Preservation of liquid semen is an important factor for breeding management in swine industry. Oxidative stress of spermatozoa during liquid preservation has a detrimental effect on sperm quality and decreases fertility. Objective of this study was to determine the effect of antioxidant, Quercetin, on capability of porcine liquid semen preservation. Freshly collected porcine semen from boars (n=3), having proven fertility was counted, diluted to 3×107/mL and divided into 5 different semen extenders. Aliquots of diluted semen with different extenders were subjected to measure the pH, motility, viability and sperm DNA structure status on elapse time after preservation for 10 days. For the first 3 days, semen preserved in all 5 different extenders maintained their initial pH and either gradually decreased or increased thereafter, indicating lipid peroxidation has started. Sperm motility (r=0.52, p=0.01) and viability (r=0.55, p=0.03) had positive correlation with semen pH. Sperm motility was maintained well (p<0.05) in especially 2 extenders containing Tris and antioxidant compared to other extenders, suggesting both Tris and antioxidant worked as pH regulator and had beneficial effects on sperm characteristic during preservation. Sperm DNA structure status accessed by sperm chromatin structure assay on elapsed time after preservation, tended to be higher in semen preserved without antioxidant. Taken together, addition of antioxidant to extender prevents the sperm from oxidative stress during storage in mechanism by which antioxidant slows the lipid peroxidation, and thus reduced the reactive oxygen species in preserved porcine semen resulted in maintaining semen pH, sperm motility and viability for 7～10 days.

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.

This study was carried out to investigate the general characteristics of semen such as semen volume, pH, sperm motility and sperm concentration of the semen collected from Shih Tzu dogs (age of 24 to 48 months, weight of 4 to 8 kg) by using the method of digital manipulation of the penis. The effect of preservation temperature and time on motility of fresh semen was also investigated in the present study. Semen was collected for 16 times from 4 male Shih Tzu dogs by multiple ejaculations (four times ejaculation per dog). The average of semen volume, semen pH, sperm motility and sperm concentration of the second fraction containing small volume of the initial third fraction per ejaculation were 2.11 ± 0.31 ml, 6.25 ± 0.07, 97.59 ± 1.03% and 2.05 ± 0.14 × 108 cells/ml, respectively. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the first fraction from the ejaculation were 1.12 ± 0.15 ml, 5.99 ± 0.14, 16.09 ± 6.18% and 5.16 ± 2.03 × 105 cells/ml, respectively. Those of second fraction were 2.07 ± 0.29 ml, 6.36 ± 0.13, 97.31 ± 1.36% and 2.15 ± 0.30×108 cells/ml, respectively. Those of third fraction were 2.60 ± 0.29 ml, 6.63 ± 0.08, 95.72 ± 1.61% and 6.03 ± 1.83 × 107 cells/ml, respectively. Sperm motility was significantly higher at 17℃ preservation temperature than at 5℃ or 36℃ during preservation period except 1 h preservation (P<0.05). When preservation temperature was 17℃, sperm motility was 96.69 ± 1.49% at 1 h, 91.38 ± 1.90% at 6 h, 88.38 ± 2.34% at 12 h, 78.13 ± 4.58% at 18 h, 58.44 ± 8.57% at 24 h and 29.56 ± 5.06% at 30 h, respectively.

The study focuses on the quality assessment of Black Bengal buck semen preserved at chilled condition. In this in vitro trial, collected semen from Black Bengal bucks was preserved at chilling temperature (4▲5줛) in tris-glucosecitrate yolk medium of 1:5 ratios for four days. Artificial Vagina (AV) method was utilized to collect semen from buck. General evaluation of semen includes the color, mass activity and density were measured by direct visual examination. However, computer-assisted sperm analysis (CASA) and phase contrast microscopy were used to figure out the motility (%), hyper-activated (HYP) motility (%) and number of abnormal spermatozoa (%) initially, and at every 24 h intervals. The result revealed that spermatozoa preserved at chilling temperature showed significantly (P<0.05) lower motility and HYP motility with the progression of preservation. The number of phenotypically abnormal spermatozoa significantly (P<0.05) increased following preservation. Although significant positive correlation (r=0.945; P<0.05) was existed between % motile and % HYP motile spermatozoa however, the % of morphologically abnormal spermatozoa was negatively correlated with % motile (r=긏0.997; P<0.05) and % HYP motile spermatozoa (r=긏0.946; P<0.01). Therefore, we concluded that the quality of chilled semen progressively losses its viability and doesn…t remain useable after certain period of preservation with respect to its motility and morphology.

The efficiency of artificial insemination (AI) for horses remains unsatisfactory. It is mainly because each process of AI causes a detrimental effect on semen quality. To sustain quality of semen properly, several factors including libido of stallions and sperm damage during sperm processing and preservation should be considered. Stallions with decent libido produce a high ratio of sperm to seminal plasma in their ejaculates, which is the ideal semen composition for maintaining sperm quality. Thus, to maximize the fertility rate upon AI, stallions should be appropriately managed to enhance their libido. Seminal plasma should have a positive effect on horse fertility in the case of natural breeding, whereas the effects of seminal plasma on both sperm viability and quality in the context of AI remain controversial. Centrifugation of semen is performed during semen processing to remove seminal plasma and to isolate fine quality sperm from semen. However, the centrifugation process can also result in sperm loss and damage. To solve this problem, several different centrifugation techniques such as Cushion Fluid along with dual and single Androcoll-ETM were developed to minimize loss of sperm and to damage at the bottom of the pellet. Most recently, a new technique without centrifugation was developed with the purpose of separating sperm from semen. AI techniques have been advanced to deliver sperm to optimal region of female reproductive tract at perfect timing. Recombinant equine luteinizing hormone (reLH) and low dose insemination techniques have been developed to maximize both fertility rate and the efficiency of AI. Horse breeders should consider that the entire AI procedure should be optimized for each stallion due to variation in individual horses for a uniformed AI protocol.

Alpha-tocopherol as an antioxidant acts in preservation of chilled semen by preserving cell membrane damage from lipid peroxidation. Optimum concentrations of α-tocopherol in egg yolk-citrate (EYC) extender need to be studied in crossbred bull’s semen. Different concentrations of α-tocopherol viz. 0, 1, 2, 4 and 6mg per ml of extender were used. Semen was collected once a week from four bulls used to regular collection, aged 4 to 7 years, weighing 320 to 450 kg, and with body condition score 4 to 4.5 and scrotal circumference 23 to 32 cm. Semen was evaluated routinely and sperm morphology was viewed under light microscope at ×1,000 magnification after fixing with buffered formal saline. Over 90% had normal head, acrosome, mid-piece and tail. Semen was diluted with egg-yolk-citrate extender to produce 15×106 spermatozoa/ml and 0, 1, 2, 4 and 6 mg/ml α-tocopherol were added. The semens amples were kept at 8℃. Sperm motility and viability were examined daily up to 5 days under light microscopy at ×200 magnification. Sperm viability was acceptable (≥40%) up to the 4th day with all concentrations of α-tocopherol and up to the 5th day with 2 mg/ml α-tocopherol. Sperm motility was acceptable (≥40%) up to the 3rd day irrespective of α-tocopherol concentration, and up to the 4th day with 2 mg/ml α-tocopherol. It is suggested that the lifespan of chilled semen may be extended up to 4 days by adding 2mg/ml α-tocopherol.

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.

This study was carried out to investigate the general characteristics, such as volume, pH, sperm motility and sperm concentration of the semen collected from Beagle dogs (age 24～48 months, weight 10～15 kg) by using the method of digital manipulation of the penis, and the effect of preservation temperature and time on motility of fresh semen. Multiple ejaculates were collected from 4 male Beagles. The average volume, pH, motility and sperm concentration of the second fraction (contained with small volume of the third fraction) per ejaculation were 2.94±0.24(SD) ml, 6.43±0.42(SD), 97.04±3.50(SD)% and 1.67±0.23(SD)×108 cells/ml, respectively. Average semen volume per ejaculate, semen pH, sperm motility and sperm concentration of the first fraction from the ejaculate were 1.24±0.20(SD) ml, 6.03±0.26(SD), 11.30±4.02(SD)% and 7.25±1.02(SD)×105 cells/ml. Those of second fraction were 2.52± 0.32(SD) ml, 6.32±0.31(SD), 96.25±3.52(SD)% and 2.35±0.35(SD)×108 cells/ml. Those of third fraction were 2.71±0.27 (SD) ml, 6.52±0.20(SD), 95.65±2.78(SD)% and 5.72±0.29(SD)×107 cells/ml. Motility of semen was higher at 17℃ preservation temperature than 5℃ or 36℃ during preservation period. When preservation temperature was 17℃, motility was 96.54±2.05(SD)% at 1 h, 90.20±3.90(SD)% at 6 h, 89.05±2.01(SD)% at 12 h, 78.21±3.50(SD)% at 18 h, 45.24±6.25 (SD)% at 24 h and 30.75±17.24(SD)% at 30 h, respectively.

본 연구는 항생제가 첨가되지 않은 돼지 혼합액상 정액을 17 정액 보관고에 보관하면서 보관일수의 증가가 따라 정자의 운동성, 정액 내 세균의 증식 여부 및 체외 수정란 생산 효율에 미치는 영향을 조사하고자 하였다. 정자의 운동성은 1일 (78.72.4%)에 비하여 3일(78.72.4%)과 5일째(64.82.4%)는 유의적으로(p<0.05) 낮은 운동성을 나타내었다. 보관일수에 따른 정액 내 세균수의 변화는 보관 5일이 Cfu로 0일과 3일의 Cfu와 C

Oxidative stress is one of the major causes of failure in in vitro storage of boar semen. Reactive oxygen species (ROS) are known to be important mediators of such stress. The present study examined the effects of pyruvate and taurine on sperm motility and expression of BAD, Cytochrome c, Caspase-3 and Cox-2 protein in in vitro storage of boar semen, and tested the effect of semen treated with antioxidant with or without hydrogen peroxide on the development of IVM/IVF porcine embryos. Semen samples were transported to the laboratory at 17℃ within 2 hr after collection and were treated with different concentration of pyruvate (1~10mM) and taurine (25~100mM) with or without 250uM H2O2 respectively. The supplementation of pyruvate and taurine increased sperm motility in boar semen during in vitro incubation at 37℃. Expression of apoptosis protein (BAD, cytochrome c, caspase-3 and cox-2) were reduced in the group of boar semen treated with pyruvate and taurine when compared to the other groups. The developmental rates of IVM/IVF porcine embryos fertilized by semen treated with pyruvate and taurine were significantly increased when compared to control (P<0.005). These results indicate that supplementation of pyruvate and taurine as antioxidants in boar semen extender can improve the semen quality and increase in vitro development of porcine IVM/IVF embryos when boar semen treated with antioxidants was used for in vitro fertilization.

한우의 유전자원을 보존하기 위하여 정액동결 시험을 수행하였으며 고급육계통은 육질에 대한 육종가 상위 10% 이내의 자손에서 선발하였고 다 유계통은 어미의 이유시 체중에 대한 육종가 상위 10%이내의 자손에서 선발하였으며 2개 계통에서 종모우 총 13두를 선발하여 공시하였다. 정액채취는 인공질법으로 실시하였고 의빈대에 수소를 계류하고 채정대상우를 승가시켰으며, 3회 가승가후에 채정하였다. 채정후 10분 이내에 실험실로 옮겨와 검사항목을 조사하고 37에서

본 연구는 돼지 액상정액을 인공수정용 100ml 플라스틱 병에 보존하면서 BF5희석액과 Butschwiler 희석액 간에 보존 온도별 차이를 조사하고, BF5 희석액에서의 글리세롤 농도의 효과를 조사하여 돼지 액상정액을 좀더 장기간 사용할 수 있는 방법을 찾고자 실시하였다. 돼지 액상정액을 5 냉장고에 보존하면서 조사한 바에 의하면, 37에서 0.5 및 2시간 배양후의 정자운동성은 전체 보존기간동안 BF5 희석액이 Butschwiler 희석액보다 유의하