The objective of this study was to investigate the influence of the addition of caffeine (alkaloid family) to the ejaculates of boar sperm quality as well as investigate their optimum concentrations for increasing the movement of sperms. Semen was collected from 9 boars by the gloved-hand technique one week interval. Semen followed by cryopreservation with egg yolk extender freezing medium using freezing protocol. The collected semen were frozen on the same day. Motility was assessed for % motile cell characteristics using computer-assisted semen analysis (CASA; SAIS SI-100, Medical supply, Korea). Frozen boar sperms were thawed in Beltsville Thawing Solution (BTS) with 5, 10, and 15mM caffeine were then incubated at 38 celsius degree for 20 minutes. In experiment 1, semen were diluted BTS and addition of different concentration of caffeine to the pre-freeze semen cryopreservation. In experiment 2, incubation of frozen-thawed sperm in BTS supplemented with different concentration of caffeine for 20 minutes improved (P<0.05) after semen cryopreservation-thawing on sperm quality. After thawing significantly increased progressive and total motility. The addition of 10 mM caffeine to cryopreserved semen after thawing significantly increased progressive and total motility compared with other treatment. These result suggest that caffeine enhanced post-thaw motility of cryopreserved boar sperm when added after thawing.

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.

For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Phospholipase C zeta (PLCz) is important enzyme in spermatogenesis, but their effect has not been confirmed in pigs yet. Therefore, this study was aimed to analyze their association with sperm motility and kinematic characteristics. DNA samples from 124 Duroc pigs with records of sperm motility and kinematic characteristics [Total motile spermatozoa (MOT), Curvilinear velocity (VCL), Straight-line velocity (VSL), the ratio between VSL and VCL (LIN), Amplitude of Lateral Head displacement (ALH)] were subjected. A SNP in non-coding region of PLCz g.158 A > C was associated with MOT (p < 0.05), VCL (p < 0.01), LIN (p < 0.01) and ALH (p < 0.05) in Duroc population. Therefore, we suggest that the intron region of the porcine PLCz gene may be used as a molecular marker for Duroc boar semen quality, although its functional effect was not defined yet. Whether the association is due to the candidate gene or not require further verification. Thus, it will be of interest to continue association studies in the regions surrounding those genes.

For evaluating the boar semen quality, sperm motility is an important parameter because the movement of sperm indicates active metabolism, membrane integrity and fertilizing capacity. Phospholipase C zeta (PLCz) is important enzyme in spermatogenesis, but the effect has not been confirmed in pigs yet. Therefore, this study was aimed to analyze their association with sperm motility and kinematic characteristics. DNA samples from 124 Duroc pigs with records of sperm motility and kinematic characteristics [total motile spermatozoa (MOT), curvilinear velocity (VCL), straight-line velocity (VSL), the ratio between VSL and VCL (LIN), amplitude of lateral head displacement (ALH)] were subjected. A SNP in non-coding region of PLCz g.158 A > C was associated with MOT (p < 0.05), VCL (p < 0.01), LIN (p < 0.01) and ALH (p < 0.05) in Duroc population. Therefore, we suggest that the intron region of the porcine PLCz gene may be used as a molecular marker for Duroc boar semen quality, although its functional effect was not defined yet. Whether the association is due to the candidate gene or not require further verification. Thus, it will be of interest to continue association studies in the regions surrounding those genes.

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.

멧돼지(Sus scrofa)는 우제목 멧돼지과 멧돼지속에 속하 는 종으로 아시아, 유럽, 북아메리카 등에 넓게 분포하고 있으며 잡식성으로 고라니(Hydropotes inermis)와 청설모 (Sciurus vulgaris) 등과 함께 환경부 지정 유해야생동물 및 수렵종으로 지정되어 있다. 우리나라에 서식하는 멧돼지는 농작물 피해에서부터 도심출현 등 다양한 분야에 걸쳐 문제 가 발생하고 있으며 국내뿐만 아니라 일본, 영국, 호주 등 전 세계적으로 개체수 증가 및 인간과의 충돌로 인해 관리 방안에 대한 연구가 지속적으로 진행되고 있다. 국외에서 오랜 기간 동안 수행된 연구 자료를 바탕으로 추정된 결과 멧돼지에 의한 피해증가의 원인은 서식지 단절 및 파괴에 따른 서식 공간 부족, 상위포식자 멸종에 따른 먹이사슬 불 균형으로 인한 개체수 증가 및 먹이부족 등의 요인이 작용 한 것으로 알려져 이에 따른 관리방안이 도출되고 있으나 국내의 경우는 멧돼지를 관리하기 위한 기초 연구가 충분히 이루어지지 않은 상태에서 피해가 지속적으로 확대됨에 따 라 포획을 통한 직접적인 개체수 조절 방안이 시행되고 있 다. 정확한 피해 원인에 대한 과학적 근거는 미비한 상태에 서의 이러한 무조건적 포획은 남획으로 인한 생태계 불균형 등의 문제를 유발할 수 있기 때문에 이러한 문제를 해결하 기 위해서는 멧돼지에 대한 행동권 분석, 서식지 환경 수용 력 평가, 개체군 역동성 등에 대한 기초생태연구가 선행되 어야 한다. 따라서 본 연구는 한려해상국립공원 에 서식하 는 멧돼지의 서식현황 및 행동생태 등의 기초생태연구를 통해 국립공원에 서식하는 멧돼지의 관리방안을 마련하기 위해 2013년 3월부터 2014년 8월까지 실시하였다. 한려해상국립공원 희망봉 일대에 서식하는 멧돼지의 서 식 현황을 흔적조사와 무인센서카메라를 이용하여 조사한 결과 멧돼지의 먹이섭식 흔적과 털, 배설물 등이 확인되었 으며 먹이로는 밤, 갈대뿌리, 칡뿌리, 곤충, 지렁이 등을 섭 식한 흔적이 확인되었다. 무인센서카메라 자료 분석 결과 멧돼지는 신봉저수지에서 희망봉 정상 하부의 사면에서 주 로 서식하고 있는 것이 확인되었으며 농경지와 묵논이 먹이 활동과 진흙목욕 장소로 이용되고 있는 것으로 조사되었고 고라니, 너구리, 노루, 청설모의 실체가 확인되었다. 멧돼지 는 주로 야간 시간인 10시 이후에 먹이활동을 하고 주간 시간인 오후 4시와 오전 8시에 서식지를 이동하는 것으로 확인되었다. 한려해상국립공원 희망봉 일대의 멧돼지 서식현황 조사 결과를 바탕으로 멧돼지 2개체를 포획하여 스마트폰 문자 로 위치 좌표를 수신할 수 있는 CDMA형 발신기를 부착하 여 현장에서 방사하였다. 2014년 5월에 포획된 HR1개체 (암컷, 70kg)는 2014년 5월 11일부터 5월 26일까지의 위치 좌표를 수집한 후 분석하였으며 HR2개체(수컷, 60kg)는 2013년 5월에 포획된 개체로 2013년 5월부터 2014년 6월 까지 총 1년 2개월간의 위치좌표를 획득하여 분석하였다. HR1개체의 행동권을 분석한 결과 MCP 100%에서 1.110㎢, 95%에서 0.980㎢, 핵심지역인 50%에서 0.360㎢ 이며 Kernel 95%에서 2.480㎢, 90%에서 2.030㎢ 그리고 핵심지역인 50%에서 0.730㎢으로 전체 행동권은 실제 좌 표 이외의 지역을 많이 포함하는 Kernel 95%의 결과에서 최대값을 나타냈다. HR2개체의 행동권은 MCP 100%에서 0.870㎢, 95%에서 0.080㎢ 그리고 핵심지역인 50%에서 0.002㎢이며 Kernel 95%에서 0.030㎢, 90%에서 0.020㎢ 그리고 핵심지역인 50%에서 0.0006㎢로 나타났다. HR1은 HR2에 비해 희망봉 일대의 서식지를 넓게 이용하고 있는 것으로 나타났으나 좌표 수집 기간이 26일로 짧고 수집된 좌표 또한 매우 적기 때문에 두 개체의 행동권 결과를 비교 하기에는 한계가 있었다. 장기간 행동권 좌표가 확보된 HR2의 서식지 이용패턴을 분석하기 위해 총 2,700여개의 위치좌표를 분석한 결과 전 체 행동직경은 약 1,150m로 매우 적은 면적을 이용하는 것이 확인되었다. 현장조사 결과 한려해상의 희망봉 지역의 HR2개체의 행동권내 서식지에는 진흙 목욕탕을과 먹이원 등 멧돼지의 주요 서식처로서의 요건이 충족되어 있으며 하부 지역은 농경지, 도로, 마을 등의 파편화로 인한 위협요 인으로 멧돼지가 서식할 수 있는 공간이 한정되어 있기 때 문에 좁은 서식지내에 안정적인 행동권을 유지하고 있는 것으로 판단된다. 멧돼지의 핵심지역(Core area)은 MCP 100%의 전체 행동권에서 위치상 중앙인 산림내부에 포함 되어 있었으며 전체 행동권의 최외각은 농경지를 포함하고 있었으며 농경지에 포함된 좌표는 전체 좌표의 약 7%로 농경지 피해는 비교적 낮은 것으로 분석되었으나 현장조사 결과 농경지 이용 흔적이 많이 발견되고 좌표 수신 간격이 2시간인 것으로 고려하였을 때 실질적인 농경지의 이용 면 적은 더 넓을 것으로 판단된다. MCP 100%의 행동권 면적 을 기준으로 HR2개체의 계절별 행동권을 분석한 결과 여름 0.400㎢(6월~8월), 가을 0.350㎢(9~11월), 겨울 0.200㎢ (12월~2월), 봄(3월~5월) 0.510㎢의 행동권을 나타냈으며 연도별 행동권 분석 결과 2013년에는 0.73㎢, 2014년에는 0.53㎢으로 유사한 행동권 크기를 유지하는 것으로 나타냈 다. 본 연구는 한정된 개체수와 짧은 연구기간으로 인하여 개체간의 행동 특성과 기간별 행동권을 비교 분석을 하는데 한계점을 갖고 있으므로 향후 개체수 확대와 장기적인 연구 를 실시하여 유의성 있는 결과를 도출할 경우 멧돼지를 관 리하기 위한 방안을 마련하는데 유익한 자료가 될 것으로 판단된다.

The purpose of this study was to examine the effects of taurine and vitamin E on sperm characteristics damaged by bromopropane (BP) in pig. We evaluated toxicity of BP on viability, membrane integrity and mitochondrial activity of spermatozoa. 1-BP (0, 2.5, 5.0, 10, and 50 μM), 2-BP (0, 2.5, 5.0, 10, and 50 μM), taurine (0, 5.0, 10, and 25 μM) and vitamin E (0, 50, 100, and 200 μM) were treated in fresh boar semen for 6 h. 10 and 50 μM of 1-BP and 2-BP inhibited sperm viability, membrane integrity and mitochondrial activity in fresh boar semen (P<0.05). 25 μM of taurine increased sperm viability and membrane integrity (P<0.05), 100 μM of vitamin E enhanced viability and mitochondrial activity of sperm (P<0.05). Finally, 10 μM of 1-BP and 2-BP was co-treated with taurine (25 μM) and vitamin E (100 μM) in the fresh boar semen. The co-treated samples did affected viability, membrane integrity and mitochondrial activity of sperm. In conclusion, taurine and vitamin E can improve and maintain sperm quality in fresh boar semen.

Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen post-thawed in boar. Recently, polymorphism (g.158T>C) of phospholipase C zeta (PLCz) gene reported to be significant association with MOT. This study was conducted to evaluate the PLCz gene as a positional controlling for motility and kinematic characteristics of post-thawed boar semen. To results, The g.158 T>C SNP of PLCz was significantly associated with frozen semen motility and kinematic characteristics. g.158 T>C SNP was high significantly associated with MOT, VCL, VSL and VAP (p<0.0001, p= 0.0002, p<0.0001 and p<0.0001, respectively). Therefore, we suggest that the intron region of the porcine PLCz, may be used as a molecular marker for Duroc boar post-thawed semen quality, although its functional effect was not defined yet. Whether the association is due to the candidate gene or not require further verification. Thus, it will be of interest to continue association studies in the regions surrounding those genes.

The aim of this study is to examine current status of swine AI and boar stud in South Korea using survey and data analysis. This survey included 48 boar studs registered as ‘semen processing business’. The survey data were collected by direct visitation, FAX and/or telephone conversation for 7 months from June through December in 2013. 48 boar studs owned a total of 3,537 boars and the Duroc breed accounted for the highest rate (75.3%) of all boar breeds. In case of ownership, agricultural management corporations was the highest (50.0%) and followed by individual ownership (33.3%). Large-scale boar studs in terms of own over 151 boar were surveyed as 4.2% and most boar studs owned less than 100 boars (77.1%). The amount of liquid semen provided by 48 boar studs were 1,889,000 doses and each boar stud provided average of 39,000 does, which is represented for 90% consumption by sows in South Korea.

The objective of this study was to evaluate the efficiency of sperm cryosurvival in boar sperm separated by Percoll containing antioxidant enzymes. The boar semen was collected into a pre-warmed (37℃) thermos bottle by gloved-hand method and was separated by 65% Percoll with superoxide dismutase (SOD), catalase (CAT) and glutathione (GSH) before freezing. The frozen sperm was thawed at 38.5℃ for 45 sec in water-bath for sperm characteristic analysis. The sperm were estimated with SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction, Rhodamine123/PI double staining for mitochondrial integrity and were analyzed using flow cytometry. In results, sperm viability, acrosome reaction and mitochondrial integrity were improved in separated sperm groups compared with unseparated sperm by Percoll (UP) group. Especially, viability was significantly higher in sperm separated by Percoll containing 400 IU CAT group compared with other groups (P<0.05). And acrosome reaction was decreased in sperm separated by Percoll with 300 IU SOD, 400 IU CAT and 0.5 mM GSH groups compared with other groups, however, there were no significantly difference mitochondrial integrity among sperm separated by Percoll with antioxidant enzymes. In conclusion, we suggest that use of Percoll containing antioxidant enzymes for sperm separation will be beneficial for sperm cryopreservation in pigs.

Cryopreservation of boar semen is continually researched in reproductive technologies and genetic resource banking in breed conservation. For evaluating the boar semen quality, sperm motility (MOT) is an important parameter because the movement of spermatozoa indicates active metabolism, membrane integrity and fertilizing capacity. Various researches have been trying to improve the quality of semen post-thawed in boar. Recently, polymorphism (g.358A>T) of cluster-of-differentiation antigen 9 (CD9) gene reported to be significant association with MOT. Also, CD9 gene was expressed in the male germ line stem cells is crucial for sperm-egg fusion, and was therefore selected as candidate gene for boar semen. This study was conducted to evaluate the pig SNP (g.358A>T) of CD9 gene as a positional controlling for semen parameters of post-thawed boar semen. To results, the g.358A>T SNP of the CD9 gene was significantly associated with the traits such as MOT, curve linear velocity, straight line velocity, average path velocity and amplitude of lateral head displacement. Particularly, the g.358A>T SNP significantly has the highest association with MOT and animals with AA genotype (p<0.001). Therefore, we suggest that the g.358A>T in the intron 6 region of the porcine CD9 may be used as a molecular marker for Duroc boar Post-thawed semen quality, although its functional effect was not defined yet.

The objective of this study was to investigate the efficiency of nicotinic acid on sperm cryosurvival and fertilization ability in frozen-thawed boar semen. Boar semen was collected by glove-hand method and was frozen using freezing solution treated to 0, 5, 10 and 20 mM of nicotinic acid. The frozen sperm for sperm characteristic analysis was thawed such as viability, acrosome reaction, and mitochondrial integrity. The frozen-thawed sperm was estimated by SYBR14/PI double staining for viability, FITC-PNA/PI double staining for acrosome reaction and Rhodamine123/PI double staining for mitochondrial integrity using a flow cytometry. The embryo was estimated in vitro development and DCFDA staining for reactive oxygen species assessment. As results, frozen-thawed sperm viability was significantly higher in 5 and 10 mM (61.1 ± 1.5%, 64.7 ± 2.0%) of nicotinic acid than other groups (0 mM, 52.1 ± 2.3%; 20 mM, 47.8 ± 5.1%, P<0.05). The live sperm with acrosome reaction was significantly higher in 5 and 10 mM of nicotinic acid (26.1 ± 1.8%, 24.9 ± 1.5%) than other groups (0 mM, 35.3 ± 0.8%; 20 mM, 36.5 ± 1.9%, P<0.05). The live sperm with mitochondrial integrity was significantly higher in 5 and 10 mM (84.2 ± 3.6%, 88.4 ± 2.3%) of nicotinic acid than other groups (0 mM, 77.3 ± 4.4%; 20 mM, 73.3 ± 3.6%, P<0.05). Blastocyst rate of in vitro development was significantly higher in 10 mM (17.0 ± 1.3%) of nicotinic acid than other groups (0 mM, 9.4 ± 0.5%; 5mM, 12.6 ± 0.8%; 20 mM, 5.0 ± 1.0%, P<0.05). Moreover, total cell number was higher in 5 and 10 mM (53.6 ± 2.9%, 57.9 ± 2.8%) of nicotinic acid than other groups (0 mM, 41.0 ± 1.4%; 20 mM, 23.2 ± 2.8%, P<0.05). Hydrogen peroxide in embryos was lower in 5 mM nicotinic acid (0.7 ± 0.1%) than other groups (0 mM, 1.0 ± 0.1%; 10mM, 0.9 ± 0.0%; 20 mM, 1.4 ± 1.0%, P<0.05). In conclusion, nicotinic acid-treated semen improves cryosurvival and quality of spermatozoa. Also, the fertilized oocytes with nicotinic acid improve quality of embryo and blastocyst formation.

The cold shock of spermatozoa is associated with oxidative stress induced by reactive oxygen species. This study was conducted to evaluate the toxicity of natural antioxidant green tea extract (GTE) in lactose-egg yolk (LEY) extender during boar sperm cooling prior to freezing. Spermatozoa were cooled to 5℃ for 3 h in LEY extender containing 0 (control), 1, 10, 100 or 1,000 mg/l of GTE, re-suspended with LEY-glycerol-Equex extender and cooled at 5℃ for 30 min. Sperm progressive motility, viability and phosphatidylserine (PS) translocation were evaluated. PS translocation was assayed by flow cytometry using Annexin V-FITC apoptosis detection kit. The sperm function including progressive motility, viability and PS translocation was not significantly different regardless of GTE concentrations (P>0.05). In conclusion, this study demonstrated non-toxicity of GTE supplement in LEY extender during sperm cooling.

The boar sperm has more lipid droplets and specialty of seminal plasma compared with other species, causing difficulties of freezing sperm and decreases for the utilization of frozen semen into the artificial insemination. However, several studies reported significant results for the recovery of sperm motility and reproductive by addition of cryoprotectants and seminal plasma after thawing. This study was designed to investigate the effects of supplementation of trehalose or glycerol in the LEY (lactose and egg yolk in BTS) solution for the conventional freezing and vitrification process. Two boars aged 16 months were used to collect semen for 2 times in a week. The samples were allotted to 3 freezing solutions (LEY + glycerol 10.5% + OEP 1.5%, LEY + trehalose 1M + OEP 1.5%, and sucrose 1.5M + trehalose 1 M + OEP 1.5%) after centrifugation at 800 g for 10 minutes. Semen was equilibrated in freezing solutions for 10 minutes and injected into plastic straws with 2∼3 air bubbles to minimize freezing damages. Vitrification was performed to locate sperm in 5 cm above LN2 for 5 minutes, and the conventional freezing was conducted with an automatic freezer. Motility and survival rates were measured by CASA (Computer assisted sperm an alyzing system) and FITC (Fluorescein isothiocyanate), respectively after thawing semen at 50℃ for 12 seconds. The results were analyzed by ANOVA with STATVIEW statistical program. The vitrificatioin solution (LEY + 10.5% glycerol + 1.5% OEP) presented higher motility (20.9%) than other solutions while the solution (LEY + 1M trehalose + 1.5% OEP) showed the lowest (motility : 5.2%). However, survival rates of vitrified sperms detected by FITC showed 1~4% live sperms in almost of dead sperms at all vitrification solutions’ groups, but survival rate of freezing solution of LEY + 1M trehalose + 1.5% OEP LEY and LEY + 10.5% glycerol + 1.5% OEP were showed 49%, and 79%, respectively. There were differences (P<0.05) survival rate of conventional freezing in LEY + 10.5% glycerol + 1.5% OEP and LEY + 1M trehalose + 1.5% OEP and the remaining showed no differences. The results suggested that vitrified boar semen was not enough to be utilized for the artificial insemination, but it showed possibility to utilize for ICSI and conventional freezing with glycerol would be useful method for artificial insemination in pig while we choose the outstanding semen against tolerance to freezing damages.

The objective of this study was to determine the effects of E. coli on boar sperm quality and reproductive performance in sows after artificial insemination. Three different levels of E. coli were artificially inoculated to semen with following concentrations; Control, 500, 5,000 and 50,000 colony forming unit (cfu)/ml. Semen samples were preserved at 17℃ for 5 days. Sperm motility was significantly decreased (p<0.05) on day 3 in the group inoculated with 5,000cfu/ml compared to control groups. In all treatment groups, sperm motility was gradually decreased as storage time increased, but the decline pattern was more drastic in the groups inoculated with 5,000 and 50,000 cfu/ml groups from day 3 (p<0.05) compared to control group. After 3 day of storage at 17℃, sperm viability in sample inoculated with the highest concentration (50,000 cfu/ml) of bacteria was less (p<0.05) than that of control group. The pH of semen sample pH was maintained 7.2～7.5 in all groups during the experimental period. No differences (p>0.05) were found for both storage time and bacterial concentration. The pregnancy rate and live born piglets tend to decrease by increasing the concentration of E. coli in semen. In particular, the rate of pregnancy was lower in the group inoculated with 50,000 cfu/ml (58.3%) compare to the other groups (81.8, 75.0, 76.5%). These results suggest that the contamination of E. coli in boar semen negatively affects fertilizing ability of boar sperm and the reproductive performance obtained from sows after artificial insemination.

Quercetin and genistein, plentifully present in fruits and vegetables, are flavonoid family members that have antioxidative function and plant-derived phytoestrogen activity. The antioxidative effects of quercetin and genistein on boar sperm characteristics and in vitro development of IVF embryo were investigated. The sperm motility was increased by addition of genistein 50 μM for 6 hr incubation compared to control (p<0.05). The sperm viability was increased by addition of quercetin 1 and 50 μM and genestein 1 and 50 μM for 3 hr incubation. In addition, the sperm viability seemed to be increased dose-dependantly by addition of quercetin or genistein 1 and 50 μM, respectively (p<0.05). The membrane integrities were not increased by quercetin or genistein treatments for 3 hr or 6 hr incubation period except for quercetin 1 μM for 3 hr incubation. In mitochondrial activities, addition of quercetin 50 μM for 6 hr incubation increased mitochondrial activity but decreased at 100 μM concentration compared with control (p<0.05). When porcine IVF embryos were cultured in PZM-3 medium supplemented with low concentrations of quercetin (1∼10 μM), the developmental rates to morula and blastocyst increased but significantly decreased at high concentrations of quercetin (25∼50 μM). The highest developmental rate to blastocysts among all concentrations of quercetin was shown at quercetin 10 μM (p<0.05). The developmental rates to morula or blastocysts at low (0.01∼1 μM) and high (5∼10 μM) concentrations of genistein were not significantly different among all treatment group and genistein did not affect on IVF embryo development. These results suggest that quercetin and genistein seem to have positive effects at certain concentrations on sperm characteristics such as motility, viability and mitochondrial activity. In addition, low concentrations of quercetin (1, 5 and 10 μM) in this experiment, seem to have beneficial effect on porcine IVF embryo development but genistein did not affect on it at all given concentrations (0.01∼10 μM).

Artificial insemination technique has been contributed immensely for production of livestock worldwide as a critical assisted reproductive technique to preserve and propagate excellent genes in domestic animal industry. In the past decade, methods for semen preservation have been improved mostly in liquid preservation method for boar semen and freezing method for bull semen. Among many factors affecting semen quality during preservation, reactive oxy-gen species, produced by aerobic respiration in sperm for survival and motility, are unfavorable to sperm physiology. In mammalian cell as well as in the sperm, antioxidant system plays a role in degradation of reactive oxygen species. Magnetized water forms smaller stabilizing water clusters, resulting in high absorption and permeability of the cell for water, implicating its application for semen preservation. Therefore, this review focuses on preservation methods of boar and bull semen with respect to improvement of extender and reduction of reactive oxygen species by using magnetized water and supplementation of antioxidants.

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℃.

The objective of this study was to investigate the effect of bacterial contamination on elapsed time after preservation on boar semen. Known numbers of Escherichia coli (E. coli) were inoculated to freshly ejaculated semen and sperm parameters such as viability, motility, agglutination, acrosome integrity and hypo-osmotic swelling test were perform-ed during 7 days of liquid preservation. Semen samples were prepared using antibiotic free BTS extender and 4 di-fferent levels of E. coli were treated to semen with following concentrations; 3,000, 5,000, 7,000, 10,000 CFU/ml of sperms. Semen samples were preserved at 17℃ for 7 days in semen storage until analyzed. Aliquots were subjected to measure the sperm viability, motility and agglutination using computer assisted sperm analysis (CASA) system, acrosome integrity was performed using chlortetracycline (CTC) staining method and hypo-osmotic swelling test was performed using hypotonic solution from day 1 (day of semen collection) to 7. Detrimental effects on sperm motility and viability were observed 3 days after preservation at the level of 5,000 CFU/ml (p<0.05). Percentage of sperm abnor-mality was higher (p<0.05) in over 5,000 CFU/ml groups. Sperm agglutination rate was also significantly higher (p< 0.05) in groups of 5,000 and 7,000 CFU/ml. The rate of acrosome reacted sperm was higher as preservation time goes in all the samples but the pattern was clearly higher among E. coli contaminated groups (p<0.05). The sperm mem-brane integrity in terms of hypo-osmotic test, E. coli affects little compared to other sperm parameters. The deleterious effects observed due to the bacterial contamination in semen suggest that importance of hygiene protocol to minimize the bacterial contamination during semen collection and processing.