Bacillus Typhosus Research Articles

An Investigation into the Agglutinating Power of Human Sera for Bacillus typhosus and Various Allied Organisms

1. Three hundred and two samples of sera from normal people, and 847 samples from cases suspected of enteric, or from contacts, have been examined for heat labile agglutinins against B. typhosus and allied organisms.2. Sera from the “not normals” show a higher percentage of agglutinins against nearly all the organisms tested than sera from the “normals.”3. In the contact group of the “not normals” the incidence of agglutinins is practically the same as in the “normal” group.4. Sera from inoculated persons show a higher percentage of agglutinins against those organisms included in the T.A.B. vaccine used during the late war, than the sera from non-inoculated persons.5. There is a higher percentage of agglutinins for B. typhosus, B. paratyphosus A and B. paratyphosus B among males than among females, especially in the age groups 20–30 and 30–40.6. The proportion of normal males whose sera agglutinate B. typhosus, B. paratyphosus A and B. paratyphosus B and the group suspension is significantly lower than corresponding proportion noted in an investigation recorded by Rosher and Fielden in 1922. The proportion of agglutinating sera for these strains among normal females is substantially the same in our findings as in theirs.7. The associations between ability to agglutinate different pairs of organisms are shown to be statistically significant in those cases which might arise from T.A.B. inoculation.8. For these reasons it seems likely that the present distribution among the adult male population of agglutinins for B. typhosus, B. paratyphosus A, B. paratyphosus B and for Salmonella strains allied to B. paratyphosus B when in the group phase, was largely determined, in the period 1925–6, by the results of T.A.B. inoculation carried out during the war. It is probable that this effect is slowly disappearing.9. Statistically significant associations were found between certain other pairs of organisms, which could not be explained as resulting from inoculation with T.A.B. vaccine, or as a result of any common antigenic stimulus. These associations were found mainly in the “not normal” group; and their meaning, at the moment, remains obscure. It may be noted that the number of sera agglutinating these particular pairs of suspensions was, in most cases, small.10. Our results, so far as they yield any information on this point, do not suggest that infection with any one of the organisms studied by us increases the probability of the patient's serum agglutinating some other bacterium, with which the infecting organism is not antigenically related.11. High titres were found chiefly in sera from the “suspected of enteric” group. The normal group usually gave low titres.

STUDIES ON BACILLUS TYPHOSUS TOXIC SUBSTANCES

It has been demonstrated that many multiples of minimal doses of Bacillus typhosus reacting factors can be neutralized by specific immune sera. The potency of a given serum can be conveniently titrated against increasing amounts of reacting factors. If the immune serum is diluted or if the amount of the reacting factors is too large for a given amount of serum, there is obtained neutralization but only irregularly. Normal and heterologous sera (therapeutic meningococcus and erysipelas horse sera) free of normal agglutinins or possessing normal agglutinins of a low titer (1:16) for Bacillus typhosus are not able to neutralize the reacting factors. There is obtained questionable neutralization with a serum possessing normal Bacillus typhosus agglutinins in dilution 1:64. The titer of the neutralizing antibodies increases in the course of immunization. Immune sera exercise a definite protection against the mortality induced by intravenous injection of Bacillus typhosus culture filtrates.

CHEMO-IMMUNOLOGICAL STUDIES ON CONJUGATED CARBOHYDRATE-PROTEINS

It has been demonstrated that many multiples of minimal doses of Bacillus typhosus reacting factors can be neutralized by specific immune sera. The potency of a given serum can be conveniently titrated against increasing amounts of reacting factors. If the immune serum is diluted or if the amount of the reacting factors is too large for a given amount of serum, there is obtained neutralization but only irregularly. Normal and heterologous sera (therapeutic meningococcus and erysipelas horse sera) free of normal agglutinins or possessing normal agglutinins of a low titer (1:16) for Bacillus typhosus are not able to neutralize the reacting factors. There is obtained questionable neutralization with a serum possessing normal Bacillus typhosus agglutinins in dilution 1:64. The titer of the neutralizing antibodies increases in the course of immunization. Immune sera exercise a definite protection against the mortality induced by intravenous injection of Bacillus typhosus culture filtrates.

Ammonium Sulphate Precipitation of Bacillus Typhosus Toxic Subtances in the Phenomenon of Local Skin Reactivity

Ammonium Sulphate Precipitation of Bacillus Typhosus Toxic Subtances in the Phenomenon of Local Skin Reactivity

Improved Method of Preparing Bacillus Typhosus Toxic Substances Necessary for Local Skin Reactivity.

ConclusionIt is possible to obtain highly potent skin preparatory and reacting factors in filtrates of washings of 20-22 hour old Bacillus typhosus cultures on plain agar. Under these conditions there is avoided much of the extraneous material present in tryptic digest broth previously employed, as well as much of the bacterial autolysis occurring in the older method. This would indicate that the skin preparatory and reacting factors are bacterial soluble substances, the production of which is independent of massive cell destruction.The filtrates prepared in this manner produce negligible primary effect upon the skin (erythema, swelling), notwithstanding their high skin preparatory potency. This is in agreement with the view previously expressed1,25 that the skin preparatory potency of a given filtrate bears no relation to its primary effect on the skin.It is of considerable interest to note that there exists a distinct reciprocal quantitative relationship between the skin and intravenous (loses. This obs...

Action of Bacteriophage upon Production of in vivo Prepared Toxic Substance of Bacillus Typhosus.

In a previous communication1 we reported the results of the inoculation of guinea pigs with the toxic material obtained from the exudate of experimental typhoid peritonitis. The filtered toxic material produced lesions closely simulating those of human typhoid fever and the animals died after 3 or 4 inoculations. The toxic substance produced in this manner was found to be of rather low potency. Single injections of from 3 to 5 cc. failed to produce death although the animals became sick and showed marked reactions to the inoculations.It was thought that the employment of typhoid bacteriophage in conjunction with this in vivo method may serve the purpose of yielding a more toxic substance. It has already been shown that the action of bacteriophage in vitro has but little if any effect upon the toxicity of certain microorganisms. It was, however, thought that since the toxin employed previously by us was prepared in vivo, the presence of this lytic agent in the field of conflict between the host and the inv...

Antigenic Analysis of Strains of Bacillus Typhosus

The antigenic composition of B:typhosus has been studied by a number of investigators. Observations range from close similarity between strains, on the one hand, to evidence that distinct differences exist. Hooker,' for example, concluded from results of agglutinin absorption and particularly complement fixation that his strains could be separated into three groups. Aside from purely bacteriological and immunological considerations, the question of the uniformity of strains of B.typhosus presents certain practical aspects. A laboratory which does any considerable amount of diagnostic work needs for Widal tests a strain of typhoid which is readily and consistently agglutinable. The search for such a strain sometimes presents difficulties. The same problem occurs in the selection of a suitable culture for the manufacture of typhoid vaccine, local strains sometimes being employed for this purpose, on the assumption that they will be antigenically similar to most other strains of the locality. Antigenic similarity or dissimilarity also affects the identification of unknown cultures.

ALLERGIC IRRITABILITY

The allergic irritability of closely inbred guinea pigs as represented by their capacity to produce hemolytic antibodies for beef and sheep corpuscles, and agglutinins for Bacillus typhosus and Bacillus abortus (Bang) differs by families and therefore is at least partly dependent on inherited characteristics. These differences show an imperfect but suggestive correlation with the differences in resistance of the same families to inoculation tuberculosis as previously determined by Wright and Lewis. The differences in antibody production also show an imperfect correlation with the differences in response in the anaphylactic reaction complex as previously determined by Lewis and Loomis. These studies suggest very strongly that the allergic irritability is one of the several inheritable characters which form a partial basis for the natural resistance to tuberculosis. The antibody-producing capacity is only satisfactorily defined when minimal or moderate amounts of antigen are used and this in single treatments. The irregularities in experimental result when repeated treatments or very large single treatments are used suggest that antibody production in the second or "acquired capacity" phase may rest on a somewhat different fundamental basis than the latent or potential natural capacity. There is some very slight evidence that production in the second phase may also be influenced by inherited qualities.

Note on the Rought and Smooth Forms of Bacillus Typhosus

Note on the Rought and Smooth Forms of Bacillus Typhosus

Note on the Agglutinogenic Properties of Washings of Cultures of Bacillus Typhosus

Note on the Agglutinogenic Properties of Washings of Cultures of Bacillus Typhosus

The Relation of the Hypophysis to Antibody Production

An operative technique was evolved permitting successful partial hypophysectomy in guinea pigs. Such animals, when immunized to Bacillus typhosus, produced specific agglutinins in the same quantity and at the same rate as unoperated and operation controls immunized at the same time and by the same method. In guinea pigs previously immunized to Bacillus typhosus and hen red blood corpuscles partial hypophysectomy had no effect on the continued production and persistence of typhoid agglutinins, hemagglutinins, and hemolysins. In guinea pigs immunized to Bacillus typhosus both the continued ingestion and the intraperitoneal injection of the whole pituitary gland extract (Burroughs Wellcome) had no effect on the subsepuent agglutinin titers as compared to that of normal animals. The experiments would appear to show either that the hypophysis does not play an important direct or indirect part in the production of and persistence in the blood of typhoid agglutinins, hemagglutinins, and hemolysins, or that the amount of hypophysis left behind in the operation in order to maintain life is adequate also to exercise the degree of functional influence on these processes which the entire hypophysis conceivably exercises.

Note on a Case of Cholelithiasis in which Bacillus Typhosus was Isolated from the Centre of a Gall-Stone

Note on a Case of Cholelithiasis in which Bacillus Typhosus was Isolated from the Centre of a Gall-Stone

THE STABILITY OF BACTERIAL SUSPENSIONS

1. The addition of proteins or serum to suspensions of bacteria, (Bacillus typhosus or rabbit septicemia) at different pH widens the acid agglutination zone and shifts the isoelectric point to that of the added substance. 2. The amount of serum required to agglutinate is much less near the acid agglutination point of the organisms. 3. The addition of immune serum prevents the salt from decreasing the cohesive force between the organisms, and agglutination therefore is determined solely by the potential, provided excess immune body is present. Whenever the potential is decreased below 15 millivolts the suspension agglutinates.

THE RELATION OF THE HYPOPHYSIS TO ANTIBODY PRODUCTION

An operative technique was evolved permitting successful partial hypophysectomy in guinea pigs. Such animals, when immunized to Bacillus typhosus, produced specific agglutinins in the same quantity and at the same rate as unoperated and operation controls immunized at the same time and by the same method. In guinea pigs previously immunized to Bacillus typhosus and hen red blood corpuscles partial hypophysectomy had no effect on the continued production and persistence of typhoid agglutinins, hemagglutinins, and hemolysins. In guinea pigs immunized to Bacillus typhosus both the continued ingestion and the intraperitoneal injection of the whole pituitary gland extract (Burroughs Wellcome) had no effect on the subsepuent agglutinin titers as compared to that of normal animals. The experiments would appear to show either that the hypophysis does not play an important direct or indirect part in the production of and persistence in the blood of typhoid agglutinins, hemagglutinins, and hemolysins, or that the amount of hypophysis left behind in the operation in order to maintain life is adequate also to exercise the degree of functional influence on these processes which the entire hypophysis conceivably exercises.

THE ORMATION OF VACUOLES DUE TO BACILLUS TYPHOSUS IN THE CELLS OF TISSUE CULTURES OF THE INTESTINE OF THE CHICK EMBRYO

The introduction of Bacillus typhosus into the hanging drop of a tissue culture of the intestine of a chick embryo leads to the rapid vacuolation of the cells of the tissues which comprise the growth of the cultures. The cells of the connective tissue, the endodermal membrane, the mesothelium, and also the clasmatocytes exhibit the ability to ingest Bacillus typhosus in the cultures.

THE SURVIVAL OF LEPTOSPIRA (SPIROCHÆTA) ICTEROHÆMORRHAGIÆ IN NATURE; OBSERVATIONS CONCERNING MICROCHEMICAL REACTIONS AND INTERMEDIARY HOSTS

1. Leptospira icterohaemorrhagiae is unable to grow in the urine, either with or without the addition of suitable culture ingredients, the acidity of the urine being detrimental to the growth. It survives less than 24 hours, unless the urine is neutralized or slightly alkalized, when the period of survival is somewhat longer. If suitable nutrient ingredients are added to the neutralized or slightly alkalized urine, the organism is able to grow for about 10 days, after which multiplication ceases. 2. Feces from normal or jaundiced persons destroy Leptospira icterohaemorrhagiae within 24 hours when a rich culture is added and the mixture allowed to stand at 26 degrees C. The addition of blood serum and corpuscles does not prevent the destruction of the organism. Autoclaved specimens and filtrates of unheated feces do not constitute a suitable medium in which to keep the organism alive for any length of time, but the addition of blood corpuscles and serum in adequate quantities renders them fairly satisfactory as media. Under natural conditions Leptospira icterohaemorrhagiae cast off in the feces cannot survive more than 24 hours. 3. Polluted water, sewage, and soil will not serve to keep Leptospira icterohaemorrhagiae alive for more than 3 days at the most. When deprived by filtration or autoclaving of their bacteria they become indifferent diluents and may be used to make up a culture medium when mixed with serum and citrate plasma of a suitable animal. Sterilized soil with a neutral reaction, when added to a culture, has an unfavorable effect upon the growth of the organism. 4. Most of the aerobic bacteria found in feces, sewage, soil, and tap water inhibit the growth of Leptospira icterohaemorrhagiae when inoculated into the same medium. Bacillus faecalis alkaligenes and many strains of non-hemolytic streptococci caused the least interference, although growth was never so vigorous or lasting in the media in which they were present as in the control media. Certain pathogenic bacteria (Bacillus typhosus, Bacillus paratyphosus, Bacillus dysenteriae, pneumococcus) are antagonistic to the growth of the spirochete. 5. Leptospira icterohaemorrhagiae is highly sensitive to the destructive action of bile, bile salts, and sodium oleate, but resists the action of saponin. In this last respect it differs from many so called spirochetes. The destructive action of these agents is counteracted by blood serum. 6. The larvae and adults of the Culex mosquito, the larvae of the house-fly and bluebottle fly, wood ticks (Dermacentor andersoni), and leeches failed to become carriers of the spirochetes when fed on infected guinea pigs or their organs; that is, they cannot play the part of an intermediary host of Leptospira icterohaemorrhagiae.

DUODENAL CULTURES IN TYPHOID FEVER

the Editor: —InThe Journal, March 10, 1917, p. 780, Dr. Durham, in his article on the isolation of the typhoid bacillus from duodenal contents, says, To the best of my knowledge and information, this is the first time that duodenal contents, secured by the use of the duodenal tube, ever gave a positive Bacillus typhosus culture. I wish to call attention to my own article published inThe Journal, Nov. 18, 1916, p. 1493.

FURTHER STUDIES ON TYPHOIDIN

This paper records the results of further studies on typhoidin made since the publication of the original article of Gay and Force. Since Gay and Force¹have reviewed the work on the application of preparations of the typhoid organism to the skin which had been published previous to their article, we shall confine ourselves to a review of observations on typhoidin which have appeared since that date. The original typhoidin employed by Gay and Force consisted of a ten-day culture of a single strain of<i>Bacillus typhosus</i>on glycerin broth evaporated to one-tenth volume. On account of deterioration of the preparation in this form, Gay and Claypole²precipitated the original typhoidin with twenty volumes of alcohol, filtered, washed with absolute alcohol and ether, and then dried on porcelain plates over sulphuric acid in a vacuum. With a freshly prepared suspension of this typhoidin powder in phenolated saline, equivalent

A Report on the use of Stock Vaccine in Infection by the Bacillus Typhosus, with an Analysis of two Hundred and Thirty Cases

A Report on the use of Stock Vaccine in Infection by the Bacillus Typhosus, with an Analysis of two Hundred and Thirty Cases

A New Culture Medium for the Isolation of Bacillus Typhosus from Stools

isolation of typhoid bacilli from feces the one at present most widely used is that devised by Endo in 1903. It allows the development of a high percentage of the typhoid organisms inoculated on it, but does not inhibit to any practical extent the growth of those fecal bacteria which develop on ordinary nutrient agar. The great value of the method lies in its sharp differentiation between the lactose-fermenting organisms and those organisms which do not ferment lactose. The colonies of the lactose-fermenting organisms are colored red after 24 hours' incubation, while the other colonies remain colorless. We have found that better results are obtained if both lactose and saccharose are added to the Endo medium, since certain members of the colon-bacillus group ferment saccharose more rapidly than lactose and hence their colonies take on the red color sooner and can no longer be regarded as slow colonies; the typhoid colony is without color in 24 hours, as on the Endo medium containing lactose alone. The chief disadvantage of the Endo medium lies in the fact that the red color is not confined to colonies of B. coli themselves, but spreads out through the medium adjacent to them. If the colonies of B. coli are close together the whole plate soon becomes red and then colorless colonies on it can no longer be distinguished from the red ones. We have tested a great number of stains separately and in combinations of varying strengths with the view of overcoming this difficulty while at the same time preserving the effectiveness of the medium for the sure growth of typhoid. In every instance where the acid or the basic fuchsin was tried, the color diffused into the medium around the red colonies, so that it was difficult, if not impossible, to recognize the typhoid colonies which may have lain in these areas. We have finally devised a medium that gives even better dif-