• 한국임상수의학회지
• /
• 제18권4호
• /
• pp.345-349
• /
• 2001

The aim of this study was to investigate the effect of sperm preservation according to the various kinds of commercially available semen extenders and to investigate the effect of sperm preservation according to the various temperature storages of fresh-extended porcine semen. To investigate the effect of sperm preservation according to the various kinds of commercially available semen extenders, porcine semens diluted in 3 semen extenders, Beltsville Thawing Solution(BTS), Androhep and Kiev, were cooled at $8^{\circ}C$ storage temperature with a controlled cooling rate of $2-4^{\circ}C$/h. Motility, progressive motility, normal sperm(%) and sperm morphology were assessed comparatively. In motility and progressive motility, Androhep extenders revealed better result than other extenders. In normal sperm(%) and sperm morphology, 3 semen extenders revealed similar results. To investigate the effect of sperm preservation according to the various temperature storages of fresh-extended porcine semen, porcine semens diluted in BTS extender, were cooled at 3 storage temperatures($8^{\circ}C$, $12^{\circ}C$ and $17^{\circ}C$) with a controlled cooling rate of $2-4^{\circ}C$/h. Motility, progressive motility, normal sperm(%) and sperm morphology were assessed comparatively. In motility and progressive motility, $8^{\circ}C$ treatment group revealed better result than $12^{\circ}C$ and $17^{\circ}C$ treatment groups. In normal sperm(%) and sperm morphology, 3 temperatures of treatment groups revealed similar results.

• Reproductive and Developmental Biology
• /
• 제39권1호
• /
• pp.7-11
• /
• 2015

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 $3{\times}10^7/mL$ 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.

• Reproductive and Developmental Biology
• /
• 제29권2호
• /
• pp.133-139
• /
• 2005

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 $17^{\circ}C$ within 2 hr after collection and were treated with different concentration of pyruvate $(1\~10mM)$ and taurine $(25\~100mM)$ with or without 250uM $H_2O_2$ respectively. The supplementation of pyruvate and taurine increased sperm motility in boar semen during in vitro incubation at $37^{\circ}C$. 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.

• Reproductive and Developmental Biology
• /
• 제35권4호
• /
• pp.479-483
• /
• 2011

The objective of this study was to determine the effects of E. coli isolated from porcine semen on sperm viability, motility, and semen pH. Semen samples were prepared using commercial extender, $Seminark^{Pro}$ (Noahbio Tech, Korea) that did not contain antibiotics. And 4 different levels of E. coli were artificially innoculated to semen with following concentrations; 4,000 of sperms with 1 of E. coli (T1), 400 with 1 (T2), 40 with 1 (T3), and 4 with 1 (T4). Semen samples were preserved at $17^{\circ}C$ for 5 days in semen storage box until analyzed by flowcytometer. Aliquots were subjected to measure the sperm viability (Live/$Dead^{(R)}$ stain), motility (mitochondrial function), and semen acidity (pH) from day 0 (day of semen collection) to day 5. Sperm motility and viability were significantly decreased (p<0.05) on day 0 (4 hrs after preservation at $17^{\circ}C$) in T3 and T4 compared to control groups and were significantly decreased (p<0.05) in all groups from day 3. Sample pH was acidic in T3 (6.90~6.86) and T4 (6.86~6.65) from day 3 to day 5 (p<0.05). On the other hand, sample pH was maintained 7.0~7.1 in control, T1, and T2 during the experimental period. Sperm motility and viability were significantly decreased from day 0 to day 5 compared to control in samples contaminated with E. coli above a value of 40:1 ($20{\times}10^6$ sperm cells/ml : $5{\times}10^5$ cfu/ml). Even on day 1 in T4 and on day 3 in T3, semen pH was acidic probably due to the acidification of dead spermatozoa. These results suggest that E. coli contamination has a concentration-dependent detrimental effect on extended porcine semen quality.

• 한국수정란이식학회지
• /
• 제25권4호
• /
• pp.267-272
• /
• 2010

The present study was to investigate the source of contamination during semen processing for in vitro uses. In the present study, frozen semen was prepared from liquid semen in our laboratory for in vitro fertilization (IVF) experiments due to lack of fresh semen. Antibiotics were added in the frozen semen extender (kanamycin and gentamicin) and in vitro culture (IVC) medium (gentamicin) for further inhibiting growth of microorganisms. Nevertheless, proliferations of microorganisms were observed in IVC culture drop during culturing of IVF embryos using frozen semen. Randomly 3 samples were taken from the liquid semen, frozen semen and egg yolk. Contaminated IVC medium, frozen-thawed semen, liquid semen and egg yolk were cultured in de Man, Rogosa and Sharpe (MRS) agar medium. Whitish colonies were detected in contaminated IVC drop, frozen-thawed semen samples and egg yolk but no colonies were formed in liquid semen samples. Gram-negative and rod-shaped identical bacteria were found in both frozen-thawed semen sample and contaminated IVC drop and egg yolk samples. Enterobacter cloacae were confirmed by API 20E kit according to manufacturer's instruction with identification value (% ID) 94.3% and T index 0.88. Antibiotic susceptibility tests were done according to Clinical and Laboratory Standards Institute (CLSI) by using ampicillin, amikacin, cephalothin, gentamicin, kanamycin, tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin test. Among them Enterobacter cloacae were resistant to ampicillin, amikacin, cephalothin, gentamicin, kanamycin but susceptible to tetracycline, oxytetracycline, sulfamethoxazole trimethoprim, norfloxacin and ciprofloxacin. From these findings it could be suggested that this contamination sources might be from egg yolk.

• Asian-Australasian Journal of Animal Sciences
• /
• 제21권3호
• /
• pp.340-345
• /
• 2008

Oxidative stress is one of the major causes of failure of in vitro storage of boar semen. Reactive oxygen species (ROS) are one of the important mediators of oxidative stress during in vitro storage of boar semen. Our study examined the effects of taurine on sperm characteristic and on in vitro developmental embryos during in vitro storage of boar semen for 7 days. Semen was randomly aliquoted into 3 centrifuge tubes and treated with different concentrations of taurine (25-100 mM). The characteristics of boar sperm were analyzed for motility by light microscopy, viability by using a Makler counting chamber and membrane integrity by a hypoosmotic swelling test (HOST). The percentages of motile spermatozoa in taurine groups after 5 days were significantly higher compared to the control. Sperm viability in the control was lower than in taurine groups after 7 days irrespective of different taurine concentration. In the hyoosmotic swelling test (HOST), significantly higher results were obtained in taurine groups after 3 days. Also, the developmental rates of IVM/IVF porcine embryos from semen treated with pyruvate and taurine were significantly increased when compared with the control (p<0.05). These results indicate that supplementation of taurine as an antioxidant in boar semen extender can improve the semen quality.

• Reproductive and Developmental Biology
• /
• 제29권3호
• /
• pp.155-162
• /
• 2005

본 연구는 돼지 정자 동결시 taurine과 $\alpha$-tocopherol의 첨가가 융해후 정자 성상과 정자 기능 활성산소계(reactive oxygen species; ROS)의 발생 정도 및 지질 산화(lipid peroxidation, LPO)에 미치는 영향을 구명하기 위하여 시행하였다. 1차 및 2차 희석액내 taurine과 $\alpha$-tocopherol이 첨가된 돼지 동결정액의 융해 후 정자 운동성 양상, 정자 생존성, 정자 기법을 적용하여 다음과 같은 결과를 얻었다. Taurine(25mM, 50mM), $\alpha$-tocopherol($500{\mu}M,\;1,000{\mu}M$ 단일처리군 그리고 taurine과 $\alpha$-tocopherol의 혼합처리군($25mM\~500{\mu}M\~1,000{\mu}M$)은 동결$\cdot$융해 후 대조군과 비해 정자 운동성과 생존성은 유의적인 차이를 나타내지 않았다. Taurine과 $\alpha$-tocopherol의 혼합처리군 $50mM\~1,000{\mu}M$에서 HOST, 점체 반응이 대조군과 비교하여 유의차는 인정되지 않았으나 다소 증가하였다. 동결$\cdot$융해 정자의 ROS 발생 억제를 위한 taurine과 $\alpha$-tocopherol의 모든 처리군은 대조군과 비교하여$\cdot\{O_2}{^-}$ 발생을 유의적으로 완화시키지 못하였다. 그러나 taurine 단일처리군(25mM), $\alpha$-tocopherol 단일처리군($50mM,\;1,000{\mu}M$)과 혼합처리군($25mM\~500mM,\;50mM\~1,000{\mu}M$은 $H_{2}O_{2}$의 발생을 대조군과 비교하여 유의적으로 완화시켰다(P<0.05). 동결$\cdot$융해 정자의 malondialdehyde의 생산은 taurine과 $\alpha$-tocopherol의 모든 처리군에서 대조군과 비교하여 유의적인 감소를 보였다(P<0.05). 이상의 결과를 종합해 보면 돼지 정액의 동결보존시 taurine와 $\alpha$-tocopherol 같은 항산화제의 처리는 ROS 발생과 LPO을 효과적으로 완화시킴에 따라 돼지 정액의 동결보존 효율을 증진시킬 수 있는 유용한 방법이라 사료된다.