(-)Rugulosin showed an antiinfluenzal effect in the allantoic cavity of chicken eggs as well as in cultured chorioallantois, proving itself to be more potent than 1-adamantanamine. Also, (-)rugulo-sin, when administered by inhalation, was as effective for protection of mice against aerosol influenzal infection as orally administered 1-adamantanamine. The effect of (-)rugulosin seemed to be mainly due to direct inactivation of influenza virus.

View full abstract

A rapid identification method of glucose nonfermentative gram-negative rods was established and 320 strains isolated were divided into five groups according to their characteristics in pigmentation, acid from glucose, cytochrome oxidase activity and motility. Further characterization of the strains in each group resulted in the identification that the strains in group I were Pseudomonas aeruginosa, strains in group II, P. aeruginosa and Pseudomonas putida. Achromogenic strains of P. aeruginosa were classified into group III, Pseudomonas maltophilia, Pseudomonas alcaligenes and Alcaligenes faecalis into group IV and Acinetobacter calcoaceticus (Acinetobacter anitratus and Achromobacter lwoffii) in group V. When fluorescent pigment production was taken as a standard, 259 out of 263 chromogenic strains were identified as P. aeruginosa and the remaining four were P. putida. Whereas forty-five achromogenic strains included twenty-four A. calcoaceticus, eight P. aeruginosa, six A. faecalis, five P. maltophilia and two P. alcaligenes. From May 1970 to June 1971, 368 strains of glucose nonfermentative rods were isolated from clinical specimens sent to the Central Laboratories of Tohoku University Hospital and three fourth (286/368) of the isolates were P. aeruginosa

View full abstract

Rifampicin was shown to be highly active against Mycobacterium leprae in mice, completely inhibiting the in vivo multiplication of this organism at a dosage of 0.1 mg per day by gavage. When drug administration was started after the establishment of M. leprae in mice (30 weeks after infection), rifampicin stupped any further bacillary increase. A second inoculation of the organisms from treated mice showed complete loss of viability. Exposure of M. leprae to rifampicin for a short period before infection significantly reduced the viability of the organism.

View full abstract

Interferon production in human leukocyte suspensions induced by Newcastle disease virus is significantly affected by the pH of the culture. The optimal pH for interferon production is higher than the physiological pH level. Human leukocyte interferon has at least three types of surface charge as demonstrated by DEAE-cellulose column chromatography. The molecular wight of these three types of interferon, as measured by gel chromatography on several kinds of Sephadex is equally 25000. The main fraction of human leukocyte interferon was purified approximately 1400-fold by a sequence of procedures which involved ammonium sulfate salting out, CM- and DEAE-cellulose column chro-matography and gel filtration on Sephadex G-50. Human leukocyte interferon of a high specific ac-tivity of about 2130000 units/mg of protein was thus obtained.

View full abstract

Mouse peritoneal macrophages cultivated in vitro were infected with Candida albicaus. The uptake of 3H-uridine and 3H-leucine as well as morphological changes in macrophages and fungus cells were studied by autoradiography. Phagocytized fungus cells remained in the yeast form and showed no activity toward incorporating 3H-uridine and 3H-leucine for 2 hr after infection. Cells not phagocytized began to produce germ tubes and actively incorporated both labeled precursors. Fungus cells within the macrophages produced germ tubes after 3 hr, some of which ruptured the host cells, then grew pseudohyphae and true hyphae. The uptake of both precursors was vigorous in candidal elements, but decreased in macrophages. Networks of long candidal filaments appeared 4 hr after infection and macrophages seemed to be degenerating with severely damaged synthesizing activities. When growth of C. albicans was compared in four different media, one with macrophages and the other three without macrophages, much more mycelial growth was obtained in the medium in which macrophages were present than in media without macrophages. From these results the role of macrophages in Candida infection was discussed.

View full abstract

A computer taxonomy was carried out as an aid to clarify the relationships between the genera Mycobacterium, and Nocardia as well as the ‘Rhodochrous’ group. The last group contained slightly acidfast organisms isolated from sputa and soil and also included strains received as "Mycobacterium rhodochrous." The group had been proposed as the genus Gordona by the present author. The rhodochrous group formed a cluster at the 87% similarity level and was clearly separated from all strains of the genera Mycobacterium and Nocardia. The cluster of the rhodochrous group was divided into seven subclusters at the 90% similarity level, and six of the seven corresponded to six species of the genus Gordona: G. bronchialis; G, rubropertincta (synonym G. rubra); G. terrae; G. aurantiaca; G. rhodochroa; G. rosea. If one accepts the concept that a phenetically well-defined cluster is a species, each of the subclusters of the rhodochrous group should be regarded as a species and the entire group as a genus. Discrepancies between the results of numerical classification and the results of lipid analyses remain as a problem to be solved in the future.

View full abstract

Bordetella bronchiseptica strains isolated from the nasal cavities of young pigs in Japan from 1969 to 1972 were surveyed for drug resistance and distribution of R factors. Of 304 strains examined, 71 (23%) were resistant to either one or more of following three drugs, streptomycin (SM), sulfadimethoxine (SA), and aminobenzyl penicillin (APC). Triple (SM. SA. APC)-resistance was most frequent among these resistant strains. Strains of double (SM. SA)- or single (SM)- and (SA)-resistance were also isolated, but were very few in numbers. Of the 71 drug-resistant strains, 61 (86%) were found to carry R factors which were capable of conjugal transfer. All of these R factors had the triple (SM. SA. APC)-resistant markers and were identified as fi- (no fertility inhibition) type. The (SM. SA. APC)-resistant strains carrying R factors had been isolated from pigs reared on various farms in different districts, and consequently the prevalence of B. bronchiseptica strains carrying R factors was considered to be relatively wide-spread in young pigs.

View full abstract

The effect of light on the pigmentation of various strains of Nocardia, Corynebacterium, Arthrobacter, Brevibacterium, and Flavobacterium was investigated. It was revealed that thirty out of fiftyseven strains of Nocardia, two out of fifteen strains of Corynebacterium, two strains of Arthrobacter, six out of thirteen strains of Brevibacterium and two out of fourteen strains of Flavobacterium were photochromogenic; i. e., these strains produced yellow or orange pigments when grown under illumination but entirely unpigmented in total darkness. From these results, it may be concluded that photochromogenicity is not a particular phenomenon limited to specific species, but a common, widely distributed phenomenon in nonphotosynthetic bacteria.

View full abstract

The optimal growth pH and conditions necessary to give a full manifestation of the effect of reductants upon the growth of Campylobacter fetus were investigated using static cultivation. The multiplication of cells was limited to a narrow range of added reductants. Thioglycollate supplemented without heating produced heavy growth in combinations of 0.05% at pH 6.80 or 0.1% at pH 6.60, and 0.2% at pH 7.00 or 7.20. Although the stimulating effect depended upon the time between the addition of the supplement and inoculation, two prominent peaks in the growth curve always appeared with both alkaline and acidic pH ranges of the medium. Autoclaving the medium with some SH compounds brought about this growth only at around pH 6.50. Consequently there was a uniform decrease in the initial pH. L-Cysteine and ascorbic acid also gave similar effects. Glucose remarkably prevented the growth promotion due to SH compounds. Optimal initial reduction potential for growth in a semisolid medium was considered to be -0.05 to -0.08 volt at an initial pH of 6.60. SH compounds supplemented without heating brought about a marked longevity of the culture in comparison with a culture in an autoclaved medium of the same composition. For the growth of C. fetus in semisolid medium, it was important to avoid decreasing the growth-promoting activity of some of the supplemented SH compounds by not heating but adding the supplement aseptically. Also a rigid regulation of the pH, the concentration of the added reductants, and the time of inoculation into the fresh medium were important.

View full abstract

The nutritional requirement of Campylobacter fetus was investigated by static cultivation at pH 6.80 in the presence of 0.05% thioglycollate, which was added aseptically to avoid reduction of the growth-promoting effect due to heating. Remarkable multiplication was recognized by the coexistence of L-cystine and thioglycollate in a vitamin-free amino acid semisolid medium. L-Cystine was very important for growth, and the composition of a minimum number of amino acids was also required for growth, such as L-aspartic acid, L-glutamic acid, L-cystine and DL-methionine. Guanine among the nucleic acid bases brought about a distinctively heavy growth. Ribonucleic acid, ribose, glutathione. pimelic acid and a certain commercial brand of glycogen clearly showed growth-promoting effects as demonstrated by the increase in additional amounts in a conditional amino acid medium. Similarly, taurocholate acted as an effective growth promoter out of four bile salts tested. Guanine and ribonucleic acid were detected by paper chromatography from the ingredients of media in common use for cultivation of C. fetus. A homemade yeast extract medium retained the typical cell form in almost all of the cultures during long term preservation at room temperature, in contrast to the production of various involution forms in commercial media and a meat extract medium. In conclusion, supplies of both thiol and disulfide groups fulfilled an important role by bringing into action some additives as nutritive substances to be utilized by C. fetus.

View full abstract

The collaborative function of some surfactants for selective cultivation of Campylobacter fetus was examined using a plain homemade yeast extract agar medium supplemented with thioglycollate as a simple basal medium. Out of the combinations of various growth-inhibiting substances with colistin used as the main selective growth-inhibiting surfactant, the combined use of colistin (250 u/ml) and either desoxycholate (0.01%) or taurocholate (0.02%) in addition to brilliant green (1:400000-500000) brought about a satisfactory result. There was no serious disorder of each selective inhibitory function for the growth of C. fetus and suppression to other bacteria, particularly to both Pseudomonas aeruginosa and Proteus vulgaris. It was presumed that this selectivity depended upon both the well-balanced combination in the quantities of the surfactants and the use of a plain basal medium such as the homemade yeast extract agar.

View full abstract

A protein-rich antigen was extracted from the cell walls of Clostridium botulinum type A strain 190L by relatively mild treatments with detergents, tryptic digestion or cell wall autolysis. The cell wall antigen was relatively labile, and destroyed by acid or formamide hydrolysis. Immunodiffusion studies with the trypsin-treated cell wall extracts from various strains of C. botulinum indicated that the antigenic cell wall polymers among the proteolytic strains of types A, B and F but not among the nonproteolytic strains of C. botulinum shared a common antigenic specificity.

View full abstract