Seasonal differences in rumen bacterial flora of wild Hokkaido sika deer and partial characterization of an unknown bacterial group possibly involved in fiber digestion in winter

Rumen digesta was obtained from wild Hokkaido sika deer to compare bacterial flora between summer and winter. Bacterial flora was characterized with molecular‐based approaches and enrichment cultivation. Bacteroidetes was shown as a major phylum followed by Firmicutes, with similar proportions in both seasons. However, two phylogenetically unique groups in Bacteroidetes were found in each season: unknown group A in winter and unknown group B in summer. The ruminal abundance of unknown group A was the highest followed by Ruminococcus flavefaciens in winter. Moreover, the abundance of these two was higher in winter than in summer. In contrast, the abundance of unknown group B was higher in summer than in winter. In addition, this group showed the highest abundance in summer among the bacteria quantified. Unknown group A was successfully enriched by cultivating with oak bark and sterilized rumen fluid, particularly that from deer. Bacteria of this group were distributed in association with the solid rather than the liquid rumen fraction, and were detected as small cocci. Accordingly, unknown group A is assumed to be involved in degradation of fibrous materials. These results suggest that wild Hokkaido sika deer develop a rumen bacterial flora in response to changes in dietary conditions.     

MytiLec,a Mussel R-Type Lectin,Interacts with Surface Glycan Gb3 on Burkitt’s Lymphoma Cells to Trigger Apoptosis through Multiple Pathways

MytiLec; a novel lectin isolated from the Mediterranean mussel (Mytilus galloprovincialis); shows strong binding affinity to globotriose (Gb3: Galα1-4Galβ1-4Glc). MytiLec revealed β-trefoil folding as also found in the ricin B-subunit type (R-type) lectin family, although the amino acid sequences were quite different. Classification of R-type lectin family members therefore needs to be based on conformation as well as on primary structure. MytiLec specifically killed Burkitt''s lymphoma Ramos cells, which express Gb3. Fluorescein-labeling assay revealed that MytiLec was incorporated inside the cells. MytiLec treatment of Ramos cells resulted in activation of both classical MAPK/ extracellular signal-regulated kinase and extracellular signal-regulated kinase (MEK-ERK) and stress-activated (p38 kinase and JNK) Mitogen-activated protein kinases (MAPK) pathways. In the cells, MytiLec treatment triggered expression of tumor necrosis factor (TNF)-α (a ligand of death receptor-dependent apoptosis) and activation of mitochondria-controlling caspase-9 (initiator caspase) and caspase-3 (activator caspase). Experiments using the specific MEK inhibitor U0126 showed that MytiLec-induced phosphorylation of the MEK-ERK pathway up-regulated expression of the cyclin-dependent kinase inhibitor p21, leading to cell cycle arrest and TNF-α production. Activation of caspase-3 by MytiLec appeared to be regulated by multiple different pathways. Our findings, taken together, indicate that the novel R-type lectin MytiLec initiates programmed cell death of Burkitt’s lymphoma cells through multiple pathways (MAPK cascade, death receptor signaling; caspase activation) based on interaction of the lectin with Gb3-containing glycosphingolipid-enriched microdomains on the cell surface.     

Effect of high intensity training on anaerobic capacity of middle gluteal muscle in Thoroughbred horses

We hypothesize that high intensity training for Thoroughbred horses that have been subjected to conventional training could further improve the metabolic properties of the middle gluteal muscle. Nine well-trained horses were subjected to high intensity (80-100% Vdot;O(2)max, 5 minx2) training for 12 weeks. Biopsy samples were obtained from the muscle before and after 4 and 12 weeks of training. Three of the 9 horses did not complete the training programme. In the remaining 6 horses, activities of succinic dehydrogenase (SDH), phosphofructokinase (PFK) and 3-hydroxy acyl CoA dehydrogenase (HAD), and the composition of myosin heavy chain isoforms were analyzed by biochemical techniques. After 12 weeks of training, a significant increase was found in PFK activity but not in the SDH and HAD activities. There were no significant changes in the composition of myosin heavy chain isoforms. The high intensity training in this study was effective at increasing glycolytic enzyme activity, indicating the possibility to improve anaerobic capacity, which potentially could contribute greatly to performance in Thoroughbred horses. This study also highlighted a fact that high intensity training should be given with the great care to prevent the skeletal muscle injuries.     

Establishment of a large-scale purification procedure for purified recombinant bovine interferon-tau produced by a silkworm-baculovirus gene expression system

We developed a procedure for the large-scale purification of bovine interferon-tau (boIFN-tau) by means of a silkworm-baculovirus gene expression system. Recombinant boIFN-tau (rboIFN-tau) was efficiently produced in the silkworm infected with boIFN-tau cDNA recombinant baculovirus and accumulated in the haemolymph. To establish a purification method suitable for mass production, we tried three crude purification methods, namely, an acidification and neutralization treatment (ANT), silica gel column chromatography (SGCC), and Blue sepharose column chromatography (BSCC) with a combination of Q-sepharose (QSC) and chelating sepharose column chromatographies (CSCC). As a result, the acidification and neutralization treatment was found to be the most efficient and cost effective. With this combination, we obtained 91% pure products. To confirm the applicability of the procedure for mass production, we inoculated 100 silkworms with the recombinant virus, and recovered about 4.55 mg (1.26 x 10(8)U/mg) of 91% pure rboIFN-tau by means of a combination of the ANT, followed by QSC and CSCC.     

Cardiovascular effects of a phosphodiesterase III inhibitor in the presence of carvedilol in dogs

The aim of this study was to determine whether dobutamine, dopamine, or milrinone (a phosphodiesterase [PDE] III inhibitor) would support cardiac function that had been attenuated by administration of the beta-blocker, carvedilol (0.2, 0.4, or 0.8 mg/kg). Hemodynamic and cardiac parameters including the heart rate (HR), left-ventricular fractional shortening (FS), and arterial pressure were measured in six healthy dogs without cardiac disease. Carvedilol did not affect FS or arterial pressure, but decreased the HR significantly. The positive inotropic and chronotropic responses to dobutamine and dopamine were attenuated by carvedilol, whereas arterial pressure was unaffected. Milrinone did not affect the HR and decreased arterial pressure, whereas FS was significantly greater both in the control and carvedilol-treated groups. Although milrinone affect the negative chronotropic effects of carvedilol, milrinone increased FS and prevented the decrease in arterial pressure. These results suggest that inhibition of PDE III preserves cardiac contractility and hemodynamic function in the presence of carvedilol.     

The 9th International Veterinary Immunology Symposium

The orchestrated migration of T lymphocytes is important for generating immunity and maintaining immunological tolerance. T lymphocytes can be divided into two populations, αβ T cells and γδ T cells, on the basis of their expression of different forms of the T cell receptor (TCR). γδ T cells represent an innate subset of T lymphocytes that play an important role in early immune response against a variety of pathogens, including bacteria and viruses. γδ T cells are abundant in the epithelial tissues. In ruminants and pigs, they constitute a major proportion of the blood lymphocyte pool, unlike in rodents and humans. Although recent studies using large animals have suggested that epithelial γδ T cells are the major source of γδ T cells in peripheral blood, and that they recirculate between epithelial tissues and blood via lymphatics, the migration pattern of these cells is largely unknown. The aim of this review is to provide an overview of the current knowledge on γδ T cell migration under steady-state conditions. A deeper understanding of γδ T cell migration may enable therapeutic modulation of innate immune responses.     

A review of seed production and stock enhancement for commercially important portunid crabs in Japan

We reviewed the present status of seed production for stock enhancement and evaluated the impact of stocking on commercial catches in Japan, of portunid crabs particularly Portunus trituberculatus and Scylla paramamosain. The mean survival rate from hatching to first-stage crabs was around 10%, and 20–30% of the larval culture trials conducted in recent years could not harvest juveniles during seed production. To achieve reliable mass seed production technologies, measures for controlling disease and the nutritional condition of larvae in seed production tanks are required to be developed. The main spawning season extends from April/May to July. Reflecting their life cycle characteristics such as high growth rates, crabs recruit to the fishery after September and largely contribute to the commercial landings until December in the hatching year. The main release season of juveniles is from June to July. Consequently, released juveniles are expected to contribute to commercial landings in the release year. Analyses of catch and release statistics in two small bays estimated the yield from released individuals (YPR, yield per release) at 2.4 g for P. trituberculatus and 3.3–7.7 g for S. paramamosain, which were similar to values reported from tagging surveys. On a major regional basis, catch and release histories of P. trituberculatus highlighted the impact of hatchery releases on commercial landings as 33.6 g YPR in the Seto Inland Sea, where catches and releases have been greatest. Although the YPR estimates were different between small bays and major regional seas, Japanese stock enhancement programmes should have had an impact on portunid crab production, dependent on the magnitude of the releases.     

Fishery biology of mud crabs <Emphasis Type="Italic">Scylla</Emphasis> spp. at Iriomote Island,Japan: species composition,catch, growth and size at sexual maturity

The fishery biology of mud crabs Scylla spp. was examined using baited traps and gill nets from September 2001 to August 2005 at Iriomote Island, Japan. To elucidate the growth of the crabs, artificially produced S. serrata juveniles were released and recaptured at the study site. The sizes at which 50% of females and males of S. serrata reached sexual maturity (SM₅₀) were estimated as an external carapace width (ECW) based on the morphology of the abdomen and the chela respectively. Two species, S. serrata and S. olivacea, were identified in the area with S. serrata being the dominant species (>95% of the catch). Changes in the mean ECW and the results of the release and recapture experiments suggested that the recruitment of young crabs to the fishery occurred from December/January to April/May. The SM₅₀ of females and males occurred at 132.4 and 150.7 mm ECW respectively. The body size composition of S. serrata revealed that immature crabs comprised approximately 40 and 65% of the catch for females and males respectively. To maintain a sustainable fishery for S. serrata, a minimum landing size based on the SM₅₀ estimates should be implemented as a fishing regulation.     

Characterization of Aquimarina hainanensis isolated from diseased mud crab Scylla serrata larvae in a hatchery

Mass mortality due to necrosis signs occurred in hatchery-reared zoea stage larvae of the mud crab Scylla serrata in Okinawa, Japan, and a causative bacterium was isolated. In this study, we identified and characterized the bacterium by genome analysis, biochemical properties and pathogenicity. The bacterium was a Gram-negative, non-motile, long rod, forming yellow colonies on a marine agar plate. It grew at 20–33°C (not at 37°C) and degraded chitin and gelatin. Phylogenetic analysis of the 16S rRNA gene sequence identified the bacterium as Aquimarina hainanensis. Genome sequence data obtained from Illumina MiSeq generated 29 contigs with 3.56 Mbp in total length and a G + C content of 32.5%. The predicted 16 chitinase genes, as putative virulence factors, had certain homologies with those of genus Aquimarina. Experimental infection with the bacterium conducted on larvae of four crustacean species, brine shrimp Artemia franciscana, freshwater shrimp Caridina multidentata, swimming crab Portunus trituberculatus and mud crab S. serrata, revealed that this bacterium was highly virulent to these species. The present study suggests that the bacterium caused mass mortality in mud crab seed production was A. hainanensis and can be widely pathogenic to crustaceans.     

High-yield SRI in West Java by decomposing straw in waterlogged paddy field

System of Rice Intensification (SRI) often achieved higher yield than conventional practice. We identified the high-yielding farmers from the yield records of 1909 paddy fields belonging to an organic farmers’ association. Farmers whose yields were from 8.4 to 10.4 t ha^?1 were interviewed and their fields surveyed. Their yields had increased by an estimated average of 40% following the adoption of SRI practices. They applied 2–12 t ha^?1 of compost. Compared to the conventional practice, they shortened seedling age at transplanting from 27.4 to 17.6 days and reduced the number of seedlings per hill from 4–6 to 2–3, while hill spacing remained unchanged. Instead of intermittent irrigation which is recommended in standard SRI, they kept shallow flooding of 1–2 cm. Although they applied a lot of compost, no correlation was found between the amount of compost application and the yields. Instead, high-yielding farmers returned rice straw into waterlogged paddy after harvest, which presumably is an ideal condition for biological nitrogen fixation. This may occur around rice straw during decomposition under waterlogged condition and might supplement the negative nitrogen balance, thereby enabling the high yield as compared with conventional practices where the fresh rice straw is removed and/or burned.