Effect of activation methods for bovine oocytes after intracytoplasmic injection

A principal nuclear transfer procedure is to inject a donor cell into the perivitelline space in an enucleated oocyte and then electric fusion is performed (cell fusion method). The effects of activation methods in reconstructed oocytes for the serum-starved somatic cell cloning procedure were investigated in this study by means of intracytoplasmic injection (i.c.i.). Bovine oocytes were enucleated at 18-22 h for in vitro maturation, and subsequently the nucleus of cumulus cell collected from Japanese Black Bulls (JBCC) after 5-7 days of starved culture was injected into the recipient cytoplast with a piezo-micromanipulator. At 1 h after i.c.i., reconstructed oocytes were stimulated with ethanol (ET) or calcium ionophore (CaI) as the first activation treatment, followed by cycloheximide (CHX) or 6-dimethylaminopurin (DMAP) treatment as the second activation. In the experiment on the first activation method, the proportion of reconstructed oocytes developing to the blastocyst stage was significantly (p<0.01) higher in the ET activation method than that with CaI (10.5% and 4.7%, respectively). And the experiment on the second activation method after ET treatment showed similar proportions of blastocyst development in both CHX and DMAP treatments (5.9% and 2.8%, respectively). The present results indicated that combined activation treatment with ET and CHX was efficient for reconstructed bovine oocytes by i.c.i.     

Clinical response of ovarian cysts in dairy cows after PRID treatment

We investigated the therapeutic effects of a progesterone releasing intravaginal device (PRID) on cystic ovarian disease (COD) and reproduction performance of cows. The possible influence of PRID on metabolic and/or health status was also examined. A total of 40 Holstein-Friesian cattle, with ovarian cystic structures, > or =2.5 cm in diameter, persisting for more than 7-14 days, without a corpus luteum (CL) were used for the study. PRID or placebos were inserted into the vagina for 12 days. Five animals lost the intravaginal device before removal and one was culled. Based on plasma progesterone concentration on the day of treatment, 20 (17 PRID and 3 placebos) of the remaining 34 cows had follicular cysts (progesterone < or =1 ng/ml) and 14 (10 PRID and 4 placebos) had luteal cysts (progesterone >1 ng/m l). Fourteen (82%) of the PRID-treated follicular cystic cows responded with formation of a CL within 14 days after treatment, and an overall conception rate of 53.8%. Likewise, 70% of the treated luteal cystic cows responded with CL formation and 71.4% conception rate. No significant differences were observed in hematocrit (Ht), white blood cell count and serum levels of glucose, blood urea nitrogen, aspartate aminotransferase, and alanine aminotransferase, between the day of PRID insertion and removal, in animals with follicular and luteal cysts. PRID treatment resulted in ovulation 2-4 days later and formation of a CL in cows that recovered.     

Percutaneous ultrasound-guided over-the-wire catheterization of the portal and hepatic vessels in cattle

A new method for catheterization of the portal and hepatic veins in cattle by means of the over-the-wire system was investigated to maintain more reliable long-term patency of catheters. Four cattle were used to evaluate the success rate, patency and safety of the procedure. The catheters, coated by urokinase were patent as long as they were in situ. In addition, the introducer was useful to prevent the catheter from being broken. No complications developed during the10 days after the procedure. Two cows were then euthanized. Post mortem findings were minimal. The results of the study reported here are promising, the benefits are significant and there is no apparent disadvantage to its use.     

Comparison of macrophage scavenger receptor-A knockout mice with wild type ones in the immune response against repeated infestation with Haemaphysalis longicornis

Using macrophage scavenger receptor-A knockout (SRKO) mice, we examined the role of macrophage class A scavenger receptors (MRS-A) on the immune response and acquisition of host resistance against repeated infestation with Haemaphysalis longicornis. Except for one batch of nymphs that infested one of the SRKO (SR-/-) mice and showed no appreciable reduction in body weight, all the other groups of nymphs manifested significant decrease in body weight. Both SR-/- and wild type (SR+/+) mice showed a sustained increase in anti-tick antibody titers, but SR+/+ mice showed significantly higher titers. The IFN-gamma assayed in SR-/- mouse immune sera was substantially less compared with that in SR+/+ mice. Immune sera from SR-/- and SR+/+ mice recognized the 51 and 44 kDa, and 44 kDa proteins, respectively, of the salivary gland antigen. The difference in the level of anti-tick resistance manifested by both groups of mice may be influenced by less efficient trapping and processing of tick antigens by macrophages in mice lacking for the macrophage scavenger receptors, and consequently affected the cascade of Th1 and Th2 responses. We have thus obtained valuable data that strongly infer the role of MSR-A in enhancing host defense against repeated infestation with H. longicornis.     

Surface deterioration of wood-flour polypropylene composites by weathering trials

The market for wood-fiber plastic composites (WPCs) is expanding rapidly in many countries including Japan, where WPCs are mainly used for exterior products. In such applications, WPCs undergo undesirable color change, chalking, and shrinkage and swelling, and accordingly there is a need to better understand the mechanisms responsible for the weathering of WPC and develop methods of improving their weathering resistance. In this study, weatherability of WPC was assessed by natural and accelerated weathering trials. Discoloration (whitening) of WPC during exposure was caused by degradation of both wood and plastic. Darker color pigments as additives improved the color stability of WPC; however, chalking on the surfaces still occurred. The color stability of WPC was improved by application of exterior coatings. Preweathering of WPC (before coatings were applied) increased the absorption of coatings by the WPC and had a positive effect on the color stability and prevented chalking of the composites. Parts of this study were presented at the IUFRO XXII World Congress Meeting, Brisbane, August 2005 and the IAWPS 2005, Yokohama, November 2005     

Pseudodynamic tests and earthquake response analysis of timber structures II: two-level conventional wooden structures with plywood sheathed shear walls

Pseudodynamic (PSD) tests were conducted on two-level timber structures with plywood-sheathed shear walls, which each had an opening of different configuration, to study the effects of the mechanical properties of the first and second levels on the earthquake response of the structure. The specimens had two-level conventional post and beam frames that were 3 m wide, 3 m deep, and 6 m high with plywood sheathings nailed on one face of the structure. The first and second levels had different opening configurations of window, door, or slit. Lateral forces were applied at the top of the first and second levels, calculating step by step the next displacement based on the North-South (NS) components of the 1940 El Centro earthquake. The test results were compared with those of the time-history earthquake response analysis using the lumped mass model and hysteresis model presented in the companion article (part I). The experimental and simulated results showed that the simulation by means of the lumped mass time-history earthquake response analysis predicted quite well the response of the first level, but tended to underestimate the response of the second level, and that the PSD tests of an individual wall system with the mass supported by that particular wall generally show a conservative estimate of the response.     

Genetic variants of the unsaturated fatty acid receptor GPR120 relating to obesity in dogs

G protein-coupled receptor (GPR) 120 is an unsaturated fatty acid receptor, which is associated with various physiological functions. It is reported that the genetic variant of GPR120, p.Arg270His, is detected more in obese people, and this genetic variation functionally relates to obesity in humans. Obesity is a common nutritional disorder also in dogs, but the genetic factors have not ever been identified in dogs. In this study, we investigated the molecular structure of canine GPR120 and searched for candidate genetic variants which may relate to obesity in dogs. Canine GPR120 was highly homologous to those of other species, and seven transmembrane domains and two N-glycosylation sites were conserved. GPR120 mRNA was expressed in lung, jejunum, ileum, colon, hypothalamus, hippocampus, spinal cord, bone marrow, dermis and white adipose tissues in dogs, as those in mice and humans. Genetic variants of GPR120 were explored in client-owned 141 dogs, resulting in that 5 synonymous and 4 non-synonymous variants were found. The variant c.595C>A (p.Pro199Thr) was found in 40 dogs, and the gene frequency was significantly higher in dogs with higher body condition scores, i.e. 0.320 in BCS4–5 dogs, 0.175 in BCS3 dogs and 0.000 in BCS2 dogs. We conclude that c.595C>A (p.Pro199Thr) is a candidate variant relating to obesity, which may be helpful for nutritional management of dogs.     

Sericin Accelerates the Production of Hyaluronan and Decreases the Incidence of Polyspermy Fertilization in Bovine Oocytes During In Vitro Maturation

Fetal bovine serum (FBS) has been widely used as a supplement in the maturation medium of bovine oocytes in vitro. However, serum contains many undefined factors and is potentially infectious to humans and animals. As a serum replacement, we evaluated the feasibility of using the silk protein, sericin, derived from the cocoons of silkworm. To examine the rates of oocyte maturation and fertilization, cumulus-oocyte complexes were cultured in TCM-199 supplemented with 0.01%, 0.05%, 0.1% or 0.15% sericin or 5% FBS. The sizes of the perivitelline space that might relate to polyspermy, the expressions of Has2 and CD44 mRNA, the amount of hyaluronan (hyaluronic acid: HA) contained in the oocytes and the rates of blastocyst formation following insemination were then compared between the oocytes cultured with 0.05% sericin and 5% FBS, because the polyspermy rates in oocytes cultured with 0.05% sericin were significantly lower than in those cultured with 5% FBS. After in vitro maturation (IVM), the mean size of the perivitelline space was significantly greater in oocytes cultured with sericin than in those cultured with FBS, although the rates of nuclear maturation, fertilization and blastocyst formation of oocytes under both IVM conditions were not significantly different. The expression of HAS2 and CD44 mRNA and the amount of HA in the denuded oocytes cultured with 0.05% sericin were significantly greater than in those cultured with FBS. These results indicate the feasibility of sericin as an alternative protein supplement for IVM in bovine oocytes.     

Is experience on a farm an effective approach to understanding animal products and the management of dairy farming?

The understanding of animal products and dairy farming is important for the promotion of dairy farming. Thus, to examine the effects of farm experience on the understanding of animal products and the management of dairy farming, the interaction between students and dairy cows was investigated in groups of first‐year veterinary nursing students in 2011 and 2012 (n = 201). These students included 181 women and 20 men. Nine items about dairy cows were presented in a questionnaire. The survey was performed before and after praxis on the educational farm attached to the authors' university. After praxis on the farm, increases occurred in the number of positive responses to the items involving the price of milk, dairy farming and the taste of milk. For these items, a significant difference (P < 0.05) was found between the scores obtained before and after training. The results of the study suggested that farm experience is useful for improving the understanding of animal products and dairy farming.