• 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.

• Journal of Embryo Transfer
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• v.26 no.4
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• pp.257-263
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• 2011

BPA, a diphenyl compound containing groups, that make it structurally similar to synthetic estrogen and is considered as one of the major endocrine disruptors. Silymarin has extensively been used to prevent and/or alleviate some human disease, especially for the treatment of adverse liver conditions. It has an antioxidative efficacy and cancer preventive efficacy. Therefore, we examined the hypothesis that silymarin can inhibit BPA-induced toxicity in boar sperm duing in vitro storage. Sperm characteristics (motility, viability, membrane integrity and mitochondrion activity) in semen exposed to BPA (10~200 uM) were sharply lowered, while it increase in a dose and time dependent manner due to silymarin addition (50~200 uM) into semen extender in the presence of BPA (100 uM). All of the evaluated characteristics were gradually improved in the groups that were treated with silymarin (50~200 uM) in the presence of BPA (100 uM) in comparison to BPA 100 uM alone group, irrespective of incubation periods (3 and 6 h). These results demonstrate that silymarin can ameliorate the toxicity of BPA on boar sperm characteristics during in vitro storage, suggesting that silymarin indirectly act as an antioxidant.

• Proceedings of the KSAR Conference
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• 2001.03a
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• pp.86-86
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• 2001

The aims of this work were to determine the acrosin activity and to evaluate the structural and functional integrity of AS boar spermatozoa. The acrosin activity of spermatozoa were 5.40, 4.10 and 3.40 mIU/10$^{6}$ sperm in raw, extended and frozen semen respectively , which differed significantly each other (P<0.05). After the raw and extended semen were exposured to cold and thermal shock, the acrosin activities of spermatozoa in the raw semen were 5.39, 5.21 and 5.29 mIU/10$^{6}$ sperm for control (non-shock), cold shock and thermal shock, and those of extended semen were 4.21, 3.98 and 4.00 mIU/10$^{6}$ sperm. This value among treatments did not differ significantly. The acrosin activities of spermatozoa in the extended and stored semen were 3.27, 3.52, 3.46 and 3.23 mIU/10/suup 6/ sperm, while hypo-osmotic test(HOST) values were 56.5%, 64.7%, 66.0% and 56.0%, following 4 days storage at 4$^{\circ}C$, 17$^{\circ}C$ , $25^{\circ}C$ and 37$^{\circ}C$, respectively. The results at 17$^{\circ}C$ and $25^{\circ}C$ appeared to be best compared with the other storage temperatures.

• Reproductive and Developmental Biology
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• v.38 no.4
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• pp.159-163
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• 2014

The objective of this study was to determine the effects of E. coli on boar sperm quality and reproductive performance in sows after artificial insemination. Three different levels of E. coli were artificially inoculated to semen with following concentrations; Control, 500, 5,000 and 50,000 colony forming unit (cfu)/ml. Semen samples were preserved at $17^{\circ}C$ for 5 days. Sperm motility was significantly decreased (p<0.05) on day 3 in the group inoculated with 5,000 cfu/ml compared to control groups. In all treatment groups, sperm motility was gradually decreased as storage time increased, but the decline pattern was more drastic in the groups inoculated with 5,000 and 50,000 cfu/ml groups from day 3 (p<0.05) compared to control group. After 3 day of storage at $17^{\circ}C$, sperm viability in sample inoculated with the highest concentration (50,000 cfu/ml) of bacteria was less (p<0.05) than that of control group. The pH of semen sample pH was maintained 7.2~7.5 in all groups during the experimental period. No differences (p>0.05) were found for both storage time and bacterial concentration. The pregnancy rate and live born piglets tend to decrease by increasing the concentration of E. coli in semen. In particular, the rate of pregnancy was lower in the group inoculated with 50,000 cfu/ml (58.3%) compare to the other groups (81.8, 75.0, 76.5%). These results suggest that the contamination of E. coli in boar semen negatively affects fertilizing ability of boar sperm and the reproductive performance obtained from sows after artificial insemination.

• Reproductive and Developmental Biology
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• v.33 no.2
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• pp.99-105
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• 2009

Curcumin is a major active component of the food flovour tumeric. It has been used for the treatment of many diseases such as inflammatory and infectious diseases, cancer and other disease due to its antioxidant properties. Curcumin is a powerful scavenger of many free radicals such as superoxide anion, hydroxyl radical and nitric oxide. The objective of this study was to investigate the antioxidative effects of curcumin against hydrogen peroxide on semen quality during in vitro storage of boar semen. The sperm treated with different concentration of curcumin (1, 5 and 10 ${\mu}M$) in the presence or absence of hydrogen peroxide (250 ${\mu}M\;H_2O_2$) were incubated for 3, 6 and 9 hr at $37^{\circ}C$ and analyzed sperm characteristics such as motility, membrane integrity (MI), lipid peroxidation (LPO), reactive oxygen species (ROS) and DNA fragmentation (DF). The sperm motility and MI in $H_2O_2$ treated group ($47.8%{\pm}6.8$ and $24.8%{\pm}2.2$) were significantly decreased when compare to curcumin treated group ($79.8%{\pm}2.7$ and $34.6%{\pm}1.0$, respectively) irrespective of incubation periods(p<0.05). The LPO of spermatozoal plasma membrane was measured by thiobarbituric acid (TBA) reactions for malondialdehyde (MDA), MDA level in control ($11.6{\pm}0.6\;nmol/L{\times}10^6$) and curcumin groups ($10.7{\pm}0.3\;nmol/L{\times}10^6$) were lower than those of curcumin plus $H_2O_2$ ($17.1{\pm}0.8\;nmol/L{\times}10^6$) or $H_2O_2$ group ($22.5{\pm}1.9\;nmol/L{\times}10^6$) from 3 to 9 hr incubation periods. The DF by sperm chromatin dispersion (SCD) test and ROS production measured by 2',7'-dichlorofluorescein (DCF) fluorescence intensity were no significantly difference through all experimental groups (p>0.05). Correlation among evaluation methods for sperm quality, motility vs MI and DF vs ROS was positively correlated while motility vs DF and ROS vs LPO were negatively correlated in all treatment groups. These results demonstrate that curcumin can effectively improve the sperm quality during in vitro storage of boar semen through its hydrogen peroxide scavenging mechanism as an antioxidant.

• Korean Journal of Animal Reproduction
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• v.8 no.2
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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.

• Korean Journal of Animal Reproduction
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• v.25 no.2
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• pp.113-118
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• 2001

The objective of this study was to investigate the effects of $\alpha$-tocopherol and selenium on the boar semen characteristics. Semen volume and pH values were not different among treatments. However sperm concerntration, total number of sperm and sperm mortility were significantly(P＜0.05) increased comparing to the control group and sperm abnormality was significantly(P＜0.05) decreased comparing to the control group. Also, sperm mortility by storage day was significantly(P＜0.05) increased comparing to the control group. The results from this experiment indicate that dietary $\alpha$-tocopherol and selenium can affect boar semen characteristics.

• Reproductive and Developmental Biology
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• v.36 no.3
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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.

• Journal of Embryo Transfer
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• v.27 no.1
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• pp.21-28
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• 2012

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.

• Journal of Embryo Transfer
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• v.19 no.1
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• pp.19-25
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• 2004

The objective of this study was to optimize the selection of sperm, optimal culture system of in vitro derived porcine embryos. The results obtained were summarized as follows: 1. When oocytes were inseminated with liquid sperm and frozen-thaw sperm, the cleavaged rate of liquid sperm (46.2%) was higher than that of frozen-thaw sperm (39.7%), however there were not show significant different each other. The blastocyst rates of liquid sperm (15.8%) was significantly higher than that of frozen-thawed sperm (9.3%)(P< 0.05). 2. When oocytes were inseminated with epididymal sperm after 1, 2 and 3 day storage, the cleavaged rate of epididymal sperm after 1, 2 and 3 day storage was 60.5, 61.0 and 56.8% respectively. The morulae (17.4, 19.9 or 17.3%) and blastocyst (8.7, 15.4, 11.3%) rate of epididymal sperm after 1, 2 and 3 day storage was no significantly respectively(P< 0.05). 3. In vitro developed to cleavaged rate of G1.3/G2.3 media used for culture was significantly(P< 0.05) higher as 62.1% compared with the results using the media NCSU23(52.8), however in vitro developed to blastocyst rate of NCSU23(11.6%) media was significantly(P< 0.05) higher than that'of G1.3/G2.3(4.7%). 4. When the fertilized oocytes were cultured with NCSU23 in addition to 1 mM glutathione(GSH), the cleavaged rate of treated groups of GSH(62.3%) was significantly higher than that of control(53.5%) respectively(P< 0.05). And in vitro developed to blastocyst rates of treated groups of GSH(15.6%) was higher than that of control(12.6%) however, there was no significant difference(P< 0.05).