scholarly journals THE PURE CULTIVATION OF SPIROCHÆTA ICTEROHÆMORRHAGIÆ (INADA)

1916 ◽  
Vol 23 (4) ◽  
pp. 557-562 ◽  
Author(s):  
Tetsuta Ito ◽  
Haruichiro Matsuzaki
Keyword(s):  
Causative Agent ◽  
Guinea Pigs ◽  
Culture Media ◽  
Biological Properties ◽  
Facultative Anaerobe ◽  
Fluid Medium ◽  
Weil’S Disease ◽  
Pure Cultures ◽  
Weil's Disease ◽  
Different Strains

Pure cultures of the spirochætal causative agent of the disease known as Weil's disease, or febrile icterus, in Japan, have been obtained by us in a solid, a semisolid, and a fluid medium. The spirochæta thus isolated remains pathogenic for guinea pigs for many generations. Up to the present time we have succeeded through the courtesy of Professor Nagayo, Dr. Konuma, and Dr. Ishihara, in cultivating three different strains. The spirochæta is a facultative anaerobe. The solid and semisolid culture media possess one disadvantage, in that they are opaque on account of the addition of red blood corpuscles; but it is hoped that this drawback may soon be overcome by further studies. We shall report later the results of investigations regarding various questions in immunity as well as further details regarding the biological properties of the spirochæta.

scholarly journals SPIROCHÆTA HEBDOMADIS, THE CAUSATIVE AGENT OF SEVEN DAY FEVER (NANUKAYAMI)

1918 ◽  
Vol 28 (4) ◽  
pp. 435-448 ◽  
Author(s):  
Yutaka Ido ◽  
Hiroshi Ito ◽  
Hidetsune Wani
Keyword(s):  
New Species ◽  
Blood Serum ◽  
Causative Agent ◽  
Guinea Pigs ◽  
Endemic Area ◽  
Etiological Agent ◽  
Field Mouse ◽  
Pure Cultures ◽  
Field Mice ◽  
A New Species

A new species of spirochete which we have called Spirochæta hebdomadis has been described as the specific etiological agent of seven day fever, a disease prevailing in the autumn in Fukuoka and other parts of Japan. This spirochete is distinguishable from Spirochæta icterohæmorrhagiæ to which it presents certain similarities. Young guinea pigs are susceptible to inoculation with the blood of patients and to pure cultures of the spirochete, and those developing infection exhibit definite symptoms suggestive of those of seven day fever in man. The blood serum of convalescents from seven day fever contains specific immune bodies acting spirochetolytically and spirocheticidally against the specific spirochetes, but not against Spirochæta icterohæmorrhagiæ. The field mouse (Microtus montebelli) is the normal host of the spirochetes, which have been detected in the kidneys and urine of 3.3 per cent of the animals examined. The endemic area of prevalence of seven day fever corresponds with the region in which field mice abound.

scholarly journals A COMPARATIVE STUDY OF EXPERIMENTAL PROPHYLACTIC INOCULATION AGAINST LEPTOSPIRA ICTEROHÆMORRHAGIÆ

1918 ◽  
Vol 28 (5) ◽  
pp. 561-570 ◽  
Author(s):  
Hideyo Noguchi
Keyword(s):  
Guinea Pigs ◽  
Lethal Dose ◽  
General Rule ◽  
Subsequent Infection ◽  
Japanese Strain ◽  
Active Immunity ◽  
American Strain ◽  
Pure Cultures ◽  
Different Strains

Guinea pigs were inoculated with suspensions of Leptospira icterohæmorrhagiæ obtained from pure cultures of several different strains, in order to determine whether or not an active immunity against a subsequent infection with virulent organisms would develop in the vaccinated animals. The experiments were so arranged as to make possible a determination of the existence of immunity against homologous strains as well as against the strains not employed as vaccine, and a brief quantitive estimation of the degree and duration of the immunity in relation to the quantities of the vaccines inoculated. Following the general rule of prophylactic inoculations with various pathogenic organisms, the inoculations were repeated subcutaneously on three consecutive occasions at intervals of 5 days. With respect to the amounts of vaccine, the experiments were divided into three groups for each vaccine, one group receiving three doses of 0.5 cc., the second three of 0.05 cc., and the third three of 0.005 cc. Four different strains were employed as vaccines, American Strain 1, American Strain 2, and one each of the Japanese and the European strains. The determination of the development, degree, and duration of the immunity was made by inoculating intraperitoneally several minimum lethal doses of each of the five following strains: American Strains 1, 2, and 3, the Japanese, and the European strains. The virulence of the different strains varied considerably, the strongest being the Japanese strain, which killed the guinea pig in a dose of 0.00001 cc., and the weakest American Strain 3, the minimum lethal dose of which was as large as 0.01 cc. The vaccinated guinea pigs were tested for immunity at the end of 2, 4, and 8 weeks after the last inoculation. The results obtained show that three successive inoculations of 0.5 cc. of the emulsions of killed cultures of Leptospira icterohæmorrhagiæ into guinea pigs rendered them completely resistant to a subsequent infection with the virulent cultures of both homologous and heterologous strains. With 0.05 cc. the protection was not so general, the animals succumbing to an experimental infection with some heterologous strains while resisting the homologous and other heterologous strains. The animals which were vaccinated with 0.005 cc. survived the infection experiments with the homologous strains in the case of American Strain 1 and the Japanese strain, but they were not protected against any other strains. The vaccines of other strains were unable to immunize the guinea pigs so highly even against their homologous strains, when the amount of each inoculation was only 0.005 cc., but 0.05 cc. conferred complete protection against the same strains. It may be concluded, therefore, that when a sufficient quantity of killed cultures of Leptospira icterohæmorrhagiæ is given, the guinea pigs will become immune to all strains of the same organism, but that smaller quantities may protect them against homologous but not against heterologous strains. A close analysis reveals the existence of group or type affinities among different strains which can be brought ' out by immunizing the animals with smaller quantities of killed cultures. In the present series of experiments American Strains 1 and 3 form one group, American Strain 2 and the European strain another, and the Japanese strain a third, which is also closely allied to the first group. In order to insure universal immunity it is wise to employ as many group or type cultures as possible in the preparation of vaccines, a polyvalent vaccine being recommended. It is not improbable that the strain recently encountered in Lorient, France, is an unusually deviated type of Leptospira icterohæmorrhagiæ, and that if successfully cultivated and used as vaccine in sufficient amount it might protect the animals against other strains of the same organism. The active immunity induced in the vaccinated guinea pigs was found to persist for at least 8 weeks after the last inoculation. It will no doubt last for a much longer period.

scholarly journals Root rot of anthracnose mother plants of garden strawberry: morphological and cultural characteristics of the pathogen and the search for effective fungicides

2020 ◽  
Vol 25 ◽  
pp. 06003
Author(s):  
Julia Kashchits ◽  
Galina Yakuba
Keyword(s):  
Causative Agent ◽  
Root Rot ◽  
Culture Media ◽  
Fungal Spores ◽  
Cultural Characteristics ◽  
Mother Plants ◽  
Pure Cultures ◽  
Krasnodar Region ◽  
High Diversity

On the territory of the Krasnodar region, anthracnose root rot, the causative agent Colletotrichum аcutatum Simmonds, is one of the most harmful diseases in the mother plants of garden strawberry. The death of affected plants ranges from 33 to 100 %. Purpose of the research was to study the morphological and cultural characteristics of the causative agent of anthracnose root rot in the mother plant of garden strawberries in the region and to assess the effectiveness of fungicides in controlling the pathogen. The studies were carried out at FGBNU SKFNTSSVV in 2018-2020 using generally accepted techniques. The object of the research is pure cultures of C. acutatum. The morphological and cultural characteristics of C. acutatum were studied on three culture media. A high diversity of the identified morphotypes was shown in terms of the size of conidia and conidiophores, the shape and color of apressoria, and the method of formation of acervules. The influence of seven fungicides of various chemical classes on the development of C. аcutatum spores has been assessed. Under in vitro conditions at the indicated concentrations, the preparations Luna Tranquility, SC (0,15 %), Sercadis Plus, SC (0,1 and 0,15 %), Strobi, WG (0,05 %), Horus, WG (0,04 %) and Skor, EC (0,04 and 0,05 %) completely prevented the formation of fungal spores.

scholarly journals THE DRUG-FASTNESS OF SPIROCHETES TO ARSENIC, MERCURIAL, AND IODIDE COMPOUNDS IN VITRO

1917 ◽  
Vol 25 (3) ◽  
pp. 349-362 ◽  
Author(s):  
Seinai Akatsu ◽  
Hideyo Noguchi
Keyword(s):  
Culture Media ◽  
Treponema Pallidum ◽  
Chemotherapeutic Agents ◽  
Inhibitory Influence ◽  
Fluid Medium ◽  
Rate Of Increase ◽  
Fluid Media ◽  
Pure Cultures ◽  
Free Media

In the foregoing experiments we attempted to determine whether or not, by subjecting several varieties of spirochetes to increasing doses of certain chemotherapeutic agents, a gradual increase of resistance to the latter could be shown. For this purpose, pure cultures of Treponema pallidum, Treponema microdentium, and Spirochœta refringens were used against the action of salvarsan, neosalvarsan, bichloride of mercury, and iodine-iodide potassium solution in vitro. For culture media, the usual ascites-broth-tissue medium as well as solid ascites-agar-tissue medium was used. After permitting the spirochetes to grow for a fortnight in media containing certain quantities of each drug, transfers were made from tubes showing various degrees of growth to the next series of tubes containing the same drug in still higher concentrations, and similar transfers repeated every 2 weeks. The results of the experiments may be briefly summarized as follows: 1. Treponema pallidum and Treponema microdentium have, within 3 to 4 months, increased their tolerance to salvarsan and neosalvarsan to five and one-half times their original mark. With Spirochata refringens the increase was about three times. 2. Against the action of bichloride of mercury, the amount of increased tolerance of Treponema pallidum was about 35 to 70 times the original, while that of Treponema microdentium was about 10 times as much and was reached within 10 weeks. Spirochata refringens resisted 30 times the original dose. 3. There was an unmistakable increase of resistance of these spirochetes to the action of the iodine-iodide solution (Lugol's solution) when they were grown for several generations in fluid media containing the iodine solution, but the rate of increase between the initial and the acquired tolerance was slight. In general, the addition of Lugol"s solution to fluid media has a weak inhibitory influence upon the growth of the spirochetes, requiring for the total suppression of growth a quantity of over 0.7 cc. to 5 cc. of the culture media. The tolerance reached was for about three times that amount. 4. A similar tolerance phenomenon has not been established when employing a solid instead of a fluid medium containing the drugs. No explanation is offered except a suggestion that the drugs held in the agar do not enter into combination with certain tissue constituents of the medium as they are able to do with tissue elements in fluid media. This may be a factor necessary for inducing drug tolerance in these organisms in vitro. 5. The increased drug-fastness in vitro has a limit beyond which no further advance can be made. This limit varies with different species of spirochetes. 6. The acquired drug-fastness in vitro gradually disappears when the spirochetes are cultivated again in the drug-free media for several generations.

scholarly journals ETIOLOGY OF YELLOW FEVER

1919 ◽  
Vol 30 (1) ◽  
pp. 13-29 ◽  
Author(s):  
Hideyo Noguchi
Keyword(s):  
Yellow Fever ◽  
Pure Culture ◽  
Guinea Pigs ◽  
Culture Media ◽  
Morphological Characteristics ◽  
Biological Properties ◽  
Freezing And Thawing ◽  
Highly Sensitive ◽  
Favorable Conditions ◽  
Aerobic And Anaerobic

By the employment of methods designed to promote the growth both of aerobic and anaerobic organisms, particularly those belonging to the class of spirochetes, it was possible to obtain a pure culture of a delicate organism, the morphological features of which place it in the genus Leptospira. On three occasions, that is, from three out of eleven cases of yellow fever, the organism was directly cultivated. These three strains were found to induce the characteristic symptoms and lesions when tested on guinea pigs. The organism was designated Leptospira icteroides. Leptospira icteroides was also obtained in pure culture from the blood of guinea pigs which succumbed to infection after being inoculated with the blood or organ emulsions from patients suffering from yellow fever. These cultures also proved to be virulent when tested on susceptible animals. The morphological characteristics and certain biological properties of the organism were considered in detail. It is invisible under translucent illumination and is difficult to stain by most aniline dyes. It is highly sensitive to the presence of bacteria and is rapidly destroyed in a medium in which certain other organisms are present. The presence of blood serum (man, sheep, horse, rabbit, etc.) seems to be essential for its growth. It grows well at a temperature of about 25–26°C. and more quickly at 37°C., though at the latter temperature it dies out within a few weeks. At 25°C. under favorable conditions and in suitable culture media it remains viable for several months without losing its virulence. Leptospira icteroides multiplies by transverse division. The virulence attained by some strains was such that 0.00001 cc. of a culture could induce typical fatal infection in guinea pigs. There exists a considerable variation among guinea pigs in their susceptibility to Leptospira icteroides. The organism is killed within 10 minutes at a temperature of 55°C. and is also destroyed by complete desiccation or freezing and thawing. Bile and bile salts dissolve it in certain concentrations, but not saponin. Leptospira icteroides passes through the pores of Berkefeld filters V and N, and there is a possibility of its having a granular phase of life under certain conditions.

scholarly journals THE PROPHYLAXIS OF WEIL'S DISEASE (SPIROCHÆTOSIS ICTEROHÆMORRHAGICA)

1916 ◽  
Vol 24 (5) ◽  
pp. 471-483 ◽  
Author(s):  
Yutaka Ido ◽  
Rokuro Hoki ◽  
Hiroshi Ito ◽  
H. Wani
Keyword(s):  
Guinea Pigs ◽  
Coal Mines ◽  
Dark Field ◽  
Point Of View ◽  
Rat Urine ◽  
Weil’S Disease ◽  
Weil's Disease ◽  
Pathogenic Organisms ◽  
The City ◽  
Highly Virulent

We have already described briefly the portals of entry and of excretion of the pathogenic spirochetes. We may mention here that we have twice prevented epidemics by disinfection of the ground and the removal of the inundated water in certain places in coal mines. In one mine 19 out of 50 workmen, and in another 9 out of 30 workmen came down with Weil's disease in about 2 weeks. We have already pointed out that the period during which the pathogenic spirochetes are excreted in the urine continues, as a rule, for 40 days, and that we must, therefore, apply disinfection for at least 40 days after the first appearance of the disease. Lately we have found that in 21 cases out of 24 the spirochetes were excreted in the urine for 40 days, in one case until the 42nd day, in one case until the 45th day, and in still another case until the 63rd day. Another important fact concerning the prophylaxis which has been brought out is that both house and ditch rats (brown) carry virulent Spirochæta icterohamorrhagæ, the causal spirochete of Weil's disease, in their kidneys. Miyajima has reported that field rats have the pathogenic organisms in their kidneys; he will report these findings in detail later. The spirochetes which he described are less virulent than ours. On his advice we have carefully examined house and ditch rats in the city and rats in the coal mines of Kyushu, where Weil's disease prevails, and found that 39.5 per cent carried highly virulent pathogenic spirochetes in their kidneys, thus confirming Miyajima's experiments. The kidneys were examined microscopically under the dark-field microscope, and in the cases in which we did not find the pathogenic spirochete, we made inoculations into guinea pigs. Thus we found Spirochæta icterohæmorrhagiæ microscopically in the kidneys or in the urine in 32.4 per cent, and by means of inoculation in 7 per cent, making a total of 39.5 per cent carrying the pathogenic organisms, out of a total number of 86 rats examined. In some instances, rats were made to bite guinea pigs and in two instances caused Weil's disease. Among fifty-five patients in our clinic, twelve were cooks; and in Europe many cases arise among butchers—indicating the relation of the disease to rats. Moreover, during the present year we observed two patients who acquired Weil's disease, one in 1 week, the other 8 to 9 days after they had been bitten by rats. These facts point to a relation between Weil's disease and rats. The infection is transmitted probably from rats to man by means of the urine of the rats, directly or indirectly. On the injection of 0.1 gm. of rat urine which contains Spirochæta icterohæmorrhagiæ into the peritoneal cavity of guinea pigs, the infection arises, while the injection of the liver or the blood of the rats into guinea pigs does not produce the typical disease. The finding that the kidneys of rats contain the pathogenic organisms of the disease is important from the point of view of prophylaxis. The large number of rats in the trenches of the European battle-fields suggests the possibility that many cases of Weil's disease may arise. We shall report on this point in more detail later.

scholarly journals THE RAT AS A CARRIER OF SPIROCHÆTA ICTEROHÆMORRHAGIÆ, THE CAUSATIVE AGENT OF WEIL'S DISEASE (SPIROCHÆTOSIS ICTEROHÆMORRHAGICA)

1917 ◽  
Vol 26 (3) ◽  
pp. 341-353 ◽  
Author(s):  
Yutaka Ido ◽  
Rokuro Hoki ◽  
Hiroshi Ito ◽  
Hidetsune Wani
Keyword(s):  
Chemical Composition ◽  
Causative Agent ◽  
Prophylactic Measure ◽  
Weil’S Disease ◽  
Weil's Disease ◽  
Field Mice ◽  
Soil And Water

1. On the basis of these findings, we conclude that the extermination of rats and field mice is a highly important prophylactic measure against Weil's disease. 2. The chemical composition of soil and water plays animportant part in the development of Spirochœta icterohœmorrhagiœ, and consequently in the spread of the disease of which it is the causative agent.

scholarly journals MOLECULAR WEIGHT, ELECTROCHEMICAL AND BIOLOGICAL PROPERTIES OF TUBERCULIN PROTEIN AND POLYSACCHARIDE MOLECULES

1938 ◽  
Vol 68 (3) ◽  
pp. 413-438 ◽  
Author(s):  
Florence B. Seibert ◽  
Kai O. Pedersen ◽  
Arne Tiselius
Keyword(s):  
Molecular Weight ◽  
Nucleic Acid ◽  
Guinea Pigs ◽  
Culture Media ◽  
Spherical Shape ◽  
Biological Properties ◽  
Natural State ◽  
Local Skin ◽  
Precipitin Reaction ◽  
Bovine Strain

Studies have been made by means of sedimentation in the ultracentrifuge, and by diffusion and electrophoresis, to determine the molecular weights and homogeneity of the tuberculin protein and polysaccharide molecules as found in their natural state in the unchanged filtrates from culture media after growth of tubercle bacilli. These results have been compared with data obtained on fractions isolated by chemical procedures from them or from old tuberculin. By means of electrophoresis in the Tiselius apparatus it was possible to separate the protein from the polysaccharide, as these two fractions occur naturally in the original culture medium filtrates of acid-fast bacilli. The protein from the bovine strain of bacillus proved to be homogeneous in sedimentation (S20 = 1.6), diffusion (D20 = 12.0) and electrophoresis, with a molecular weight of about 10,000. The tuberculin polysaccharide isolated in electrophoresis appeared to be practically the same in sedimentation and in precipitin reaction as the polysaccharide isolated by chemical procedure. The latter proved to be homogeneous in sedimentation (S20 = 1.6) and diffusion (D20 = 11.0) with a molecular weight of about 9000. A practically homogeneous protein was isolated from the culture filtrate of the human tubercle bacillus H 37 by fractional ammonium sulfate precipitation, with a molecular weight of 32,000 (S20 = 3.3; D20 = 8.2). It was electrochemically homogeneous, with an isoelectric point at pH 4.3 and an isoionic point at pH 4.7. It could be dried or frozen with no loss in homogeneity. It was highly antigenic in the precipitin reaction and produced the anaphylactic type of local skin reaction in tuberculous guinea pigs, in contrast to the true tuberculin type of reaction caused by a purified PPD fraction. Furthermore death resulted in tuberculous guinea pigs from intracutaneous injection of exceptionally small amounts. A protein with molecular weight of about 17,000 was isolated from the filtrate from cultures of the timothy bacillus. The nucleic acid originally occurring in old tuberculin (OT) seems to be responsible for the high electrochemical mobility observed. From OT and the PPD made from it, potent but non-antigenic molecules of 16,000 and 9000 weight and with a low content of nucleic acid were isolated. With increase in size these deviated much from the normal compact spherical shape, and aggregation was evident from the tendency toward gel formation. The smallest molecule (9000) was homogeneous (S20 = 1.0; D20 = 10.0) and had lost some tuberculin potency.

scholarly journals A COMPARISON OF THE SPIROCHETE OF YELLOW FEVER (LEPTOSPIRA ICTEROIDES NOGUCHI) WITH THE LEPTOSPIRA OF WEIL'S DISEASE

1928 ◽  
Vol 47 (2) ◽  
pp. 255-260
Author(s):  
Erich Martini
Keyword(s):  
Drinking Water ◽  
Yellow Fever ◽  
Guinea Pigs ◽  
Natural Conditions ◽  
Weil’S Disease ◽  
Weil's Disease ◽  
Set Up ◽  
Rule Out

The most noteworthy point observed in our studies is the extraordinary duration of life and the relatively meagre requirements for sustenance of Leptospira icteroides. It is conceivable that under natural conditions opportunities might arise for the prolonged existence of Leptospira icteroides, so that possibly after a lapse of years, the disease might reappear without introduction from outside. At any rate, the great viability of Leptospira icteroides must be considered in this connection. A decrease in the virulence of the leptospira does not rule out the possibility of a renewed outburst of yellow fever, since Uhlenhuth and Zuelzer have shown that it is possible to set up Weil's disease in guinea pigs by means of apparently saprophytic spirochetes obtained from drinking water, when the virulence of these organisms has been artificially heightened.

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