Cryopreservation of canine spermatozoa affords potential exchange of genetic material, and thus may lead to improvement in the breeding management. However, canine spermatozoa undergo many damages such as, cold shock, ice crystal formation, oxidative stress during cryopreservation. In this study used the CASA for investigating the effect of various trehalose concentrations and thawing temperatures on the sperm viability. In addition, the efficacy of the most optimal of the tested cryopreservation protocols in this study was verified by AI as the in vivo test. Also, this study evaluates the variation of frozen- thawed canine spermatozoa during different incubation condition. The addition of trehalose 25 mM was optimal concentration and frozen-thawed semen quality was significantly higher better than control (Glucose) and other concentration groups. In effect of thawing temperature on frozen-thawed sperm movement and intact acrosome evaluations, which result enhance the sperm motility and movement value depending on increase temperature condition at 36, 54 and 72℃. Also, in the effect of different incubation condition on frozen-thawed sperm after thawing at 36℃ for 60 sec, that the results trehalose 25 mM was significantly better (p<0.05) than glucose in general as well as, the post-thawed sperm motility and intact acrosome was reduced depending on increase the incubation time. Especially, incubation at 4 to 8 hour was rapidly depreciation of movement value and the rate of intact acrosome was dropped similar tendency. Thus, incubation 17℃ was better than other incubation groups on sperm motility and acrosome integrity. For the in vivo evaluate of spermatozoa survival and is the most definitive test of sperm function, we performed artificial insemination in estrous bitch. The semen was prepared for intrauterine insemination using the 25 mM trehalose freezing extender and thawing at 36℃, and 2 bitches were inseminated with 1×106 motile spermatozoa by surgical method. The results of AI, the pregnancy rates, mean litter size and oocyte fertilization rate were 16.6% (1/6), and 50% (2/4), respectively. In conclusion, based on the results of these experiments, the effect of addition of trehalose on extender improves the movement and intact acrosome of frozen-thawed semen. In particular, trehalose 25 mM groups was higher than other different concentration group on movement value and acrosome integrity of frozen-thawed sperm. Also, through incubation condition, this study identify the optimal incubation temperature after thawing was 17℃. Furthermore, the information will be contributed to develop the canine ART including AI, IVF and canine ICSI. * This research was supported by iPET (Grants 110056-3), Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.

In this work, the silver nanoparticles have been synthesized by electrical explosion of wire in three liquid mediums: deionized water (DIW), polyvinylpyrrolidone (PVP) and sodium dodecyl benzene sulfonate (SDBS) solutions. Absorption in the UV-visible region of these suspensions was measured in the range of 300-800 nm. A surface plasmon peak was observed at ~400 nm in all suspensions in measured wavelength range. Particle size was analyzed by transmission electron microscope. It showed that the particles had nearly spherical shape in all samples. The average particle sizes prepared in DIW, PVP and SDBS solution were 37, 31 and 27 nm, respectively. Stability of the suspensions was estimated by multiple light scattering method. The presence of PVP and SDBS surfactants in the exploding medium resulted in enhanced stability of the silver suspensions.

Cell transplantation therapy using adult stem cells has recently been identified as a potential treatment for spinal cord injury (SCI). But, recovery after traumatic SCI is very limited. As dogs are physiologically much more similar to human compared with other traditional mammalian models in disease presentation and clinical responses, a number of researches demonstrated canis familiaris is a suitable model for human diseases. This study investigated the effect of transplantation of canine Mesenchymal Stem Cells (cMSC) and neural-induced cMSC (nMSC) to understand how these cells improve neurological function in canine SCI model. The differentiation of cMSC into neural precursor cells was induced in dulbecco’s modified eagle’s medium supplemented with N2-supplement, dibutyryl cyclic adenosine monophosphate, and butylated hydroxyanisole. SCI was induced between T1 and T2 by surgical hemi-section in adult dogs, and then assigned to two groups according to the applied cell types (cMSC vs nMSC). Pelleted cMSC or nMSC were transplanted directly into the injured site after SCI, respectively. Analysis of motor function after transplantation was evaluated by modified Olby score. Magnetic resonance imaging (MRI), histological and immunohistichemical analysis were also performed. Functional recovery in group of cMSC was increasing gradually after transplantation and was higher than nMSC. In MRI, we could not confirm any difference between the cMSC and nMSC experimental groups. Immunohistochemically, beta3-tubuline and nestin were observed in injury site of two experimental groups with the expression level close to non-injured groups. Transplantation of mesenchymal stem cells could promote neuronal reconstruction and repair motor function in SCI. These showed mesenchymal stem cells could be a great candidate as a therapeutic tools in degeneration disease, and dogs could be used to explore human regenerative medicine as a promising animal model. This research was supported by iPET (Grants 110056032CG000), Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.

Bacterial contamination reduces the semen quality, semen preservation, and cause of disease spread as well. Sperm fertility is essential factor of reproductive performance in swine. Sperm fertility is affected by semen quality such as sperm motility, abnormality, morphology, and rate of bacterial contamination. This study was conducted to determine the relationship between elapsed time after semen preservation on the changes of bacteria and semen quality. Semen was diluted with BTS extender without antibiotic for 7 days and sperm parameter and fertility were measured. Sperm motility was measured by CASA and total bacteria number was counted after 22 24 hr incubation from counting agar plate in which sperm dilute to 10 106 in 0.9% saline solution and inoculate to agar. Acrosomal integrity was measured by Chlortetracycline (CTC) staining. CTC patterns were uniform fluorescence over the whole head (pattern A), characteristic of uncapacitated acrosome-intact spermatozoa; fluorescence-free band in the post-acrosomal region (pattern B), characteristic of capacitated acrosome-intact spermatozoa; and almost no fluorescence over the whole head except for a thin band in the equatorial segment (pattern C), characteristic of acrosome reacted spermatozoa. Total number of bacteria was significantly increased (p<0.0001) 3 days after preservation. Sperm motility, viability, and morphological abnormality on elapsed time after preservation were lower from 5 (77.24±6.47, p<0.001) and 7 days (77.24±6.47, p< 0.001) after preservation compared to 1 (15.71±7.18) and 3 days(18.39±7.22) after preservation, respectively. Sperm viability was significantly lower (53.25±35.03, p<0.0001) at 7 days after preservation. Mohological abnormality of sperm was lower (p<0.001) at 1 (15.71±7.18) and 3 (18.39±7.22) days compared to (5 21.84±7.91) and 7 (22.59± 9.93) days after preservation. Acrosomal integrity and capacitation rate (pattern A) were significantly lower (p<0.001) from 5 days after preservation.

Acteoside (verbascoside) is a typical phenylethanoid glycoside, extracted from various plants. It has various biological functions such as anti-oxidant, anti-inflammation, and anti-hypertension. Specially, it was powerful anti-oxidants either by direct scavenging of reactive oxygen and nitrogen species, or by acting as chain-breaking peroxyl radical scavengers. We examined the role of acteoside in IVM medium on the morphological progress of meiosis, developmental competence, and ROS in porcine oocytes. And we investigated effect of acteoside on the oocytes condition represented by cytoplasmic maturation by homogeneous distribution and formation of cytoplasmic organelles and regulation of apoptosis-related genes. The selected COCs were cultured in TCM-199 with various concentration of acteoside: 0 (control), 10, 30, and 50 μM. After 22 h of maturation with hormones, the oocytes were washed twice in a fresh maturation medium before being cultured in hormone-free medium for additional 22 h. The oocytes maturation rates of supplemented with acteoside were no significantly different compared with control group (71.13, 75.96, 72.95 and 73.68%, respectively). Level of ROS was significantly decreased in acteoside treated group. Furthermore, the parthenogenetic blastocyst rate was significantly improved in 10 μM acteoside treated group compared with control group (40.03 vs. 22.95%). During IVM, 10 μM acteoside treated oocytes showed that the mitochondria and lipid droplet were smaller and homogeneous distribution in cytoplasm compare with non-treated control oocytes. And reverse transcription polymerase chain reaction (RT-PCR) witarthenogenetic blstocysts revealed that acteoside increased the anti-apoptoticgenes, otherwise reibued pro-apoptotic genes. In conclusion, our results represents that addition of acteoside to the IVM medium has a beneficial effect in physiology of porcine oocytes such as viability and activation, providing a improved method for porcine oocytes in vitro.

The green peach aphid(Myzus persicae) and sweet-potato whitefly(Bemisia tabaci) biotype Q are serious pests in hot pepper and cucumber. To optimize the formulation type of Clothianidin 1.8% granule, coated and extruded granule formulation types were tested against green peach aphid in hot pepper and sweetpotato whitefly in cucumber. Clothianidin 1.8% granules were incorporated in soil before transplant of hot pepper and cucumber. At recommend dose(540g ai/ha), the efficacy of Clothianidin 1.8% coated and extruded granules were 83.3% and 94.4% and continued until 50 days and 70 days against green peach aphid, respectively. Against sweet-potato whitefly, the efficacy of Clothianidin 1.8% coated and extruded granules were 65.7% and 96.3% at 32 days in cucumber, respectively. Clothianidin 1.8% extruded granule was safe on crops (hot pepper, cucumber, melon, cabbage etc.) at 540g ai/ha (recommend dose) and 1080g ai/ha. The results suggested that Clothianidin 1.8% extruded granule is promising as a best insecticide against green peach aphid and sweet-potato whitefly, because of the high efficacy and low phytotoxicity.

The Cu nanofluid in ethylene glycol was prepared by electrical explosion of wire, a novel one-step method. The X-ray diffraction, field emission scanning electron microscope and transmission electron microscope were used to study the properties of Cu nanoparticles. The results showed that the nanoparticles were consisted of pure face-centered cubic structure and near spherical shape with average grain size of 65 nm. Ultraviolet-visible spectroscopy (UV-Vis) confirmed Cu nanoparticles with a single absorbance peak of Cu surface plasmon resonance band at 600 nm. The nanofluid was found to be stable due to high positive zeta potential value, +51 mV. The backscattering level of nanofluid in static stationary was decreased about 2% for 5 days. The thermal conductivity measurement showed that Cu-ethylene glycol nanofluid with low concentration of nanoparticles had higher thermal conductivity than based fluid. The enhancement of thermal conductivity of nanofluid at a volume fraction of 0.1% was approximately 5.2%.

In present work, amorphous TiCuNi powders were fabricated by mechanical alloying process. Amorphization and crystallization behaviors of the TiCuNi powders during high-energy ball milling and subsequent microstructure changes were studied by X-ray diffraction and transmission electron microscope. TEM samples were prepared by the focused ion beam technique. The morphology of powders prepared with different milling times was observed by field-emission scanning electron microscope and optical microscope. The powders developed a fine, layered, homogeneous structure with milling times. The crystallization behavior showed that glass transition, , onset crystallization, , and super cooled liquid range were 628, 755 and 127K, respectively. The as-prepared amorphous TiCuNi powders were consolidated by spark plasma sintering process. Full densified TiCuNi samples were successfully produced by the spark plasma sintering process. Crystallization of the MA powders happened during sintering at 733K.

In this work, effect of various process-control agents (PCAs) on the mechanical alloying of amorphous alloy of has been investigated. The dependence of the particle shape, size and crystallization behavior of the amorphous alloy powders on the type of PCAs and their concentrations was investigated by using X-ray diffraction, field-emission scanning electron microscopy and differential scanning calorimetry. It was found that the additive of toluene could affect positively the amorphization and thermally induced crystallization processes, as well as the size refinement, morphology and particle-size distribution of as-milled powders in comparison with alloy obtained without PCA.

A method of estimating the lower bound of coronal magnetic field strength in the neighborhood of an ejecting plasmoid is presented. Based on the assumption that the plasma ejecta is within a magnetic island, an analytical expression for the force acting on the ejecta is derived. The method is applied to a limb coronal mass ejection event, and a lower bound of the magnetic field strength just below the CME core is estimated. The method is expected to provide useful information on the strength of reconnecting magnetic field if applied to X-ray plasma ejecta.

TiCuNiAl quaternary amorphous alloy was prepared by high-energy ball milling process. A complete amorphization was confirmed for the composition of TiCuNiAl after milling for 30hrs. Differential scanning calorimetry showed a large super-cooled liquid region (T = T T, T and T: glass transition and crystallization onset temperatures, respectively) of 80 K. Prepared amorphous powders of TiCuNiAl were consolidated by spark-plasma sintering. Densification behavior and microstructure changes were investigated. Samples sintered at higher temperature of 713 K had a nearly full density. With increasing the sintering temperature, the compressive strength increased to fracture strength of 756 MPa in the case of sintering at 733 K, which showed a 'transparticle' fracture. The samples sintered at above 693 K showed the elongation maximum above 2%.

In this research, it is studied whether an abandoned tunnel can be used for a warehouse to keep rice. Today's agricultural industry especially rice farming in our nation is put in the bad condition such as not sufficient man power, not enough capital and so on, and moreover dose not have competitive power as well-known against products from other nations through FTA. But there are many ways to increase competitive power we can guess. Development of technique to keep quality of the agricultural product by controlled atmosphere is one of the reasonable ways. From these backgrounds, it has been studied whether an abandoned tunnel is suitable for rice as storage place or not. In the result, the inner condition of the abandoned tunnel was measured that temperature is 15℃ in average and relative humidity is 55% in average and it is therefore found that to use an abandoned tunnel as a place to storage rice is reasonable in the consideration of thermal and cost-saving aspect.