• Asian-Australasian Journal of Animal Sciences
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• v.19 no.4
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• pp.486-494
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• 2006

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.

• Applied Microscopy
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• v.39 no.2
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• pp.89-99
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• 2009

To investigate the morphological characteristics of spermatozoa of the grey red-blacked vole (Clethrionomys rufocanus) belongings to the subfamily Cricetinae, subgenus Clethrionomys were examined by scanning and transmission electron microscopes. The sperm head of C. rufocanus was an ax or hatchet in shape with a curved single dorsal hook. The total length of C. regulus sperm was 95.8 ${\mu}m$. The length of sperm head was 7.8 ${\mu}m$, and the tail (88.0 ${\mu}m$) consisted of four major segments: the neck (1.0 ${\mu}m$), middle piece (22.0 ${\mu}m$), and principal piece plus end piece (65.0 ${\mu}m$), respectively. The segmented columns were about 10~12 in number. The number of gyres of mitochondria ranged from about 170 to 178. The post-nuclear cap occupied about a half of nucleus. The equatorial segment is located between the post-nuclear cap segment and acrosomal cap on the nuclear surface. Nos. 1, 5 and 6 of the outer dense fibers were larger than the others. A fibrous sheath and longitudinal column of the principal piece were in evidence, but the fibrous sheath was not seen at the end piece. In conclusion, the morphological structures of sperm head and tail may be useful information to patterning of sperm evolution and classifying of species.

• Journal of Embryo Transfer
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• v.29 no.2
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• pp.163-169
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• 2014

The objective of this study was to evaluate the characteristics of Korean Native Cattle sperm frozen-thawed with L-cysteine and/or catalase. The semen from bulls was collected by the artificial vagina method, and Triladyl containing 20% egg-yolk and/or L-cysteine (L), catalase (C) and L-cysteine + catalase was added to the diluted semen for cryopreservation. The results showed that sperm viability was significantly higher in the L-cysteine + catalase ($69.49{\pm}3.16%$) group than in the control ($60.5{\pm}3.94%$) group (p<0.05). Acrosome damage was significantly lower in the L-cysteine ($17.12{\pm}1.08%$) group than in the control ($21.46{\pm}1.14%$), catalase ($20.54{\pm}0.76%$), and L-cysteine + catalase ($19.29{\pm}0.65%$) groups (p<0.05). In addition, the level of intact mitochondria in the spermatozoa was significantly higher in the L-cysteine ($58.65{\pm}1.39%$) group than in the control ($50.63{\pm}2.37%$) group (p<0.05). The hydrogen peroxide level in the frozen-thawed sperm was significantly lower in the L-cysteine ($3.74{\pm}1.66%$), catalase ($4.65{\pm}1.87%$), and L-cysteine + catalase ($8.11{\pm}2.15%$) groups than in the control ($13.22{\pm}1.6%$) group (p<0.05). The glutathione level was significantly higher in the L-cysteine ($1.33{\pm}0.03%$) group than in the control ($1.08{\pm}0.06%$), catalase ($1.05{\pm}0.02%$) and L-cysteine + catalase ($1.11{\pm}0.03%$) groups (p<0.05). In conclusion, L-cysteine and catalase could protect the membrane of Korean Native Cattle sperm from damage during sperm cryopreservation. Especially, L-cysteine was more effective for keeping acrosomes and mitochondria intactness during sperm cryopreservation.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.193-198
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• 2013

This study was designed to evaluate the effect of L-cysteine on sperm characteristics and oocyte cleavage in vitro in Korean native cattle. For this study, the freezing of diluted semen were added with Triladyl containing 20% egg-yolk and/or 0, 5, 10 and 20 mM L-cysteine before cryopreservation. The viability in frozen-thawed sperm were estimated by SYBR14/PI double stain, acrosome damage with FITC-PNA, mitochondria intact with Rhodamin123 and hydrogen peroxide($H_2O_2$) level with carboxy-DCFDA by flow-cytometry. The developmental capacity was also assessed with cleavage rates in oocytes fertilized in vitro by frozen-thawed sperm. In results, the sperm viability was significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). In addition, acrosome damage was significantly decreased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). The mitochondria intact was also significantly increased in 10 mM and 20 mM concentrations of L-cysteine than other groups (p<0.05). On the other hand, the cleavage rates were significantly increased in 0 mM, 5 mM and 10 mM groups than 20 mM concentration of L-cysteine (p<0.05). The oocyte degeneration of oocytes were significantly decreased in 0 mM, 5 mM and 10 mM groups than in 20 mM L-cysteine group (P<0.05). However, there are no significantly differences among the L-cysteine treatment groups. We suggest that concentration of 10 mM L-cysteine have beneficial impact for sperm cryopreserved in Korean native cattle. This result also could be recommended for artificial insemination program if supported by an improvement in the fertility results and required further study.

• Korean Journal of Animal Reproduction
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• v.26 no.3
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• pp.229-234
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• 2002

This study was carried out to investigate the general characteristics such as concentration sperm motility and abnormality of sperm on the whole epididymal semen(EWS), RSP-S(removed seminal plasma by saline) and RSP-T(removed seminal plasma by tris-buffer) semen and survival rates after freezing on motility of whole and RSP-S and RSP-T semen and extender containing 2~8% glycerol, and ability of frozen-thawed sperm to penetrate homologous oocytes. 1. The concentration, motility and abnormality of epididymal WES, RSP-S and RSP-T sperm were 4.25 $\pm$ 0.25($\times$10$^{6}$ Cells/$m\ell$), 3.85$\pm$0.20($\times$10$^{6}$ Cells/$m\ell$), 4.05 $\pm$ 0.28($\times$10$^{6}$ Cells/$m\ell$), 50.55 $\pm$ 2.75%, 67.25 $\pm$ 2.55%, 78.75 $\pm$ 3.55 and 9.45 $\pm$ 2.25%, 37.75 $\pm$ 2.10%, 24.25 $\pm$ 1.55%, respectively. 2. The survival rates of slow and rapid frozen epididymal RSP-S and RSP-T sperm were 35.00 $\pm$ 2.35%, 45.50 $\pm$ 2.15％ and 16.50 $\pm$ 3.55%, 22.55 $\pm$ 3.95%, respectively. The survival rate of epididymal WES and RSP-T sperm after freezing following dilution with tris-buffer containing 2~8% glycerol were 9.25 $\pm$ 1.55%~17.50 $\pm$ 2.50%. 3. The percentage of capacitated and acrosome-reacted sperm prier to culture for fresh and frozen -thawed epididymal RSP-T semen were 45.25 $\pm$ 5.75%, 7.06 $\pm$ 0.25%, 48.20 $\pm$ 6.80% and 13.00 $\pm$ 2.35%, 3.55 $\pm$ 0.85%, 15.50 $\pm$ 1.90%, respectively. The penetration rate the number of sperm per penetrated for fresh and frozen-thawed epididymal RSP-T sperm were 39.25 $\pm$ 4.72%, 34.24 $\pm$ 3.93% and 1.30 $\pm$ 0.33, 1.10 $\pm$ 0.50., respectively.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.1
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• pp.29-34
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• 2008

The aim of the study was to investigate the effect of dietary fish oil supplementation on the semen characteristics of the Markhoz buck. Sixteen bucks were randomly allocated into 4 groups and received four different diets: unsupplemented control diet, supplemented with fish oil at 2.50% dry matter (DM), supplemented with fish oil (2.50% DM) and vitamin E (0.30 g/kg DM), and supplemented with vitamin E (0.30 g/kg DM). All experimental diets were formulated according to AFRC (1998). Semen was collected at 14 d intervals from June 17, 2006 to September 2, 2006. Semen characteristics were evaluated. Significant effects (p<0.05) of the week (sampling time) were observed for all parameters except semen volume. Also a significant effect (p<0.05) of dietary treatment was observed for all parameters except for percent sperm with normal morphologies and semen volume. Fish oil supplementation with excess vitamin E had a significant effect (p<0.05) on total number and sperm density, motility and progressive motility, percentage viability and dead sperm. The interaction between fish oil feeding and sampling time was significant (p<0.05) for all of the parameters. The bucks that received fish oil in association with vitamin E, effect fish oil showed the greatest improvement in semen characteristics compared with the other groups (p<0.05). This study showed that fish oil supplementation with vitamin E may have a beneficial effect on the semen quality and fertility of Markhoz bucks.

• Korean Journal of Animal Reproduction
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• v.24 no.2
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• pp.209-216
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• 2000

This study was carried out to investigate the effects of breed, age of boar, season, parity and mating system on boar semen characteristics and fertilizing capacity. A total of 4181 sows and 199 boars of Durocs (D), Landraces (L), and Yorkshires (Y) were used for this experiment at Darby Artificicial Insemination Center from 1996 through 1999. Semen volume per ejaculate was largest in Landrace (266.8 $m\ell$), followed by Yorkshire, and was smallest in Duroc. Sperm motility did not show significant differences among the above breeds. Sperm concentration was lowest in Landrace (4.7$\times$10$^{9}$ sperm/$m\ell$) and was highest in Duroc (5.7$\times$10$^{9}$ sperm/$m\ell$). Semen volume per ejaculate according to the age of boars was largest at the age of 2 years, followed by the age of 4 and 3 years, and was smallest at the age of I year. Semen volume per ejaculate according to the season in boars was largest in winter (228.6 $m\ell$), followed by autumn and summer, and was smallest in spring. Sperm concentration was highest in spring (5.9$\times$10$^{9}$ sperm/$m\ell$), followed by summer and winter, and was lowest in autumn. The average litter weight at birth did not show any differences according to the mating type. But the number of pigs born alive per litter was largest (9.5 pigs) in the natural mating ＋ artificial insemination group, followed by the artificial insemination group (9.2 pigs), and was smallest (8.9 pigs) in the natural mating group (P＜0.01). The average litter weight at birth and number of pigs born alive per litter did not show any differences between the natural mating and artificial insemination. The L (♀)$\times$Y (♂) and L (♀)$\times$L (♂) matings show $\varepsilon$ d higher average litter weight at birth and number of pigs born alive per litter than the Y (♀) $\times$ Y (♂) and Y (♀) $\times$ L (♂) matings. The pigs in the 2~6th parities had higher average litter weight at birth and number of pigs born alive per litter than those in the 1 st and 7~9th parities.

• Journal of Embryo Transfer
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• v.33 no.1
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• pp.17-22
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• 2018

This study was conducted to determine the effect of pentoxifylline levels on sperm motility, survival rate, sperm membrane integrity of frozen semen and fresh-extended equine semen in Jeju cross-bred horses. As a result of sperm characteristic comparison depending on pentoxifylline levles at 30 minutes post-thaw, the progressive motilities were $53.25{\pm}2.87$ (4mM pentoxifylline) and $50.28{\pm}2.14$ (8mM pentoxifylline) and significantly higher compared to the control group($40.09{\pm}5.15$) and other treatment group (16mM pentoxifylline, $41.27{\pm}2.82$). The progressive fast motility were $22.44{\pm}1.62$ (4mM pentoxifylline,) and $22.74{\pm}3.07$ (8mM pentoxifylline) and significantly higher compared to the control group ($13.47{\pm}1.48$) and other treatment group (16mM pentoxifylline, $14.66{\pm}3.68$) (p<0.05). As a result of sperm characteristic comparison depending on pentoxifylline levles at 30 minutes post-thaw were $68.96{\pm}1.64$ (4mM pentoxifylline) and $67.90{\pm}6.72$ (8mM pentoxifylline) and significantly higher compared to the control group ($53.48{\pm}4.84$) and other treatment group (16mM pentoxifylline, $58.14{\pm}2.65$) (p<0.05). In conclusion, these results suggest that treatment groups with 4mM and 8mM pentoxifylline were higher compare to equine seperm mobility and the control group and treatment groups with more than 16mM pentoxifylline has a negative effect on sperm characteristics. After thawing, the total motility in post-thawed equine sperm has increased by 10 percent for 1 hour. these results suggest that pentoxifylline contributes to the improvement of the equine sperm motility and characteristics in post-thawed semen.

• Journal of Embryo Transfer
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• v.31 no.3
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• pp.293-297
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• 2016

For evaluating the boar semen quality, sperm motility is an important parameter because the movement of sperm indicates active metabolism, membrane integrity and fertilizing capacity. Phospholipase C zeta (PLCz) is important enzyme in spermatogenesis, but the effect has not been confirmed in pigs yet. Therefore, this study was aimed to analyze their association with sperm motility and kinematic characteristics. DNA samples from 124 Duroc pigs with records of sperm motility and kinematic characteristics [total motile spermatozoa (MOT), curvilinear velocity (VCL), straight-line velocity (VSL), the ratio between VSL and VCL (LIN), amplitude of lateral head displacement (ALH)] were subjected. A SNP in non-coding region of PLCz g.158 A > C was associated with MOT (p < 0.05), VCL (p < 0.01), LIN (p < 0.01) and ALH (p < 0.05) in Duroc population. Therefore, we suggest that the intron region of the porcine PLCz gene may be used as a molecular marker for Duroc boar semen quality, although its functional effect was not defined yet. Whether the association is due to the candidate gene or not require further verification. Thus, it will be of interest to continue association studies in the regions surrounding those genes.

• Journal of Embryo Transfer
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• v.21 no.2
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• pp.95-100
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• 2006

Thirty heads of proven bulls were used to identify the effects of mounting conditions (dummy cow vs. teaser bull) on semen characteristics in Hanwoo. Semen was collected from bulls daily two times with 1 h interval every $3{\sim}6$ days for 6 months. Bulls mounted dummy cow (BDC) had higher both the $1^{st}$ and the $2^{nd}$ sperm concentrations than in bulls mounted teaser bull (BTB), resulting in more total sperm number (p<0.05). The total sperm number in the $1^{st}$ collection was the highest in BDC with collection interval of 5 days. Total sperm number in the $1^{st}\;and\;2^{nd}$ collections tended to be more in the BDC of $4{\sim}5$ years old and BTB of $6{\sim}7$ years old. These results indicate that semen collection using dummy cow has a better effect than teaser bull on semen characteristics.