• Reproductive and Developmental Biology
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• 제36권3호
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• pp.163-166
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• 2012

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(Beltsville Thawing Solution) 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 $10{\sim}10^6$ 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 F), characteristic of incapacitated 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 AR), 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 ($77.24{\pm}6.47$, p<0.001) and 7 days ($77.24{\pm}6.47$, p<0.001) after preservation compared to 1 ($15.71{\pm}7.18$) and 3 days($18.39{\pm}7.22$) after preservation, respectively. Sperm viability was significantly lower ($53.25{\pm}35.03$, p<0.0001) at 7 days after preservation. Morphological abnormality of sperm was lower (p<0.001) at 1 ($15.71{\pm}7.18$) and 3 ($18.39{\pm}7.22$) days compared to 5 ($21.84{\pm}7.91$) and 7 ($22.59{\pm}9.93$) days after preservation. Acrosomal integrity and capacitation rate (pattern F) were significantly lower (p<0.001) from 5 days after preservation. Based on the data we obtained from this study suggested that semen preserved more than 5 days without antibiotic would not recommend use for artificial insemination.

• Clinical and Experimental Reproductive Medicine
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• 제23권3호
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• pp.311-318
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• 1996

Mycoplasmas have long been suspected of contributing to involuntary infertility in couples. However considerable disagreement exits concerning the role of genital mycoplasma infection in human infertility. Several investigators have noted abnormalities in the semen analysis of men with positive mycoplasma cultures, and early epidemiologic studies indicated that Ureaplasma urealyticum was linked to human reproductive failure on the basis of higher frequencies of isolation from infertile versus fertile couples and successful pregnancies in infertile couples after doxycycline therapy. However, subsequent investigators have questioned these findings because there are many studies in which treatment for mycoplasma in the male or female did not demonstrate an improved pregnancy rate, and semen samples from unexplained infertile men containing ureaplasmas have not revealed poorer motility, fewer spermatozoa and more aberrant forms. The objective of this study were to investigate the incidence rate of mycoplasma in semen and to investigate whether the presence of mycoplasma in semen makes significant difference to the semen volume, sperm motility and sperm counts. The results were that the rate of isolation of mycoplasma species was 70.3%. Semen volume is $2.84{\pm}1.01ml$ for culture negative and $3.15{\pm}1.42ml$ for culture positive group. Sperm motility is $46.23{\pm}15.80%$ for culture negative and $50.09{\pm}15.69%$ for culture positive group, and sperm count is $95.47{\pm}47.14({\times}(P)10^6/ml)$ for culture negative and $86.73{\pm}47.59({\times}10^6/ml)$ for culture positive group. In conclusion, we suggest that the presence of mycoplasma in semen makes no significant differences to the sperm parameters.

• 한국가축번식학회지
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• 제8권2호
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• pp.79-86
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• 1984

This cepseiment was carried out to cerify the effect of thawing methods and preservative temperature on the sperm motility and fertility after thawing semen with plastic straws in fresh and warm water. Sperm motility in vitro stored at room temperature after thawing were conducted by the various storage hours. A field trial after thawing semen with warmed water in straws from Cheju native cows involving 4 technicians and 800 cows first (or second) services gave the following results. The thawing methods of warmed water for one minute in semen motility were considerably higher than that in iced water during 12 hours after thawing semen, however, the sperm survival index of ice-water shwed a better results according as the time passed away, but not significant differences. Preservative temperaure at 5$^{\circ}C$ (iced water) after thawing gave significantly better results than that of thawed at 3$0^{\circ}C$ (warmed water). The N R rate to 175 inseminations with semen thawed at 15-2$0^{\circ}C$ (fresh water) was 82.8%, 80.9% for 610 inseminations thawed in warm water. Conception rate ofthe semen thawed in warm water for 10-60 secs gave no significant difference among storage hours, because the semen used to be inseminated within one hour almost, but in decreased when semen thawed at the period of one minute over.

• 한국가축번식학회지
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• 제25권3호
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• pp.201-206
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• 2001

본 연구는 요크샤 종모돈의 정액성상, 동결-융해후의 정자생존성 및 혈청 중 테스토스테론의 농도에 미치는 영향을 봄 (3~5월)과 여름 (6~8월)으로 나누어 실시한 바, 그 얻어진 결과는 다음과 같았다. 1. 요크샤 종모돈의 정액량, pH, 희박정자 부분의 정자농도는 봄과 여름철에 큰 차이가 없었다. 그러나 봄철 농후정자 부분의 정자농도는 여름철 농후정자 부분의 정자농도보다 높았다 (P＜0.05). 2. 원정액 정자에서는 운동성 및 정상첨체에서 봄과 여름철간에 차이가 없었다. 그러나 동결-융해정자의 운동성 및 정상첨체는 봄철의 종모돈에서 생산한 정자가 여름철의 종모돈에서 생산한 정자보다 우수하였다. 3. 요크샤 종모돈의 혈청 중 테스토스테론의 농도 변화는 봄과 여름에 각각 4.04 ng/$m\ell$과 2.85ng/$m\ell$l으로, 봄철 테스초스테론의 농도가 여름철의 농도보다 높았다(P＜0.05). 4. 이상의 결과를 종합해 보면, 요크샤 종모돈에 있어 정자의 내동성은 혈청 중 테스토스테론의 농도가 높을수록 우수한 것으로 나타났다.

• 대한수의학회지
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• 제40권2호
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• pp.403-414
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• 2000

To improve the semen production, the selenium(Se) and vitamin E(Vit. E) were administrated into Hanwoo young sire for intensification an antioxidant system and the taurine were supplemented into semen extender for improving the semen characteristics. The 16 heads ranging from twenty to thirty two months of age were randomly assigned to control group, Se-admistrated group(Se-group), Vit. E-administrated group(Vit. E-Group) and Se and Vit. E administrated group(Se and Vit. E-group). Se and Vit. E dministrated 3 times every 30 days by intramuscular injection. The administration of Se, Vit. E, and Se and Vit. E didn't affect on semen volume, sperm concentration, and total sperm number among all groups(p>0.05). Adiministration of Se improved sperm motility and viability. Motility of Se-group and control were 26.01% and 19.20%, respectively(p<0.05). Viability of Se-group(47.07%) was higher than control group(35.73%), Vit. E group(36.55%)(p<0.05). The administration of Se and Vit. E didn't affect sperm capacitation and acrosome reaction. The 100mM taurine supplement into semen extender increased the motility of frozen/thawed semen in the Vit. E-group(p<0.05) and had a beneficial effects on decreasing abnormality of frozen/thawed semen in all groups(p<0.05). These results indicate that Se administration improve sperm motility and viability, and the taurine supplement into semen extender decrease abnormality in Hanwoo young sire.

• 한국임상수의학회지
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• 제24권4호
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• pp.577-583
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• 2007

It is important to obtain semen with good quality for efficient fertilization and pregnancy. To obtain these semen, various methods have been developed but most of these methods are time consuming and require costly equipment. Therefore, the objective of this research is to investigate the usability of column filtration system as quick and simple method to get sperm with better quality. Ejaculates were obtained from 5 dogs and analyzed with basic quality parameters before each filtration. Sperm concentration was adjusted to $5{\times}10^7/ml$ after dilution. The experimental groups were divided into non-filtered group(control) and filtered groups(glass wool, Sephadex 5% and Sephadex 20%). Ejaculates were filtered through each filter system and assessed by recovery rate of sperm, motility, normal morphology, CFDA/PI stain and plasma membrane integrity(hypo-osmotic swelling test, HOST). The lowest recovery rate of spermatozoa was recorded in glass wool filtration group, followed by 20% Sephadex filtration group(p<0.05). There was no significant difference between control(non-filtered) and 5% Sephadex filtration poop. Also, there was no significant difference of sperm motility assessed under light microscope among experimental groups. Morphological normality of canine spermatozoa was the highest in the glass wool filtration group and the lowest in the 5% Sephadex filtration group with no significant differences versus 20% Sephadex filtration and control group, respectively(p<0.05). Viability of canine sperm assessed by CFCA/PI staining was the highest in the glass wool filtration poop with no significant difference versus the control group, and the lowest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, respectively(p<0.05). HOS values of canine sperm was the highest in the 20% Sephadex filtration group with no significant difference versus 5% Sephadex filtration group, and the lowest in the control poop with no significant difference versus glass wool filtration group, respectively(p<0.05). Therefore, these results indicated that filtration treatment for extended canine sperm would be useful method to get sperm with better quality by trapping the damaged sperm, consequently filter would be physical barrier against injured or immotile sperm.

• 한국수정란이식학회지
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• 제15권1호
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• pp.85-94
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• 2000

These experiments were conducted to examine the effects of theophylline, pentoxifylline and heparin on frozen-thawed Hanwoo sperm for enhancing motility and viability of sperm. Frozen-thawed semen collected from one bull was treated in TALP(tyrode-albuminlactate-pyruvate) containing varous concentrations of theophylline and pentoxifylline. After incubated at 5% CO2 in air atmosphere for 6 hours, the motility of sperm after the treatments was characterized by CASA(computer aided semen analysis) system. When monitored notility(MOT) and curvilinear velocity(VCL), theophylline and pentoxifylline exerted their optimal action at the concentration of 30 mM and 3 mM, respectively. No difference of sperm motility was observed when the sperm was treated with both substances compared with a single treatment of each substance. Comparison was then made for evaluating the effect of theophylline and / or pentoxiophylline on the motility and viability of significant treatment effects of each substance, high MOT and VCL values were detected in sperm treated with theophylline. In the case of sperm viability examined by an eosin-nigrosin staining, however, a significant decrease was found after the combined treatment of theophylline+pentoxyphilline than after the treatment with heparin alone or no treatment(P<0.05). In conclusion, theophylline, pentoxiphylline or heparin can be used for enhancing the motional characteristics and viability of frozen thawed Hanwoo semen. Considering characteristics of these substances, theophyline may be useful in the artificial insemination system, which requires vigorous sperm motility. While, heparin supporting sperm viability in vitro can be effectively used for improving in vitro-fertilization system.

• Clinical and Experimental Reproductive Medicine
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• 제29권2호
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• pp.149-157
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• 2002

Objectives: To assess the scavenging effect of carnitine derivatives on oxidative damage to sperm during sperm processing, cryopreservation and thawing. Materials and Methods: Fresh semen samples from 20 normal healthy volunteers were collected by masturbation after at least 48 hours abstinence. After liquefaction of semen samples at room temperature, the specimens were diluted with sperm wash media (Ham's F-10, Life technologics) to a uniform density of $20{\times}10^6/ml$. L-carnitine or acetylcarnitine were added with various concentration of $0{\mu}M$, $10{\mu}M$, $30{\mu}M$ in semen sample or cryoprotectant. All specimens were cryopreservated at $-196^{circ}C$ $LN_2$ for 3 days. Sperm motility, vitality, fertilizing capacity, reactive oxygen species formation and the level of lipid peroxidation were analyzed by computer assisted semen analyzer, eosin-nigrosin stain, hypoosmotic swelling test, chemiluminescence and thiobarbituric acid method, respectively, during sperm processing, cryopreservation and thawing. Results: The sperm motility was only increased in proportion to the concentration of acetylcarnitine with no statistical significance (p>0.05). The sperm vitality was also significantly improved in proportion to the concentration of acetylcarnitine with statistical significance (p<0.05). The sperm fertilizing capacity was significantly increased in proportion to the concentration of L-carnitine and acetylcarnitine and reactive oxygen species generation and lipid peroxidation were significantly decreased with same fashion (p<0.05). On comparison of effects between L-carnitine and acetylcarnitine, acetylcarnitine was superior to L-carnitine on the improvement of sperm motility and vitality as well as the suppression of reactive oxygen species generation and lipid peroxidation. Conclusions: These results suggest that carnitine derivatives have a scavenging effect against oxidative damages during sperm processing, cryopreservation and thawing. Therefore, carnitine derivatives may be useful as an oral antioxidant in patients with male infertility due to increased ROS generation.

• 한국가축번식학회지
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• 제3권1호
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• pp.30-35
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• 1979

A, B and C dilutors were used to make Ka (A plus B (1 : 1)) and Na (B plus C(1 : 1)) dilutors in this experiment. Three aliqots of semen were respectivly diluted 1 : 1 and 1 : 2 (semen: dilutor) with Ka, Na and C dilutors and stored at 5$^{\circ}C$ for 7 days in order to study their livability during storage. Fertility was checked for the diluted semen with Ka, Na and C dilutors. Whole semen and extended semen with Na dilutos with and without DMSO were cold shocked at various temperatures for 10 min. Effects of different 1st and 2nd dilution with A, B, C and Na dilutors and of vessels on freezability of spermatozoa were investigtigated. 1. Extended semen 1 : 2 with Na and C dilutors showed highest live sperm index during storage for 7 days at 5$^{\circ}C$. 2. The components of Na dilutor per 100$m\ell$ were skim milk 2.5g, trisaminomethane 0.54g, citric acid 0.265g, glucose 2.835g, fructose 1.5g, sodium lauryl sulfate, 0.08g, penicillin 0.06g, streptomycin 0.075g, and egg yolk 10$m\ell$. 3. Fertility of diluted semen was higher than that of whole semen. Ka dilutor showed higher fertility than Na and C dilutors, and there was no difference in the fertility between Na and C dilutors. 4. Na dilutor with DMSO showed slightly higher livability than Na dilutor without DMSO during storage for 7 days at 5$^{\circ}C$. 5. Cold shock at 1$0^{\circ}C$ for 10 min. decreased greatly the sperm livalility of whole semen but not of extended semen with Na dilutor. Addition of DMSO to Na dilutor has no effect in prevention of cold shock. 6. The extended semen with C. C dilutor (1st and 2nd dilution with C and C dilutor) showed higher post-thawing sperm livability than A.A and Na. B dilutors. Na. B dilution shwed higher post-thawing sperm livability than A.A dilution. There was no difference in the post-thawing livability between semen in 1$m\ell$ straw and 10$m\ell$ aluminium package.

• Clinical and Experimental Reproductive Medicine
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• 제26권2호
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• pp.257-263
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• 1999

It is well known that discard of seminal plasma from the semen and separation of motile sperm should be preceded before insemination for IUI or IVF. Till now, more than ten kinds of semen treatment methods have been developed. Of those, swim-up and Percoll methods have been used widely in ART laboratories as a routine semen treatment methods because of its advantages. However, there are reports that Percoll can make a genetic trouble because of its chemical structure and therefore the necessity has been arisen to substitute Percoll for other equivalent materials. This study was performed to evaluate the effects of three different sperm preparation methods (swim-up, Percoll and Sil-Select) on sperm motility, sperm recovery rate and fertilization rate. Also, the feasibility of using Sil-Select instead of Percoll in ART was evaluated. Each semen samples were divided into three fractions and motile sperm were recovered by swim-up, Percoll and Sil-Select gradient centrifugation methods. Normal and sub-normal criteria of fifteen semen samples and seventeen IVF cycles were included in these study. As results, no significant difference was found in sperm recovery rate in normal semen treated by a Swim-up, Percoll and Sil-Select method ($13.2{\times}10^6,\;17.5{\times}10^6\;and\;17.7{\times}10^6$ respectively). The initial sperm motility was 61.9% and this increased to 87.1%, 92.6% and 89.5% through Swim-up, Percoll and Sil-Select treatment, respectively. Higher motility was observed in Percoll and Sil-Select treated groups (81.5%, 79.2%, respectively) than swim-up group (66.8%) after incubation for 24hrs. In sub-normal group, sperm recovery rates were higher in Sil-Select group $(2.9{\times}10^6)$ than Percoll gradients group $(1.8{\times}10^6)$. In IVF cycles, the outcomes of fertilization using sperm treated by swim-up and Sil-Select group were similar (82.2%, 79.7% respectively). In conclusion, our results indicate that Sil-Select can be used as a substitute material for sperm preparation instead of Percoll.