So, Kyoung-Min;Sa, Soo-Jin;Kim, Hyo-Jin;Chung, Ki-Hwa;Jung, Byeong-Yeal;Son, Jung-Ho;Kim, In-Cheul
Reproductive and Developmental Biology
/
v.35
no.4
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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.
Lee S. H.;Kim T. S.;Cheong H. T.;Yang B. K.;Kim C. I.;Park C. K.
Reproductive and Developmental Biology
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v.28
no.4
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pp.261-265
/
2004
The present study was conducted to assess sperm characteristics in miniature-pig. The semen samples were transported to the laboratory at 17℃ within 3 hours after collection. The extended semen was stored at 17℃, and sperm quality was evaluated at 0, 1, 3, 5 and 7 days after storage. The semen volume of miniature-pig (62±22㎖) was significantly (p<0.05) lower than that of Duroc (155±25㎖) and Yorkshire (154±23㎖). Significant differences were also observed in sperm concentrations. During 3 days of storage, sperm viability did not differ among miniature-pig, Duroc and Yorkshire. However, the viability was significantly (p<0.05) lower in miniature-pig than in Duroc and Yorkshire semen after Day 3 of storage. In abnormality, acrosome intactness and intensity, there were no differences among miniature-pig, Duroc and Yorkshire semen. On the other hand, the viability of frozen-thawed sperm in miniature-pig was significantly (p<0.05) lower than in that of Duroc and Yorkshire. This study also examined CTC patterns in frozen-thawed spermatozoa. The rates of AR pattern were higher in miniature-pig than in Duroc and Yorkshire. However, no difference was found in F, B and AR patterns. The results of present study suggest that further research is necessary to develop of semen extender and freezing methods to improve sperm quality in miniature-pig.
Oxidative stress is a critical factor affecting the quality and viability of sperm during boar semen storage. Oxidative stress is also a significant concern during the process of freezing semen. The process of semen storage involves exposing the sperm to various stressors, including temperature changes, cryoprotectants, and extended periods of incubation. In addition, oxidative stress can lead to the production of reactive oxygen species (ROS) within the sperm, resulting in oxidative damage to cellular components, such as lipids, proteins, and DNA. Striking a balance between ROS production and the antioxidant defense system is crucial for maintaining sperm viability and functionality during semen storage. Moreover, the prolonged storage of boar semen leads to an increase in ROS levels, which can impair sperm motility, membrane integrity, and DNA integrity. ROS-induced lipid peroxidation affects the fluidity and stability of sperm membranes, leading to decreased sperm motility. Moreover, oxidative damage to the DNA can result in DNA fragmentation, compromising the genetic integrity of the sperm. In conclusion, oxidative stress is a significant challenge in maintaining sperm quality during boar semen storage. Understanding the mechanisms underlying oxidative stress and their impacts on sperm function is crucial for developing effective strategies to minimize oxidative damage and improve sperm storage outcomes.
Bacteroiospermia is a frequently finding in fresh raw and extended porcine semen and can results in detrimental effects on semen quality and longevity. This study aims to evaluate the type of bacterial contaminants in raw and extended porcine semen and the reducing effect of antibiotic test. To investigate bacterial contaminants, out of 387 sample (raw semen 201, extended semen 186) were collected from 6 artifical insemination centers in Gyeongsangnam-do, were inoculated onto blood agar and MacKonkey agar, respectively. Bacterial colonies were selected after culturing for 48 hours, at $37^{\circ}C$, followed by Gram staining, KOH test, oxidase test, catalase test and eventually identified using VITEK System. Total 15 genus and 24 species of bacteria were isolated from these semen samlpes. In raw semen, the most prevalent contaminants were Pseudomonas aeruginosa, Escherichia coli, Proteus mirabilis, Staphylococcus auricularis, Delftia acidovorans, Acinetobacter lowffii, S. aureus and others. And in extended porcine semen, A. lowffii, S. aureus, S. auricularis and other bacteria were identified. Most of them was G(-), which is nonpathogenic bacteria. It seems that bacterial contaminants in fresh raw and extended porcine semen originated from multiple sources at the farms/stud, and were from animal origin and non-animal origins. Whereas, the 7 virus which is known to be detected in porcine semen in 75 cases was not detected. This results showed that removal of bacterial contamination in raw and extended porcine semen is essential and farms were kept for biosecurity and individual hygienes.
The purpose of this study was to select high-quality boar semen after the glass wool filtration of extended boar semen. After collecting boar semen, its concentration, morphology, viability, and motility were examined according the glass wool's height and time. After glass wool filtration, the sperm concentration decreased, but the proportion of normal sperms and the sperm viability increased. Nevertheless, the sperm motility showed no changes. The above results showed that the glass wool filtration of boar semen is a method of obtaining sperms with relatively low abnormal rates and high viabilities.
A study was conducted on four crossbred bulls, used as artificial insemination (AI) sires, to correlate their semen quality with their non return rate (NRR). Semen was collected once a week via an artificial vagina, diluted in egg yolk-citrate and maintained at $+7^{\circ}C$ for three days. It was evaluated for sperm motility, viability, morphology immediately after collection and was examined daily for sperm motility, viability and morphology of acrosome, mid piece and tail for a total of three days. A total of 2016 cows were inseminated by two AI technicians. The proportions of sperm with normal heads were 83.4% (63.7~91.7%), the proportion of spermatozoa exhibiting normal morphology (acrosome, mid piece and tail), motility and viability were 89.2% (82.3~92.0%), 71.3% (61.7~75.0%) and 76.7% (65.7~85.0%), respectively in fresh ejaculates. Sperm motility and sperm viability was significantly ($p$ <0.05) lower in Holstein-Friesian ${\times}$ Local bull than in other bulls during all three days of storage. The overall NRR for four bulls was 82.7% (72.9-87.5%). Bulls with higher sperm motility, viability and normal morphology of spermatozoa of individual bull had significantly (each $p$ <0.05) higher NRR. The highest ($p$ <0.01) NRR (87.5%) was observed in a Red Chittagong bull whose semen qualities were significantly ($p$ <0.05) higher than Holstein-Friesian ${\times}$ Local bull (NNR 72.9%). The results of the present study concluded that NRR at 56 days post AI is related to parameters of semen quality. Therefore, semen evaluation may allow the discarding of bulls with poor fertility in an AI program.
The study had two objectives, namely, to estimate the andrological disorders leading to disposal of Karan Fries (KF), Sahiwal cattle and Murrah buffalo bulls and to study the effect of various factors (species/breeds, season of birth and period of birth) on male reproductive parameters. Records on occurrence of subfertility problems and disposal pattern of bulls maintained at the National Dairy Research Institute herd were collected for 15 years (1991 to 2005). Percentage of bulls producing freezable semen was less in the crossbred cattle (58.46%) as compared to Sahiwal (81.69%) and Murrah bulls (81.05%). Various subfertility traits like poor libido and unacceptable seminal profile were found to be the significant reasons (p<0.01) for culling of the breeding bulls. Inadequate sex drive was the main contributing factor for bull disposal in Sahiwal (22.55%) and Murrah bulls (15.12%) whereas poor semen quality and freezability were most frequently observed in KF bulls (24.29 and 7.29 percent, respectively). Least squares analyses of different male reproductive parameters showed that species/breeds had significant effect (p<0.05) on all traits except for frozen semen production periods (FSPP). Periods of birth were significantly different (p<0.05) for all traits except for semen volume. Age at first semen collection (AFSC), age at first semen freezing (AFSF) and age at disposal (AD) were highest in Murrah, while frozen semen production period (FSPP) and semen production period (SPP) were highest in KF and lowest in Sahiwal. The age at first semen donation and breeding period could be reduced by introducing the bulls to training at an early age. These results revealed a declining trend in AFSC, AFSF, FSPP, SPP and AD, thereby indicating an improvement in reproductive performance over the years. The age at first semen donation in bovines can be reduced by introducing the young male calves to training at an early age, which could increase the dosage of semen obtained from each male.
Jang H. Y.;Cheong H. T.;Kim C. I.;Park C. K.;Yang B. K.
Reproductive and Developmental Biology
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v.29
no.2
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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.
In order to protect the spermatozoa against cold shock, hen egg yolk is widely used as a cryoprotective agent in semen freezing extenders for domestic animals. The protective action of yolk is largely presumed to be due to low density lipoproteins (LDL). The effects of LDL on sperm quality of bull and northern pike (Esox lucius) after freezing-thawing have been reported, but no study has been made to evaluate the effect of LDL on boar sperm motility and other characteristics. The experiment was carried out to investigate the effect of LDL on the freezing of boar sperm in 0.25 ml straws. The aim was to evaluate the quality of boar spermatozoa cryopreserved in the presence of LDL. Motility of semen cryopreserved in LDL was analyzed and compared to semen cryopreserved with Tris-citric acid-glucose (TCG) and Tris-citric acid-fructose (TCF), two basic freezing extenders containing egg yolk. Similarly, acrosome and plasma membrane integrity were also evaluated and compared to semen cryopreserved with TCG and TCF. Analysis of sperm quality after freeze-thaw showed that the motility, acrosome and plasma membrane integrity were improved with LDL in the extender, as compared to the TCG and TCF. The highest post-thaw integrity of acrosome and plasma membrane and motility were obtained with 9% LDL (w/v). Consequently, the optimum LDL concentration in the extender was 9%. It is also suggested that the concentration of LDL addition is important for the effect on boar sperm protection during freezing and thawing. The percentage of motile spermatozoa was significantly higher after freezing in 9% LDL than in TCG and TCF 54.4% versus 30.4% and 30.1% (p<0.05), respectively. The integrity of acrosome and plasma membrane were also significantly higher at 70.3% and 50.5% respectively with semen frozen in 9% LDL extender compared to TCG at 37.8% and 30.3% and TCF at 36.4% and 29.9%, respectively (p<0.05),. In conclusion, we propose that extender containing LDL extracted from hen egg yolk could be used as a cryoprotective media with a better efficiency than TCG and TCF. LDL improved boar semen quality, allowing better spermatozoa motility, acrosome and plasma membrane integrity after the freeze-thaw process. Furthermore, we found out that the extender with 9% LDL concentration significantly enhanced motility, acrosome and plasma membrane integrity of boar sperm after freezing and thawing.
Park, Yong-Seog;Lee, Sun-Hee;Han, Sang-Chul;Koong, Mi-Kyoung;Kim, Jong-Woo;Seo, Ju-Tae
Clinical and Experimental Reproductive Medicine
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v.30
no.2
/
pp.165-169
/
2003
Objective: To evaluate the results of CASA systems and to compare its results. Methods: Fifty semen sampales were analysed. Concentration, motility and forward progression were evaluated simultaneously on the same semen samples using Cell Soft System-3000 (CS system) and Sperm Quality Analyzer-V (SQA system). Results: Mean semen volume was $2.8{\pm}1.2\;ml$. Mean value of sperm concentration, motility, forward progression using CS system were $83.4{\pm}45.7{\times}10^6/ml$, $52.3{\pm}16.4%$ and $48.6{\pm}13.4%$, respectively. And mean value of sperm concentration, motility, forward progression using SQA system were $78.2{\pm}42.9{\times}10^6/ml$, $57.0{\pm}24.0%$ and $50.6{\pm}21.9%$, respectively. There were no statistical significancy of sperm concentration, motility, forward progression between the two devices. Conclusion: SQA system variables well correlated with the CS system. As a screening test for semen quality, CS system and SQA system is considered as useful in the management of male infertility.
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