In-straw cryoprotectant dilution for bovine embryos vitrified using Cryotop

The aim of this study was to develop an in-straw dilution method suitable for 1-step bovine embryo transfer of vitrified embryos using the Cryotop vitrification-straw dilution (CVSD) method. The development of embryos vitrified using the CVSD method was compared with those of embryos cryopreserved using in-straw vitrification-dilution (ISVD) and conventional slow freezing, outside dilution of straw (SFODS) methods. In Experiment 1, in vitro-produced (IVP) embryos cryopreserved using the CVSD method were diluted, warmed and exposed to the dilution solution at various times. When vitrified IVP embryos were exposed to the dilution solution for 30 min after warming, the rates of embryos developing to the hatched blastocyst stage after 72 h of culture (62.0-72.5%) were significantly lower (P<0.05) than those of embryos exposed to the solution for 5 and 10 min (82.4-94.3%), irrespective of supplementation with 0.3 M sucrose in the dilution solution. In Experiment 2, the rate of embryos developing to the hatching blastocyst stage after 48 h of culture in IVP embryos cryopreserved using the SFODS method (75.0%) was significantly (P<0.05) lower than those of embryos cryopreserved using the CVSD and ISVD methods (93.2 and 97.3%, respectively). In Experiment 3, when in vivo-produced embryos that had been cryopreserved using the CVSD, ISVD and SFODS methods and fresh embryos were transferred to recipient animals, no significant differences were observed in the conception and delivery rates among groups. In Experiment 4, when IVP embryos derived from oocytes collected by ovum pick-up that had been cryopreserved using the CVSD and ISVD methods and fresh embryos were transferred to recipient animals, no significant differences were observed in the conception rates among groups. Our results indicate that this simplified regimen of warming and diluting Cryotop-vitrified embryos may enable 1-step bovine embryo transfer without the requirement of a microscope or other laboratory equipment.     

Thermal stimulation at 39°C facilitates the fusion and elongation of C2C12 myoblasts

The aim of this study was to determine the effect of thermal stimulation at 39°C on the fusion and elongation of skeletal muscle cells. During a 5 day differentiation process of C2C12 cells, nine groups subjected to varying lengths of thermal stimulation at 39°C were established. Afterward, all groups were immunostained using anti‐muscle heavy‐chain antibody to test for myotube formation. Quantification of the myotube area demonstrated a significant increase in the group subjected to thermal stimulation at 39°C during the latter half of the differentiation compared with the control group, but the fusion index was significantly higher in the group that received hyperthermic treatment during the first half of the differentiation period. Moreover, the longitudinal length of myotubes was significantly increased in the groups that were subjected to thermal stimulation at 39°C during the latter half of the differentiation period. The distance between the center of myotubes and the nucleus farthest away from the center was substantially extended in the group receiving thermal stimulation at 39°C only on the fourth day of the differentiation. Together, these results demonstrate that thermal stimulation at 39°C facilitates myoblast fusion and elongation.     

Structure-based mechanistic insights into DNMT1-mediated maintenance DNA methylation

DNMT1, the major maintenance DNA methyltransferase in animals, helps to regulate gene expression, genome imprinting, and X-chromosome inactivation. We report on the crystal structure of a productive covalent mouse DNMT1(731-1602)-DNA complex containing a central hemimethylated CpG site. The methyl group of methylcytosine is positioned within a shallow hydrophobic concave surface, whereas the cytosine on the target strand is looped out and covalently anchored within the catalytic pocket. The DNA is distorted at the hemimethylated CpG step, with side chains from catalytic and recognition loops inserting through both grooves to fill an intercalation-type cavity associated with a dual base flip-out on partner strands. Structural and biochemical data establish how a combination of active and autoinhibitory mechanisms ensures the high fidelity of DNMT1-mediated maintenance DNA methylation.     

Damping-off of cabbage plug seedlings caused by Pythium ultimum var. ultimum in Japan

In October 2004, Pythium ultimum var. ultimum was isolated from rotten stems of cabbage plug seedlings in a commercial nursery in Mie Prefecture (Japan). The isolated fungus was then used to inoculate seedlings and subsequently reisolated from the seedlings with the damping-off disease, showing that P. ultimum var. ultimum is a new pathogen causing cabbage seedling disease.     

Possibility of quantitative assessment of the contribution of paddy irrigation and caldera lakes to river water in Bali Island using water isotopic physics

Paddy and Water Environment - This study aims to investigate a possibility of quantitative assessment of the relationship between return flow from paddy fields and river regime by using hydrogen-...     

Bacterial wilt of bellflower caused by Ralstonia solanacearum in Japan

A sudden wilt of bellflower (Campanula lactiflora) was observed in Japan in 1997. A bacterium that formed white fluidal and mucoid colonies resembling those of Ralstonia solanacearum was isolated from the infected plants. The bacterium was bacteriologically identified as biovar 3 of R. solanacearum. This is the first report of R. solanacearum affecting a plant species of the Campanulaceae family.     

Light-chain multiple myeloma in a cat.

A diagnosis of light-chain multiple myeloma was made in an 11-year-old male American Shorthair cat. The cat showed atypical plasma cell infiltration in the bone marrow, biclonal gammopathy caused by polymerization of myeloma protein (M-protein), and Bence-Jones proteinuria. The M-protein in the serum of the cat was analyzed by using 12% sodium dodeyl sulfate (SDS) polyacrylamide gel electrophoresis with Coomassie brilliant blue staining. An intense band with a size of 27 kDa, the size of the immunoglobulin light chain, was clearly observed, whereas the band corresponding to the immunoglobulin heavy chain (59 kDa) was undetectable. The 27-kDa band was confirmed to be an immunoglobulin light chain by Western blotting by using antibodies for feline immunoglobulin. These data suggested that the neoplastic plasma cells produce light chain only, leading to the diagnosis of light-chain multiple myeloma in the cat.     

Knowledge,attitudes, and practices (KAP) related to brucellosis and factors affecting knowledge sharing on animal diseases: a cross-sectional survey in the dry zone of Sri Lanka

Farmers’ lack of knowledge is assumed to have affected the presence of brucellosis in Sri Lanka for decades. This study, carried out in the Ampara district in the dry zone of Sri Lanka, revealed that there is a significant knowledge gap for brucellosis compared to foot and mouth disease (FMD) (p?<?0.001). Only 8.3% of farmers knew that brucellosis causes cattle abortions. Only 2.6% knew that it is zoonotic. The difference in knowledge of the symptoms and transmission of brucellosis and FMD was significant (p?<?0.001). Farmers’ attitudes and practices related to the spread of the disease were poor. Farmers’ education and spoken language had a negative influence on knowledge. Young people and those with strong social relationships were efficient in knowledge sharing. It can be concluded that brucellosis knowledge, attitudes, and practices are poor; thus, there is a need for more attention in disease control policymaking. Backward farmer groups should be the focus in animal health extension programs.