This study was conducted to investigate the effects of different levels of soybean curd residue as an ingredient of TMR on growth performance and carcass characteristics of Holstein steers. Twenty four Holstein steers of 7 months age(average body weight 274.6 kg) were randomly allocated to 3 experimental groups, which contained 8animals per group during 15-months feeding trial. The treatments composed of the group fed concentrate and rice straw separately (Control), the group fed TMR containing low levels of soybean curd (T1), the group fed TMR containing high levels of soybean curd residue (T2). Body weight and average daily gain were not significantly different among the treatment groups during the feeding trial periods. Feed intake was not differ among treatment groups, but feed conversion ratio was significantly higher in T2 group compared to T1 group during late fattening stage (p<0.05). However, it was not different in overall average. According to the yield traits, carcass weight of control, T1 and T2 were 466.4, 451.1 and 460.1 kg, respectively. Back fat thickness, rib eye area and yield index were not different among the groups. The marbling score, meat color, texture and maturity in quality traits were not different among the treatment groups, but fat color was significantly higher in T2 group compared to T1 group (p<0.05). The quality grade (1+:1:2:3, NO.) tended to be higher for T1(0:2:1:5) and T2 (0:1:4:3) compared with the control (0:0:3:5). Therefore, these results indicated that TMR with soybean curd residue may be appropriate the growth performance and carcass characteristics of Holstein steers.

Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. This study was designed to determine effect of glycerol and ethylene glycol as cryoprotectant in extender on improve the freezability of Jeju horse semen. The semen was cryopreserved with glucose-EDTA extender containing each 5% glycerol, 5% ethylene glycol, 8% glycerol or 8% ethylene glycol, respectively. Post-thawed sperm were evaluated motility, viability, Membrane integrity and acrosome integrity. Post-thawed sperm motility were not significantly differences among treatments. However, sperm viability were significantly higher (p<0.05) in 8% glycerol (39.85% ±11.41) than in 5% glycerol treatment (18.08%±1.61). In membrane integrity, swelling sperm ratio was significantly higher (p<0.05) in 8% glycerol (34.12%±11.02) than other treatments. In the percentage of capacitated sperm assessed by CTC staining, F pattern was significantly higher in 8% ethylene glycol than 5% glycerol and 5% ethylene glycol (p<0.05). B pattern ratio was significantly increased in 5% ethylene glycol compared with 8% glcerol and 8% ethylene glycol (p<0.05). Moreover, 8% ethylene glycol treatment was significantly decreased AR pattern ratio compared with other treatments (p<0.05). It is concluded that treatment of 8% glycerol was improved the sperm viability and 8% ethylene glycol was improved the sperm ascrosome integrity after thawing. However, they were not significantly difference between 8% glycerol and 8% ethylene glycol on post-thawed sperm viability. Therefore, 8% ethylene glycol was more effective sperm cryoprotectant than 8% glycerol in Jeju Horse.

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.

This study was designed to determine whether low-density lipoproteins (LDL) from egg yolk and taurine, hypotaurine and trehalose as antioxidant in extender improve the freezability and fertility of Korean Jeju Black Bull semen. The semen was cryopreserved with tris egg yolk extenders containing 7% glycerol and treated 4% LDL, 20 mM taurine, hypotaurine and trehalose. Frozen-thawed sperm were evaluated motility, viability, membrane, and acrosome integrity and sperm penetration ability. The results were compared to semen cryopreserved in tris egg yolk extender only as control. Frozen-thawed semen evaluation cleary indicated that the addition of LDL and LDL-antioxidants (taurine, hypotaurine and trehalose) combination were significantly improved (p<0.05) the viability (%; with staining test using eosin-Y) compared to control spermatozoa. Also, in membrane integrity (%; with supravital hypo-osmotic swelling test), not only LDL-antioxiants combination but also LDL were significantly increased (p<0.05) the swelled sperm using HOST compared to control. Sperm acrosome integrity state was classified by CTC (chlortetracycline) staining test. F pattern was significantly increased in LDL-antioxidant combination than control (p<0.05) and B pattern was not significantly differences among all treatments and control. However, AR pattern was significantly decreased in LDL-antioxidants combination than control (p<0.05). Pronucleus formation and sperm penetration index (SFI) were significantly increased in LDL and LDL-antioxidants combination than control (p<0.05). Especially, LDL-taurine significantly improved pronucleus fomation and SFI than LDL (p<0.05). It was concluded that LDL and LDL-antioxidants in extender improved the freezability and fertility of Korean Jeju Black bull spermatozoa.

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).

This study was carried out to establish most suitable freezing condition, to evaluate the different glycerol concentration of freezing and thawing rates on motility, viability, membrane integrity and acrosome intecrity of frozen Korean Jeju Black Bull spermatozoa, Semen was collected from a Korean Jeju Black Bull using an artificial vagina and transported to the laboratory. The semen was extended gradually 1:5 then cooled slowly for 2 hrs to 4. The semen was diluted 1:1 with cryoprotectant extenders (3%, 5% and 7% glycerol) and equilibrated for 2 hrs at cold chamber and packed to 0.5 ml straws. The semen straws were located above 3 cm of liquid nitrogen for 5 minutes, above 5 cm for 10 min and above 8 cm for 10 min. And then the frozen straw was plunged into LN. The presented straws were examined the viability and motility after thawed at 37 water bath. The viability and membrane integrity immediately post-thawing were significantly higher in samples frozen in 7% glycerol than 3% and 5% glycerol (p<0.05). After CTC staining to assess acrosome integrity, F pattern was significantly increased, but B pattern was significantly decreased in 7% glycerol (p<0.05). Freezing distance of 5 cm from liquid nitrogen and pre-cooling for 10 min yield better survival and membrane integrity, but not significant difference. However, AR pattern according to CTC staining was significantly decreased in 3 cm for 5 min.

Endocrine disruptors bind to hormone receptors on sperm membrane, therefore spermatozoa are potentially a useful model for examining estrogenic activities of endocrine disruptors. The objective of this study was to compare the effects of two xenoestrogenic compounds [genistein (Gen) and 4-tert-octylphenol (OP)] to those of two steroids [estrogen (E2) and progesterone (P4)] on boar sperm % motility and motion kinematics of in vitro. Porcine spermatozoa were incubated with various concentrations (0.001～100 μM) of each chemical for 15 or 30 min, and then assessed % motility and sperm motion kinematics using computer assisted sperm analyzer (CASA). Each chemical decreased sperm % motility, and OP decreased VSL and VAP compared with untreated control(p<0.05). E2 stimulated the motion kinematic changes except VCL. Moreover, Gen had effects on VCL and VAP alterations after 30 min incubation. In summary, since all chemicals studied effectively altered sperm % motility and motion kinematics, it was concluded that porcine spermatozoa could be a useful model for in vitro screening of potential endocrine disruptors.

본 연구는 정액의 보존 기간 동안 정액의 질적 변화를 알아보고자 시행하였다. 돼지 정액을 Beltsville Thawing Solution (BTS)에 희석한 후 17'C 에서 5일 동안 보존하였다. 보존 기간 동안 정자의 운동성(%)과 linearity는 3일째부터 유의하게 감소하였으나, 다른 운동 역학 변수에서는 유의적 변화를 나타내지 않았다. 또한, 5일 동안 정액을 보존할 경우 첨체의 온전성에도 변화가 없었다. 그러나 제 4일째부터 첨체 변화가 야기된 정자는 유의적으로 증가하였으나, 수정능 획득이 일어난 정자는 유의적으로 감소하였다. 정액의 보존 기간 동안 첨체의 온전성의 유의적 변화가 없었다. 즉, 보존 기간 3일동안 정자의 질적 운동성 및 첨체 온전성에는 유의적인 변화가 없었으므로 상업용 돼지 액상정액은 17'C 에서 적어도 3일간 수정능력을 만족스럽게 유지함을 보여준다.

본 연구는 정자 내의 phsopholipid hydroxide glutathion peroxidase (PHGPx) mRNA 발현 수준, heparin-binding protein (HBP) mRNA 발현 수준, 수태율 그리고 산자수 사이의 관계를 조사하고자 실시하였다. 수태율과 산자수 사이에 있어서 상관관계는 나타나지 않았다. PHGPx mRNA 발현 수준에 있어 산자수가 10두 이상 군에서 (2,414.7±400.7) 8두 미만 군보다 (1,875.8±311.2) 높게 나타났으나 유의적인 차이는 없었다. HBP mRNA 발현 수준에 있어서도 산자수 10두 이상 군에서 (2,255.9±360.8) 8두 미만 군보다 (2,155.4±378.0) 약간 높은 결과를 보였으나, 유의적인 차이는 인정되지 않았다. PHGPx mRNA 발현 수준과 산자수 사이의 관계는 (r=0.206) 정의 상관관계를 보였으나, 유의한 상관관계는 나타나지 않았다. 본 실험의 결과, PHGPx와 HBP의 발현수준이 수태율, 산자수와 유의한 상관관계를 보이지 않았기 때문에 PHGPx와 HBP가 정자의 수정능력을 예측하기에는 미흡한 면이 있다.

경제동물에 있어 정자의 수정능력의 예측은 매우 중요한 일이다. 그러나 현재까지 이용되고 있는 정자의 기능적 분석은 정자의 운동성, 활력, 농도 및 일반적인 형태적 분석 등과 같은 양적 형질의 평가만을 수행할 뿐 수정능력 예측을 위한 객관적이고 정확한 도구는 개발되지 못하였다. 즉, 인반적인 정자의 분석은 웅성의 수정능력을 진단하는데 있어 그 한계를 가지고 있는 실정이다. 우수 동물 및 농가의 생산력을 증진시키고 안정적인 생산체제를 구축하기 위하여 새로운 정자의 수정능력 예측법의 개발이 시급한 실정이다. 최근 보다 정확하고 객관적인 수정능력을 예측하기 위하여 정자의 hyperactivation, 수정능획득과 같은 intact-acrosome, 정자의 투명대 결합능력, 정상적 DNA의 상태 등과 같은 다양한 방법이 수행되고 있다. 그러나 이러한 방법들이 정자의 기능과 수정능력 사이의 상관관계가 매우 적기 때문에 정확한 수정능력의 예측에 있어 많은 의문이 제기되고 있다. Proteomics는 postgenome 시대에 새로운 연구로서, 생물학적 과정에 관련된 세포 mechanism을 식별하는 proteome의 질적, 양적 비교로 정의할 수 있다. 정자는 transcription이 불활성화 되어 있는 세포이므로 proteomics를 이용하여 정자의 분자적 기능을 보다 정확히 이해할 수 있다. 또한 정자 proteome의 비교 분석은 정자가 수정을 위한 수정능획득의 방법과 정자가 서로 다른 수정 능력을 가지는 원인을 이해하는데 있어 매우 중요하다. 본 연구의 목적은 돼지의 체내 수정능력을 예측하는 방법의 최적화를 통하여 새로운 예측방법을 개발하고, 나아가 정자의 수정능과 관련된 프로테옴을 동정하여 정자의 수정능력을 예측하는 바이오 마커로의 이용 가능성을 입증하고자 하였다. 본 실험의 분석 결과의 민감도를 높이기 위하여 모든 과정을 최적화 하는 과정을 거쳤으며, quality control을 실시하여 안정적인 결과를 도출하였다. 투명대를 제거한 햄스터 난자와 미성숙 돼지 난포란을 이용한 정자의 침투능력검사에 있어 heparin을 처리한 실험구에서 가장 높은 침투율을 나타냈다. 정자 침투능력검사에 있어 투명대를 제거한 햄스터 난자를 공시하였을 때 돼지의 미성숙 난포란을 이용한 결과보다 유의적으로 높은 결과를 나타냈다. 소의 동결정액을 이용하여 quality control을 실시하여 실험의 변이를 제거하였으며, 또한 암컷의 변이 요인 또한 가능한 모두 제거하였다. 최적화된 SPA (sperm penetration assay using zona-free hamster oocytes)는 돼지의 평균 산자수와 유의적인 상관관계를 나타냈으나, 수태율과는 어떠한 상관관계도 나타내지 않았다. 산자수 8마리 이하를 예측하기 위하여 Sperm Fertility Index (SFI)을 1.2로 cutoff 하였을 경우 정확성은 92%이며, 산자수 10마리 이상을 예측하기 위하여 SFI를 2.5로 정하였을 경우 96%의 정확성을 나타내어 수정능력 예측을 위한 방법으로 유용성이 입증되었다. 산자수가 낮은 group과 높은 group의 정자로부터 발현되는 단백질의 수준을 분석한 결과, 총 108개의 단백질 spot을 찾을 수 있었다. SigmaGel programe을 이용하여 spot의 농도를 분석한 결과, 25개의 단백질 spot이 서로 다르게 발현하였으며, 산자수가 낮은 정자에서 보다 높은 농도로 발현하고 있음을 알 수 있었다. 25개의 단백질 spot을 LC-MS/MS 방법을 이용하여 분석한 결과, 총 20개의 단백질을 동정할 수 있었으며, 이들 20개 단백질 중 mitochondrial trifunctional proteins이 산자수가 높은 group의 정자에서 높은 발현을 나타냈으며, PHGPx와 Arginine Vasopressin Receptor 2가 산자수가 낮은 group의 정자에서 높은 발현을 나타내었다. 이러한 결과들은 이들 방법들이 단지 정자의 수정능력을 분석하는데만 이용할 수 있는 것이 아니라 biomarker로서 정자의 수정능력을 조절하는 인자와 관련된 유전자를 발굴하는데도 유용할 것으로 사료된다.

본 연구는 소의 미성숙 난포란의 체외성숙시 의 첨가가 체외성숙, 체외수정 후 웅성전핵의 형성 및 세포질 내의 GSH 수준에 미치는 영향을 알아보고자 실시하였다. 체외성숙시 과 의 를 첨가한 경우 대조구에 비하여 성숙율이 증가하는 것으로 나타났으며(p<0.05), 모든 실험구에 있어서 12시간 체외성숙보다 24시간 체외성숙에서 높은 성숙율을 나타냈다(p<0.05). 체외수정 후 웅성전핵 형성에 있어서는 와 농도의 첨가구에서 대조구에 비하여 높게 나타났으나