The changes in proximate compositions and glycogen contents in the dorsal ordinary muscles of the full-cycle cultured Pacific bluefin tuna Thunnus orientalis occurring with growth

ABSTRACT:   Using full-cycle cultured (FC) Pacific bluefin tuna (body length [BL], 42.6–66.4 cm; body weight [BW], 1.66–7.40 kg, n  = 15), the changes in chemical compositions and histological structure of the cephalal parts of the dorsal ordinary muscles (DOM) occurring with growth were investigated. A positive correlation ( r  = 0.9644, P  < 0.05) was observed between BL and BW with growth. The protein, lipid and ash contents of DOM and condition factor did not change with growth. However, the glycogen content of DOM increased ( P  < 0.05) from approximately 55 cm (BL) in this study. Using optical microscopic photographs, the various shapes of muscle fibers were observed and it was noted that the muscle fiber diameter increased ( P  < 0.05) with growth. Using transmission electron microscopic observation, many glycogen granules were observed in muscle fibers (especially, side of connective tissue) of DOM throughout the growth stage in this study. These results indicate that the glycogen content of DOM of FC Pacific bluefin tuna increases before the lipid with growth.     

Effective Synthesis and Antifouling Activity of Dolastatin 16 Derivatives

Some derivatives of dolastatin 16, a depsipeptide natural product first obtained from the sea hare Dolabella auricularia, were synthesized through second-generation synthesis of two unusual amino acids, dolaphenvaline and dolamethylleuine. The second-generation synthesis enabled derivatizations such as functionalization of the aromatic ring in dolaphenvaline. The derivatives of fragments and whole structures were evaluated for antifouling activity against the cypris larvae of Amphibalanus amphitrite. Small fragments inhibited the settlement of the cypris larvae at potent to moderate concentrations (EC₅₀ = 0.60-4.62 μg/mL), although dolastatin 16 with a substituent on the aromatic ring (24) was much less potent than dolastatin 16.     

Growth Complementation of hrpXo Mutants of Xanthomonas oryzae pv. oryzae by Virulent Strains in Rice Cultivars Resistant and Susceptible to the Parental Strain

Xanthomonas oryzae pv. oryzae (X. o. pv. oryzae) T7174 is virulent on rice cultivar IR24 and avirulent on IR-BB2. From recent reports, some virulence and avirulence factors of plant pathogenic bacteria are transferred to plant cells through the hrp-dependent type III secretion system. In this study, we investigated the involvement of hrp genes in the compatible and the incompatible interactions between rice and X. o. pv. oryzae after co-inoculation with hrpXo mutants derived from T7174 and virulent strains. Growth of the mutants, named 74ΔHrpXo and 76ΔHrpXo, was repressed in IR24 when the mutants were applied alone. However, growth of the mutants was complemented by co-inoculation with virulent strains. Growth of bioluminescent hrpXo mutant 76ΔHrpXo in IR24 and its growth in IR-BB2 after co-inoculation with T7133, which is virulent on both cultivars, was equally complemented, as detected by bioluminescence from the mutant. On the other hand, only partial complementation of growth of T7174L76, which is a bioluminescent and pathogenic derivative of T7174, by T7133 was observed in IR-BB2. Thus, growth of the hrpXo mutant of X. o. pv. oryzae was complemented by virulent strains in both susceptible and resistant rice leaves with the parental strain. Received 21 July 2000/ Accepted in revised form 26 October 2000     

Generation of canine dendritic cells from peripheral blood mononuclear cells

Dendritic cells (DCs) are the most potent antigen-presenting cells that are expected to be therapeutic agents for tumor immunotherapy. In this study, we generated DCs of sufficient number for DC-based immunotherapy from peripheral blood mononuclear cells (PBMC) in dogs. PBMC were cultured in the presence of phytohemagglutinin (PHA). On day 6, large adherent cells with dendrite-like projections were seen, and the number of these large cells with projections increased on day 8. These cells were positive for esterase staining. They expressed MHC class II, CD11b, CD8 and weakly CD4 on their surface. They tended to make contact with lymphocytes under culture conditions. We obtained about 2-5 x 10(6) of DCs from 10 ml of peripheral blood. These DCs phagocytosed HEK-293 cells by overnight co-culturing. These cells generated from PBMC are possible canine DCs and are applicable to clinical trials of DC-based whole tumor cell immunotherapy in dogs.     

Detection of canine distemper virus antigen in canine serum and its application to diagnosis

Canine distemper virus (CDV) antigen was detected in the serum of dogs by an ELISA and the results of this assay were compared with an anti-CDV immunoglobulin M (IgM) antibody test. In paired sera from 26 naturally infected dogs, the antigen-positive rate was 26.9 per cent at the first examination and 11.5 per cent at the second examination two to three weeks later. The antigen was detected in three of the 10 dogs which were negative for anti-CDV IgM antibody at the first examination. It could also be detected in the serum of between eight and two of 40 specific pathogen-free dogs vaccinated against CDV, for up to four weeks after they were vaccinated.     

Hyaluronidase is not essential for the lethality of Erysipelothrix rhusiopathiae infection in mice

To investigate the role of hyaluronidase in the pathogenicity of Erysipelothrix rhusiopathiae, transposon Tn916 was transferred from Enterococcus faecalis CG110 to a virulent strain of E. rhusiopathiae, and hyaluronidase-deficient mutants were isolated. A virulence assay in the mice showed that of the seven hyaluronidase-deficient mutants tested, six mutants were avirulent, but that one mutant, designated AST121, was as virulent as its parental strain. Western immunoblotting with a monoclonal antibody specific to the capsule, a major virulence factor of the organism, revealed that all of the avirulent mutants had lost the capsular antigen, whereas the mutant AST121 did not. These results suggest that the lack of virulence of the six hyaluronidase-negative mutants could be due to a loss of the capsule and that hyaluronidase does not contribute to the lethality of E. rhusiopathiae infection in mice.     

Successful treatment of two dogs with allergic dermatitis by anti-allergic peptides (MS-antigen)

The effects of non-specific immunotherapy with anti-allergic peptides extracted from the urine of human allergic patients (MS-antigen), in two dogs with allergic dermatitis (AD) have been described. Clinically, severe pruritus accompanied by secondary bacterial pyoderma did not respond to conventional therapy with systemic antibiotics. The first clinical change appeared as a significant reduction in pruritus within 3 months, around the time of the 15th injection in both cases. The clinical condition was stabilized after 5 months, allowing the gradual withdrawal of concurrent therapies and an increase of injection intervals. The correlation between the results of intradermal skin tests before and after treatment and the improvement of clinical signs was not obvious.     

Detection of DNA restriction fragment polymorphisms in Streptococcus equi

Large-restriction-fragment (LRF) polymorphisms in Streptococcus equi (S equi subspecies equi) were studied by pulsed-field gel electrophoresis. Five or six chromosomal fragments of between 194 and 915 kb were separated by digestion with the restriction endonuclease Notl. All 20 isolates of S equi, including 12 from independent Japanese outbreaks, four from independent American outbreaks, two from a single Irish outbreak, us vaccine strain F43, and type strain NCTC 9682 were successfully typed. Seven distinctive, reproducible and stable types were identified. The 12 Japanese isolates collected between 1992 and 1998 were of LRF type II suggesting that they were derived from the same source. The remaining eight isolates were of six types. The results indicate that LRF typing should be a useful technique for investigating the source and transmission of S equi.     

Ultrastructural study on scab resistance expressed in epidermal pectin layers of pear leaves

Proliferation and collapse of subcuticular hyphae of Venturia nashicola race 1 were studied ultrastructurally, after inoculation of susceptible Japanese pear cv. Kousui, resistant Japanese pear cv. Kinchaku, resistant Asian pear strain Mamenashi 12 and nonhost European pear cv. Flemish Beauty leaves, to understand the nature of the resistance mechanism. After cuticle penetration by the pathogen, the hyphae were observed at lower frequency in epidermal pectin layers and middle lamellae of leaves of the three resistant plants than in those of susceptible ones. This result suggested that fungal growth was suppressed in the incompatible interaction between pear and V. nashicola race 1. In the pectin layers of all inoculated plants, some hyphae had modifications such as breaks in the plasmalemma with plasmolysis, necrotic cytoplasm and degraded cell walls. More hyphae had collapsed in the leaves of the three resistant plants than in those of the susceptible cv. Kousui. In collapsed hyphae, the polymerized cell walls broke into numerous fibrous and amorphous pieces, showing that the scab resistance might be associated with cell wall-degrading enzymes from pear plants.