Some bacterial strains of anaerobic coryneforms and propionibacteria were examined for effects on such biological activities as RES-stimulation, lysosomal enzymes in macrophages, humoral or cell-mediated immune responses and anti-tumor activities in mice. The results obtained are as follows. 1) With regard to ability to increase the weights of spleen and liver and to enhance carbon clearance from the blood circulation, the strains could be arranged in the decreasing order as follows: Corynebacteriumanaerobium 578 (Serol. group II), C. parvum 0208 (Serol. group I), Propionibacteriumgranulosum 0507 (Serol. group III), P. avidum 0575 (Serol. group IV), and C. parvum 11829 (Serol. group III). 2) The intracellular contents of β-glucuronidase and acid phosphatase in macrophages activated with each strain were reversely correlated with the above-mentioned order of RESstimulation and carbon clearance. It is of interest to note that the enzyme contents became smaller and smaller in accordance with the elevation of the grade of activation of, macrophages, probably as a result of the extracellular excretion of both enzymes. 3) Various degrees of enhancing effect on hemolytic plaque-forming cells in the spleen and suppressive effect on delayed-type hypersensitivity to SRBC were observed. Neither effect, however, was strictly parallel with such ability to stimulate RES as estimated by the carbon clearance test. 4) The strains were arranged in the decreasing order of anti-tumor activity against Ehrlich ascites tumor as follows: P. granulosum 0507>C. anaerobium 578>C. parvum 0208>P. avidum 0575>C. parvum 11829. They were listed in the decreasing order of anti-tumor activity against Sarcoma 180 as follows: P. avidum 0575=C. parvum 0208=C. anaerobium 578=P. granulosum 0507≠C. parvum 11829. From the above-described results, it wasascertained that there was a considerable difference in the biological activities examined among five strains of coryneforms and propionibacteria. Furthermore, discussion was made on the possible implication of these results in the light ofmacrophage activation.
Lipopolysaccharide (LPS) antigens were purified from heated cell extracts of Pseudomonas aeruginosa types A, B, D, E, and I. Indirect hemagglutination (IHA) with a given type of them showed specific reactivity to anti-Pseudomonas rabbit serum immunized with the same type of antigen. As a result of absorption of heterotypic antibody, type-specific antibody in human serum was titrated successfully by using LPS-coated sheep erythrocytes. Sera from healthy adults and patients infected with P. aeruginosa were studied subsequently and revealed to possess antibodies against P. aeruginosa. IHA antibody against type E wasresistant to 2-mercaptoethanol (2-ME) and classified as IgG in patients infectedwith type E of Pseudomonas. It was however, sensitive to 2-ME and classified as IgM in patients infected with any other type of P. aeruginosa, as well asin healthy adults. The titer and positive incidence ofantibody against type-E Pseudomonas were higher than those of antibody against any other type of Pseudomonas examined.
Many pleiotropically carbohydrate-negative mutants lacking components of the phosphoenolpyruvate: sugar phosphotransf erase system (PEP. PTS or simply PTS), i. e., pleiotropic PTS mutants, of Vibrio parahaemolyticus were isolated by the methyl-α-D-glucoside screening method described previously (J. Bacteriol. 119: 632-634, 1974). As expected from the selecting procedure, all the mutants isolated were deficient in the utilization of glucose as the carbon and energy source. Their patterns of pleiotropy for the utilization of the other eight carbohydrates, however, were strikingly different from one another. Some of the metabolic defects of the mutants could be overcome by supplementing cyclic adenosine 3', 5'-monophosphate (cAMP) to the medium. Therefore, such metabolic defects might be due not to any defect in the PTSmediated phosphorylation of carbohydrates, but to an insufficient supply of cAMP to induce certain enzymes involved in metabolism of the sugars. A similar finding has been reported. in PTS-mutants of Escherichia coli. On the other hand, the pleiotropic patterns of the V. parahaemolyticus mutants were still heterogeneous even in the presence of exogenous cAMP. Therefore, at least three different types, B, C, and D, of mutants were recognized. Mutants, of type B were defective in the utilization of five carbohydrates, glucose, trehalose, fructose, mannose, and mannitol, whereas mutants of type D could utilize fructose normally, and mutants of type C were lacking only in the utilization of glucose and trehalose when cAMP was present in the medium. A possible interpretation for this phenomenon is that the PEP·PTS of the organism has at least three protein components, which are common to the PTS-mediated phosphorylation reaction for more than two carbohydrates.
Scanning electron microscopy was proved to be useful for observation on the cell arrangement in bacterial colonies. Satisfactory results were obtained by employing such preparative procedures as cultivation on the cellulose dialysis membrane and the CO2 critical-point drying technique. The double fixation with osmic acid vapor and 3% glutaraldehyde on colonies was also effective, although optimal fixation time varied from one species to another. Studies were made on several species of the genera Lactobacillus, Bacillus, Corynebacterium, and Salmonella. It was concluded that there was an intimate relationship between the macroscopic appearance of colonies, especially the regular or irregular outer edge and the smooth or rough surface, and scanning electron microscopic findings, such as regular or irregular cell arrangement, and the chain formation and angular apposition of daughter cells after cell division within colonies.