IMAGE EVALUATION TEST TARGET (MT-3) 1.0 I.I 11.25 ia|21 125 itt lii 122 1^ lag ■" 2.0 111 Ki U 14.0 lli& u 1^ 1 w ■'I FhotogFaidiic Sciences Corporalion 23 WiST MAIN STRICT WIMTIR,N.Y. 14SS0 (716) •72-4503 ,*^' CIHM/ICMH Microfiche Series. CIHIVI/ICIVIH Collection de microfiches. Canadian Institute for Historical Microroproductions / inatitut Canadian da microraproductions hiatorlquaa Tachnical and BtbliograpMe NotM/NotM taehniqtMS at bibliographiquM Tha Inatituta haa attamptad to obtain tha baat oriflinal copy availaWa for fHming. Faaturaa of thia copy which may ba bibliographically uniqua, %vhieh may altar any of tha imafpaa in tha raproduction. or which may aignificantly change tha uaual mdthod of filming, ara chackad balow. □ Coloiirad covaro/ Couvartura da eoulaur r~1 Covara damagad/ D D D D Couvartura andommagAa □ Covara raatorod and/or laminatad/ Couvartura raataurto at/ou paUicuMa □ Covar titia miaaing/ La titra da couvartura manqua □ Cdourad mapa/ Cartaa gtographiquaa an eoulaur r*n Coiourad inic (i.a. othar than Mua or biaeic)/ Encra da ooulour (i.a. autra qua blaua ou noiral Coiourad plataa and/or illuatrationa/ Planchaa at/ou illuatrationa an eoulaur Bound with othor matarlal/ RaM avae d'autraa documanta Tight biniing may cauaa ahadowa or diatortion Ki>ng intarior margin/ Larviiura aarria paut cauaar da i'ombra ou da la di ator al on la lono ,-J,- . »ok1>m:ode Bt JOHN M«CRAE, B. A., M.B. (Tor.), Fellow in Pathology, McOUl ITntvertity. {From the J. IT. S. Mohon Pathological Laboratory, MeOill University, Montreal.) While carrying out some studies upon agglutinations by the usual method of injecting bacilli, living or dead, into the animal tissues, I was led to try a method of enclosing the bacilli in capsules, which were inserted into the abdominal cavity of thfe animal, and I found that by this method also the serum gained agglutinative power. The extent and the other characters of such agglutinations will be dealt with in later paragraphs, but before recording my results, it is well, I think, that a few w^ords should be said with regard to the preparation of the capsules. For although Nocard and Roux's * work on the micro- organism of pleuro-pneumonia of cattle, and Nocard's' experiments on the transformation of human into avian tubercle bacilli, by placing them in sealed capsules within the fowl's peritoneal cavity, have brought increased attention to this method of growing bacteria within the body, free from the intervention of the body cells, there does not exist, to my knowledge, in bacteriological literature, any detailed account of a satisfactory method of preparation of the same, and if I mistake not, the difficulty in making capsules which will not rupture nor leak, has been found so considerable that the method has by 4kiny been taken up only to be abandoned. ^ The idea of using collodion or celloidin capsules is, bacteriologically 8peakingjj|||l comparative antiquity; quite early in the nineties, a Bomewhil^Hpiitive capsule was employed in the Pasteur Institute by ^Ann. de MBWlK Paateur, 1898, xii, p. 270. > Ibid., 1898, xii, p. S61. *if. Obtained by Use of Celloidin Capsules •■Vai^-'mktn. ThM 'was formed by taking a glass rod or H into colkiiicai until the desired thicknees of coating I6d| the capsule "WM then stripped off the rod and the culture ilHeited, the mouth tied^ and finally, the interstices at the neck were cemented over with collodion. The procedure was first mentioned in an article by Hetchnikoil, Boux and Salimbeni.' The defects of this method are obvious: namely, the amount of manipulation required, the long time the capsule has to be kept exposed to contamination while the mouth is being sealed up and the liability for the new coat of collodion at the neck not to adhere thoroughly to the previously dried material. Gradually, it would seem, the method of making these capsules was improved in Paris, though I can find no clear statement of how the capsules were made by Metchnikoff, Eoux, and Nocard in their later work. A distinct step in advance was made upon this side of the Atlantic some two years back, so far as I have been able to trace, by Dr. Prudden and others in the Laboratory of the College of Physicians and Surgeons of New York. It consists in employing the gelatin cap- sules now obtainable at any druggist's, and used for the administration of unpleasant drugs. These capsules are taken as a framework and coated with celloidin; next, the gelatin is dissolved out in a sterile test tube, the cap being then luted upon the body of the capsule by means of painting with thin celloidin. We obtained a knowledge of this method from Drs. Trudeau and Baldwin at the Saranac Lake Laboratory, and, if we are not mistaken, yet further advance was there made by replacing the gelatin cap with a length of glass tubing to which the body of the capsule was luted, the tube being sealed in the flame after filling the capsule. At Saranac Lake they employ a bulb-shaped capsule, and from what I learn from Dr. Baldwin, some little di^culty has been experienced owing to the tendency ^i capsules so mr ..o to undergo rupture within the body and thus set up a general infection. The method described below has proved so simple an|nithd same time so successful that a detailed account of it may fur^^K means to sToxine et antitoxine cbol^rique. Ann. de Vlnstitut Pasteur, 1896, x, p. 257. r'jpKr'rlVf'-'t.ll '^■li^^ John McCrae Others of adopting this manner of paidilg iMiioKil^ < body without coming into direct conttet with the ti/MMp. methods have been touched upon lest I be thought to cl«llii fil^ slightest degree the credit due to originality. Celloidin is especially adapted to this work, as it preyents the escape of the organisms while it allows osmosis of the fluids to go on freely. Enclosed in tho celloidin capsule the bacilli lie exposed to the body fluids, their soluble products have free egress to the tissues, and, should observations upon the bacilli themselves be desired, the capsule can be removed with a certain knowledge that it contains the form that was originally introduced. 1. Gelatine capsule, natural size. 2. Diagram of glass tube with adherent capsule. 3. Method of filling the capsule. 4. Capsule filled and sealed ready for introduction into the peritoneal cavity ; natural size. A piece of glass tubing, 1 cm. in external diameter and 6 to 8 cm. long, is taken and a narrow and rather abrupt neck is drawn upon it about 3 cm. from one end. It is well, as a matter of precaution, to round off the edges of this end in the flame, lest any sharp edge should later, on manipulation or movement of the capsule within the body, cut through the celloidin. Then over this end, after heating it slightl^^Wie body of a gelatin capsule is fitted, the top being dis- carded; t^piot glass melts the gelatin and there is immediate adhe- sion. The accompanying diagram 1 gives the exact shape and -«aaDiKK hy Use of Celloidin Capsules ,^e advantage of this form over .Ijonns with neck is that in the t^t lieok joins the body, and il dvhich is most obvious is that agglutination, however produced, would appear to be strictly associated with the existence of the bacteria in a living state within the body, for other- wise we cannot explain the fact that removal of the encapsulated bacteria fvoni the peritoneal cavity is followed by the steady dis- appearance of the agglutinating property of the serum of the animal. So far as these observations go, they would appear to explain the continued existence of the reaction for months and years in some individuals following upon an attack of typhoid and the rapid dis- appearance of the reaction in other cases, and this by the continued existence of the bacteria within the tissues in one set of cases and by their complete destruction in the other series. A few years ago this conclusion would have seemed impossible, but now-a-days we are, I think, prepared to accept it, for it is now a familiar experience that typhoid bacteria may be obtained from the gall bladder many months after the patient has apparently wholly recovered from the acute disease, while similariy, long months after the patient has been sub- jected to the disease we occasionally encounter the bacilli in pure culture in abscesses in the neighborhood of joints and other lesions, observations which prove absolutely that the bacilli may continue for long periods, either lying latent or proliferating very slowly in some one or other region of tlie body. In attempting to form a satisfactory theory of agglutination, wo possess data which lead us to suppose that the bacteria in culture form certain agglutinms which unite with certain other agglutinins which are^^he product of the tissues. (The normal serum will, in certain ca|H produce in low dilutions an agglutination; but this power is probably not an inherent quality of tissues or serum, but is Um of Ctlloidin Caimilen by a bMillufl nearly related it made. So rarely does occur that it may be .viacted upon by an lnfaetion» mpofiA by the pirodiiifllltiiin|P9|||p|||k By the oapaule method we are ablo to assert that the agglutfain produced by the tissues is not a reaction to the ba^illary bodies, nor yet a reaction of the kind called inflammatory, but a reaction to the chemical bacillary products or a combination of the serum with those chemical products. SUMMARY. 1. Capsules made as described above allow dialywH, when placed in the peritoneal cavity. 2. The normal tissuw, unstitnulnted, do not possess the power of causing agglutination; they do not require to be stlniulati-d hy the presence of the bacterial boflies, but will produce their share of the agglutinins when acted upon by the bacillary protlucts. 3. Agglutination follows the insertion, in the peritoneal cavity, of "capsuled" bacilli; it gradually increases in degree, and on the removal of the capsule containing the bacilli, begins to disappear. 4. Varieties of bacilli, related closely in morphology and cultural ronotiona, do not, ns a rule, produce serums which iiiter-agglutinate.