[Evaluation of recovery methods of Shigella species from fresh marine fish and shellfishes].

Recovery experiments of Shigella strains from fresh marine fish and shellfishes, including fresh sea urchin, which have been artificially contaminated with the strains, were performed using the improved Shigella broth-enrichment method and the culture method reported by Mehlman et al. All of the 43 Shigella stock cultures strains tested were recovered easily by the enrichment method from sea urchin individuals inoculated with a small number of viable cells of each strain. That is, a total of 24 strains (56%) were recovered from sea urchin individuals inoculated with less than 10 viable cells per one individual, and the other 19 strains were also recovered when 10 to 1,000 cells of each strain were inoculated. Recovery of Shigella strains from fish and shellfishes by the enrichment method was hardly affected by the number of contaminated bacteria (SPC, standard plate counts) in these materials. In order to confirm reliability of the enrichment method, similar experiments were performed using S. flexneri strain B as the inoculum and more fish and shellfishes as the samples (24 specimens of fresh sea urchins, 11 specimens of fresh oysters and 5 other specimens including prawns). Except for one oyster specimen which showed an especially high SPC value, the inoculum was able to be recovered from most of the materials inoculated with less than 10 viable cells, and all of the tested samples became Shigella positive when they were inoculated with up to 1,000 viable cells.(ABSTRACT TRUNCATED AT 250 WORDS)

Purification and partial characterization of fimbriae of Vibrio cholerae O1.

Fimbriae of Vibrio cholerae O1 were purified from a strain of the classical biotype, Inaba serotype (Bgd 17), and a strain of the El Tor biotype, Ogawa serotype (K23), grown on TCG agar medium by the following procedure; homogenization of the cell suspension to detach fimbriae, ultracentrifugation to remove remaining cells and their debris, concentration of the supernatant containing fimbriae with ultrafiltration, and 20 to 40% sucrose linear gradient centrifugation of the concentrated material. The fimbriae in both preparations were flexible, long fibres readily distinguishable under the electron microscope from those of CFA/I, CFA/II seen in ETEC strains. Their structural subunit was a protein of 16 kdaltons. Fimbriae isolated from both serotypes and biotypes shared antigenic determinants.

Depression of protective mechanisms against microörganisms in tumor-bearing mice and its restoration by adjuvants.

Bacterial growth within 72 hr after an intravenous inoculation with Listeria monocytogenes was enhanced strikingly in the liver of mice, when viable cells of sarcoma-180 were injected subcutaneously into ddN, C3H/He, and BALB/c mice 5 hr before the inoculation. Such an enhanced bacterial growth appears to be attributable to a depressed ability of macrophages to digest engulfed bacteria. Pretreatments with zymosan, killed BCG, or viable BCG prevented such depression in tumor-bearing mice and increased the bactericidal activity in the liver of normal and tumor-bearing mice above the level of non-treated normal mice. Such adjuvants may be useful not only for augmentation of antitumor activity but also for augmentation of antimicrobial activity in tumor-bearing hosts.

Cellular mechanisms in the protection against infection by Listeria monocytogenes in mice.

Listeria monocytogenes, in doses of 2-0 X 10(3) to 3-0 X 10(3) viable organisms, was injected into athymic nude mice, irradiated mice and mice treated with reticuloendothelial system-blocking agents. Viable counts on liver and spleen homogenates were made at intervals after infection. In both nude mice (nu/nu) and normal littermates (nu/+) of BALB/c background, the bacteria grew rapidly for 24 h but increased only slowly thereafter, to reach a plateau of about 10(5) per organ at 72 h. In nu/+ mice, the number of viable bacteria began to decrease after 6 to 9 days, with complete elimination by day 12. In nude mice, the number of Listeria remained at a stable level of approximately 10(5) per organ during the observation period of 21 days. In lethally irradiated nu/+ mice, bacteria grew progressively and extensively to reach 10(7) per spleen and 10(9) per liver by 72 h. Bacterial growth during the first 72 h was markedly enhanced by treatment with carbon particles, dextran sulphate 500 or silica. These enhancing effects were also observed in nude mice and in AKR, C3H/He and C57BL/6 animals. We conclude that both non-immune phagocytes and T cell-dependent mechanisms contribute to the resistance of mice to Listeria infection.