• Journal of Animal Reproduction and Biotechnology
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• v.39 no.3
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• pp.179-193
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• 2024

Background: Because oxidative stress can induce decreased quality of caprine semen during the storage, there has been limitation for the use of stored semen in the assisted reproductive technologies. The present study, therefore, assesses the potential of Annona muricata (A. muricata) to reduce semen storage associated-damages. Methods: Semen was collected by electro-ejaculation from ten bucks, and extended with Tris-egg yolk (TEY) supplemented with A. muricata leaf aqueous extract (SAE) at 20 (SAE20), 40 (SAE40), and 80 (SAE80) ㎍/mL. Sperm variables including motility, motion characteristics, viability, membrane functionality, and DNA integrity were assessed at different storage periods (6, 24, 48, and 72 hr). In addition, oxidative stress indicators in the extender supplemted with SAE were also assessed for each group. Results: By adding SAE at 80 ㎍/mL in TEY, the storage of goat buck semen was improved, resulting in reduced loss of sperm motility, viability, DNA fragmentation, and membrane integrity during chilled storage at 4℃ for up to 72 hr. In addition, enrichment of TEY extender with SAE significantly (p < 0.05) reduced malondialdehyde, an indicator of oxidative stress, compared to the negative control. Conclusions: Supplementation of SAE in TEY extender can reduce buck spermatozoa liquid storage associated damages due to oxidative stress.

• Korean Journal of Animal Reproduction
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• v.21 no.1
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• pp.25-30
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• 1997

This study was done to find out the preservation possibility of liquid boar semen at variabel room temperature of 9 to 16$^{\circ}C$. The percentages of sperm motility and NAR acrosome were highest in B tschwiler extender compared to B tschwiler＋Hepes, Andro＋Hepes and Andro extenders. The extenders with Hepes buffer showed detrimental effect for preservation of liquid boar semen. The pH of ejaculated sperm-rich fraction was 7.5. The pH of B tschwiler＋Hepes, B tschwiler, Andro＋Hepes and Andro extenders was 6.9, 7.5, 7.1 and 8.1, respectively. The pH of liquid boar semen with B tschwiler＋Hepes, B tschwiler, Andro＋Hepes and Andro extenders was 6.6, 6.9, 6.7 and 6.9 at 1st day of storage, and 5.5, 5.7, 5.6 and 5.8 at 7th day of storage, respectively. Gilts and sows were inseminated twice with liquid boar semen stored at 9~16$^{\circ}C$ in B tschwiler extender for 3~4 days. Farrowing rate, litter size and average pig weight at birth between AI and natural service did not differ significantly in gilt and sow, respectively. However, sow showed higher farrowing rate and litter size compared to gilt both in AI and in natural service. As a result of this study, we found out that liquid boar semen can be stored for 5~7 days at uncontrolled room temperature of 9~16$^{\circ}C$ in B tschwiler extender.

• Reproductive and Developmental Biology
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• v.32 no.1
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• pp.59-64
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• 2008

The objective of this study was to estimate modification of semen quality during storage. Liquid boar semen samples extended in Beltsville Thawing Solution were stored at $17^{\circ}C$ up to 5 days. While % motility and linearity significantly decreased eon day 3 in extender, the qualitative motility patterns were maintained satisfactorily. Also the storage of boar semen up to 5 days before insemination did not significantly changed the acrosome intactness. However, acrosome changed sperm significantly increased and capacitated sperm significantly decreased from day 4. No significant modifications in acrosome integrity were showed during sperm storage; these results suggest that liquid boar semen may keep the quality in extender for 3 days.

• 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.

• Korean Journal of Poultry Science
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• v.30 no.2
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• pp.129-134
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• 2003

This study was conducted to investigate the effects of liquid rooster semen on reproductive ability in chicken. Raw and diluted semens were stored at 5$^{\circ}C$ cold temperature for 6, 30, and 54 hours after semen collection. There was no statistically difference in sperm motility throughout the 6 hours period of storage among raw semen and diluted semen groups with skim milk glucose solution (SM), egg yolk glucose solution (EY), and saline. But there was decrease in those throughout the period of 30 and 54 hours of storage. Sperm motility and normal sperm for the period of 30 and 54 hours of storage were significantly better in SM and EY diluted groups (P<0.05). Fertilization rates of rooster semen diluted with SM were 90.77, 87.70, and 59.46% for 6, 30, and 54 hours stored groups, respectively, those proved to be higher in SM-diluted group than other groups.

• 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.

• 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.

• 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 $17^{\circ}C$.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.11
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• pp.1501-1508
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• 2004

In this study the effect of extenders and temperatures on sperm viability and fertilizing capacity of boar sperm during long-term storage was investigated. Acrosomal integrity, membrane integrity, motility and hypo-osmotic resistance were evaluated by fluorescence and light microscopy. An in vitro fertilization test was performed to assess the fertilizing capacity of stored spermatozoa. The five diluents tested were ranked according to their ability to maintain sperm functional parameters and Zorlesco (ZO) extender with BSA or with PVA instead of BSA produced the best results. Zorlesco extender substituted with PVA (ZO+PVA) was found to maintain motility both at 15 and 20$^{\circ}C$. within 5 days of storage, but the quality of semen stored at 15$^{\circ}C$ decreased thereafter as compared to semen stored at 20$^{\circ}C$ Semen stored at 5$^{\circ}C$ demonstrated rapid loss of motility already within 24 h. Both fertilization and cleavage of semen stored at 20$^{\circ}C$ in ZO substituted with PVA instead of BSA did not change significantly until day 8 of storage. It is therefore concluded that PVA can be used to substitute for BSA and 20$^{\circ}C$ was more suitable than 15$^{\circ}C$ for boar semen storage, and in vitro fertilizing capacity of spermatozoa was maintained for at least 8 days in ZO+PVA at 20$^{\circ}C$.

• Korean Journal of Animal Reproduction
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• v.20 no.2
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• pp.101-109
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• 1996

This study was done to find out the methods of long-term use of liquid boar semen in 100 ml plastic bottle for artificial insemination and to investigate differences between Lactose-Egg yolk and Biitschwiler diluents according to storage temperature, and effect of final glycerol concent ration in Lactose-Egg yolk diluent. Liquid boar semen diluted with Lactose-Egg yolk diluent showed significantly higher sperm motility (p<0.05) after 0.5 and 2h incubation at 37$^{\circ}C$,than Butschwiler diluent at all storage length when it was preserved in the 5$^{\circ}C$ refrigerator. The NAR acrosome in Lactose-Egg yolk diluent a after 0.5 and 2h incubation at 37$^{\circ}C$, respectively, during preservation periods was similar to that in Biitschwiler diluent. When liquid boar semen was preserved at 15$^{\circ}C$, liquid boar semen in the Butschwiler diluent showed significantly higher percentages of sperm motility and NAR acrosome from third day to seventh than that in Lactose-Egg yolk diluent. In the effect of final glycerol concentration of liquid boar semen in the Lactose-Egg yolk diluent, the final glycerol concentration of 2% showed higer percentages of sperm motility and NAR acrosome than that of 0, 1, 3, and 5%. Farrowing rate, litter size and average pig weight at birth did not differ significantly between Lactose-Egg yolk and But schwiler diluents. As a result of this study, we found out that liquid boar semen can be stored for 6-7 days at 5$^{\circ}C$ in Lactose-Egg yolk diluent and at 15$^{\circ}C$ in Butschwiler diluent.