The objective of this study was to compare the effect of semen extenders on the sperm motility, viability, acrosome integrity and functional integrity of plasma membrane (HOST: hypo-osmotic swelling test) during liquid preservation of Korean Native boar semen. In this experiment, semen was diluted in Androhep plus, Beltsville Thawing Solution (BTS), ModenaTM, Seminark and Vitasem LD. Sperm-rich fractions were collected from three Korean Native boars and sub-samples were diluted (30×106 spermatozoa/ml) in different semen extenders. Semen samples were stored at 17℃ for 96 hours. On everyday (0, 24, 48, 72, 96 h) after storage, the sperm characteristics relevant for fertility, such as sperm motility, viability, acrosome integrity and HOST positive were evaluated. The motility of spermatozoa stored in different extenders was no significantly different among other extenders (P>0.05). Also, no difference was observed among samples processed with different extenders in the percentage of sperm viability, acrosome integrity and HOST positive. All extenders maintained a high percentage (70%) of sperm motility, viability and acrosome integrity through 96 h of storage. The result of this study show that there was no significant differences among extenders in their capacity to preserve motility, viability and membrane integrity of spermatozoa from normal, fertile Korean Native boars for 96 h of liquid preservation at 17℃.

이 연구에서는 참깨 과립종자의 저장 시 활력손실 정도를 파악하여 적정한 저장 방법을 규명하기 위하여 저장고의 온도 를 4, 15, 25oC로 하고 상대습도를 40, 60, 80%로 달리하여 1개월 간격으로 4개월간 저장기간별로 종자의 수분함량 및 활력유지 정도를 비교하였다. 1. 무처리 종자의 수분함량이 가장 높게 나타났으며 점토 단 독처리이거나 점토성분이 함유된 과립에서는 수분함량이 많았 고, 활석처리에서는 종자수분함량이 가장 낮게 나타났다. 2. 상대습도가 80%일 때는 4oC에서 15oC와 25oC일 때보다 종자수분함량이 높게 나타났다. 3개월째는 종자수분함량이 가 장 높았으며 이때 발아율은 가장 낮았다. 3. 종자활력검사결과를 종합해보면 점토+질석의 조합으로 과 립화할 경우 15oC이하이면서 상대습도 80%이하로 저장할 경 우 50%이상의 활력을 3개월 이상 유지할 수 있었다. 그러나 25oC로 저장할 경우 2개월까지만 가능하였다. 점토 +피트모 스 +활석의 조합으로 과립화할 경우 15oC이하이면서 상대습 도 80%이하의 조건에서 2개월까지 50% 이상의 활력유지가 가능하였지만, 25oC이하와 상대습도 80%이하의 조건에서는 1 개월까지만 저장 가능하였다. 활석 단독으로 과립화할 경우 25oC이하 및 상대습도 80%이하의 조건에서 1개월까지만 저장 가능하였다. 따라서 점토+질석의 조합으로 과립화하였을 때 종 자의 저장수명이 가장 긴 것으로 나타났다.

This study was designed to evaluate the effect of L-cysteine on sperm characteristics and oocyte cleavage in vitro in Korean native cattle. For this study, the freezing of diluted semen were added with Triladyl containing 20% eggyolk and/or 0, 5, 10 and 20 mM L-cysteine before cryopreservation. The viability in frozen-thawed sperm were estimated by SYBR14/PI double stain, acrosome damage with FITC-PNA, mitochondria intact with Rhodamin123 and hydrogen peroxide(H2O2) level with carboxy-DCFDA by flow-cytometry. The developmental capacity was also assessed with cleavage rates in oocytes fertilized in vitro by frozen-thawed sperm. In results, the sperm viability was significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). In addition, acrosome damage was significantly decreased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). The mitochondria intact was also significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). On the other hand, the cleavage rates were significantly increased in 0 mM, 5 mM and 10 mM groups than 20 mM concentration of L-cysteine (p<0.05). The oocyte degeneration of oocytes were significantly decreased in 0 mM, 5 mM and 10 mM groups than in 20 mM L-cysteine group (P<0.05). However, there are no significantly differences among the L-cysteine treatment groups. We suggest that concentration of 10 mM L-cysteine have beneficial impact for sperm cryopreserved in Korean native cattle. This result also could be recommended for artificial insemination program if supported by an improvement in the fertility results and required further study.

여우는 한국에서 1980년대 이후로 절멸된 것으로 취급하고 있으며, 현재는 환경부에서 멸종위기 1급 동물로 지정되어 소백산국립공원에서 복원사업이 이루어지고 있다. 본 연구에서는 여우복원사업에 필요한 적정 모델링과 방사 전략을 수립하기 위하여, VORTEX 프로그램을 이용한 개체군 생존력 분석(Population viability analysis: PVA)을 수행하였다. 초기 개체군 0에서 매년 10개체(암5, 수5)를 향후 10년간 지속적으로 방사할 경우, 개체군의 이입으로 50년간 개체군 성장률은 0.018±0.204였으며, 절멸확률은 0.354로 나타났다. 최초 방사 후 16년차에 116.34로 최대 개체군 크기를 보였으나, 17년차부터 매년 1.22의 감소율을 나타내었다. 방사된 여우의 지속적인 성장과 절멸을 방지하기 위해서는 최초 방사 17년 이후 추가적인 방사가 필요하며, 장기적인 여우 존속을 위해서는 매년 12개체 이상을 방사하는 것이 필요한 것으로 분석되었다. 반면 연간 6개체 이하일 경우 50년 후의 절멸 확률은 80% 이상으로 나타났으며, 최소한의 개체군 성장을 위해서는 8개체 이상의 추가 방사가 필요한 것으로 나타났다. 설정된 조건값에서 매년 10개체씩 암수의 성비를 변화하여 6:4 로 10년간 방사할 경우 1:1 비율보다 안정적인 개체군 성장률을 보였으나, 암컷의 비율이 7:3이상으로 높아질 경우 부정적인 결과를 보였다. 재난요인으로 설정된 road-kill과 밀렵은 여우복원사업의 성패에 중요한 요인으로 확인되었다. 초기 적용값을 기준으로 각각의 요인이 3% 감소할 경우에는 절멸 확률은 30%로 낮아졌으며, 3% 이상 증가할 경우 50년 후의 절멸 확률은 약 90%에 이르는 것으로 분석되었다.

In this study, we conduct the economical analysis about the floating tracking PV generation structure manufactured by steel, aluminum, and GFRP (glass fiber reinforced polymeric plastic) structural member. The structural safety of floating PV generation structure has been proved through numerous previous researches. Moreover, the generating efficiency of tracking PV generation system can be more larger than immobile system. In this study, structural analysis using the FEM method has been performed to establish the safety of the floating tracking PV generation structure and commercial viability evaluation has been performed through the cost of materials.

Melanism is one of the most marked phenotypic variations that naturally occur in a wide range of organisms. In this study, we established a homozygous melanism mutant strain with black pupae spontaneously occurring within a wild-type population of Spodoptera exigua. The S.exigua pupal melanic strain showed several viability advantages. The melanism is associated with faster development, heavier pupa weight and higher fecundity after eating seven different host plants. Female adults of the two strains both tend to attract xenogeneic male adults for mating, and the fecundity of the melanic strain is significantly higher than the wild-type strain. However, the melanism is associated with slower mean flight seed, shorter mean flight duration and distance. The melanic strain adults have weaker flight capacity in different ages. The viability advantages above will contribute in more generations per year, increasing population and more serious damage. Meanwhile, based on the mating competition results, the melanic strain will be able to interfere with the reproduction of wide-type strain and replace it. However, decreased flight capacity will influence the long-distance migration ability of the melanic strain and limit its range of damage

This study investigated viability variation of airborne bacteria in indoor environments. The survival in air as a temporal function of bioaerosol viability was reported for Escherichia coli (KCCM 12119, ATCC 11775). Bacteria suspended in distilled water were aerosolized and entered the vertical duct oriented downward. After measurement of number concentration and colony forming unit (CFU) of the bacteria at different locations of the duct, the viability function was calculated. It was found that the bacteria viability(%) decreased with time after aerosolization, 28.454e^-0.132x (x:time, min). This study demonstrated the potential application of viability function of airborne bacteria to studies of exposure assessment and infection risk analysis.

LIVE/DEAD® BacLight™ and alamarBlue® are fluorescent materials used for the enumeration of live and dead bacteria. LIVE/DEAD® BacLight™ is generally used for confocal microscopy applications to differentiate live from dead bacteria in a biofilm or planktonic state. AlamarBlue® has also been used widely to assay live and dead bacteria in a planktonic state. Whilst these materials are successfully utilized in experiments to discriminate live from dead bacteria for several species of bacteria, the application of these techniques to oral bacteria is limited to the use of LIVE/DEAD® BacLight™ in biofilm studies. In our present study, we assessed whether these two methods could enumerate live and dead oral bacterial species in a planktonic state. We tested the reagents on Streptococcus mutans, Streptococcus sobrinus, Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans and Enterococcus faecalis and found that only LIVE/DEAD® BacLight™ could differentiate live from dead cells for all five of these oral strains. AlamarBlue® was not effective in this regard for P. gingivalis or A. actinomycetemcomitans. In addition, the differentiation of live and dead bacterial cells by alamarBlue® could not be performed for concentrations lower than 2 × 106 cells/ml. Our data thus indicate that LIVE/DEAD® BacLight™ is a more effective reagent for this analysis.

The Oriental White Stork (Ciconia boyciana) is a representative wetland species distributed across East Asia. The species has been declined to face the threat of species extinctions with estimation of at about 3000 individuals. In order to re-introduce the endangered storks in the field, we developed a baseline model using the program VORTEX, performed sensitivity test, and finally suggested an ideal model based on results of the sensitivity test. The baseline model predicted 12.5% extinction probability with mean time to first extinction of 82.0 year. Sensitivity test revealed that two demographic variables (first-year mortality and percent of adult female breeding) had the greatest impacts on population persistence. Thus, corrected model improved the population persistence, where the extinction probability decreased to 1.0% in 100 years by changing values of two variables within a range of applicable to the population. Our models for stork re-introduction suggest this population will be stable by improving first-year mortality and adult female fecundity.

This study was designed to determine whether low-density lipoporoteins (LDL) extracted from egg yolk in extender improve the function of Korean Jeju Black Bull semen. The semen was cryopreserved with 5% ethylene glycol (EG) or 7% glycerol (G) extenders containing 10% egg yolk (EY), 4% LDL and 5% EY or 8% LDL. Frozen-thawed sperm were evaluated sperm motility, viability, membrane integrity and acrosome integrity. Post-thawed sperm motility has been significantly higher (p<0.05) in 4% LDL + 5% EY (; EG and ; 7% G) than 8% LDL (; EG and ;G). Treatment of 4% LDL + 5% EY-EG () has been significantly improved sperm viability compared to other treatments except 10% EY - EG. Moreover, in membrane integrity, swollen sperm ratio has been only significantly increased (p<0.05) in 4% LDL + 5% EY - EG () among all treatments. In assess to detect acrosome integrity, especially, AR pattern ratio has been significantly decreased (p<0.05) in 4% LDL + 5% EY - EG among all treatments. In sperm viability as time passes, between 4% LDL + 5% EY and 10% EY, there was no significant difference, but 8% LDL was significantly decreased sperm viability in EG (1 and 2 hrs) and G (30 min, 1, 2, 5 and 12 hrs) extender. However, there were no significant differences among all treatments except 8% LDL-G in sperm membrane integrity. 8% LDL-G has been significantly decreased swollen sperm ratio at 5 hrs after thawed. It is concluded from these results that 4% LDL + 5% EY to the freezing extender showed more positive effect on the frozen-thawed spermatozoa in Korean Jeju Black bull.

For reconstituting genetic resource(Korean Native Chicken: KNC) with grem-line chimeric chicken made with cryopreserved biastdermal cells, the experiments were carried out to optimize cryopreservating conditions. Stage X biastdemal cells were collected from KNC embryos and dissociated. Cells were susupended in medium containing cyopretectant and fetal bovine serum(FBS), and distributed into plastic ampules. Cell susupensions were seeded to induce ice formation at — 7℃ to — 35℃ at in the experiments, the effect of modification of dissociation way, concentration of FBS and cell density on the vaibility of frezen-thawed cells were investigated by trypan blue exclusion. Then change the way of cell dissociation from pipetting to short time vortexing, viability of frozen- thawed cell tended to be increaced from 29 % to 52 %. Increase concentraition of FBS in frozen medium from 20 % to 80 % made viability of thawed cell from 28 % to 35 %. The viability of thawed cells were 33.9% frozen at 2 embryos/ 0.5 ml, and 43.6 % frozen at 20embryos/0.5 ml. Furthermore, combination of three modifications make big improvement. The viability of frozen-thawed cell was 60 % for combinated method, and 41 % for general method. This result means the advance to practical cryoreservation of blastdermal cell of the KNC(Ogolgye breed).

For reconstituting genetic resource(Korean Native Chicken: KNC) with grem-line chimeric chicken made with cryopreserved biastdermal cells, the experiments were carried out to optimize cryopreservating conditions. Stage X biastdemal cells were collected from KNC embryos and dissociated. Cells were susupended in medium containing cyopretectant and fetal bovine serum(FBS), and distributed into plastic ampules. Cell susupensions were seeded to induce ice formation at —7 ℃ to —35 ℃ at in the experiments, the effect of modification of dissociation way, concentration of FBS and cell density on the vaibility of frezen-thawed cells were investigated by trypan blue exclusion. Then change the way of cell dissociation from pipetting to short time vortexing, viability of frozen-thawed cell tended to be increaced from 29 % to 52 %. Increase concentraition of FBS in frozen medium from 20 % to 80 % made viability of thawed cell from 28 % to 35 %. The viability of thawed cells were 33.9% frozen at 2 embryos/ 0.5ml, and 43.6 % frozen at 20 embryos/0.5 ml. Furthermore, combination of three modifications make big improvement. The viability of frozen-thawed cell was 60 % for combinated method, and 41 % for general method. This result means the advance to practical cryoreservation of blastdermal cell of the KNC(Ogolgye breed).

The objective of this study was to investigate the motility and kinematics of boar sperm that while stored at 4C. The samples of fresh boar semen were place into an extender, Androhep, and stored at . In three of these samples, cryoprotectants were added. The sperm's motilities and kinematics were evaluated by using microscope () and the viability status was evaluated by using with eosin staining method. The 5 sample groups are; Goup A:Androhep (extender), stored at . Group B:Androhep (extender), stored at . Group C:Androhep (extender), + 3% glycerol (cryoprotectant), stored at . Group D:Androhep (extender), + 3% DMSO (cryoprotectant), stored at . Group E:Androhep (extender), + 3% ethylene glycol (cryoprotectant), stored at . In group A, the sperm's motility was reduced. On day one the sperm's motility was () and day 5 the motility was (). In group B, C and D the sperm's motility were reduced to 0 on day 5. In group E the sperm's percentage of motility decreased. On day one the sperm's motility was () and day 5 the motility was (). When comparing cryoprotectant in samples of boar sperm there is a slight improvement in the results when the use of Androhep Lite (extender), + 3% ethylene glycol (cryoprotectant), stored at are used. Based on these results, ethylene glycol can protect sperm from heat shock at , but not satisfactory level. However, it showed the possibilities of liquid semen preservation at by using cryoprotectant.

The objective of this study is to estimate the effect of adding TES to LEY and FGE freezing extender for the sperm viability, acrosomal morphology and DNA fragmentation from miniature pig sperm, we evaluated sperm characteristics in TFGE, TLE and LEY with various thawing condition ( for 20 sec, 45 sec and for 5 sec, respectively), and in different concentration of glycerol at 1%, 1.5%, 3%. The sperm viability and normal acrosome intact(NAI) in TFGE (Viability : , NAI : ), TLE (, ) extender significantly(p<0.05) increased than that in LEY (, ) extender thawed at for 5 sec. According to the results from glycerol concentration, the viability and NAI of miniature pig sperm in 1.5% glycerol TLE (, ) was highest among the experimental groups. In accordance with this, DNA fragmentation rates was the lowest in TLE () while that in LEY () is the highest. Therefore, these results suggest that TLE extender method for freezing- thawing of miniature pig sperm increased the viability after thawing.

The objective of this study was to evaluate the effect of Tea-N-tris medium on the sperm viability and acrosomal morphology for semen of normal and miniaure pig by type of freezing extender. The present study was to determine of Tea-N-tris (0.02 g/ml) effect to freezing extender LEY(Lactose 11% + Egg yolk 20%) and FGE(Fructose 3%+Glucose 7%+Egg yolk 20%) for the spermatozoa viability, acrosomal morphology and DNA fragmented analysis from normal and miniature pig semen, were evaluated freezing extender TFGE, TLE and LEY during thawing at 37℃ for 45 sec and 75℃ for 5 sec, respectively. Interestingly, the result that sperm after addition of Tea-N-tris extender(TFGE, TLE) during 15 4℃ cooling significantly increased the viability(p<0.05), as compared to than of sperm cooling in LEY extender, but lower the percentage of AR(acrosome reacted spermatozoa) pattern than LEY extender. The sperm viability and AR pattern after freezing was appeared like sperm cooling method pattern. And treatment spermatozoa during freezing after addition of Tea-N-tris extender significantly (p<0.05) increased the viability and AR to miniature pig sperm, than normal pig sperm, but most highly percentage of viability and AR pattern to normal pig sperm during freezing in LEY extender. Chromosomal DNA fragmentation increased from LEY extender to sperm of normal and miniature pig, but decreased from the Tea-N-tris extender. Therefore, suggest that Tea-N-tris freezing extender method for freezing of miniature pig sperm is required for increasing viability. This Study was supported by Technology Development Program for Agriculture and Forestry, Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.

In mammals, the meiosis division in testes produces equal numbers of two different types of gametes: X chromosome-bearing sperm (X-spermatozoa) and Y chromosomebearing sperm (Y-spermatozoa), which have equal potential to fertilize the oocytes. Therefore, the expected 1: 1 sex ratio is observed. However, under some conditions like endocrine disruptors (EDs) exposure the sex ratio is deviated than the expected with more males or more females. And recently many hypotheses have been postulated to explain the mechanism of sex ratio deviation; however none of them introduced a proven experimental explanation. To solve this enigma, we hypothesized that the differences between X- and Y-spermatozoa survivability under specific conditions due to differences in their chromosome contents are the key leading to the sex ratio alteration. To examine our hypothesis, we combined two techniques; first, hypo-osmotic swelling (HOS) test that was applied to test viability of spermatozoa and second, fluorescence in situ hybridization that was applied on HOS-treated spermatozoa to define sex chromosome composition. In the present study, human spermatozoa were incubated with a group of EDs represent a widespread chemicals in the environment bisphenol A (BPA, 100 μM), nonylphenol (NP, 10 μg/ml), 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD, 2.5 μg/ml), genistein (Gen, 100 μM), and the following pesticides, dibromochloropropane (DBCP, 10 μg/ml), atrazine (Atraz, 500 μM), and diazinone (Diaz, 500 μM) for 6 hr at 37℃ in 5% CO2. Then, the viability of spermatozoa and their sex chromosome contents were evaluated simultaneously. Among seven chemicals studied only four chemicals (Atraz, DBCP, TCDD, and Diaz) significantly decreased Y-sperm viability when compared to those of X-spermatozoa in the same treatment group and viability of Y-spermatozoa when compared to those in the negative and positive (DMSO) control groups (p<0.05). Also, in these four treatment groups the sex ratio of live sperm population was significantly lowered compared to the control groups (p<0.05). Otherwise, Gen, BPA, and NP did not show any significant effect on viability of Yspermatozoa or decreasing sex ratio in live sperm population as compared to the control groups. It has been proven that TCDD, DBCP, and the pesticides decrease the sex ratio, but the same effect was not observed in case of Gen, BPA, and NP. From the present findings, there is no doubt that the EDs may alter sex ratio via decreasing Y-spermatozoa viability.

The objective of this study was to examine effect of ethylene glycol for semen cryopreservation in Korean Jeju Black Bull. The semen was cryopreserved with extenders containing cryoprotectants (7% glycerol and 3%, 5%, 7% ethylene glycol) and packed to 0.5 ml straws. The semen straws were located above 3 cm of liquid nitrogen for 5 min, 5 cm for 10 min and 8 cm for 10 min. And then frozen straw was plunged into LN2. Post-thawed sperm motility, viability and membrane integrity were significantly higher in 5% ethylene glycol (72.5±5.00%, 54.88±0.66% and 46.00±2.40%; p<0.05). Motility and viability were similar between 7% glycerol and 5% ethylene glycol. However, the membrane integrity was significantly higher in 5% ethylene glycol (34.69±4.64% vs 46.00±2.40%; p<0.05). The viability and membrane integrity were significantly higher in 5 cm for 10 min and 8 cm for 10 min than 3 cm for 5 min (viability: 55.81±2.94, 55.19±3.34 vs 47.94±3.48%; p<0.05 and membrane integrity: 44.94±3.51, 46.06±2.25 vs 40.38±1.03%; p<0.05). The percentage of capacitated sperm assessed by CTC staining, percentage of F pattern was higher in 7% glycerol, 5% and 7% ethylene glycol, and AR pattern was significantly higher in 3% ethylene glycol. F pattern was significantly increased in 5 cm for 10 min and 8 cm for 10 min (p<0.05), but AR pattern was significantly increased in 3 cm for 5 min (p<0.05).

The objective of this study was to determine the effect of semen extenders on the motility, viability and fertility in vitro of spermatozoa during storage of fresh boar semen diluted in different commercial extenders used for pig artificial insemination (AI). In this experiment, semen were diluted in Androhep plus, Beltsville Thawing Solution (BTS), Modena, Seminark and Vitasem LD. Five ejaculates were collected from three Duroc boars and sub-samples were diluted (30×106 spermatozoa/ml) in different extenders. Semen was stored at 17℃ for 10 days. Sperm motility and viability was assessed using Computer-Assisted Semen Analysis (CASA) and flow-cytometry on 1, 3, 5 and 10 day post collection. The motility of spermatozoa stored in different extenders was gradually decreased by increasing the duration of storage of semen. However, there was not significantly different in the sperm motility and viability among other extenders. On the other hand, the in vitro-matured oocytes were fertilized and cultured in vitro to assess the fertility of boar spermatozoa stored for 3 and 10 days in different extenders. The percentage of morula and blastocyst were taken as indicators of fertility in vitro of spermatozoa. Therefore, there were no differences in the rate of embryos developed to the molular and blastocyst stage. There were no differences in the motility and fertility in vitro among 5 kinds of commercial boar semen extenders.