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f\^e>osje,7t<iWW. 
 
 NOTES OF THREE HUNDRED AGGLUTINATION TESTS WITH 
 
 B. COLI €0MMUN1S. 
 
 BT 
 
 JOHN MoCBAB, M.B., (Tor.) 
 Fellow in Pathology, MoGill University. 
 
 Reprinted from the Montreal Medical Journal, May, 1900. 
 
 .^r-\ 
 
 
NOTES OF THREE HUNDRED AGGLUTINATION TESTS 
 TRIED WITH B. COLI COMMUNIS. 
 
 HY 
 
 John McCrae, M.B., (Tor.) 
 
 Fellow in Pathology, McGill lJiilver8ity. 
 
 (From the Molson Pathologicrvl Laboratory, McGlll University,) 
 
 In the absence of sufficient data upon the agglutinative action of bfood 
 serum upon B. coli, it was thouglit well to try this action in a routine of 
 cases taken at random in the various wards of the hospitals ; and, in addi- 
 tion, some healthy cases were observed. Many of the cases noted, were 
 examined with a view to their Widal reaction, and those which gave the 
 reaction are referred to as typhoid. 
 
 Technical Method. 
 Three forms of B. coli were isolated, selected from a number, and 
 named I., II. and III. 
 
 1, Showed slight motility, slight fermentation and very slow formation 
 of acid. It was isolated from the healthy bowel. 
 
 II., from the stomach of a case of pernicious anaemia (and proved to 
 be B. coli), was of very slight motility, moderate acid-forming and 
 slight coagulating ability, and of moderate gas-producing power. 
 
 III., wias isolated from the bowel of a case of chronic diarrhoea, waa of 
 slight motility, strong acid-forming and coagulating power, and strong 
 gas-producing power. 
 
 Tlic limit of reaction fixed at one hour; and the dilution was in case 
 of fresh serum, 1 in 10. Where the specimen was dried blood, a dilution 
 uas made as near as possible to 1 in 10, though of necessity this could 
 be only estimated. Whenever a specimen was positive at 1 to 10, it was 
 tried at 1-20, 1-30, etc., until reaction failed. 
 
 A complete reaction was decided to be present in those cases in which 
 cessation of motion occurred, together with clumping and decided clear- 
 ance or tendency to clearance of the field between the clumps. \Vhere 
 there was a distinct clumping without the field being cleared of free 
 bacilli, the reaction was called " suggestive." These semi-reactions 
 wero fairly frequent, and no doubt indicate the cases in which a true 
 reaction would occur if the dilution were 1-2, 1-3, etc., instead of 1-10. 
 
 ]']vcry hanging-drop was examined as a control previous to the serum 
 being added to the culture, to prevent pseudo-reactions. This was 
 especially necessary in the case of Colon iii, which at times would show 
 a pseudo-reaction 3 to 3 hours after the bouillon had been inoculated. 
 
It was found necessary in this case to use one of two expedients, \it. ; 
 (1) use the culture at a very early moment after inoculation, or (2) use 
 a bouillon culture 3 to 5 days old, when the greater number of the bacilli 
 settled as a sediment, and those in the supernatant fluid showed little, if 
 any tendency to clump. At the same time, the question arises if, from 
 some cause, such bacilli would not have lost their power of clumping to 
 ordinary stimuli (such as that of bloom serum). 
 
 Results (300 Agglutination Tests). 
 
 Positive 10 
 
 Suggestive 18 
 
 Negative 278 
 
 Of 10 positive reactions, the distribution was: 
 
 With bacillus 1 1 
 
 II 1 
 
 Ill 8 
 
 
 it 
 
 Of 18 suggestive reactionp, the distr »ution was: 
 
 With bacillus I. 
 
 .. II. 
 
 III. 
 
 
 (( 
 
 tt 
 
 .1 
 
 .2 
 
 .16 
 
 Of the 10 positive in 1-10 dilution, one showed a positive reaction in 
 1-15 dilution; the others no higher than 1-10. One case only (a case 
 of undiagnosed chronic stomach trouble, (gastritis) reacted to bo+' ii 
 and iii. 
 
 Of 10 positive reactions, the diseases were as follows : 
 
 Bronchitis 
 
 Typhoid (convalescent) 
 
 Anaemia 
 
 Gastric ulcer , 
 
 Gastritis? (v. sup.) 
 
 Undiagnosed 
 
 Of 18 suggestive reactions, the diseases were as follows: 
 
 Appendicitis 
 
 Acute rheumatism 
 
 Bronchitis (acute) , 
 
 Exoph. Goitre , 
 
 Typhoid , 
 
 Gastritis (v. sup.) , 
 
 Organic heart disease 
 
 Undiagnosed 
 
 Normal 
 
 3 
 ] 
 1 
 1 
 3 
 1 
 1 
 7 
 1 
 
8 
 
 Of cases, possibly of B. coli infectioo, which gave no reaction, the 
 following were tested : 
 
 Cholecystitis 1 
 
 Appendicitis 3 
 
 Typhoid 11 
 
 Probably typhoid, no certain data 8 
 
 Results, iNFiiBENOES, Etc. 
 
 1. The greater liability to reaction of III, renders it very desirable 
 that different races of B. Coli should be used, each one with a shorter 
 series of cases than 100. 
 
 2. As has been frequently stated, the liability of a strongly acid and 
 gas-producing bacillus to clump (so far as this series shows) is demon- 
 strated. 
 
 3. It is desirable that the virulence of such a strongly acid and gas- 
 producing bacillus, be tested; as it is probable that from its growth and 
 character on different media ^11. would have proved to be the most 
 virulent of the three forms selected. 
 
 4. It is also desirable that a dilution of at least 1 in 10 and a time limit 
 not exceeding 1 hour be adopted, that the clumping power of normal 
 blood serum be avoided. 
 
 5. It is desirable that a series of cases of such acute infections as are 
 liable to be caused by B. Coli, should be tested. So far as the above 
 series goes there appears to be no increased reaction observed in the 
 cases of appendicitis, cholecystitis, etc. 
 
 From the number of postitve typhoids which gave no reaction (in 
 which the Widal test was made the crucial point), it, so far, does not 
 differ from Ihe observations already made by Drs. Johnston and Mac- 
 Taggart, that a decided reaction of B. Typhosus is generally accompanied 
 by a low degree of reaction to B. Coli. 
 
 6. If, as the researches conducted here by Drs. 1 dami, NichoUs and 
 Ford seem most clearly to demonstrate, the colon bacilli and other 
 bacteria are constantly being taken in small numbers into the system, 
 and are to be found in the tissues in various stages of degeneration, then 
 this mere taking up, even if constant, is not, under ordinary conditions 
 of health, sufficient in man to develop any pronounced serum reaction 
 against the colon bacillus. This would seem to indicate that the mere 
 act of arrest of growth and destruction of bacteria by the tissues, does 
 not in itself lead to the development of any large quantity of aggluti- 
 nating substance in the circulating blood and may indicate that there 
 must ,be an active proliferation of the bacteria in the tissues before any 
 recognisable amount of agglutinating material is produced. 
 
 7. Lastly, remembering how abundantly the colon bacilli proliferate 
 
within the intestine under normal conditions, it would not havo been 
 surprising to find that human blood constantly exerted a very definite 
 agglutinating action upon the colon bacillus. The very fact that the 
 reaction is so relatively uncommon is of interest as showing that the 
 •presence of toxine-bearing and toxine-producing bacteria within the 
 intestines does not result in the circulating blood containing what I may 
 term, broadly, reactive substances. It is true that like those of the 
 typhoid bacillus, the toxines of the colon bacillus only diffuse out with 
 difficulty. It is difficult, however, to imagine that none of these tox- 
 ines are discharged from the bodies of the bacilli when they arc growing 
 within the intestinal canal and that none are absorbed. Indeed, the 
 results here presented, so fur as they go, seem to indicate that for the 
 development of agglutinating substances in recognisable amounts within 
 the blood, there must be some very intimate reaction between bacilli 
 which have invaded the tissues and the tissues themselves.