It is generally thought to be extremely difficult to isolate and identify causative Shigella from drinking water in the course of dysentery epidemics.
Despite the fact that there were not a few cases of dysentery outbreaks in this country in the last several decades in which waterborne infection was suspected as its cause, well legitimated reports of success in isolating the causative organisms from the imputable drinking water were scarce.
To our knowledge, the first successful case was reported by Meguro et al. including the authors in 1966 in which the causative Shigella was isolated from the well water by the use of Membrane Filter Method (MF) in a dysentery outbreak occurring in Higashi Murayama city, Tokyo, in that year.
Since then, such successful cases with MF have been accumulated by the authors and the number of them reached eight by May, 1969.
This paper describes the details of the experiments with MF comparing with some other ordinary methods and its clinical application to eight waterborne-suspected dysentery outbreaks.
1) Experimentally, excellent results were obtained with MF, compared with ordinary methods such as enrichment broth culture method and centrifuged sediment culture method. MF could recover the Shigella from the Shigella-suspended physiological saline so diluted as 3-5 bacillus per 500 ml fluid, whereas enrichment broth culture and centrifuged sediment culture required 30-50 and 300-500 bacillus per 500 ml saline for the isolation, respectively.
2) Clinical applications of MF to eight waterborne-suspected dysentery outbreaks occurring within Tokyo Metropolis during 1966-1969 were all successful. Shsgella sonnei were isolated in 6 of them, and Shigella flexneri 2a and 3a in the other two cases, respectively.
3) In all instances, the isolates from the drinking water were examined for the serotype, colicine type and antibiotic sensitivity patterns, and compared with those of the isolates from the patients. They were found identical, then consequently, the causality of the isolates from the drinking water was confirmed in all these eight incidences.

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Efficacy of recently introduced oral antibiotic preparation, Rifampicin (RFM), to bacillary dysentery in children was assessed with our both laboratory and clinical data which were epitomized as follows:
1) MIC of RFM against a series of recently isolated strains of Shigella was found to range from 3.12 to 12.5 mcg/ml.
2) MIC of RFM against pathogenic E. coli and Salmonella strains was found to range from 12.5 to 25.0 mcg/ml.
3) RFM plasma levels reached the peak 1 to 3 hours after an initial RFM administration and remained demonstrable up to 8 hours after that.
4) Continued administration of RFM had a tendency to promote the fecal RFM concentration to increase; For instance, a two-day medication to school children with the dosage of 0.3 gm t.i.d. saw their fecal level rose as high as 170.0 mcg/gm, the level being far over the aforementioned MIC and thought apparently to suffice both bacteriostatic and bactericidal effects to those intestinal pathogens in feces.
5) Eighteen children with bacillary dysentery were treated with RFM of 30.0 to 40.0 mg/Kg/day for 5 successive days. Within 2 to 5 days, the patients ceased to yield purulent bloody stool which had been frequently seen before and began showing normal stool. In sonne-dysentery cases of them, the. organisms disappeared from the feces within 3 to 5 days and reappearance did not occur in any of them.
6) Any significant adverse effects on the gastrointestinal system, blood system, liver or renal function were encountered in this series. Only harmless reddish discoloring of the urine and feces was seen in the course of medication.
(This paper was read in part before the 17th Eastern Regional General Meeting of the Japanese Association for Infectious Diseases held in October 1968).

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We have been tackling for many years the etiological problems of rheumatic fever mainly from the standpoint of Host-Parasite relationship. Terawaki, one of the authors, previously published a part of the studies in this magazine (Vol 42 No 3, June 1968) in which reproduction of human rheumatic fever was attempted on monkeys by the use of Streptococcus hemolyticus (S.H.) group A type 12. He obtained some evidence of hope in it. However, it was by no means considered fully satisfactory.
After some preliminary experiments to reassess the suitability of experimental animals as the hosts and injecting materials as antigens or stressors, we decided this time to use the same species of monkey as above of 2.5 to 3.0 Kg body weight and adopt two types of group A S.H., type 12 and type5, in the form of live organisms, M protein fraction and lipopolysaccharide extracts. LD50 of live S.H. suspension fluid was 0.25 to 0.35 mg to dd-mice of 20 gm. M protein was prepared by Lancefield's method, and lipopolysaccharide by Westphal's method. In the case of M protein and lipopolysaccharide experiments, we combined the use of Antigen-Antibody-Complex prepared by Ouchterlony method in the final phase of the experiments. In some groups of the animals, we sensitized them with the cow serum in advance.
The period of experiments were mostly within from January 1968 to March 1969. The intervals of every injections varied from 3-5 days, one, two, four weeks to one month. Amount of the injecting materials varied from 1-5 mg to relatively large amount, 30-50 mg, at one time.
The animals were divided into 12 categories according to administration mode and the kind of injecting materials, which are briefly summarized as follows:
1) Live S.H. administration (i.v.) to cow-serum-sensitized monkeys (type 12 to 3 monkeys (1), type 5 to 3 monkeys (2)).
2) Live S.H. (s.c.)(type 12 to 2 monkeys (3), type 5 to 2 monkeys (4), alternate use of type 12 and 5 to 2 monkeys (5)).
3) Intra-tonsillar injection of live S.H. (type 12 to one monkey (6), type 5 to one monkey (7), alternate use to 2 monkeys (8)).
4) M protein mixed with Freund's complete or incomplete adjuvant (combination of s.c., i.m. and i.v.) combined with Antigen-Antibody-Complex (intra-myocard or s.c.)(type 12 to 4 monkeys (9), type 5 to 4 monkeys (10)).
5) Lipopolysaccharide extracts mixed with Freund's complete or incomplete adjuvant (combination of s.c., i.m. and i.v.) combined with Antigen-Antibody-Complex (intra-myocard or s.c.)(type 12 to 4 monkeys (11), type 5 to one monkey (12)).
6) Two monkeys were used as contrast.
With all the cases, the following symptoms and data were checked: fever, joint swelling, limping, cardiac murmur, PCG, ECG, sludge phenomenon of the blood and eye grounds vessels, chest X-ray, ASL-0, total protein and r-globulin level, complement value of the serum and anti-myocard antibody level.
Conclusively, several cases with prolonged live S.H. injections, type 12 or 5, were found much more resembling to human rheumatic fever in their symptoms and data than those of the previous experiments. And, in one case with M protein and Antigen-Antibody-Complex injections, we believe that we could reproduce clinically strikingly identical findings to human rheumatic fever in the animal. It thereby demonstrated fever, positive sludge phenomenon and most importantly the apical systolic murmur both in auscultation and PCG just characteristic of mitral insufficiency of human rheumatic carditis. Altogether with the fact that it was given the component of S. H., the changes are in agreement with revised Jones Criteria (1965) for human rheumatic fever.
Additionally, based on our data, several hypothesis concerning the development of rheumatic carditis and M protein were rendered in this paper.

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Jasomycin (JM), a new antibiotic, was tried orally in the treatment of scarlet fever and hemolyticstreptococcal tonsillitis and evaluated with the following laboratory and clinical data:
1) Sensitivity of 60 hemolytic streptococcal strains against JM was investigated. The value of MIC was found distributed between 0.09 and 0.39γ/ml.
2) Relationship between oral dose and bacteriological effects in vivo was studied employing the findings of pharyngeal hemolytic streptococcus as a marker. One-time dose of 5 mg/Kg was almost ineffective in eradicating the cocci from the pharynx, but 8 mg/Kg or more yielded the effects in 4-12 hours.
3) Twenty cases of scarlet fever in acute stage were given about 10 mg/Kg JM every 8 hours for 5 days. In all the cases, there found marked clinical effects such as remission of fever, recovery of pharyngeal symptoms and disappearance of the cocci from the pharynx. After the haltof the medication, however, reappearance of the cocci in the pharynx was observed in 40% of them and fever recurrence in 15%.
4) Ten cases of hemolytic-streptococcal tonsillitis were treated in the same manner as in the case of scarlet fever. The results were excellent in all the cases. Reappearance of the cocci was noted in three cases, fever recurrence in one case.
5) As to side effects, vomiting was seen only once in one case who, however, tolerated the further medication. Blood, urine and liver functions were tested in 5 cases at the end of the medication course but no particular changes were revealed.
From these results, it is concluded that JM, used at the one-time dose of 10mg/Kg or more, is as effective to scarlet fever and hemolytic-streptococcal tonsillitis as the other antibiotics such as penicillin or erythromycin which are already authorized to these diseases.

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