1) Five kinds of basic antagonistic substances, which resembled streptothricin in nature, were obtained in a form of hydrochloride, crystal-line helianthate and reineckate, and the identification between these sub-stances was performed. 2) All of these substances had rather lower potencies than strepto-mycin, in a dilution titer against E. coli, while the substances No. 259 and No. 120 (Seki) were higher inhibitory against Staphylococcus (Terashima strain) than streptomycin. 3) Six substances, including streptomycin, were arranged in the order of increasing toxicities as follows: -36, streptomycin, 120 (Seki), 24, 39, and 259. The substance No. 36 was able to be injected both, intravenously and intramuscularly into mice in an amount of 20mg without showing any toxic signs. In this respect, the substance No. 36 has advantages over streptomycin. 4) The decomposition points of helianthates or reineckates of 4 streptothricin-Tike substances were found to be in the vicinities of that of streptothricin except No. 36 (Roseomycin). In reference to the other biological and chemical properties, however, the 5 substances seem quite different from streptomycin and from each other. 5) Maltol and Sakaguchi reactions were found to be positive in streptomycin alone, while glucosamine and other sugar reactions were positive in both streptomycin and streptothricin-like 5 substances. The chemical reactions positive in streptothricin group only, were not found yet. 6) A strain of E. coli, which was made resistant to the hydrochlorides of 4 substances (36, 20, 259, and 120) respectively was resistant to each of 5 substances and somewhat less resistant to streptomycin too, while the same strain of E. coli, which was made resistant to streptomycin, was not resistant to each of 5 substances. In this respect it seems very probable, that each of 5 substances resembles closely in their mechanisms of the anti-biotic functions.
1) Five kinds of basic antagonistic substances, which resembled streptothricin in nature, were obtained in a form of hydrochloride, crystal-line helianthate and reineckate, and the identification between these sub-stances was performed. 2) All of these substances had rather lower potencies than strepto-mycin, in a dilution titer against E. coli, while the substances No. 259 and No. 120 (Seki) were higher inhibitory against Staphylococcus (Terashima strain) than streptomycin. 3) Six substances, including streptomycin, were arranged in the order of increasing toxicities as follows: -36, streptomycin, 120 (Seki), 24, 39, and 259. The substance No. 36 was able to be injected both, intravenously and intramuscularly into mice in an amount of 20mg without showing any toxic signs. In this respect, the substance No. 36 has advantages over streptomycin. 4) The decomposition points of helianthates or reineckates of 4 streptothricin-Tike substances were found to be in the vicinities of that of streptothricin except No. 36 (Roseomycin). In reference to the other biological and chemical properties, however, the 5 substances seem quite different from streptomycin and from each other. 5) Maltol and Sakaguchi reactions were found to be positive in streptomycin alone, while glucosamine and other sugar reactions were positive in both streptomycin and streptothricin-like 5 substances. The chemical reactions positive in streptothricin group only, were not found yet. 6) A strain of E. coli, which was made resistant to the hydrochlorides of 4 substances (36, 20, 259, and 120) respectively was resistant to each of 5 substances and somewhat less resistant to streptomycin too, while the same strain of E. coli, which was made resistant to streptomycin, was not resistant to each of 5 substances. In this respect it seems very probable, that each of 5 substances resembles closely in their mechanisms of the anti-biotic functions.
1) Among 258 strains of our laboratory collection, (most of them being antagonistic ones), 52 strains (20.2%) could grow well on Krainsky's agar con taining 10u/cc.streptomycin.But, when a dose of 100u/cc.was used, besides 3 streptomycin producing strains only 3 strains showed a relatively good growth.One of these 3 strains was found to produce a new specific antibiotic, which is quite different from streptomycin and now under investigation.The other two strains, chromogenic and resembling Str.roseochromogenus, showed a strepto thricin-like antibiotic spectrum when tested by the streak plate method.When these culture filtrates, however, were tested by the cup assay method, they showed a spectrum characteristic of streptomycin.This fact suggests, that these strains.may probably produce streptomycin like substance besides other antibiotics.(2) Other antagonistic actinomyces, which produce some of the strepto thricin-like .substances, became to grow abundantly on the agar containing 800u/cc. of streptomycin after being successively transferred to the agar con taining gradually increasing amount of streptomycin.After this procedure, there were noticed neither morphological nor biochemical changes as well as.no tendency to produce streptomycin itself.(3) Among several non-antagonistic strains, it was able to obtain.some antagonistic colonies after transferring to streptomycin containing agar, as.Kelner showed after the treatment by X ray or ultraviolet light.Although these colonies were not confirmed to be streptomycin producing ones, they were so characteristic in their stronger inhibitory action on B. anthracis, because accord ing to our experimental results almost all low antagonistic strains.were more highly inhibitory to B. subtilis, whereas streptomycin-producing strains were found more inhibitory to B. anthracis than B. subtilis.(4) For industrial purpose such a simple procedure, i.e. the transferring cultivation on streptomycin-containing agar is useful for preventing "running down" of the potency of streptomycin production.
1. Several strains of antagonistic and non-antagonistic streptomyces were tested upon their abilities to grow on the streptomycin containing agar. Beside streptomycin producing ones, many strains were able to grow on the agar containing 10u/cc streptomycin, while only a limited number of streptomyces strains including streptomycin producing ones were capable to grow on the agar containing 100u/cc of streptomycin. Such a concentration of streptomycin, therefore, appeared to be specific and suitable for isolation and preservation of streptomycin producing strains.2. Even if the streptomyces strains producing other kind of antibiotics than strptomycin became to grow luxuriantly on streptomycin containing agar, there was seen no tendency to produce streptomycin.3. It was able to select some, even if weakly, antibiotic colonies from the non-antagonistic strans of streptomyces by serial passages through streptomycin containing agars, although it was impossible to decide whether they were truely streptomycin producing ones, or not. The antibiotic strains thus obtained inhibited B. anthracis or Myco. phlei more strongly.4. It was confirmed, that the streptomycin producing strains were reduced in colony numbers, became homogeneous in morphological properties and partially increased in inhibitory activities of culture filtrates, when they were cultivated on streptomycin containing agar through several generations.
1) A streptothricin-like substance, belonging to streptothricin type I was isolated from the culture filtrate of Str. roseochromogenus No. 24.2) The in vitro activity was nearly the same with that of streptomycin, but 5 mg. injected intravenously into mice, represented a delayed toxicity.3) Though the crystalline helianthate and reineckate of this salt differ from those of streptothricin type I (roseomycin) in their decomposition points, these compounds seem to be quite similar to the latter in other respects.
1) Suitable conditions to isolate the strains of actionmyces from soils were studied which were collected from many places throughout all over the territories of Japan, and the asparagine glucose agar of Krainsky was found to be a most suitable isolation medium, because more actionmyces and fewer bacteria grew in it than in the others. The distributions of actinomyces were closely related to the kinds of soil, to their pH, and to the seasons of the sampling. 2) 1223 strains of actinomyces isolated from soils collect-ed from 188 places were cultivated in several kinds of fluid culture media and the culture filtrates were tested for their inhibitory activities, using mainly Sta. aureus (Terashima strain) as test organisms, and 157 (13%) strains were found to be antagonistic. The above method is well reliable in finding inhibitory strains whose activities were detectable in their culture filtrates. It was not so sensitive, however, to reach a highest detection rate of effective strains. 3) By the so-called “Cross streak method” 144 (44%) active strains were selected from 261 actinomyces. This method is very simple and sensitive to find many antibiotic strains and to examine a specific “anti-bacterial spectrum.” The development of this task will be reported in the next report. 4) The representative strains were classified, according to Krainsky and Waksman, into several species by the cultural properties. on several kinds of media. Although a difinite identification of all strains was scarcely possible, many strains were found to belong to Str. roseochromogenus.
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