonsillar lymphatic glands were the size of sniall walnuts, and contained a number of caseous nodules.

The mesenteric glands were not obviously enlarged or tubercuIous, and no lesions could be found in the gut. The Peyer's patches were conspicuous and in,jected. The liver, spleen, and kidneys appeared normal. The Iungs were slightly injected. Bronchial glands not enlarged. The axillary and inguinal glands were not markedly enlarged.

MrcRoscoPici\LLu.-The tonsillar glands were typically tuberculous, and showed tubercle bacilli.

The feeding began on 3rd October 1901.

A deep cervical gland was similarly affected.

No other tuberculous lesions could be found.

480 GEORGE DEAN AND CHARLES TODD.

~. ~ ..... ..- ~ ~

FEEDISG WITH SPUTUM.

CONTROL PIGS Nos. 11, 12, AND 13.

As already stated, these three animals were of the same litter as pigs Nos. 7, 8, 9, and 10. At the time of removal from the sow their average weight was 33 Ib. The material used for feeding the animals was the same as that used for pigs Nos. '7, 8, 9, and 10, except that in the case of the controls no tuberculous sputum was added. On the 29th November their average weight was 53 Ib., whereas the average weight of pigs Nos. S, 9, and 1 0 was 27 Ib. The control animals remained perfectly healthy, and when killed a t the age of seven months were found to be free from disease, their average weight being 8 2 lb.

TABLE II.--Sliozoing Result of Experinienfs on Pins. ~ . - -~ __ - - . . - - ._

I ISOCULATED SUBCUTANEOUSLY WITH SPUTUM.

No. Weight. Sputum.

294 Ib. 1 Patient S.

1 Died. 'Killed.

...

+

,

227

137

I . _ _ ! 3 i

... ' 102

j i + . i I " " I I

35 ,, 31 I

__ - - - - - ___

56 Ib. Tubel-les, local ; bron- ~ chial and mediastinal ~ glands, lungs, liver.

24 , , Tubercles, local ; abscesses, inguinal and lumbar glands, lung, liver, spleen.

1Gt ,, Tubercles, local ; lumbar I nnd bronchial glands

iO ,, '2 , I

24 I ,

'

- I ~ .~~

lungs, liver, spleen, kidneys.

Tubercles, local ; lumbar glands, lungs.

Tubercles, local ; inguinal and lumbar glands, ~

lungs, liver, spleen.

and lumbar gc~%: ' Titbercles, local ; in

lung. I - _____- _ _

I

SUBCUTANEOUSLY WITH EMULSION KODULE LIVER, PIG No. 3. I

~ ~

9 30 ,, 10 36 ,,

I I I I '

- _ _ - _ _

Tubercles, tonsillar and

Tubercles, tonsillar glands. Tubercles, tonsillar and

cervical glands.

cervical glands. I I

EXPERIMENTS ON TUBERCULOSIS. 481

-

Teniperature Three Hoiirs Six Hours Nine Hours before Injection. after. after. after.

102"'4 j 102"'6 103"'4 102O.4

APPENDIX OF TEMPERATURE CHARTS AND TABLES.

C H A R T S OF REJECTED A?JI;\fALS.

.pt. 5 6 7 8 9 10 111 I 1

__-_ .

Twelve Hours Fifteen Hours nfter. after.

102O.2 I 102""2

CH.4RT 2.-White Star. Rejected. Rejected.

ected.

CALF No. 1. The temperatures taken on the two days before the injection of tuberculin

September 10- On September 11, at 10.30 A.M., 0.5 C.C.

on September 9 were :-Morning, 101"-6 ; evening, 103O.2. Morning, 101O.8; evening, 103".4. of tuberculin was injected.

First Test.

482 GEORGE DEAN AAW CHARLES Tom.

The temperatures takeii before the second test were :-On October 10- Morning, 101'.2 ; evenirig, 102'. Octuher Z l-Morning, 102" ; evening, 1W.6. Test was done as above on October 13.

Second Test. - I__-.

Temperature before Injection.

.__

1 OO"'8

- - .. ._.-- _I__I_ --^- IXL

after.

. _ Three Honrs 1 Sir Hours

oftcr. aqzpr. after. [ after. I

The teiuperatures taken before testing after inoculation were :-On April 7- April 6-Morning, 100" ; evening, 100"*8. Morning, 100" ; evening, IOl'2.

The test was dorie as above on April 9.

S'est ajfer Inocadatio,r. ^_-__.- - -

I I

Teinperoture Thr;;w~~re

100".8 1 99'

belore Injection. - - .--

I !

II--_ .__---I -_I-

I

A'ote.--TT)iis nnimsl has iiot been iiwluded in the tinrl yesults. I t w8s inoculated s i th materiel from a cat sripyosed to b tuberculous, but subxqnent itwestig&ion of the meterial made it doubtful ivhetber it contained any living tubercle bacilli. The caif remained healthy, and, as ~ 3 1 be seen from the above figize3, gave no reaction.

CX I AP ;I' 4.--Celf 2, ~pt. 9 months. Before inoculation. Admitted, 4th Septemb<

CHART 5.--Calf 2. After inocnlation.

:r $901.

EXPERIMENTS ON TUBERCULOSIS. 483

Temperature Three Hours Six Houm belore Injection. nlter. after,

101"% 101"'8 103"4

-- ~~ .- .. . . I ._l_

CALF No. 3.

-_- IOr.2 1 102"'S

The temperatures taken on the two days before injection of tuberculin September 10-

On September 11, 0 6 C.C. tuberculin was were :-On September 9-&rning, 102"-2 ; evening, 703". Homing, 102"; evening, 103".6. injected.

Firat Teea.t.

I Temperature nreeHours Sm Hours Nme Hours Twelve Hours , hefore Inlectioa after. utter. after / ":-- ____._ - I__

103" I 02" '6 102"'6 ' - r r 101"'2 I I

Fifteen Hours after.

I_--

102"

The temperatures taken before the second test were: October lo-- Morning, 100' ; evening, 102"4. October 1 I-Norning, 101" ; evening, 102".4. Tested with 0.5 C.C. tuberculin on October 12, 6 A.M.

Second Test.

hefore Injection. ~. -

I 101" 1

after

103'2 I 103" I

102",4 101O.4

after. after. ---- -

104"

Temperatare before Injection. 1 Thr%~?Un 1 utter.

100".4 100"$

Fifteen Hours nlter.

104"

CALF No. 4. The temperatures taken on the two days before the injection of tubercuIin

were :-On November 13-Morning, 103O ; evening, 103"*6. Xovember 14- Morning, 102'.8; evening, 102".6. On November 15, 0-5 C.C. of tuberculin was injected at 6.30 A.M.

First Ted.

Twelve Hours Fifteen Hours helore injection alter after. after alter.

1

- l_l_

Tbe temperatures taken before testing after inocuIation were :-April 21-

Fifteen Hours alter.

1 03" '2 -I_

484 GEORGE DEAN AND CHARLES TODD.

-

Temperature Three Hours Six Hours Nine Hours Twelve Hours before Injection. after after. after. 1 after.

_ _ _ - -

i 101"'2 102O.8 102'.2 103'4 i 102"'6

Fifteen Hours after.

103"

Temperature Three Hours before Injection. after. .. . - .

101"'8 102'

CALF No. 6.

I - ~ - ~ _ _ _ - - _ _ _ 102" 104"'-1 i 102"'4

I

CHART B.-Calf 6. Before inoculation. After inoculation.

CALF No. 7. The temperatures taken on the two days before injection of tuberculin

November 14- First test was applied 6.30 A.M., November

First Test.

wcre :-On November 13-Morning, 103" ; evening, 103". Morning, 102O.6 ; evening, 103". 15, with 0.5 C.C. tuberculin.

Nine Hours after.

103O.2

Tempernture Three Hours before Injection. nfter.

Twelve Hours after.

102"

~ .. I

Six Hours after.

lOO"8

Fifteen Hours after.

_____

103O.2

EXPERIJflZW" O N TUBERCULOSlS. 485

The temperatures taken before testing after inoculation were :-April 21- Morning, 103"+3 ; evening, 103"*4. Test was applied as above, April 33, 9 A.N.

____. - - - -_-_____--___I__ _ _ I

Temperature Three Hours ' Six Hours Hine Hours 'helve Houra j Fifteen Hour8 I before Injection. after I after. after. after. after , !

102" I 102O.6 102"%

.-

102O.4 1 102O.6

Teiirperature j Three Hours hclore Injection. after.

-.

1 0L3

CALF No. 8.

- Six Horirs Nine Hours Twelve Hours

after. after. after. after.

I 102"'S ; 103"

1

. ._ ~ ~ _ _ _ _ _ _ _

The temperatures taken on the two days before injection of tuberculin November 14-

On Xovember 15, at 6.30 A.M., 0.5 C . C . of

Test .

were :-On November 13--Morning, 103" ; evening, 101".6. Morning, 102"*4; evening, 103"*2. tuberculin was injected.

The teniperatures taken before testing after inoculation were :-On May 14- The test was administered as above a t 9 6 .31 . Morning, 101"; evening, 102".

on May 15. Test afler Itwedation.

I I ' Temperature Six Hour8 Kine Houm I Twelve Houm Fifteen Houn , I after. 1 nfter. nfter. I

) 102" 1 102" I 105"'4 1 104"'6 1 103"*6 102O.6

I I I

CALF No. 9.

The teinperatures taken on the two days before the injection of tuberculin November 14-

0.5 C.C. of tuberculin was injected Kov- were :-On November 13--i\.forning, 102' ; evening, 103".2. Moruing, 102"; evening, 102"*4. ember 15, 6.30 A.M.

Firsf Test. I -__-- .-

I 1 before Injection. I after. 1 -after. 1 after. 1 after. I after Temperature Three Hours Six Hours Nme Hours Twelve Hour8 Fifteen Hours

i I ____

The temperatures taken before testingafter inoculation were:-On M a y 14- The test was done as above a t 9 A.M., Morning, lOl".4; evening, 103"-2.

I lay 15.

486

I Tempernture Three Hoiiry Six Hour8 ' Kine Hours Twelve Hours I before Injection. I after. j after. 1 after. 1 after. _ _ _ _ . ~ - . __- -

102O.2 103".8 1 1 0 1 O . 8 I 102"'8 I I

GEORGE DEAN AND CHARLES TODD.

Fifteen Hours after.

__ 102O.6

Test after Inoculation.

j Temperature before Injection.

100' I

Three Hours Six Hours Kine Hours 1 T\\el\e Hours Fifteen Hours after. I after. after after.

lOl"'8 j l o l a 1 102"*8 102O.6

~-

The temperatures taken before the second test were :-On December 11- On December 12, a t 6.15 A.M., 0.5 C.C. Xorning, 103"; evening, 103".4.

tuberculin was injected. Second Test.

Temperature Three Hours Six Hows N i y f E m s 1 Twelve Hours Fifteen Hours 1 hefore Injection. 1 after. I after. after. after.

The temperatures taken before testing after inoculation were :-On June 3 -Morning, 102O.4 ; evening, 103"*3. 6 A.M.

The test was done as above on June 4

Test after Inoculation.

Temperature Three Hours Tnelbe Iiours Fifteen Hours

_ _ _ _ _ _ ~ -

Injection. after. after. after. ' 104" I 10Y.1

-~

99"'8 101"'1

CALF No. 11.

The temperatures taken on the two days before the injection of tuberculin were :-On October 30-Morning, 103" ; erening, 102"*2. October 31- Morning, 102; evening, 102O.2. Test applied, Soveniber 1, 6 A.M.

First Test. -

i Temperature Three Hours I Six Hours Kine Hours Twelve Hours Fifteen Hours 1 before Injection. after. i alter.

~ 101"*4 I

after. I after. I Piter. - _____ ... _._ ~ .~ -. I

102" 1 102O.2 1 102"'6 _. I ..

EXPERIMENTS ON TUBERCULOSIS.

' Temperature Three Hours Six Hours Nine Hours j before Injection. after. after. nfter.

____ . .- . -. - - -

487

I Twelve Hours Fifteen Hours

after. after. . - I - . - - - -.. . . . .

-_ - ___ -.

101O.2 I I ! I 1 101O.4 i 102"'2 i , 0 2 Y 102"

__ ___ __I__

BLACK CAW.

The temperatures taken on the two days before inoculation with tuberculin October 31- were :-On October 30-Morning, 102'.8 ; evening, 102".6.

3Iorning, 103; evening, 102'*4. Test applied, November 1, 6 A.r,r.

102O.2 j

Test. I- -. -

before Injection. after. after. after. after. . - ~____ I_ ___ -

103"'4 1 10Y.6 .~ .---_-___-I

1 Temperature 1 Three Hours before Injection. ' nfter.

I

SPOTTED CALF. The temperatures taken on the two days before injection with tuberculill

October 31--Xlorning, were :-October 30->%0rning, 103" ; evening, 102".6. 102O.6; evening, 102".2. Test applied, Xovember 1, 6 A.X.

Tesf .

i Six Hours Nine Hours Twelve Hours 1 Fifteen Hours after. 1 after. after. 1 after.

I -- --_ -

i Temperature before Injection.

Rejected on account of bad condition and being one of an infected batch

Three HOUR Six Hours Nine Hours Twelve H o u n I Fifteen HOUR after. after. after. after. I after.

RED CALF.

The temperatures taken on the two days before iiijection with tuberculin October 31-Mornbp, were :-October 30-Morning ,101".3 ; evening, 104".

102"*2 ; evening, 102".4. Teat applied, 6 A.M., Xovember 1.

102"'6 1 , 102O.4

--___

I 9 9 ° ' ~ i 100O.6 102".2 102O.4 J

488

I Teniperature Three Hours Six Hours 1 Nine Hours I before Injectlon. I nfter. I nfter. after. I

104O.2 i

GEORGE DEAN AND CHARLES TODD.

Test.

Twel\ e Hours after. - ___._

Temperature Three Hours before Injection. nfter.

~ 102"'4 102"'4 I

ROAN CALF. The temperatures taken on the two days before injection with tuberculin

were :-September g-Morning, 102"*8 ; evening, 103O.2. September 10- Morniiig, 102'3 ; evening, 104". Test applied, September 11, 11.30 A.M.

Six Hours Nine Hours Twelve Hours Fifteen Hours nfter. after. after. nfter.

104" I 103" I 103" 1 103"

-- -____

Test.

_-__ Temperature

before Injection. _ _ _ _ - --

lO'L"'4 -

-

Three Hours Six Hour8 Nine Houn after. nfter. nfter. after. after.

_ _ _

104" 103"'6 1 0 5 O . 4 104O.6 104O.6

BIG RED AND WHITE CALF.

The temperatures taken on the two clays previous to injection with tuberculin were :-October 30-Morning, 103O.8 ; evening, 104"-4. October 31 --Morning, 103"; evening, 102O.2. Test applied, 6 A.M., November 1.

- - Temperature Three Hours

before Inlection. after. _____

102O.8 10p.2

Six Hours Eine Hours ' Tnel\e Hours Fifteen Hours after. after. j after. nlter.

--

102" 102"'2

YOUNG RED CALF. The temperatures taken on the two days before injection with tuberculin

October 31--hlorning, were :-October 30--hIorning, 102" ; evening, 101".2. 102".8 ; evening, 102O.6. Test was applied on Noveinber 1 a t 6 A.M.

Test.

This animal died of gastro-enteritis and malnutritioii, Decenber 5, 1901. No tubercle at post-mortem.

EXPERIMENTS O N TUBERCULOSIS. 489

before Injection. after. nfter. ~- ~

102" I

WHITE STAR CALF. The temperatures taken on the two days before inoculation with tuberculin

were :-October 30--Morning, 102O.8 ; evening, 102O.8. October 31-Mornh16, 101O.4 ; evening, 102O.8. The test was done on Kovember 1, 6 A.M.

Twelve Houra Fifteen Houre after. after.

._

103"% 104"

Test.