The purpose of this study was to establish an optimal storage temperature and characteristic analysis after frozen-thawed dairy goat sperm. Sperm was collected at Chojeong dairy goat farm using an electric stimulator and dilluted with semen washing media. The egg yolk-triladyl frozen solution was used for the freezing of Saanen dairy goat sperm and the freezing concentration was set to 1×108sperm/ml. The frozen sperm were thawed in water bath at 37.5℃ for 45 seconds and motility was measured after preservation for 0, 30, 60, 90 and 120 min at 4℃, 17℃ and 37℃, respectively. Sperm characteristic analysis was conducted by flow cytometry. In results, sperm motility at 30, 60, 90 and 120 min after thawing was significantly higher in 17℃ than 4℃ and 37℃ (p<0.05). On the other hand, the frozen-thawed sperm motility were gradually decreased with storage periods increased (30, 60, 90 and 120 min) at 4℃, 17℃ and 37℃. Viability(42%), acrosome damage(24%), mitochondrial damage(28%) and ROS level(4%) were analyzed by flow cytometry in frozen-thawed spermatozoa in 7 male dairy goats. In summary, the motility of frozen-thawed spermatozoa in Saanen dairy goat was more efficient for storage 17℃. The average of viability 42%(30%~54%), acrosome damage 24%(16%~33%), mitochondrial damage 28%(20%~54%) and ROS level 4%(3%~6%) were arranged as standard value by 7 male dairy goats. However, more researches are needed to establish the optimal conditions or proper supplementation for sperm preservation. This study was carried out with the support of research project on feasibility study of the research & development projects for activating the hillside livestock farming and the development of goat grazing program of Rural Development Administration by Korea government (2018PJ013546).

During the freezing and thawing process, fatty acids in the plasma membrane of sperm are released, which results in a functional damage of sperm. Sperm with functional loss due to cryo-damage result in a decrease in fertility. Previous studies have shown that the addition of one of the fatty acid alpha-linolenic (ALA) with carrier proteins improves the stability of plasma membrane and reduces the damage. In this experiment, we focused on the functional aspects of the plasma membrane of sperm and experimented with motility and morphology. For preparation of ALA-carrier protein complex, 3 ng/ml ALA was mixed with 0.7 μg/ml bovine serum albumin (BSA) or 14 ng/ml methyl-β-cyclodextrin (MBCD) in distilled water. The boar semen was purchased from GUMBO Company. Boar semen was cryo-preserved in 20% egg yolk freezing extender containing ALA, BSA, MBCD, ALA+BSA, ALA+MBCD. The frozen boar sperm was thawed at 37.5 ℃ for 45 sec in water-bath. The sperm motility and morphological abnormalities were evaluated under a phase-contrast microscope at 200 × magnification and randomly counts of 200 sperm each sample. In results, motility of frozen-thawed sperm was increased in all treatment groups. In particular, there has been significant improvement in ALA+BSA and ALA+MBCD treatment groups than control (p<0.05). However, there was no significant difference in ALA, BSA and MBCD treatment groups. Morphological normalities in frozen-thawed sperm was reduced in complex treatment groups (p<0.05). However, there was no significant difference in single treatment groups. In both motility and morphology characteristics, ALA+BSA and ALA+MBCD treatment group was higher than all treatment groups. In conclusion, the addition of ALA with carrier proteins during cryopreservation has a positive effect in its functional aspect. This work was supported by the National Research Foundation of Korea (NRF) grant funded by the Korea government (Ministry of Education) (2016R1D1A1B03931746).

본 연구에서는 약용식물 추출물을 함유한 숙취해소 음료가 항산화 및 알코올 분해에 미치는 영향을 조사하기 위하여, 약용식물 추출물의 총 페놀성화합물 함량, 총 플라보노이드 함량 및 DPPH radical 소거활성 및 ALDH(aldehyde dehydrogenase) 활성을 측정하였고, 약용식물 추출물을 함유한 숙취해소 음료의 복용이 음주 후 호흡 중 알코올 농도에 미치는 영향을 조사하였다. 항산화활성은 빼빼목 추출물 에서 높은 수치(total phenolic compound 97.5.6mg/g, total flavonoid 306.5mg/g, DPPH radical 소거능 80.1%)를 나타냈으며 Acetaldehyde(ALDH)에 대한 활성은 헛개 및 울금 추출물에서 306% 및 292%로 가장 높은 활성을 나타내어, 헛개 및 울금 추출물이 ALDH 제거에 효과적인 것으로 나타났다. 헛개 및 울금 추출물이 함유된 숙취해소 음료를 이용하여, 임상실험을 실시한 결과, 시간경과에 따라 대조군에 비해 숙취해소음료 섭취군에서 알코올 농도가 감소되는 경향을 나타냈으며 흡연자에 비해 비 흡연자가, 남성에 비해 여성에게 숙취해소 음료의 섭취가 더 효과적인 것으로 확인되었다.

The oocyte undergoes various events during In vitro maturation (IVM) and subsequence development. One of the events is production of reactive oxygen species (ROS) that is a normal process of cell metabolism. But imbalances between ROS production and antioxidant systems induce oxidative stress that negatively affect to mammalian reproductive process. In vitro environments, In vitro matured oocytes have many problems, such as excessive production of ROS and imperfect cytoplasmic maturation. Therefore, In vitro matured oocytes still have lower maturation rates and developmental competence than in vivo matured oocytes. In order to improve the IVM and In vitro culture (IVC) system, antioxidants, vitamins were added to the IVM, IVC medium. Antioxidant supplementation was effective in controlling the production of ROS and it continues to be explored as a potential strategy to overcome mammalian reproductive disorders. Based on these studies, we expect that the use of antioxidants in porcine oocytes could improved maturation and development rates.

This study was carried out to investigate the characterization of iron oxide nanotubes (INTs) by anodization method and applied adsorption isotherms and kinetic models for phosphate adsorption. SEM analysis was conducted to examine the INTs surface formation. Further XRD and XPS analysis were performed to observe the crystal structure of INTs before and after phosphate adsorption. AFM analysis was conducted to determine of Fe foil surface before and after anodization. Phosphate stock solution for adsorption experiment was prepared by KH2PO4. The batch experiment was conducted using 20 ml phosphate stock solution and 40 cm3 of INTs in 50 ml conical tube. Adsorption isotherms were applied Langmuir and Freundlich models for adsorption equilibrium test of INTs. Pseudo first order and pseudo second order models were applied for interpretation of adsorption rate by reaction time. The determination coefficient (R2) values of Langmuir and Freundlich models were 0.9157 and 0.8876 respectively.

This study was conducted to evaluate effect of α-linolenic acid (ALA) and bovine serum albumin (BSA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved using freezing extender containing 3 ng/mL ALA and/or 20 μg/mL BSA. Cryopreserved boar sperms were thawed in 37°C water-bath for 45 sec to analysis. Viability, acrosome reaction, and mitochondrial intact were analyzed using flow cytometry. In results, viability of frozen-thawed boar sperm was significantly higher in only ALA+BSA supplement group than control group (p<0.05), whereas there was no difference either in ALA or BSA supplement. However, acrosome reacted sperm in both of live and all sperm population were significantly decreased in all treatment groups than control (p<0.05). Interestingly, mitochondrial intact of boar sperm was enhanced in ALA and ALA+BSA groups compared with control (p<0.05). In this study, we showed that supplementation of ALA and BSA in freezing extender enhanced the sperm viability, mitochondrial intact and decrease acrosomal membrane damage. In conclusion, our findings suggest that quality of frozen-thawed sperm in mammalians could improve by using of ALA and BSA.

The aim of this study was to evaluate effect of α-linolenic acid (ALA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved in 20% egg yolk freezing extender containing ALA (0, 3, 5, and 10 ng/mL) with 0.05% ethanol. The frozen-boar spermatozoa were thawed at 37.5°C for 45 sec in water-bath. The spermatozoa samples were evaluated the plasma membrane integrity, acrosome reaction, and mitochondrial integrity using flow cytometry. In results, population of live sperm with intact plasma membrane was significantly higher in control and 3 ng/mL ALA treatment group than ethanol group (p<0.05). In contract, dying sperms were higher in ethanol group than 3 ng/mL ALA treatment (p<0.05). Acrosomal membrane damage in all sperm population was reduced in 3 ng/mL ALA groups compared with ethanol treatment (p<0.05). However, acrosome damage in live sperm population was no significant difference among the all treatment groups. Mitochondrial integrity was not influenced by ALA treatments in both of live and all sperm population. In conclusion, this results show that supplement of ALA during the cryopreservation process could reduce the membrane damages including plasma and acrosomal membrane, whereas ALA did not influence to mitochondria in boar spermatozoa. Therefore, these results suggest that ALA can protect against the membrane damage derived cryo-stress, and cryopreservation efficiency of boar semen would be improved by use of ALA.

This study was carried out for characterization of MIO synthesized in our laboratory by co-precipitation method and applied isotherm and kinetic models for adsorption properties. XRD analysis were conducted to find crystal structure of synthesized MIO. Further SEM and XPS analysis was performed before and after phosphate adsorption, and BET analysis for surface characterization. Phosphate stock solution was prepared by KH2PO4 for characterization of phosphate adsorption, and batch experiment was conducted using 50 ml conical tube. Langmuir and Freundlich models were applied based on adsorption equilibrium test of MIO by initial phosphate solution. Pseudo first order and pseudo second order models were applied for interpretation of kinetic model by temperature. Surface area and pore size of MIO were found 89.6 m2/g and 16 nm respectively. And, the determination coefficient (R2) value of Langmuir model was 0.9779, which was comparatively higher than that of Freundlich isotherm model 0.9340.

본 연구는 계방산 가문비나무 및 전나무 임분의 산림식생유형분류와 정량적 분석을 위하여 Z-M 식물사회학적 방법으로 식생구조의 유형분류를 실시한 결과, 군락단위에서 가문비나무군락, 전나무군락으로 분류되었으며 가문비나무군락은 군단위에서 흰인가목군과 부게꽃나무군으로 세분되었으며, 전나무군락은 복장나무군과 생강나무군으로 세분되었다. 분류된 식생단위를 기준으로 중요치를 산출한 결과, 가문비나무는 식생단위 1과 2의 교목층에서 각각 30.73%, 20.25%로 비교적 높게 나타나 당분간 가문비나무의 우점이 계속될 것이라 판단되었다. 또한 종다양도를 분석한 결과 식생단위 4의 종다양도 지수는 0.6976으로 가장 낮았으며, 식생단위 2의 종다양도 지수는 1.1256으로 가장 높게 나타났다. 군락간의 유사도는 식생단위 1과 4, 2와 4가 각각 0.2880과 0.3626으로 낮게 나타났으며 식생단위 1과 2, 3과 4의 군락유사성은 각각 0.5411과 0.5041로 나타나, 군락 간 구성종의 차이가 크지 않은 군락으로 판단되었다. 종간연관에 대한 Chi-square matrix와 성좌표를 각각 분석한 결과, 크게 2개의 유형으로 나뉘어졌는데 Ⅰ유형의 식물 종들은 대부분 식물사회학적으로 분류된 가문비나무군락에서 주로 출현하는 식별종과 표징종이었으며, Ⅱ유형의 식물종은 전나무군락에서 주로 출현하는 식물종으로, 비교적 습한 곳에 나타나는 종들로 나뉘어졌다. 이러한 결과는 각 수종들이 선호하는 생육환경이 비슷한 종들끼리는 정의 상관이 인정되고, 선호하는 환경이 다른 종들끼리는 부의 상관을 보이는 것으로 판단된다.

The aim of this study was to determine the effect of additional alpha-linolenic acid (ALA) supplementation during in vitro maturation (IVM) and culture (IVC) on nucleic maturation and embryo development of pigs. Cumulus-oocyte complexes (COCs) were incubated in IVM medium containing different concentration of ALA (25, 50 and 100 μM) for 44 h. After in vitro maturation, nuclear maturation of oocytes were evaluated by aceto-orcein stain. Mature oocytes with 50 μM ALA were fertilized and cultured in IVC medium with ALA (25, 50 and 100 μM) during early-embryogenesis (48 hours after fertilization). Then, embryos were cultured with 25 μM ALA during early embryogenesis and/or late embryogenesis (120 hours after early-embryogenesis). In results, oocyte maturation were significantly increased by 50 μM ALA treatment groups compared with control groups (p<0.05). Treatment of 25 μM ALA during early-embryogenesis enhanced cleavage rate of embryo compared with other groups (p<0.05), whereas formation and total cell number of blastocyst had no significant difference. Similarly, cleavage rate of embryos were increased by 25 μM ALA supplement during early- or late-embryogenesis than ALA treatment both stage of embryogenesis (p<0.05), but did not influence to blastocyst formation. Interestingly, total cell number of blastocyst were enhanced in ALA treatment group during early-embryogenesis. These findings indicated that ALA supplement enhance the nuclear maturation of oocyte and embryo development, however, excessive ALA could negatively influence. Therefore, we suggest that ALA is used for improvement of in vitro production of mammalian embryo and further study regarding with functional mechanism of ALA is needed.

The efficiency of regeneration of callus and explants from leaf and stem disks of Epimedium koreanum was examined on the MS media containing 2,4-D, NAA, Kinetin, BA and TDZ. Calli were formed on the 2mg/l 2,4-D media at the rate of 32% from leaf discs and 52% from stems. No callus was produced on the media which are containing BA or TDZ alone. The combination of 2,4-D and BA showed the effect on the formation of callus. The combination of 2mg/l 2,4-D and 0.lmg/l BA in the MS media had produced the highest percentage of callus formation, 50% from leaf discs and 40% from stems, respectively. The combination of 2mg/l 2,4-D and 1mg/1 BA in the MS media had affected the formation of callus in the rate of 40% from leaf discs and 25% from stems. The combined plant growth regulators of 2,4-D and BA increased the formation of calli from leaf discs, but single treatment of 2,4-D showed the highest callus formation from stems. Multiple shoots from leaf discs were formed on the media containing NAA, BA, kinetin, and TDZ. The highest number of multiple shoots were obtained 0.1mg/l NAA combined with 1mg/l kinetin. As a result, leaf discs or stems can be used for the mass propagation of Epimedium koreanum, but stem elongation of shoots from calli was not easy.