• Title/Summary/Keyword: Liquid Semen

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Studies on the Properties of Charolais Semen (Charolais의 정액 성상에 관한 연구)

  • 고광두;손봉환;변명대;김선환
    • Korean Journal of Animal Reproduction
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    • v.4 no.1
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    • pp.20-27
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    • 1980
  • This experiment was to study semen properties of Charolais for the a, pp.ication ot artificial insemination. The result obtained were summarized as follows: 1. In the preservation of liquid semen for 6 days, the survival rates of Charolais semen averaged 57.14% in skim milk solution and 58.17% in tris buffer solution. There were not differences. 2. Recovery of semen after thawing was vigorous in the semen that was diluted and frozen in 48 hrs. 3. The real rates of survival sperm for Charolais averaged 83% after living sperm was diluted and stained for 6 days. 4. Methylene blue reduction test diluted semen was fresh when it was diluted within 48 hrs. 5. If the diluted semen was preserved below 5$^{\circ}C$ in Charolais, the pH decreased by 0.2 in a day. 6. Diluted semen was more resistant to the cold shock than fresh semen. 7. In resistance against hot shock, sperm was almost dead in 20 minutes in 46.5$^{\circ}C$ in diluted semen, while it was dead in 30 minutes in 42.5$^{\circ}C$ in diluted semen. 8. In examination of morphological changes of sperm acrosome for 6 days, normal sperm in skim milk solution and tris buffer solution was 80% and 76.97% respectively, swelling sperm 12.8% and 15.27%, deficient sperm 0.6% and 0.97% abnormal staining 3.07% and 5.25%, immature sperm 0.28%, and 0.23%, whereas other abnormal sperm was 1.28% and 1.42%.

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Effects of L-Carnitine and Nicotinic Acid on Sperm Characteristics in Miniature Pigs

  • Lee, Yeon-Ju;Lee, Sang-Hee;Kim, Yu-Jin;Hwangbo, Yong;Lee, Seunghyung;Cheong, Hee-Tae;Yang, Boo-Keun;Park, Choon-Keun
    • Reproductive and Developmental Biology
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    • v.40 no.1
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    • pp.1-5
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    • 2016
  • This study investigated the effects of L-carnitine (LC) and nicotinic acid (NA) on sperm viability during liquid storage at $18^{\circ}C$ in miniature pigs. $10{\mu}M$ LC and 30 mM NA, combined LC and NA (LN) were treated in fresh semen for 3, 7, and 10 days. In results, sperm survival increased in NA- and LN-treated semen on 7 and 10 days (p<0.05), mitochondrial integrity of live sperm increased in LN-treated semen on 7 days (p<0.05), but not NA-treated semen. In addition, we examined the acrosome reaction of sperm in miniature pigs. LC and NA did not influence on acrosome reaction of boar sperm. In conclusion, LC and NA effectively maintained the viability and quality of sperm during long-term storage in miniature pigs, suggesting that the combined LN may be useful for improving the semen extender for long-term liquid storage in pigs.

Studies on the Liquid and Freezing storage of Boar Semen III. Effects of Dilutor on Livability and Fertility of Liquid Semen and Effects of Dilution and Vessel on Livability of Frozen Semen (돈정액의 액상 및 동결보존에 관한 연구. III. 보존액이 액상정액의 정자생존성과 수태율에 미치는 영향과 희석방법과 용기가 동결정액의 생존성에 미치는 영향)

  • 임경순;정장용
    • Korean Journal of Animal Reproduction
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    • v.3 no.1
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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.

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Effects of Storage in Different Commercial Semen Extenders on Sperm Motility, Viability and Membrane Integrity of Korean Native Boar Spermatozoa

  • Sa, Soo-Jin;Kim, In-Cheul;Choi, Sun-Ho;Hong, Joon-Ki;Kim, Du-Wan;Cho, Kyu-Ho;Kim, Young-Hwa;Chung, Ki-Hwa;Park, Jun-Cheol
    • Journal of Embryo Transfer
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    • v.28 no.4
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    • pp.349-353
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    • 2013
  • 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 ($30{\times}10^6$ spermatozoa/ml) in different semen extenders. Semen samples were stored at $17^{\circ}C$ 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 $17^{\circ}C$.

Effect of Matruation Media and Liquid Boar Semen on Maturation and Fertilization of Pig Oocytes In Vitro (돼지 난모세포의 체외성숙 및 체외수정시 배양액과 액상정액의 효과)

  • Park, C.S.;Lee, K.S.;Park, B.K.;Zhang, X.K.;Lee, Y.H.;Xu, Z.
    • Korean Journal of Animal Reproduction
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    • v.21 no.1
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    • pp.19-23
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    • 1997
  • This present study was carried out to examine the effect of maturation media and liquid boar semen on in vitro maturation and feritilization of pig oocytes. The results obtained were as follows : When the oocytes were cultured for 36∼42 hours in mTCM-199, Waymouth MB 725/1 and mTLP-PVA medium, the maturation rates were 90%, 92% and 88%, respectively. The sperm penetration rates of pig oocyte matured in vitro were 87%(mTCM-199), 90%(Waymouth MB 725/1) and 86%(mTLP-PVA), respectively. The rates of nuclear maturation and fertilization of pig oocytes among three different media did not differ. However, the rate of male pronucleus formation of pig oocytes was significantly higher in pig oocytes matured in Waymouth MB 725/1(91%) than oocytes matured in mTCM-199(66%) and mTLP-PVA(62%) medium (P<0.05). When the collected sperm-rich fraction without diluent was used fro in vitro fertilization in mTCM-199 fertilization medium, the fertilization rate was 87.9%. However, when the liquid boar semen diluted with B tschwiler diluent was used at day 3 and 5 after dilution, the fertilization rate was 40.8% and 0.0%, respectively.

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Effects of Sugar Type on Viability of Frozen-Thawed Canine Spermatozoa

  • Lim, Y.H.;Son, J.M.;Shin, Y.J.;Kim, Y.S.;Lee, D.S.;Yoon, K.Y.;Shin, S.T.;Cho, J.K.
    • Journal of Embryo Transfer
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    • v.23 no.4
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    • pp.239-243
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    • 2008
  • This study was conducted to investigate the effects of type of the sugar supplemented to the extender on the vigor, viability and intact acrosomal rates of frozen-thawed dog spermatozoa. The ejaculated semen was diluted with TRIS-citric acid extender containing 200mM TRIS, 73mM citric acid, 6% (v/v) glycerol, 20% (v/v) egg yolk, 1% (v/v) antibiotics (streptomycin/penicillin), 44 mM sugar, which was either glucose, fructose or glucose-fructose combination, and distilled water to make the final volume of 100ml. Extended semen samples were cooled at $4^{\circ}C$ for an hour, packaged in 0.25ml straws, equilibrated for 10 minutes in liquid nitrogen vapor, and frozen in liquid nitrogen. Samples were thawed by placing straws into $37^{\circ}C$ water for 120 seconds. After thawing, vigor, viability and intact acrosomal rates of frozen-thawed semen were compared according to type of sugar. No significant differences were observed between glucose and fructose groups. In addition, combination of the 2 sugars also did not show any significant differences in the vigor, viability and intact acrosomal rates. In conclusion, glucose and fructose were equally efficient as sugar supplements for freezing extender.

Intra-uterine Insemination with Frozen Canine Semen Using Vaginal Endoscope (개에서 동결정액의 경관내 주입을 통한 인공수정)

  • 정동희;최윤주;임상현;김용준
    • Journal of Veterinary Clinics
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    • v.18 no.1
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    • pp.44-47
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    • 2001
  • Semen were collected from 9 male dogs and frozen by liquid nitrogengas. Frozen semen were thawed at 7$0^{\circ}C$ for 8 seconds. About $2{\timss}10^8$ sperm per insemination were inseminated to 10 bitches (3 Retrievers, 4 Chihuahuas, 1 Yorkshire Terriers, 1 Maltese, and 1 Poodle) at three and six days after the estimated peak of luteinizing hormone. For small breed dogs, uretero-renoscope (Kahl Storz, Germany, 12.5 Fr) was used for trans-cervical insemination, whereas cystoscope(Kahl Storz, Germany, 22Fr) was used for large breeds (Retrievers). Pregnancy was diagnosed by ultrasonography at 30 days after insemination. All of 3 Retrievers (100.0%) and 3 bitches of 7 small breed dogs (42.9%) were conceived (60.0% in total). This result indicated that trans-cervical insemination using endoscope is an effective method for AI with frozen semen not only for large breed dogs such as Retriever but also for small breeds.

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Effect of Short-term and Long-term Preservation on Motion Characteristics of Garole Ram Spermatozoa: A Prolific Microsheep Breed of India

  • Joshi, Anil;Bag, Sadhan;Naqvi, S.M.K.;Sharma, R.C.;Rawat, P.S.;Mittal, J.P.
    • Asian-Australasian Journal of Animal Sciences
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    • v.14 no.11
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    • pp.1527-1533
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    • 2001
  • Garole is a prolific, rare, less known and small size Indian sheep breed found in low and humid Sunderban region of West Bengal. Although information on stored Garole ram liquid semen upto 24 h is available, but there is a need to further investigate the short-term and long-term preservability of Garole ram semen for extensive utilization of this valuable germplasm by artificial insemination. The aim of the present study was to apply computer-assisted sperm analysis technique for assessing the motion characteristics of Garole ram semen stored (i) in liquid state at refrigeration temperature for short-term preservation upto 48 h and (ii) in frozen state at $-196^{\circ}C$ for long-term preservation after packaging in mini straws. Short-term preservation had a significant effect on motility (p<0.01) as the motility progressively decreased from 90.1% at 0 h to 85.5% and 73.2% after 24 and 48 h of storage, respectively. Although the decline in rapid moving sperms was also significant (p<0.01) on storage but the decrease was more pronounced at 48 h as compared to 24 h of storage period. Storage of chilled semen had also a significant effect on % linearity (p<0.05), % straightness (p<0.01), sperm velocities (p<0.01), amplitude of lateral head displacement (p<0.01) and beat frequency (pO.Ol) of spermatozoa. The replication had a significant effect for all the variables except average path and straight line velocity. However, the interactions of short-term storage and replication were non-significant for most of the variables except % of medium moving sperms, sperm velocities and beat frequency. On long-term preservation of Garole ram spermatozoa under controlled conditions the mean post-thaw recovery of 70.4 and 71.4% motile spermatozoa was achieved having 48.8 and 48.9% of rapidly motile spermatozoa, respectively in both the replicates. The effect of replication on cryopreservation was significant (p<0.05) on amplitude of lateral head displacement and beat frequency, but there was no significant effect on motility, rapidly motile spermatozoa, linearity, straightness and sperm velocities of frozen-thawed spermatozoa. It can be concluded from these results that an average 70% motility can be achieved on storage of Garole ram semen in chilled liquid state upto 48 h or in liquid nitrogen after freezing under controlled conditions in straws. However, further studies are required to evaluate the fertility of short-term and long-term preserved Garole ram semen for extensive use of this prolific sheep breed.

Post-thaw Thermal Resistance Test on Motility and Acrosomal Integrity of Filtered and Non-filtered Frozen Semen of Murrah Buffalo Bulls

  • Maurya, V.P.;Tuli, R.K.
    • Asian-Australasian Journal of Animal Sciences
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    • v.16 no.10
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    • pp.1424-1428
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    • 2003
  • Present investigation was conducted to determine the post-thaw sperm motility and acrosomal damage of filtered and non-filtered frozen semen of Murrah buffalo bulls. Twenty semen ejaculates (from four Murrah buffalo bulls collected at weekly interval) were diluted in Tris egg yolk glycerol extender and divided into two parts. One was filtered through sephadex G-100 column and the other portion was kept as such (non-filtered). Both fractions were frozen in liquid nitrogen ($-196^{\circ}C$) by the standard method developed in the laboratory. After 24 h of freezing, non-filtered and filtered semen samples were thawed at $37^{\circ}C$ for 1 min. These samples were incubated at $37^{\circ}C$ in a water both. The different seminal characteristics i.e. percent progressive sperm motility, live and abnormal spermatozoa and spermatozoa with damaged acrosome were assessed at hourly interval till they remained motile. The filtered frozen and thawed semen showed significantly (p<0.05) high sperm viability and acrosomal integrity as compared to non-filtered semen.

Studies on Liquefaction of Semen (정액(精液)의 액화(液化)에 관한 연구(硏究))

  • Kim, Suk-Hee;Lee, Hee-Yong
    • Clinical and Experimental Reproductive Medicine
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    • v.3 no.2
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    • pp.35-42
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    • 1976
  • The human semen ejaculated in a form of liquid state, coagulates immediately after ejaculation, and then liquefies again. However, the mechanisms of neither coagulation and liquefaction of semen have not been explained clearly so far, and very limited numbers of report are available, although the spermatology and andrology made rapid progress. This clinical study has been undertaken to investigate the liquefaction phenomena and practicability of the results might be applied to fertility and infertility problems. As a preliminary study, in this report the liquefaction time of various semen groups is measured and analysed. The following results are obtained: 1. An average liquefaction time of semen of a total of 60 subjects: 25 minutes. 2. An average liquefaction time of semen according to sperm count: 1) Normospermia group (20 cases): 34 minutes. 2) Oligospermia group (20 cases): 21 minutes. 3) Azoospermia group (20 cases): 20 minutes. 3. An average liquefaction time of semen according to abstinence period: 1) Less than 3 days group (30 cases): 22 minutes. 2) More then 5 days group (30 cases): 28 minutes. In conclusion: 1. The liquefaction time of semen of the normospermia group is longer than oligospermia group or azoosermia group. 2. The liquefaction time of semen may not be greatly influenced by the various factors such as abstinence period, semen volume, semen pH, age of the subjects and so on. 3. In routine semen analyses, it is recommended to begin the analysis at least 25 minutes after the ejaculation. 4. Further studies are required in conjunction with practical application of liquefaction mechanism in infertility and fertility control.

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