We can now extract all kinds of exotoxins except diphtheria toxin from various bacterial bodies. The common principle through these cases is to collect young and active cells in the early stadium of their growth, because it is demonstrated at each bacterium that young one is heavily toxic and on the contrary, old one has little toxin within its body.
As far as diphtherial toxin concerns, however, this is not the case. At first, it must be said that even the young cell has few toxin within its body and the toxin never fails to exist in the medium. Secondly, it must be pointed out that the toxin production in the medium is entirely paralell with the growth of the organism. Such a complete r paralell between the toxin production and the growth can not be seen in any other case. Even the tetanus toxin was proved to be excreted at the stadium of cytolysis. Thus diphtheria toxin nowadays is thought to be different from other exotoxins in the mechanism of excretion. The one is a metabolite in physiological youth and the others are products in cellular lysis. Pappenheimer's studies seem to promote this assumption to more reliable working hypothesis. This discrimination between these exotoxins, however, seems to be rather improbable. Therefore, author designed to analize once more the growth curve of this organism. Numerous works on its growth curve and toxin production up to date show almost the same results and does not seem to bring any other available clue to scrutinize the problem. In order to know the new aspects of the problem we must use other techniques. A series of studies were now undertaken on the biochemical points, from which the organism had never been studied.

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The first identification of S. narashino was reported by Nakaguro et al. in 1938, who isolated it from blood and feces of a patient suffered from typhoidlike fever. Subsequently, Yamada et al. isolated the same organism from the submaxilliary lymph nodes of a swine at the Metropolitan Slaughter Hause, in 1942. No other report, so far, on the isolation of S. narashino in animals had been found.
Recently, an epizootic outbreak of a disease with diarrhea occurred in guinea pigs in the premises of the National Institute of Health, Japan, and the etiological analyses have shown that S. narashino was the causative agent. In this paper, the results of these analyses will be described.

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Many species of Salmonella have long been described as the cause of epizootic outbreaks in rodents. In Japan, as for guinea pigs, S. enteritidis, S. typhimurium, S. blegdam, S. moscow, S. amersfoort, etc. have been reported. And our previous report presented an epizootic outbreak in guinea pigs caused by S. narashino at the National Institute of Health, Japan. As it was pointed out in the paper, another outbreak concurrently occurred, and the etiological analyses resulted in isolation of several species of Salmonella. The findings will be given here.

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More than 70 years have passed since the prophylactic treatment of rabies with dried attenuated vaccine was first reported by L. Pasteur. But the treatment of rabies to-day still remaines on the same therapeutic principles of giving active immunity by repeated injections of an emulsion prepared of the nervetissue of animals infected with fixed virus before the street virus invades into and proliferates in the central nerve-tissue of the injured. Thus, clinicians are still troubled with the postvaccinal paralysis. Moreover, as it was detected by Nitsch and Dodero, the vaccination is useless or even harmful as it shortens the incubation period when the injury is severe.
On the other hand, in nearly all instances, the locality infected and the time of infection are quite evident indicating the availability of immune serum. Following the report by Babes and Lepp in 1889 on a dog survived the subdural injection of street virus after it had received 6 successive daily injections of immune serum obtained from a dog immunized with rabies vaccine, numerous reports have been published on rabies immune serum treatment some stating that it is highly benefitlal and the others useless, But most of those studies were not satisfactory on the points that the number of the animals used was too small, the neutralizing antibody titer of immune serum used was not precisely determined, the method of infection and the dosage used were not adequate etc. While, A. Hoyet, et al. in 1935 statistically proved the effectiveness of immune serum against the experimental infection attempted on mice with fixed virus. In the year 1945, K. Habel, being based on the experimental findings of animal protection tests (guinea pigs, mice and monkeys) by the use of rabbit immune serum and street virus or fixed virus as challenge virus, stated that the protective effect was observed on the animals received immune serum and practically no difference was observed in the prophylactic effect between the vaccination started immediately after and that started 3 to 6 days after serum injection, and he concluded that the first application of immune serum and then to start vaccination upon positive diagnosis of the suspicious animal are quite sufficient for the protection of the injured.Following the above, in 1950, H. Koplowski reported that the inoculation of an adequate amount of rabbit or goat hyper immune serum on hamsters and guinea pigs within 24 to 72 hours after infection with the street virus recovered from the salivatry gland of a rabid dog, could lower the number of fatal cases, and that the combined application of this immune serum and Semple's vaccine showed no antagonism nor synergism of the antigen and antibody, and further, he recommended the use of this combined therapy presenting 2 evidences of successful treatment both injured by positively rabid animals.
In our country, the incidence rate of rabies has remarkably increased since the end of the last world war and a considerable number of postvaccinal paralysis cases have occurred. Thus, efforts have been excised on the improvements of the vaccine as well as on the prophylactic use of immune serum. As the result of animal experiment, in which rabbit immune serum highly immunized with fixed virus was used, the application of immune serum within 6 hours after infection was statistically proved to protect mice infected with fixed virus. The experiment in which street virus was used as infecting virus also confirmed the above finding.
Reported in the following are the details of similar experiments carried out by using immune serum obtained from the horses highly immunized with the fixed virus. Further, the prophylactic effect of the treatment with horse hyperimmune serum alone was compared with that of the combined treatment of the same immune serum and the vaccine inactiveted by ultra-violet irradiation.

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In a previous publication evidence was presented for the conclusion that biochemical activity of C. diphtheriae during the growth phase should be differenciated from those of other bacilli. It seems to be rather contradictorious to say that chemical activities of this organism fall to the minimum lebel during its growth phase. This paper develops further the experimental and theoretical aspects of the above facts by reporting quantitative deta relative to the viable counts of the bacilli at each stage of the growth and by checking morphological findings at the same time by light and electron microscopes.
Earlier study by Eisler reported that the greater part of the pellicle consists of dead cells of C. diphtheriae. This was an excellent opinion but it is a pity that he did not show his experimental method and deta, because enumeration of the viable units of the suspension is considerablly controversial owing to the clumping characteristics of the organism. Author tried to overcome the difcuties by following technique.

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All the exotoxins except diphtheria toxin nowadays can be extracted successfully from the bacterial bodies, Common principle for success through these experiments is to be collect the bacilli in early stage of cultures. Exact and detailed studies recently demonstrated that appearance of the toxin in media was not owing to secretion of physiologically young cells but to lysis of degradated cells. This was proved to be true even for the tetanus toxin by Seki and his colleagues. As far as diphtheria toxin concerns, however, this principle seems to be unable to be applied, because the exotoxin always exists in the medium and not within the cells, however early they may be harvested.
It has long been postulated, but never demonstrated up to date that the exotoxin is formed within the cells. In a series of preceding papers, however, author claimed that there is no reason for differenciation between the diphtheria toxin and the other exotoxins in regard to the site of toxin formation. According to author's conclusion the growth curve of C. diphtheriae is very close to the accumulation curve of dead cells of the degradated cells and consequently to obtain physiologically young cells of this organisms is not so easy as to obtain those of other bacilli. From this viewpoint it seems to author that there has been no one who collected really young cells of C. diphtheriae. As it was demonstrated in preceding paper trials to obtain them seem impossible as far as static culture is used. Present experiment was undertaken to find another method available for this aim. It seems to be adequate for author to use shaking culture, because the organism in shaking culture shows nearly logarithmic growth, in which all the bacilli are physiologically young and active. This was demonstrated by many authors, such as Lingood and Fenton, Woodward, Tallman, and Perrotta, and Mitsuhashi and Kojima, If physiologically young cells can be obtained by this method, it is a next object for author to clarify whether they contain the exotoxin within the cells or not.

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Botulism, very rare disease as it is, has long been reported in Europe and America, although no report referring to occurrence of this disease had been presented in Japan prior to 1952, when Nakamura, Iida and Saeki reported an outbreak of type E botulism at Iwanai, Hokkaido, due to herring-izushi. In the following year, another two outbreaks of type E botulism due to izushi involving fish were reported by the same authors. Again, early in October, 1953, the authors learned of the fourth outbreak in Hokkaido.
Incidentally, concurrently with the fouth outbreak in Hokkaido, four persons of a family at Tenno, a small town upon the coast near the Lagoon Hachiro, Akita, developed symptoms of botulism and two of them died. The incliminated foodstuff, gilthead-izushi, was shown to possess type E botulinal toxin and therefrom a strain of Cl, botulinum type E was isolated. This occurrence adds up the total outbreaks of botulism in Japan, all of which were due to izushi involving fish and type E organisms, to five with thirty-two cases and nine fatalities. The data relating to the known outbreaks of botulism in Japan are shown in the table.

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