The mechanisms of the heterologous immunity of phenol-killed BCG treated mice against staphylococcul infection was not clear as yet. As a help for elucidating this mechanism, the behaviors of Staphylococcus aureus in the kidney of phenol-killed BCG treated and untreated mice were compared. At first, the course of experimental infection with Staphylococcus aureus in the tissue of untreated mice was studied. In the lungs, liver and spleen, the population of culturable staphylococci showed a gradual decline when sampled at various times after infection. However, the fate of this organism in mouse kidney showed a sharp increase until the first two days. From the results of these experiments, the behaviors of Staphylococcus aureus in the kidney of phenolkilled BCG treated mice, the population of staphylococci showed an increase. Neverthless, in the kidny of phenol-killed BCG treated mice, the growth of this organism was suppressed during the observation period. This finding suggested that the resistance of phenol-killed BCG treated mice against staphylococcal infection was due to the growth suppression by this organism in the kidney.
1) The inhibitory action of 17 compounds of acridine derivatives and its related compounds on the multiple-drug-resistance transfer from the resistant donor Shigella flexneri 3a strain MZ17R to the sensitive recipient E. coli strain K-12 C25S and the inverse transfer from the resistant donor E. coli strain K-12 C17R to the sensitive recipient Shigella flexneri 3a strain MZS were examined and the following results were obtained. Among the compounds tested, proflavine, acetylproflavine, acridine orange, acridine yellow, acriflavine and thioflavine T inhibited the multiple-drug-resistance transfer in a low concentration. The minimal concentration of each compound required for complete inhibition on the both transfer system (C17R→MZS and MZ17R→C25S) were respectively as follows: proflavine 5 and 20μg/ml., acetylproflavine 10 and 20μg/ml., acridine orange 40 and 80μg/ml., acridine yellow 10 and 40μg/ml., acriflavine 5 and 10μg/ml. and thioflavine T 10 and 80μg/ml. The minimal concentration required for the inhibition of the multiple-drug-resistance (factor) transfer from C17R to MZS was lower than that in the transfer from MZ17R to C25S. 2) A new evaluation method that excluded the antibacterial activity was devised. According to this method, acrinol was proved to have a superior antibacterial activity to the inhibitory one on the multiple-drug-resistance transfer. Hence, acrinol was considered not to have a specific inhibitory effect on the multiple-drug-resistance transfer from MZ17R to C25S but to affect slightly the transfer system from C25R to MZS. 3) From the observation of the chemical structures and the inhibitory activity of acridine derivatives on the multiple-drug-resistance transfer system, NH2 or N (CH3)2 in position 3 and 6, CH3 in position 2 and 7, and CH3 or H and chlorine in position 10 of acridine nucleus were necessary for the activity. The acridine itself and its N-oxide, propionamido or benzamido in position 3 and 6 and ethoxy or methoxy radicals in position 2 or 7 of acridine nucleus such as N, N-dipropionylproflavine, N, N-dibenzoyl-proflavine, acrinol and acrinxamine (atabrine) showed a reduced activity. Neutral red, thionine and methylene blue, in which carbon in position 9 of acridine was substituted with nitrogen or sulfur, thioflavine S, and acridine red presented also reduced activity. In the comparison of thioflavine T and thioflavine S, it might be of interest that the former had the inhibitory activity on the multiple-drug-resistance transfer system, but the latter did not.
1. Each frequency of the resistant strains to PC or SA of Staphylococcus aureus collected in 1961-1963 was 90 to 100 per cent, that to SM or TC was about 60 per cent; both of the rates were not yearly increased. That to CP or EM was below 50 per cent, was increased considerably year after year. 2. Each frequency of the resistant strains to PC, TC, CP or SM of α or β Streptococcus collected in 1961-1963 was low. 3. With regard to E. coli the resistant strains to SM, TC or CP were augmented obviously for the same three years; with respect to Klebsiella each rate of the resistant strains which was higher than that of E. coli was not increased. 4. With respect to E. coli or Klebsiella four-drugs-resistant strains were found most frequently, and one-drug-resistant strains followed, and two-and three-drugs-resistant strains were fewest. Regarding Pseudomonas aeruginosa or Proteus, four-drugs-resistant strains were found most frequently, and followed by threedrugs-resistant strains, two-and one-drugs-resistant strains followed in order. 5. Antibiotics resistant strains of Bac. tuberculosis investigated in 1956-1963 were augmented to SM or PAS after 1959 or 1961 respectively, but that to INH was increased gradually year after year. 6. E. coli isolated from the urine investigated in 1957-1963 were not increased yearly, and another Enterobacteriaceae and Pseudomonas auruginosa isolated were augmented. 7. Staphylococcus and Enterobacteriaceae isolated from pus were increased yearly, whereas Streptococcus were not increased and the detection rate was low. 8. Enterobacteriaceae especially Staphylococcus isolated from throat swab were augmented yearly. 9. Of bacteria from bile oral type were more frequent than fecal type.
1) The oral administration to a mouse of antibiotics as SM and others gives changes to the fecal bacterial flora, and Yeast-like bacteria having resistive power to these drugs extraordinarily increase, and the susceptibility to the oral infection of salmonella enteritidis enhanced. 2) This phenomenon above is remarkably suppressed with the administration of drug resistant Str. fecalis. 3) On the suppressive power of Str. fecalis over Yeast-like bacteria according to the result of the examination in vitro of the antagonistic phenomenon of the two, it is considered that the mechanism of the suppresion of Str. fecalis over Yeast-like bacteria is not so much due to acid generated by the former as to the difference of the quantity of their consumptive power of glucose in media.
Since Koyama and Akiba noticed the inhibitory effect of sodium deoxycholate on the transfer of multiple drug resistance (R) factor from E. coli to Shigella, substances such as bile, bile salt, cholic acid and dodecyl sulfate as surface active agents and chloramphenicol, 8-azaguanine, gramicidin J and mitomycin C as inhibitors of biopolymer synthesis were described to inhibit the transfer of R factor. However, the works so far appeared were not so precautious in testing whether the used concentration was lethal or not on the fate of both donor and recipient. In this study, several inorganic salts and antibiotics were tested for their inhibitory effect on the transfer of R factor, by estimating the inhibitory concentration on the transfer together with bacteriostatic concentration on both donor and recipient. As a result, a series of zinc compounds such as ZnCl2, Zn(CH3COO)2 and ZnSO4 were found to inhibit the transfer of R factor at the concentration without affecting the growth of both donor and recipient. Among them, ZnSO4 was most powerfull, and the site of action of this substance was further analysed. Pretreatment of donor or recipient did not affect the transfer, and did not reveal any eliminating effect of R episomes. Whereas the presence of ZnSO4 in the culture in which specific pairings of donor and recipient are proceeding was a necessary requisite for obtaining the inhibition. In those inhibited cultures, whenever ZnSO4 was removed from the medium, instantaneous recovery of the transfer was observed. Further analysis on the action of ZnSO4 was made by pursuing the transfer after blendor treatment for the pairing, and it appeared that ZnSO4 inhibited the specific pairing of donor and recipient like Puck's finding in which Zn++ ion inhibited the irreversible binding of T1 phage to E. coli B. As a matter of fact, concentrations of Zn++ ion effective for the inhibition of these two biological systems were almost the same.
Rabbits were treated by live culture, vaccine or toxoid of staphylococcus. Then the dermal reaction by toxoid or vaccine, antitoxic antibody and agglutinin were tested. Promotion of dermal reaction proceeded the increase of antibody when rabbits treated by live cocci or vaccine whereas the latter prceeded by toxoid treatment. The serum of high antitoxic titer failed to protect mice from death caused by the intraperitoneal infection of the cocci. Those immunized rabbits rarely showed protection to the intravenous challenge of cocci, although they were resistant for the α-toxin.
A new classification method was proposed from the view point of relationship among microorganisms, which was based on the Adanson's thinking, Sheath's similarity value and the author's concept of center species. According to this method, some reorientation for the classification of the family Micrococcaceae was made and the following results were obtained. 1) The classical genus Gaffkya should be integrated to the genus Staphylococcus. 2) The five, non-fermentative species, M. colpogenes, M. cryophilus, M. denitrificans, M. morrhuae and M. halodenitrificans should be excepted from the genus Micrococcus, because of their lower similarity values, while the other eleven, fermentative species constituted the genus Micrococcus. 3) The assignment of the species in the genus Peptococcus was found to be proper, and the Peptostreptococcus glycinophilus indicated a higher similarity value to the genus Peptococcus rather than to the genus Peptostreptococcus. 4) Amoung ten species which have been included in the classical genus Sarcina, the five species such as S. ventriculi, S. maxima, S. methanica, S. barkeri and S. litoralis should be omitted from this genus, because they have a lower similarity value to the center species, if all of these ten species were considered to be in the same genus. It was proper that the residual five, aerobic and tetrad species constituted the genus Sarcina, because of their higher similarity values to the center species, S. flava. 5) The two, microaerophilic, fermentative and large tetrad species, S. ventriculi and S. maxima should be elevated to the genus Zymosarcina from the original subgenus. 6) The two, strictly anaerobic, methane-fermentative and relatively small tetrad species, S. methanica and S. barkeri should be included in the genus Metanococcus. 7) S. litoralis should be excepted from the genus Sercina and belonged to any other genus. Various, originally heterogenous species have been included together in the classical genus Sarcina, probably owing to their morphological character of packet. According to the author's method, each of these species was consequently placed in its proper genus respectively, and moreover, they had the specific correlated features to the genus. 8) The classical family Micrococcaceae should be reduced to a lower rank as the following two tribes, namely, the tribe Micrococceae including Staphylococcus, Micrococcus and Peptococcus, and the tribe Sarcineae including Sarcina, Zymosarcina and Methanococcus.
The authors tried to elucidate the therapeutic effect of hyperbaric oxygenation on the experimental infection of the anaerobic pathogens, using mice and rabbits as experimental animals. 1) Cl. novyi was rather resistant to the action of oxygen at high pressure. It was presumably due to the spore formed. 2) In order to prevent the death of the mice inoculated with Cl. novyi, the animals should be repeatedly exposed to the oxygen at moderate pressure. The excessive oxygenation was toxic to the animals, rendering them less resistant to the infection. 3) Toxin of Cl. novyi was not decomposed by hyperbaric oxygenation. It has also no effect on the detoxication process of the organisms or on the toxin neutralization in vitro and in vivo. 4) In case of gas gangrene hyperbaric oxygenation therapy should be combined with antitoxin treatment, as the former suppresses the growth of pathogens and the latter neutralizes toxin produced. 5) In the experiment with with rabbit, repeated hyperbaric oxygenation was proved markedly effective to the infection with Cl. norvyi, but less effective to Cl. tetani. In the latter case, the excessive oxygenation was unfavorable to the infected animals.