Part 19

_Agglutination tests with serum of recovered cases, May 17, 1908._ ──────┬─────────────────────────┬────────┬───────────────────────────── Case. │ How infected; symptoms. │Strains.│ Serum dilutions. „ │„ │„ │¹⁄₂₀ │¹⁄₄₀ │¹⁄₆₀ │¹⁄₁₀₀│¹⁄₂₀₀ ──────┼─────────────────────────┼────────┼─────┼─────┼─────┼─────┼───── 1 B. │(From this case stool 2.)│A │ + │ + │ + │ ± │ 0 K. │ Laid rat poison │ │ │ │ │ │ │ 5–2,5–5; some days │ │ │ │ │ │ │ later diarrhea; later │ │ │ │ │ │ │ vomiting; convalescence│ │ │ │ │ │ │ and recovery May 10. │ │ │ │ │ │ „ │„ │B │ + │ + │ + │ ± │ 0 „ │„ │C │ + │ + │ + │ ± │ 0 „ │„ │D │ + │ + │ + │ + │ ± ──────┼─────────────────────────┼────────┼─────┼─────┼─────┼─────┼───── 2 K. │Ate three pieces of │A │ + │ 0 │ 0 │ 0 │ 0 E. │ infected bread May 2; │ │ │ │ │ │ │ mild diarrhea several │ │ │ │ │ │ │ days. │ │ │ │ │ │ „ │„ │B │ + │ ± │ 0 │ 0 │ 0 „ │„ │C │ 0 │ 0 │ 0 │ 0 │ 0 „ │„ │D │ + │ ± │ 0 │ 0 │ 0 ──────┼─────────────────────────┼────────┼─────┼─────┼─────┼─────┼───── 3 H. │Brought the virus April │A │ + │ 0 │ │ │ B. │ 28; perhaps touched it;│ │ │ │ │ │ │ next day diarrhea and │ │ │ │ │ │ │ vomiting; recovery │ │ │ │ │ │ │ after several days. │ │ │ │ │ │ „ │„ │B │ + │ 0 │ │ │ „ │„ │C │ + │ 0 │ │ │ „ │„ │D │ − │ − │ − │ − │ − ──────┼─────────────────────────┼────────┼─────┼─────┼─────┼─────┼───── 4 G. │Father of man that died; │A │ + │ + │ + │ + │ 0 S. │ laid virus; two days │ │ │ │ │ │ │ later mild diarrhea for│ │ │ │ │ │ │ one or two days. │ │ │ │ │ │ „ │„ │B │ + │ + │ + │ + │ 0 „ │„ │C │ + │ + │ + │ 0 │ 0 „ │„ │D │ + │ + │ + │ + │ ± ──────┼─────────────────────────┼────────┼─────┼─────┼─────┼─────┼───── 5 J. │On April 27 laid virus; 2│A │ ± │ 0 │ 0 │ 0 │ 0 K. │ days later diarrhea for│ │ │ │ │ │ │ several days. │ │ │ │ │ │ „ │„ │B │ ± │ 0 │ 0 │ 0 │ 0 „ │„ │C │ ± │ 0 │ 0 │ 0 │ 0 „ │„ │D │ − │ 0 │ 0 │ 0 │ 0 ──────┼─────────────────────────┼────────┼─────┼─────┼─────┼─────┼───── 6 J. │Laid virus April 27; two │A │ 0 │ 0 │ 0 │ 0 │ N. │ days later had diarrhea│ │ │ │ │ │ │ for several days. │ │ │ │ │ │ „ │„ │B │ + │ + │ + │ ± │ „ │„ │C │ 0 │ 0 │ 0 │ 0 │ „ │„ │D │ + │ + │ + │ ± │ ──────┼─────────────────────────┼────────┼─────┼─────┼─────┼─────┼───── 7 G. │“Held” the virus April │A │ + │ + │ + │ + │ ± I. │ 28. Next day diarrhea │ │ │ │ │ │ │ for four days. │ │ │ │ │ │ „ │„ │B │ + │ + │ + │ + │ ± „ │„ │C │ + │ + │ + │ + │ ± „ │„ │D │ − │ − │ − │ − │ − ──────┼─────────────────────────┼────────┼─────┼─────┼─────┼─────┼───── 8 H. │Ate and associated with │A │ + │ + │ 0 │ │ R. │ persons who handled │ │ │ │ │ │ │ virus; diarrhea several│ │ │ │ │ │ │ days. │ │ │ │ │ │ „ │„ │B │ + │ − │ 0 │ │ „ │„ │C │ + │ ± │ 0 │ │ „ │„ │D │ − │ − │ − │ │ ──────┼─────────────────────────┼────────┼─────┼─────┼─────┼─────┼───── 9 K. │Associations as case 8; │A │ + │ + │ + │ + │ S. │ headache several days │ │ │ │ │ │ │ and loss of appetite. │ │ │ │ │ │ „ │„ │B │ + │ + │ + │ + │ „ │„ │C │ + │ ± │ 0 │ │ „ │„ │D │ − │ − │ − │ − │ − ──────┼─────────────────────────┼────────┼─────┼─────┼─────┼─────┼───── 10 │Associations as case 8; │A │ 0 │ 0 │ 0 │ 0 │ │ diarrhea for eight │ │ │ │ │ │ │ days; vomiting several │ │ │ │ │ │ │ days. │ │ │ │ │ │ „ │„ │B │ + │ + │ + │ ± │ „ │„ │C │ 0 │ 0 │ 0 │ 0 │ „ │„ │D │ + │ + │ + │ ± │ ──────┴─────────────────────────┴────────┴─────┴─────┴─────┴─────┴───── A = Strain from Loeffler. B = Strain from market virus. C = Strain from stool 1 (fatal case). D = Strain from stool 2 (case 1 of table). − = No test made. 0 = Negative. ± = Slight. Controls = Serum of five normal persons tested as above; gave in no case agglutination higher than 1 : 20.

The attending physician, noting that most of his patients had come into recent contact with rat virus (_B. typhi murium_) and suspecting that to be responsible, sent specimens of stools to the Hygienic Institute at Munich, where they were carefully examined with reference to this subject.

Organisms identical with Loeffler’s _B. typhi murium_ were isolated from the two stools examined and these cultures were compared with and conformed with a culture of _B. typhi murium_ obtained from Loeffler and also a culture from the virus on the local market.

Two guinea pigs injected with cultures from the two stools gave, after the second injection, serum which agglutinated all the above organisms 1:200. A mouse typhoid serum obtained from Loeffler agglutinated all the above strains distinctly in dilutions 1:640 and slightly in 1:1280.

In conclusion, the author considers three possibilities: 1. The mouse typhoid bacillus was the cause of the illness. 2. The bacillus was accidentally present, having no part in the production of the symptoms. 3. The bacillus was able to multiply only in case pre-existing intestinal trouble; then, however, causing the inflammation.

The case of Mayer, who became infected during the course of some laboratory experiments, is particularly instructive.

During an epidemic of mouse typhoid among his laboratory mice, evidently spread from some inoculated mice by ants, Mayer[BI] who had personally handled the infected mice and their cages, became sick July 15, just seven days after the first appearance of the ants and after the observed rise in virulence of the mouse typhoid among the mice. His clinical history is as follows:

Footnote BI:

Mayer, Georg: Ueber die Verschleppung typhöser Krankheiten durch Ameisen und die Pathogenität des Loeffler’schen Mäusetyphusbazillus für den Menschen. Münch. med. Woch., vol. 52, 1905, p. 2261.

July 15: Weakness, epigastric pain, obstipation, temperature 37.7, pulse 90.

July 16: Slight diarrhea, increase of pain in region of trans-colon, temperature 38.3, pulse 98.

July 17: Diarrhea continued, pains increased—severe, chill in evening, temperature 39.1, pulse 102.

July 18: Obstipation, symptoms worse, chill again in evening, temperature 39.4, pulse 104.

July 19: Symptoms better, stools from purgative, evening temperature 36.9, pulse 68.

July 20: Left bed. Temperature and pulse normal, but weakness and slight epigastric pains continued till August 7.

Patient’s serum agglutinated as follows:

──────────────────────────┬──────────────┬──────────────┬────────────── │ Typhoid. │ Paratyphoid. │Mouse-typhoid. ──────────────────────────┼──────────────┼──────────────┼────────────── July 23 │ 1–50+│ 1–50+│ 1–250+ August 7 │ 1–50+│ 1–50+│ [BJ]1–100+ August 16 │ 1–50+│ 1–50+│ 1–50 + ──────────────────────────┴──────────────┴──────────────┴──────────────

Footnote BJ:

Slight

The bacillus of mouse typhoid was isolated from the patient’s stools July 21 and 23. Negative results thereafter. Examinations continued a month.

Same organism isolated from urine July 21. Negative thereafter.

The author concludes that the _B. typhi murium_ is able to cause in man a rather severe acute illness of short duration.

Shibayama[BK] gives the following report of outbreaks of human infection that have come to his knowledge in Japan, where mouse-typhoid virus has been used in considerable quantities.

Footnote BK:

Shibayama, G.: Ueber Pathogenität der Mäusetyphusbazillen für den Menschen. Münch. med. Woch., May, 1907, p. 979.

_Outbreak 1._ In April, 1905, in a village of the Province of Saitama, 30 people became ill and 2 died with severe gastro-intestinal symptoms. Outbreak investigated by Dr. H. Sezuki, district medical officer, formerly of the Tokio Institute for Infectious Diseases.

It was found that all the 30 people had partaken of a dish of cooked vegetables served at a meeting of the town council, and that for application of sauce to these vegetables (after cooking) a wooden vessel had been used which two days before had been used for mixing mouse-typhoid virus with meal, without subsequent cleansing or sterilization.

The symptoms came on within twelve to forty-eight hours thereafter (usually twenty hours), chill or chilly sensations, rise of temperature to 38° or 39° C., or even to 40° C.; face flushed; pulse accelerated; great weakness; thirst, nausea, colicky pains in abdomen followed by severe diarrhea and vomiting. In general, the fever and diarrhea lasted two or three days; but malaise, anorexia, weakness, and mucous stools persisted for several days. The more severe cases showed choleraic symptoms of collapse. Two persons died in spite of medical treatment—a 6-year old boy and a man of 43, on the second and third day, respectively.

From the intestinal contents of these two cases, from the stools of several other cases, and from the remnants of the dish of vegetables in the wooden bowl an organism was isolated, which was demonstrated to be identical with the bacillus of mouse typhoid.

These results were confirmed at the Tokio Institute for Infectious Diseases by Shibayama, by biological and immunizing tests.

_Outbreak 2._ On December 7, 1905, a peasant of a village in the Province of Miyaki brought home some mouse virus mixed with meal in cakes. This being mistaken for “mochi” was eaten about 2 p. m. the next day by two little girls, 3 and 8 years old, respectively, and their grandfather, 61 years old.

The man and the 8-year-old girl became sick at 9 p. m. the same day and the other child at about 3 p. m. on December 9. The symptoms in all cases were those of severe gastro-enteritis, as described under outbreak 1.

The man died December 12, the 8-year-old child died on the 10th; the 3-year-old child recovered after several days’ illness. These three alone ate of the virus and no other persons in the house became sick.

No bacteriological examination was practicable.

_Outbreak 3._—In a village of the Province of Iskawa, on April 22, 1906, a lot of rat poison was prepared by mixing agar cultures of the mouse-typhoid bacillus with meal and water in a large wooden bowl.

On April 24 there was a festival in the village at which about 170 persons were served with 240 pounds of rice, which, after being cooked, was kneaded into cakes in a wooden bowl. About 80 pounds of this rice was so kneaded in the bowl previously used for preparing the rat poison. Twenty to twenty-four hours later 120 people who had eaten of the rice became ill with the already described symptoms of gastro-enteritis, of mild type among the strong but severe among the children and old people. Eighty-nine cases came under medical treatment. There were no deaths, but a number of cases were confined to bed for a week or more; mild cases recovered in one to three days.

No bacteriological examination was made, but the physicians and town officials were unanimously of the opinion that the rat virus was the cause of the outbreak.

_Outbreak 4._—A peasant of the province of Niigata brought home on May 14, 1906, some rat virus (cultures of mouse-typhoid bacilli mixed with meal) which he laid away. Two of his grandchildren—a boy of 5 and a girl of 7—together with the 4-year-old daughter of a neighbor, found and ate the rat virus. The next day all three children became ill with severe gastro-enteritis, of which the 4-year-old child died on the third day. The others recovered after several days of medical treatment.

_Outbreak 5._—On May 16, 1906, a peasant in the province of Jamagata brought home some rat virus (6 c. c. cultures of mouse-typhoid bacillus mixed with meal), which was accidentally mixed with the feed given to a healthy horse next morning. The same evening the horse showed loss of appetite and appearance of sickness. Within two days he developed a severe enteritis, of which he died on the seventh day. The body was buried, but was dug up in the night by a laborer who cut off the hind quarters, took them home, and distributed the meat among friends and neighbors.

Within three days 34 persons who had eaten of this meat became ill with symptoms of severe gastro-enteritis. A 72-year-old man died after five days; the others recovered in three to eleven days.

This outbreak was investigated by Dr. H. Segawa, a medical officer of the province and former member of the institute at Tokyo, who isolated from the remains of the horseflesh by plate cultures and animal inoculations, an organism identical with the bacillus of mouse typhoid. A culture was sent to Shibayama, who carefully verified it (details not given).

Shibayama concludes: In all cases the close relationship between the bacillus of mouse typhoid and the illness was established; and he thinks this organism must be accepted as the direct cause of the outbreaks.

Referring to Loeffler’s uniformly negative human experiments, he calls attention to known cases where men have taken virulent cultures of typhoid, diphtheria, etc., without infection. According to many bacteriological investigations, _B. typhi murium_ is identical with the bacillus of enteritis. If it is proven that the latter is a cause of acute gastro-enteritis then the conclusion is likewise justified that the _B. typhi murium_ is frequently pathogenic for man, causing an acute gastro-enteritis.

Fleischanderl[BL] a reports six cases of illness—three severe and three mild—occurring in his practice in the latter part of April, 1908, presenting the following symptoms: Onset with rapidly increasing body pains, followed in a few hours by diarrhea, rise of temperature, and general prostration; in the next two or three days aggravation of the symptoms, fever (39° to 40° C.), copious diarrhea, vomiting (in one case), severe body pains, vertigo, and considerable prostration. Symptoms abated quickly in a few days, leaving considerable prostration, convalescence requiring two weeks in one case. In the less severe cases there was no fever, and the other symptoms were generally milder.

Footnote BL:

Fleischanderl, Fritz: Mitteilung über einige Krankheitsfälle, hervorgerufen durch Mäustyphusbazillen. Munch. med. Woch., vol. 56, Feb., 1909, p. 392.

The simultaneous appearance of these and other similar rumored mild cases among the neighbors (about 20 in all) pointed to a common cause. It was found that three of the six cases were in people who had handled mouse-typhoid cultures the day before their illness, taking no precautions to avoid infection.

The other three occurred in a family which, on the day before the onset of the illness, had drunk raw milk obtained from a house where the rat virus had been used shortly before, and only three members of the family who drank the milk became ill.

In order to prove the etiology of these cases Fleischanderl, who had never suffered any intestinal troubles, had had nothing to do with any case of typhoid fever for a year, and was in excellent health, took a culture of the mouse-typhoid bacillus as used in the neighborhood, rubbed a glass rod over the surface, washed it off in a glass of water, and drank this before breakfast on the morning of May 3.

In twenty-two hours he experienced mild, increasing body pains, followed within a few hours by diarrhea, and a few hours later by slight chill, rise of temperature to 38.2, pulse 106, severe pains in body, and feeling of great weakness.

May 4, 9 p. m.: Temperature 39.2° C., pulse 120. Height of symptoms.

May 5: Temperature 38.2° to 38.5° C., pulse 106 to 120. Other symptoms continued.

May 6: Temperature and pulse normal. All symptoms disappeared except weakness, which lasted two days.

Bacteriological investigations conducted by Herbert Berger in the K. K. Serotherapeutischen Institut and by Doctor Reichel, assistant in the Hygienic Institute of the University of Vienna, follow:

From the stools of one of the patients infected from milk an organism was isolated which, injected into mice (1 c. c. emulsion of forty-eight-hour culture), killed them in two to five hours. Mice infected by eating these dead mice died in thirty to forty-eight hours.

Control mice inoculated similarly with a culture of the market mouse-typhoid virus died in twenty to thirty hours, while the mice infected through eating these died after three to four days.

The following strains were used for cultural agglutination tests:

A. From stools of patient infected from milk.

B1. Market virus used in injecting mice.

B2. Market virus taken by author.

C25. From stools of author twenty-five hours after infection.

C55. From stools of author fifty-five hours after infection.

LL. Stock culture of Loeffler’s mouse-typhoid bacillus.

LP. Stock culture of para-typhoid bacillus.

All organisms (A-C55) were demonstrated as motile bacilli, not liquefying gelatine, not forming lactic acid, and forming gas from dextrose.

The serum of a rabbit after two injections of LL agglutinated LL and LP in dilution of 1:1280, did not agglutinate A, B1, B2, C25, and C55.

Serum of rabbit after one injection of B2 agglutinated in 1:320 dilution A, B1, B2, C25, C55, and LL; did not agglutinate LP.

Serum of rabbit after one injection of C25 gave exactly similar results.

Doctor Reichel considers it proven that the organisms A to C55 are undoubtedly identical with Loeffler’s bacillus of mouse typhoid, and distinct from para-typhoid bacilli. The author considers it proven that this bacillus was the sole cause of the cases of enteritis observed.

Recently Mallory and Ordway[BM], in a paper read before the American Association of Pathologists and Bacteriologists held in Boston, reported that lesions analogous to the early stages of typhoid lesions may be produced in rats by the use of Danysz virus.

Footnote BM:

Mallory, F. B., & Ordway, T.: Lesions produced in the rat by a typhoid-like organism (Danysz virus). Journ. Am. med. assn., vol. 52, May 1, 1909, p. 1455.

In view of these facts the statements of some of the advertising matter of certain rat viruses call for revision.

REFERENCES TO THE LITERATURE.

Loeffler,[BN] 1889, gives an account of two spontaneous outbreaks among the mice kept at the Hygienic Institute at Griefswald. It was from these animals that he obtained and described the original _B. typhi murium_. He determined that the infection was by ingestion and that the organism was especially virulent for field mice. He described the organism in detail and also the lesions.

Footnote BN:

Loeffler, F.: Ueber Epidemieen unter den im hygienischen Institute zu Griefswald gehalten Mäusen und über die Bekämpfung der Feldmäusplage. Centblt. f. Bakt., Orig., vol. 11, 1898, p. 129.

Laser,[BO] 1892, reports that on the morning of February 6, 1892, 70 of the 76 field mice (_Mus agrarius_) used as experiment animals in the Hygienic Institut at Königsberg were found dead. A small bacillus twice as long as broad, displaying a very lively specific motility, was isolated from the spleen. It was tested upon animals and all the results compared with Eisenberg’s tables and found to be closely allied to the bacillus of ferret plague (Ebert-Schummelbusch), to the bacillus of American swine plague (Billings), and to that of French swine plague (Chantamesse and Cornil).

Footnote BO:

Laser, Hugo: Ein neuer für Versuchsthiere pathogener Bacillus aus der Gruppe der Frettschen-Schweinseuche. Centblt. f. Bakt., Orig., vol. 11, 1892, p. 184.

Mereshkowsky,[BP] 1893, isolated an organism at the Royal Bacteriological Institute at St. Petersburg from a stock of Zisel (_Spermophilus musicus_) among which a spontaneous epizootic had occurred. The author found this culture to be virulent for domestic and field mice.

Footnote BP:

Mereshkowsky, S. S.: Ein aus Zieselmäusen ausgeschiedener und zur Vertilgung von Feld-resp. Mäusen, geeigneter Bacillus. Centblt. f. Bakt., Orig., vol. 17, 1895, p. 742.

Zupnik,[BQ] 1897, states that Joseph, of the Agricultural Institute of Breslau, in 1882 originated the use of favus fungus for the destruction of mice. Zupnik tested _B. typhi murium_ and Danysz virus upon mice. No experiments with rats.

Footnote BQ:

Zupnik, Leo: Ueber die pratische Verwendbarkeit der Mäuse bacillen inbesondere des Loeffler’schen _Bacillus typhi murium_. Centblt. f. Bakt., Orig., vol. 21, 1897, p. 446.

Issatschenko,[BR] in 1898, described briefly a bacillus obtained by him from gray rats. Recent investigation showed this bacillus to be very virulent for rats and mice, but harmless for the different species of domestic animals. Four hundred and forty-three experiments were made upon rats with pure cultures of the bacillus combined with dough and fed to the rats. He gives a table showing that the mortality occurred in 431 rats at an average of ten and one-half days. The greatest mortality occurred during the first fifteen days (84.2 per cent), with the greatest number on the seventh day (20.1 per cent).

Footnote BR:

Issatschenko, B.: Untersuchungen mit dem für Ratten pathogenen Bacillus. Centblt. f. Bakt., Orig., vol. 31, 1902, p. 26.

Danysz,[BS] 1900, isolated a cocco-bacillus during an outbreak of spontaneous disease amongst field mice which presented the general characteristics of the colon bacillus and to this extent resembled Loeffler’s bacillus (_B. typhi murium_), and which from the beginning exhibited some pathogenicity for gray rats (_M. decumanus_). Of ten such rats fed upon a culture of this organism, two or three died, while others that had fallen sick recovered and the same remained well. This small mortality offered some hope that it would be possible to increase the virulence of the bacillus by ordinary methods; that is, passing it from rat to rat. It was found, however, that the opposite was true; the virulence was always weakened by this process regardless of the method of administration. Thus in every series the first culture killed the animals in seven to twelve days; occasionally after one or two passages five to seven days; but subsequent passages decreased the virulence so that none died.

Footnote BS:

Danysz, J.: Un microbe pathogène pour les rats (_Mus decumanus_ et _Mus rattus_) et son application à la destruction de ces animaux. Ann. Inst. Pasteur, 1900, vol. 14, p. 193.

The general result is that it is difficult to maintain the virulence of the cocco-bacillus of the rat or to increase it when it is found to be small. It can only be effected by constantly making a large number of experiments and frequently testing the virulence of the culture. Danysz succeeded in keeping up a supply of cultures of sufficient strength for eight years. In 60 per cent of the operations where this culture has been used it has been successful in causing the absolute disappearance of the rats. In 15 per cent the result was entirely negative, and in the remaining 25 per cent there was a large diminution.

Oettinger,[BT] in 1903, increased the virulence of the Danysz bacillus by growth in an egg rendered alkaline after the method previously introduced by Wiener.

Footnote BT:

Oettinger, M.: Ueber die Wienersche Methode zur Virulenzsteigerung der Danysz Bazillen. Munch. med. Woch., vol. 1, 1903, p. 324.