C14-Labeled proteins obtained in the incorporation of C14-leucine into proteins by cell-free preparations from Pasteurella multocida, were analyzed and the following results were obtained. 1) The radioactivity of C14-labeled proteins was not so greatly lowered by the treatment with alcali. 2) More C14-leucine was released from C14-labeled proteins in acid hydrolysis of them with the increasing rate of hydrolysis. 3) Many sorts of radioactive peptides were detected in the tryptic digest of C14-labeled proteins. 4) The radioactivity of N-terminal or C-terminal amino acids of C14-labeled proteins was 0.5 or 30.5 per cent of the total, respectively. From these results it is concluded that the incorporation of amino acid into proteins by cell-free preparations from Pasteurella multocida was the incorporation into the internal linkage of peptide chains, namely due to the synthesis of peptides.
Further studies were carried out to investigate the 2-mercaptoethanol (2ME)-sensitivity of JE virus antibody of pigs. The results obtained are as follows: 1. In the natural infection cases, 2ME-sensitive antibodies were detected during the first 2 to 4 weeks of antibody response; thereafter the antibodies were found to be resistant to 2ME treatment. 2. Five pigs have been subjected to the observation of 2ME-sensitivity of the sera for 2 or 3 consecutive years. Sera obtained from one pig at the third oversummering were found to be sensitive to 2ME treatment, i.e., the hemagglutination inhibition (HI) titers of the sera were reduced to a quarters of the original titers by 2ME treatment. However, the 2ME-sensitivity of antibodies of other cases which experienced the second or third oversummering, were found to be 2ME-resistant as before. 3. In the experimental vaccination cases, the first antibodies produced after vaccination were found to be 2ME-sensitive and thereafter they became 2ME-resistant. 4. Three pigs possessing 2ME-sensitive HI antibodies were detected in 173 cases possessing antibodies during the interepidemic period. Those were found in March, April and October in 1965, respectively.
Antibiotic-resistance of coagulase positive staphylococci isolated from various animals including human was tested by disc method“Eiken”. Of the six kinds of antibiotics tested PC, TC and SM were relatively resistant, but EM, CM and KM were fairly sensitive. Multiple antibiotic-resistances were seen in these strains isolated from dogs (4 antibiotics), human (4 antibiotics), fowls (3 antibiotics), bovine (2 antibiotics) and pigs (2 antibiotics). After all the resistant strains ratios to each host-derived strains tested were as follows: human (91 strains of 100 strains, 91%), pigs (19 strains of 23 strains, 83%), fowls (59 strains of 100 strains, 59%), bovine (26 strains of 100 strains, 26%), dogs (39 strains of 160 strains, 24%), horses (1 strain of 55 strains, 2%).
With formalinized erythrocytes (FRC) from chickens, men and guinea-pigs, hemagglutination and hemagglutination-inhibition test for myxoviruses with Salk pattern method were studied and the results obtained are as follows. 1) Spontaneous agglutination of FRC was prevented with 0.05 per cent bovine serum albumin in the suspending medium. 2) There was no or little if any differences between HA- or HI-titers obtained with FRC and those with native red cells. 3) FRC was available for HI test after the lyophylization. 4) Hemadsorption test in HVJ-infected HeLa cells was successfully performed with FRC without substitution of bovine serum albumin. 5) HVJ-adsorpting capacity of FRC was identical with that of native cells, but the eluting capacity was decreased in FRC. 6) The ABO-antigenicity of human red cells was partially destroyed by the formalinization.
Acetone dried cells of Saccharomyces cerevisiae were treated with 45% phenol and further with autoclaving at 135° in water, and then the cell residue was extracted with 3% NaOH under N2 gas at 70° for 40 minute. An alkali-soluble glucan was precipitated from the extracts in a cellophane bag which was dialyzed against running tap water. No nitrogen and phosphorus was found in the gelatinous solid glucan. On the other hand, Fehling-Cu++ solution was added in the above mentioned hot-water extracts, and after removing mannan-Cu complex non-Cu polysaccharide (Fr I, II) was fractionated with DEAE-Sephadex column chromatography. Sugar components of the Fr I and II contained mannose and glucose, and a large quantity of glycogen was estimated in the Fr I but not in the Fr II. Fr-G resulted from dilute HOAc treatment of alkali-soluble glucan contained glucose and a trace of mannose as sugar components. Anti-whole cells serum of S. cerevisiae which was absorbed with a complete purified mannan showed extremely weak precipitin titers (×200∼400) against these glucan fractions. Therefore, it appears that the antigenicity of glucan which located on internal wall of the whole cells may be almost neglected in case of agglutination test.