I90a.— No. 9. DEPARTMENT OF THE iNTERtOR. BUREAU OF GOVERNMENT LABORATORIES. BIOLOGICAL LABORATORY. A REPORT ON HEMORRHAGIC SEPTIC/EMIA IN ANIMALS IN THE PHILIPPINE ISLANDS. By PAUL G. WOOLLEY, M. D., and J. W. JOBLING, M. D. MANILA: BUREAU OF PUBLIC PRINTING. 1904. 9940 5 190^.— No. 9. Department of the Interior. BUREAU OF GOVERNMENT LABORATORIES. BIOLOGICAL LABORATORY. A REPORT ON HEMORRHAGIC SEPTICiEMIA IN ANIMALS IN THE PHILIPPINE ISLANDS. By PAUL G. WOOLLEY, M. D., and J. W. JOBLING, M. D. MANILA: BUREAU OF PUBLIC rRlNTIiN(; 1904. 9940 LETTERS OF TRANSMITTAL. Department of the Jnteimok, buheau of g0v]*]iinment lvboratories, Ol^rFICE OF THE SUP]^]RINTENDENT% Manila, Orloher 19, 1908. Sir: I have the honor to submit for publication n report on ^Tlemorrhagic septicaemia of animals in the Philippine Islands/^ by Dr. Panl G. Woolley^ Pathologist in the Biological Ijaboratorj, and Dr. James W. Jobliiig, Director of the ScM'iim Ijaboratory. Very respectfully, Paul C. Freer, Superintendent Governni ent Laboratories. Hon. James F. Smith, Acting Secretary of the Interior, Manila. P. L Department oi*^ thI'] Lnterlor, Bureau of Government IjAiiORATOiaES, B 10 L0(H C A L T J ABO R ATO RY,, Manila,, October H, 190S. Sir: In reply to your communication regarding the occurrence of a recent epidemic among the Government carabao in and a])out Manila, I have the honor to transmit the in(^h)sed rc^port from this Laboratory on hemorrhagic septicaemia in tlie Philippine Islands, by Dr. Paul G. Woolley, Pathologist, Biological Ijaboratory, and Dr. Jauies W. Jobling, Director- of the Serum Ijaboratory. A^ery r espectf u 1 ly , JJiOHARi) P. Strong, Director Biological Labora lory. Dr. Paul C. Freer, Sirperintendent Governm ent lAiboratorics, Manila, P. I. :^ A REPORT ON HEMORRHAGIC SEPTIC/EMIA IN ANIMALS IN THE PHILIPPINE ISLANDS. [By Faiil G. Woolley, M. I)., Pathologist, Biological Laboratory, and J. W. .lobling, M. I)., Director of the Serum Laboratory.] The appearance of lieniorrliagic septiea^mia in the Philippine IslandvS was a serious coinplication of the local catth' ])robieni. llinderpest, which had destroyed tliousands of carabaos and wliich liad brought agriculture nearly to a standstill^ was just beginning to be controlled by vigorous work with })rotectiv(^ sera, and cattl(» were being imnmnized in China for iin]){)rtation into these Islands. This work had been progressing very satisfactorily, when suddenly and without warning hemorrhagic septicaemia a])peare(I in a lierd of cattle arriving in Manila from Shanghai. At about the same time that the disease was recognized here reports w(*re received from Hongkong giving accounts of a virulent disease })r(n^alent there which very much resembled bul)onic plague in human beings, and which was called ''cattle ])lague.''^ Tims far these reports have been so lueager that we are unable to state whether or not the disease is endemic at that ])ort, as it seems to b(^ in other ])arts of the world, and whether or not it corresponds witli the disease studied here. That hemorrhagic septicannia existed in the Phili])])ine Islands ^ Since the above notes were Avritten, the report of the (U)vernnient bac- teriologist at Hongkong lias been received. In this there is a brief sum- mary of the nior])hologic and cultural characteristics of the organisms found in the animals that died in the epidemic there. 'Phis germ grew leadily on the "ordinary culture media." It was a bacillus that stained more deeply at the ])oles, and which did not stain by Oram's method and which was nonmotile. Its appearance on culture media was similar to that of /^ coli. All inoculated animlas died after t\venty-fo\u' to forty-eight hours with symptoms of septiciiemia. From his facts Hunter concluded that he is dealing with a form of hemorrhagic septicaemia. But whatever the disease may be, it is not the same one that we are studying, judging from the description of the organism. 5 6 previous to the present epideruic can not be stated with any (k^gree of deliniteness. Since our preliminary i-eport^ of t1ie jjresent out- break some Philippine pliysicians have stated tliat they luxd pre- viously seen aiiinuils dying in the same manner as the ones lately affected, but wlietlier or not these cases were identi^^.al with ours can not be determined. (Jaral)aos are sometinu^s affcK'ted with a con- dition similar to heat stroke, and if one of these animals should die in such an attack the clinical fcuitures presented by it might remarkably resemble those of tlu^ acute form of liemorrliagic sep- ticannia. One of us had recently seen such a case, in which a cara- hao lately landed from a vessel had been di'iven thi'ough tlie town and was suifering for lack of water. It suddenly stagger(Ml and fell by the roadside, frotliing slightly at the mouth, and was unable to rise for at least an hour, and not until it had been drenched fre- quently with water. If the animal had died in this attack acute liemorrliagic septicasmia might have been suspected and a correct diagnosis could have been nuide only after an autopsy. But the above-mentioned statements of the previous occurrence of hemo]'- rhagic septica'uiia must be taken cum grano sails, for there is a proneness among many to recognize any syuiptom com])lex as the disease under discussion at tlu; time. This tendency is well shown by the fact that ])S(uido-farcy is taken for surra by nu^n w(ill acquainted with horses. It is, however, possible tliat hemorrhagic septicaemia has been seen here before and that the disease has thus far evaded the detection of labcn^atory jnen, even though these have a wide experience with cattle diseases. The disease is a widespread one and lias been noted in almost every quarter of tlie globe. The first cases were in Germany and were cited by Bollinger. Later reports of epidemics in Germany have come from Kitt, .lakoby, Buch, and others; from Fram^', Nocard and Ijeclainche have reported cases, as have also Galtier and others; P>osso, Oreste, and Armanni, and others have detailed descriptions of epidemics in Italy; Pools repoi'ts cases from Hol- land; Jensen from Denmark; Piot from Egypt; Van Eecke Fischer and Ilubenet from Java ; Peischig, Sequens and v. Patz from Ilungary ; Janson froui Japan; Sanfelice, Loi, and Malato fi'om Sardinia; Pease from Ih'itish India; Carrougean and Blin and Carre from Indo-Cliina; Lignieres from South Auierica, and Smith, Wilson and Brimhall, Peynolds, Fennimore, and Nocard VRead at a nieetin^^ of the Manila Medical Society July, 190;^. from ihe United States. It is easy to see liow tlie spread may have ineliided tlic Philippine Islands if the disease is a contagious one and not endemic in that Archipelago. There is a possibility that China is not to blame for the cases recently discovered here. It may be tliat the disease is endemic in the Philippine Islands and that by food or water the organisms gained access to tliese animals through woimds of tlie gastro-intestinal tract (or even witliout such wounds), or through wounds on the surface of the body, or pos- sibly by way of the respiratory tract. It is now settled beyond doubt tliat ojganisms resembling those of hemorrhagic septicemia are found in waters and soils. It has also been shown that similar ones are present on tlie mucous mem- l)ranes of healthy animals. Moore, of 11 le Depnrtnu^nt of Anim^al Industry at Washington^ for instance^ has demonstrated that organ- isms resembling those of hemorj'hagic se})ticeuiia and capable of producing the disease ocurred in 80 per cc^nt of the cattle, 48 per cent of the hogs, 50 per cent of the sheep, I() per cent of the liorses, :]0 per cent of the dogs, and 90 ])cv cc^nt of the cats examined by him. Davaine, Pasteur, Gaffky, Gauudeia, S. Mayr, and Kitt luive found organisms in waters, soils, sputum, and broncliial secretions wdiich, injected into the circulation of animals, caused se])ticemia which agreed with the usual disease in all its details. Since this disease so closely resembles bubonic plague, it might be supposed that it corresponds with it also in the sources and means of infection, but these points aw awaiting explanation. Since the organisms occur so conrmonly in healthy aniuuils, it i-s probahle that a lowering of resistance to disease, whether brought about by trauma or by abnormal conditions of environment, rela- tively increases the virvdence of the organisms and bi'ings about conditions fi-om which infection and disease result. This is cer- tainly true in the sporadic pneunu)nias of cattle, described by Theo- bald Smith, who believes that the organisms, present in such con- ditions, are secondary invaders. Tlie history of the present epidemic of hemorrhagic septicemia in the Philippine Islands is as follows: A sliipment of cattle arrived in Manila Inay from Shanghai on May 28, 1903, and was kept on board until June 1, when the ani- mals were sent to the Perez estate, in Paco. On the following day two of the animals M^ere noticeably ill, and were sent to the serum laboratory for observation. 8 Case I. — The first animal to die was very weak when liist observed, bnt in fair physieal eondition otlierwise. The eonjnnetiva* were somewhat congested, respiration was rapid, and tlie feces normal. Temperatnre, ^^{).2° C. When taken off tlie truck at the Laboiatory it staggered a few stej)s and fell on its side. There were numerous bruise)^ on the body, j)robably the result of a rough voyage across the China Sea. It ate food when placed near it and also drank, although it did not, apparently, suffer from thirst. It had no cough. During the next few days it became a little brighter and somewhat strongei-. On June 6 it was again weak and could not stand up, the hind legs seeming to be especially feeble, ft gradu- ally became weaker and diarrhea developed, but with no traces of blood or nuK'Us. Death occurred on the ninth day after landing. The post-moitem examination showed a few patches of subcutaneous edema on ttie sides. There were a few small pericardial hemorrhages about the base of tlie heart. The lungs showed a number of subpleural-nodules, which on section exposed granular areas similar to those seen in broncho- pneumonia, in the stages of red and giay hepatization and suppuiation. The suppurating areas were filled with a thick, granular, greenish-yellow% sticky material. Cultures were nuide from the lung abscesses on agai' and blood-serum. After twenty-four hours at 37° C the agar tubes showed a growth of small, trans})arent, grayish, round colonies. The blood-serum showed a very scanty growth of small colonies. Transfers were made from these tubes to various other media, and plates were also made. After a careful study of its morphologic and cultural characteristics, it appeared that the organism under consideration was a short bacillus with rounded ends, and nonmotile. Its measurenu»nts varied between 1 and 2, microns in length and 0.3 and 0.5 of a micron in thickness. The largest forms were seen in glucose media, the snuillest on potato. From the animal body it showed well-marked polar staining, although this was not so distinct in organisms grown on artificial media. It was stained easily with the usual watery aniline stains, but was not stained by Gram's or Weigert's methods. The rods, as a rule, occurred singly, often in pairs, occassionally in chains of five or six individuals. The appearance of the growths on tlie usual culture media was in no way chaiacteristic. The colonies on agar were small, grayish, transparent, and well circumscribed, with little or no tendency to spread. On all tlie solid media approximately the same appearance was noticed. In gelatin no liquefaction was caused. \n bouillon a granular de])Osit was formed on the sides and bottom of the tube; during the first few hours of tlie growth the whole medium was faintly clouded, but as the sediment was deposited the liquid became clear. After a few days the sediment became viscid, as could be show^n by shaking the tube, when the ]>recipitate rose, not in floccules but in threads. In Dunham's peptone solution the same general characteristics were observed as in broth, but the growth was not so abundant. Indol (cholera-red) was produced rapidly so that at the end of twenty-four to thirty-six hours the addition of pure sulphuric acid (free from nitrites) produced a well-marked pink color. No phenol could be appreciated. No gas was produced in solid media t'ont [lining sugars nor was the leaction clianged. Milk remained unad'eeted even after tvu days; no acid was piodueed, no coagulation occurred, and there was no reduction of litmus. Stab cultures in solid media showed nothing renu\rkal)le. The growth followed tlie line of inoculation closely with no tendency to s])read. anrum Laboratory on a truck, and on arriving there was unable to walk more than thirty feet. Its eyes were clear, the res})iration quickened, and the body covc^rtMl with numerous l)iuises. On the 4th of dune it was able to walk and graze, and seenuHl to be in good condition. On dune 14 it was again dull and weak, but still able to walk, the greatest weakness being in the hind quarters. Nevertheless, in spite of ji good a])petite tind in the absence of other symptoms, the animal gradually became gieatly emaciated. On rjune 21 a swelling was noticed on the inner and outer sides of the left ankle. At this time the animal was hardly able to stand, although the appetite was still fail-. The swelling on the ankle was inc'ised by Dr. Slee and the contents evacuated. The abscess cavity was a loculated one* and the in- cised tissue was quite edennitous. On dune 25 diarrhea develoi)ed and two days later, wdien just about to die, the animal was killed and an autopsy done immediately. The findings were as follows: Besides the incision ovei- the left ankle there w^ere dried scars on both sides of the body, and a small opening, dis- charging a greenish-yellow pus, in the left Hank over the qmirter. Dissec- tion of the left ankle showed that the abscess cavity extended around the joint without involving it, but reaching far up the leg. An opening into the abscess of the flank led into a nniss of sui)j)urating glands, which looked much like the broken-down ones of bubonic plague, save that the contents wer(» more granular. The tissues about these sup})urating areas were infiltrated with a gelatinous exudate of a distinct yellow color. in the intermuscular and surrounding tissues there were more or less well-marked hemorrhages. On skinning the aninnil a widespread subcutaneous gelatinous edema was discovered, with large and snmll hemorrhages. In the stomach walls and beneath the pericardium and the jdeura hemorrhages were present, some of which were well circumscribed and others irregular in outline. There was a very large extravasation of blood into the mediastinal ])ortion of the ])arietal pleura, and another similar one along the descending aorta. The lymph glands were generally enlarged, pale, and edematous, but some were smaller and hemorrhagic. There were ])etecchiae in sonic of the edematous glands. The lungs were for the most ])art crepitant, although there were aieas in the left one which resembled the stage of red hepatiza- tion of bi-oncho-])neumonia. The si)leen was of about noi'mal siz(\ with no hemorrhages into its substance but with nniny subcapsular ones. The liver showed nothing remaikable. The kidneys w(»re of fair size and the peri- renal tissues were edematous: theii- ])elves were filled with a gelatinous material. There were no hemorrhages into mesentery. In the abscesses from the left ankle and from the hind quarters small 10 polar-staining bacilli were found in almost pure culture. Smears from the lymph glands showed similar organisms, but the heart's blood was apparently free from them. Cultures showed an organism that corresponded with that from Case I. Case in. — This animal^ a full-grown carabao, was apparently perfectly well until one morningj when it looked stupid, its gait was clumsier than usual, and its eyes appeared to be sunken. It did not care to use the mud bath, but wandered about aimlessly. (The native overseer said that it was insane.) On the third day weakness across the loins and in the forelegs was evident. Its appetite was still fair. On the morning of the fourth day it fell over and was unable to rise again, and died in the after- noon. Smears showed a few organisms that were polar-staining. Culture on placenta fluid showed organisms that resembled diplococci. Culture lost in moving. Gxise IV. — This animal was stupid and weak when first noticed, and as nearly as could be determined had been sick for about three hours. It kept on its feet and moved from place to place for several hours, eating occa- sionally. Death took place suddenly. Smears from organs showed a few polar-staining organisms. Cultures not made. Case y.-^This carabao lived about three days after the appearance of the first symptoms, the principal one of which was weakness, especially in the forelegs. At autopsy the base of the heart was edematous and there were scattered areas of subcutaneous edema with a few hemorrhages. Case VI. — ^Death was very sudden, occurring twenty-four hours after the beginning of the illness. Autopsy revealed edema and blood-stained infiltrations about the inguinal and prescapular glands. The latter were much swollen and showed areas of necrosis and a few small hemorrhages in their substance. Hemorrhages, varying in size, were present in the inner and outer surfaces of the intestines, under the visceral and parietal pleura, under the parietal and visceral pericardium, and under the endo- cardium. Petechiae were present on the surface of the liver and were especially well marked on the inner surfaces of the branches of the portal vein. A few small hemorrhages were found under the capsule of the spleen and under the surface af the diaphragm. Smears from lymph glands, liver, spleen, and lungs showed polar-staining bacilli. Case VII. —The duration of the illness was two and one-half days, dur- ing which the animal could scarcely stand, although the appetite remained good. The almost characteristic edemas were well marked in this case and polar-staining bacilli were found in smears. Case VIII. — ^This animal, one of the finest in the herd, was alive and well on the morning, of June 7 ; the following morning it was found dead. The post-mortem examination showed hemorrhagic edema under the skin along the spine, about the prescapular glands, and around the base of the heart. There were eechymoses on the surface of the heart, under the capsule of the spleen, in the inguinal and prescapular glands, in the pancreas, in the gall bladder, and under the serous and mucous surfaces of the intestines. Smears from heart and spleen showed polar-staining bacilli, some of which showed evidences of having been a capsule or pseudocapsule. Blood serum 11 from the case had no agglutinative reaction on the bacilli previously obtained from other cases. Gases IX, X. — ^These were calves. Their clinical histories and the details of the autopsies are wanting. All that is known is that in neither case was the spleen enlarged, that there were subcutaneous gelatinous edemas, and that the usual edema was about the base of the heart in each. Case XL — In this case also the clinical history is wanting, but the same pathological changes that were observed in the two previous cases were present. Smears from the blood showed a number of polar-staining bacilli occurring singly or in chains of three or four individuals. Cultures were made from the heart's blood on agar and an organism was obtained in pure culture which corresponded to that isolated from Case I. Case XII. — This animal died within twenty-four hours after the appear- ance of the first symptoms. It was large and well nourished, and when examined was lying on the ground, the legs stiff and the head drawn back. The agonal stool was bloody. There were hemorrhages along the spine into the subcutaneous tissues, as well as on the surfaces (inner and outer) of the small intestine. The prescapular glands showed areas of hemorrhage and necrosis. Case XIII. — This was a fairly well-nourished carabao which had arrived from Shanghai three days previous to the apearance of illness. It was well on the evening of July 22. On the following morning it was found deadi In this case the lesions were confined to the heart and lungs; the subcutaneous gelatinous edema seen in the other cases was absent. The lungs were not collapsed and contained air in only a few patches. The costal pleura was intensely inflamed and showed numerous ecchymoses and was covered with a thick stratum of fibrino-purulent exudate. The visceral pleura was in a similar condition. The mediastinum was filled with a mass of yellow gelatinous material composed of serum and fibrin. On section the lung tissue was red, in places very dark, and divided into large and small lobules by white fibrous and fibrinous bands intersecting in all directions. These bands varied from b to i an inch in width and were studded with loculi containing a serofibrinous and fibrino-purulent material. In the pericardial cavity was a large amount of serous fluid, containing floccules; both layers of the pericardium were covered with fibrinous shreds. The serous membrane under this exudate was inflamed and dotted with ecchymoses. The auricular appendages were thickly mottled with hemorrhages. Smears from the heart's blood, lungs, liver, and lymph glands showed a considerable number of polar-staining bacilli, and cultures showed a similar organism in imcontaminated growths. This case followed closely the description of sporadic pneumonia as given by Theobald Smith. Besides the cases of hemorrliagic septicemia seen in carabaos, a native horse was found to be suffering from a malady which was caused by the same organism. In this case the chief lesions were found in the lungs, and these corresponded perfectly with those of 12 Case I. There was also a well-marked gelatinous edema about the base of the heart. The organism was recognized in smears and cultures and identified with that from Case I. In summarizing the chief clinical features of these cases we can say that there have been three fairly well differentiated types — one^ intensely acute^ in which the animals died within a few hours after the onset of symptoms; one less acute^ in which the animals suffered chiefly from weakness in the limbs^ and more especially in the hind ones; during the course of this form of the disease the appetite was little or not at all affected. Lastly, one in which the course was prolonged^ accompanied by great weakness and emacia- tion and occasionally by suppuration. These are not distinctly defined types, for rarely does the disease affect the animals in the same way, except in the acute form, in which the course is so rapid that there is little time to observe symptoms. The pathological types are as indefinitely defined as the clinical types. In a general way, however, there is a pulmonary type in which the lesions resemble those of broncho-pneumonia, sometimes with abscess formation, and with or without pleuritis. Combined with these changes there may be marked changes in the interlobular tissues, with the formation of fibrous bands and with emphysema. Another type is the rapidly fatal septicemic form, in which there are few macroscopic changes in the organs beyond incipient paren- chymatous degenerations. The third type is a glandular and sup- purative one which terminates in a general infection. In all these forms there have been more or less wide-spread hemorrhagic lesions combined with gelatinous edemas. This last-named condition was especially frequent about the base of the heart, and in some cases it was the only macroscopic lesion. In but few cases have complete temperature charts been kept — this because most of the animals have come from herds regarded as healthy and have died too suddenly to allow records to be prepared. The temperature charts of Cases I and II will, however, be ap- pended. These curves show very little except a primary rise of temperature, after which there is a fall, with the subsequent curve, running either near the base line or very irregularly. The clinical cases given above agree in many points with those described in the literature, but in many details, too, they do not. Wide variation clinically and pathologically seems to be one of the most common features of the disease caused by i?. plurisepticus. 13 In Bollinger's report the cases are divided into an exanthematous and pectoral type. The former is characterized by fever^ swellings of the face and neck^ inflammation of the month and tongue^ with ecchymoses under the mucous membranes, and bloody diarrhea. Such cases we have not seen, nor have we observed any cases resem- bling them. This may be because the hide of the animals that we meet with here is thick enough to prevent the edematous swellings from becoming apparent, for certainly in the majority of cases there have been subcutaneous edemas, some of quite considerable size. The latter type showed, principally, pulmonic lesions. We have studied three cases that followed this type — two in carabaos and one in a horse. In one carabao and in the horse the chief lesion was a broncho-pneumonia without pleuritis ; in the other carabao it was a pleuro-pneumonia. Lignieres divides the cases he has seen into three classes — first, a diarrheic one, the acute form, in which death occurs in from one to two days after the onset of the diarrhea. In this, pectoral lesions are common (i. e., pneumonia, pleurisy, etc.). The second class he calls Enteque. The course of this form is more chronic, and the animal may live for three to four weeks with more or less fever and fetid diarrhea, culminating in anemia and death. The third or cachectic form is marked by wasting, articular localiza- tions, and profound anemia, followed by death. Speaking generally the commonest types in the literature are the exanthematous and pulmonary. Oreste and Armanni described cases in which edemas and hemorrhages were the chief lesions. Many of Kitt's animals showed pneumonia and pleuritis, Jensen's showed phlegmonous edemas, fibrinous pleuritis and pericarditis, and gastro-enteritis ; Galtier described hepatization of the lungs, subendocardial hemorrhages, and peritonitis; Guillebeau and Hess found fibrinous pleuritis and pericarditis, hemorrhagic infarcts in the lungs, and small hemorrhages into the mesentery and on the surface of the kidney ; Bosso described hemorrhages into the serous membranes and into the mucus lining of the intestine; Pease observed cases in which the chief lesions were ecchymoses, widely disseminated in the bodies; Fennimore remarked inflammation of the mucus membrances of stomach and intestines, pleuritis, peri- carditis, and edema of the lungs ; and Smith carefully studied and minutely described peculiar lung lesions in which interlobular changes were an important feature. 14 From a summary of the facts given in the literature we may con- clude that the animals attacked are of all ages, the onset is usually sudden, the course is rapid and death occurs in from 85 to 98 per cent of the cases. In the less rapid forms of the disease the symp- toms may be refusal of food, cessation of rumination, and lacta- tion. The initial rise of temperature may be followed by a drop to normal or subnormal. The respirations may be rapid or labored, bloody material may be discharged from the nostrils, bowels, or bladder, and noncrepitant swellings may occur on face, neck, or about the back and ankles. From all these varying types of disease similar organisms have been isolated. In the outbreak which we have studied the charac- ters of the microbe have been as follows : Short bacilli with rounded ends, polar staining, occasionally encapsulated, and nonmotile ; nonliquefyng, non-Gram staining, growing invisibly on potato, not producing gas, not coagulating milk or reducing litmus, producing indol and nitrites, and not forming spores. It is an organism which corresponds closely with the bacilli of hemorrhagic septicsemia of Hueppe, and especially with that member of the group called B. hovisepticus (KruBe), B. hovisepticuin (Kruse-Migula), B. plu- risepticus (Kitt), B. bipolare inuUocidum (Kitt), B. der huff el- seuche (Oreste-Armanni), etc. The chief differences between the present bacillus and the ones described in the literature are its indol production, which is invariable, and its invisible growth on potato, whifch also appea^rs constant. ANIMAL EXPERIMENTS.^ The organism has been injected into guinea pigs, rabbits, mon- keys, small birds, a dog, a chicken, and a calf. In all but the last three animals death has followed in from nine to twenty-four hours after intrapleural injection, in five to eighteen hours after intra- peritoneal inoculation, and in* five to fourteen hours after intra- venous injection of virulent material. Subcutaneous injection with a culture kept at 60° C. for ten minutes killed a rabbit in thirty-six hours ; intravenous injection with the same material killed in eigh- teen hours. In both these two last cases the organism was recovered from the hearts blood. The lesions in animals killed with virulent cultures varied with *In these experiments Mr. Clegg, Assistant Bacteriologist in the Biologi- cal Laboratory, has given us much valuable assistance. 15 the length of the disease and with the animal used. In all there was edema and some hemorrhagic infiltration or ecchymosis at the site of inoculation. In addition to this two or three cases were noticed in which there were large collections of pus surrounded by tissues in a state of coagulation necrosis in the immediate neigh- borhood of the wound of inoculation. The more resistent the ani- mal and the more attenuated the organism (within certain limits) the greater was the tendency to suppuration. The protocols of the animal cases are as follows : Rahhit S21. — Inoculated subcutaneously with 0.5 c. c. of an emulsion of a twenty-four-hour-old agar growth of the organism froni Case I. Dead in eighteen hours. Large hemorrhage at site of inoculation, and a few small ones in axillary and inguinal glands. Lungs congested, vessels of the heart distended with blood. Liver soft and showed diffused pale areas of fatty or necrotic tissue. Adrenals congested. No changes in gastro-intestinal tract. Small amount of fluid in peritoneal cavity. Smears from the liver, spleen, heart's blood, and site of inoculation showed the typitjal organisms which were recovered in pure culture from the blood, Rahhit ^2^.— Inoculated subcutaneously with 1 c. c. of a suspension of a twenty-four-hour-old agar growth of the organism obtained from Case XIII. Dead in five hotirs. No w^ell-marked lesions. Some edema about the wound of entrance. Liver and kidneys eongested. No hemorrhages or necroses. Smears from heart's blood, spleen, kidney, and liver showed the typical organisms, and these were obtained in pure culture from the blood. Rahhit 355.— Inoculated intravenously with 1 c. c. of a bouillon culture of the organism from . Case I, which had been kept at 58° C. for ten minutes. Dead twenty- four hours later. Autopsy showed focal necroses in the lower lobes of both lungs, in liver, and spleen. Smears from spleen, kidney, and blood showed a few small polar-stained bacilli, which, however, were recovered in pure culture. Monkey 256. — ^On June 16 1 c. c. of a bouillon culture was injected under the skin of the right side. On June 18 1 c. c. of a culture in placenta fluid was injected into the peritoneal cavity. Death occurred six hours after the second inoculation. About the points of inoculation were areas of edema. Surrounding the point of inoculation On the right side was also a large intermuscular abscess filled with a pale, greenish-yellow, gelatinous pus. Practically the whole right side of the animal was the seat of a subcutaneous gelatinous edema. The peritoneal cavity contained a large amount of sanguinous fluid, and the luster of the peritoneum was dulled and it was injected. The other lesions were cloudy swelling of the parenchymatous organs. Smears from the spleen, liver, and peritoneum, and from the pus at site of inoculation showed polar-stained organisms; none seen in blood. In several leucocytes from the peritoneal cavity polar- staining bacilli were seen. The heart, peritoneum, and liver showed pure cultures of this organism. 16 Monkey 297. — Inoculated intraperitoneally, with 1 c. c. of a broth culture of the organism from the horse mentioned above. Death in thirty-six hours. The abdominal wall about the wound made by the needle was edematous. The scrotum was distended and edematous. There was well-marked sub- cutaneous ecchymosis and gelatinous edema with a certain amount of purulent infiltration. Coils of intestines were adherent, and there was a quantity of sanguino-purulent exudate in the peritoneal cavity. The pus extended down along the spermatic cords into the scrotum. The left testicle was much enlarged and contained some purulent and occhymotic areas. The right testicle was small and surrounded by a purulent exudate. The intestines showed a few small hemorrhages under the serous layer. The other organs showed no marked change. Smears from the abdominal cavity, liver, spleen, subcutaneous abscesses, and testicular abscesses showed polar-stained bacilli. These were recovered in pure culture. These ease reports of animals dying spontaneously and those dying after inoculation will give some idea of the ability of the organisms to cause widely varying types of disease. It is as Key- nolds has said: *^^For the present^, at least, we must consider the term hemorrhagic septica9mia as quite inclusive — a sort of generic name, which must cover a multitude of varying types of disease." When we encountered the first cases of the present epidemic we feared that the outbreak might take on as dangerous proportions as it had in some of the epizootics first reported, and as time has gone on this fear has been partially realized, but we still hold to the idea that the cases have been the direct result of trauma, in many of these cases, and the indirect, of infection with an organism present in the mouths of healthy animals. It does seem very probable now that a pasture may become infected and that new animals placed on such infected ground may die of the disease in one of its forms, especially if the animal is in any way predisposed to disease. For this reason it seems most unwise, in the present state of our knowledge of the disease, to move animals from place to place, and so risk infecting new ground to be used by well animals. In one of the corrals in Manila a few animals in one herd had died. Following these deaths came heavy rains, so that the corral became a great mud hole. Into this place a new herd of carabaos were driven after they had crossed the China Sea from Shanghai. Following the advent of these animals deaths occurred rapidly, and in all but a few of the dead the lesions of hemorrhagic septicemia were found. It is our opinion that the disease was the result of the rough sea 17 voyage^ change of diet;, the preliminary inoculation against rinder- pest;, and finally the filthy surroundings in Manila. Under these circumstances disease might be expected. The question concerning the source of the infection by which death resulted remains to be answered. Were the organisms in the animals themselves? Were they in the mud of the corral.^ Neither of these questions can be answered positively as yet. We have not been able to find the bacilli in the mud. But just as long as there is any doubt the animals must be treated as though they could scatter the germs of the disease with their excreta. Up to the present time there has been no evidence that the dis- ease here is directly contagions^ but Carrougeau and Blin^ Bollin- ger, Friedberger and Frohner, and others considered the disease which they studied was. On the other hand ISTocard and Leclainche think that the cases are sporadic and that the disease is enzootic, and Wilson and Brimhall, and Eeynolds believe that it is not con- tagious. For the purpose of eliminating all or most chances of contagion we suggest that each animal in a suspected herd should be isolated or staked out separate from the rest of the herd for at least ten days after the occurrence of the last case in the herd to which it belongs and until any abrasions on the surface of the body have healed. (Brimhall and Wilson ]^ave suggested that skin lesions may play a part in the spread of the disease.) While the herd is under sus- picion none of the animals should be allowed to use the mud baths. After an animal has died the ground upon which it has stood should be cleansed by fire or antiseptics, and if it has had access to the mud bath this should be disinfected in the most thor- ough way with lime or crude carbolic acid and then flushed out with water. There seems to be some difference of opinion as to whether the organism is able to live any length of time in contami- nated water, but since similar ones have been found in rivers and in soils it is wiser to be on the safe side and use all precautions. Theobold Smith states that the organism is very vulnerable, and Nocard and Leclainche say that it is a facultative parasite. The result is that for the present it must be dealt with as though its resistance were great. Kitt, in Kolle and Wassermann's Hand- buch, recommends that it be treated as though it were the bacillus of anthrax. 9940 2 18 Up to the present time we have been unsuccessful in obtaining any serum or protective substance that will modify or ameliorate the course of the disease in experimental animals. Work along these lines is, however, being done and will be made the subject of a later report. Wilson and Brimhall report that they were able to produce a fairly high degree of immunity. In the book on ^Mjcs Maladies Microbiennes des Animaux/' by ISTocard et Leclainche, it is stated that the belief of Lignieres — ^that the organism of bovine pasteurellosis can be attenuated and used as a vaccine — is possible. The details of his method are not given, and thus far we have been unable to obtain them. ISToTE. — We have been interested to hear, since the notes for this report have been prepared, that there have been epidemics of an unknown nature among chickens and swine in the provinces about Manila. The repc^l-ts make it seem probable that these diseases are related to the hemorrhagic septica3mias of fowls and hogs. We have received from a gentleman in Manila a chicken that had died sud- denly. From this we have isolated an organism that resembles the bacillus of chicken cholera, which, when inoculated into small birds in minimal quantities, gives rise to a rapidly fatal septicaemia that corresponds exactly with the disease chicken cholera. In inoculat- ing with this organism a needle whose tip was covered with a culture of the organism was plunged through the skin and into the subjacent muscles. The disease prevalent among swine remains to be determined. However, hog cholera has been previously reported here. These observations make it seem probable that chicken cholera is endemic here, and, if there is any value in analogy, that other forms of hemorrhagic septicaemia may be. LITERATURE. 1. Wilson and Brimhall. — Rept. Minn. State Board of Health. St. Paul, 1901. %. Eeynolds.— Amer. Vet. Rev. 1902, XXVI, 807. lUd. 1903, XXVI, 912. 3. Brimhall.— I6M. 1903, XXVII, 103. 4. Cakeva.— Cent. f. Bakt. 1891, IX, 557. 5. Affanasieff.— -Arb. a. d. Path. Inst. Tubingen. 1893, I, 263. 6. Bunzl-Federn.— Cent. f. bakt. 1891, IX, 787.^ 7. Davaine.— Bull. Acad. Med. 1879, VIII, 121. 8. Friedberger and Frohner. — ^London, 1898. 9. Galtier.— Compt. Rend, de FAcad. Sc. Paris, 1889, CVIII, 626. 19 10. GALTiEE.—IUd. 1889, CVIII, 822. 11. Kruse. — Fluegge's Die Mikroorganismen, Liepzig, 1896. 12. MiGULA.— System der Bakterien. Jena, 1901. 13. KiTT. — Kolle and V/asserman's Handbiieh der pathogenen Mikroorganis- men, Jena, 1903. 14. Leclainche.— Rec. de Med. Vet. 1888, 280. 15. LiGNiERES.-— Rev. Vet. Buenos Aires. 1900, 260. 16. LigniI:res.— Bull Soc. Cent. Med. Vet. 1898, 836. 17. NocARD et Leclainche. — Les Maladies Microbiennes des Animaux. Paris, 1903. 18. PiANA.— Moderno Zoojatro. 1894, 23. 19. V. Ratz.— Cent. f. Bakt. 1896, XX, 289. 20. YoGES.—Ibid. 1896, XX, 906. 21. VoGES.— Zeit. f. Hyg. 1898. 22. VoGES u. Proskauer.— /5i4. 1898, XXVIII, 20. 23. Blin. ET. Carrougeau.— Ree. de Med. Vet. 1902, IX, 571. 24. Krueger.— Berl. Tierarzt. Woch. 1903, 261. 25. RuDOVSKY.— Zeits f. Tiermed. 1901, V, 142. 26. Tanzer.-— Tierarzt. Cent. 1901, XXIV, 277. 27. Sanfelice, Loi, and Malato. — Cent. f. Bakt. 1897, XII, 33. 28. V. Ratz.— Zeits. f. Tiermed. 1896, XXII, 329. 29. Janson.— Arch. f. wiss. u. Pract. Tierhk. 1894, XX, 274. 30. HuEPPE.— Berl. Klin Woch. 1886, 753, 794. 31. Moore.— Rept. Bur. Animal Industry. Bull. No. 10. 1896. 32. Sternberg. — ^Manual of Bacteriology. 1893. 33. ScHEiN.— Rev. de Med. Vet. 1902, IX, 345. 34. Blin et Carrougeau.— Z&w^. 1902, IX, 107. 35. VoGES.— Cent. f. Bakt. 1902, XXXI, 645. 36. Galeotti.— Sperimentale, 1902, LVI, 545. 37. Carr^.— Rec. de Med. Vet. 1902, IX, 467. 38. Serra.— Rev. de Med. Vet. (Lisbon). 1902, I, 161. 39. Bollinger. — ^Ueber eine neue Wild-und Rinder-seuche. Muenchen, 1878. 40. Kitt.— Sitzb. d. Gesell. f. Morph. u. Phys. i. Muenchen. 1885, I, 140. 41. Kitt. — Jahresber. d. k. Tierarzt. i. Muenchen. 1887-88. 42. Bosso.— Cent. f. Bakt. 1898, XXIII, 318. 43. Oreste and Armanni. — Atti del R. Inst. Sc. Nat. 1887, VI. 44. Jensen.— Maandsh. f. Dry. 1889, I, 183. 45. Jensen. — ^Monatsch. f. Prakt. Tierheilk. Bd. II, p. 1. 46. PoELS.— Fortsch. d. Med. 1886, 388. 47. Jakobi.— Berl. Tierarzt. Woch. 1892, 39. 48. BucH.— Monats. f. Pract. Tierheilk. 1892, III. 49. PioT.— Bull. d. ITnst. Egyptian. 1889. 50. Van Eecke.— Thierarzt. Blatt. f. Niederliind Indie. 1891, V, 290. 51. Van Eecke.— Veeartsen. blad. f. Neder. Ind. 1895, 300. 52. FiscKEU.— Ibid, 1895, 153. 53. Fischer.— /6ic?. 192. 54. HuBENET. — Ibid. 248. 20 65. Reischiq. — ^Veterinarius. 1891. 56. Sequens.— /6w2. 1894. 67. V. .Ratz.— Deutsch. Zeitz. f. Tiermed. u. verg. Path. 1896, XXII, 329. 68. Pease.— Veterinarian. 1898, LXXI, 278. 59. LiGNifiBES.— Rec. de Med. Vet. 1898, 761. 60. NocARD.— Bull, de la Soc. Cent, de Med. Vet. 1891, XLV, 424. 61. Smith.— Rept. Bur. Anim. Ind. 1895-96, 119. 62. Fennimore.— Jour. Comp. Med. and Vet. Arch. 1898, XIX, 629. 63. Guillebeau u. Hess. — Schweis Arch. f. Tierheilk. 1894, XXXVI, 49. 64. GuiiXEBEAU. — Ann. de Microgr. 1879, 193. 65. Pasteur.— Compt. Rend. 1880, 239, 315, 673, 952, 1030. 66. Gamemia.— Centr. f. Bakt. 1888, IV, 161. CASE I. 84 m m : 37 m m) 40 41 42 Date. 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