• Asian-Australasian Journal of Animal Sciences
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• v.14 no.8
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• pp.1196-1202
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• 2001

In boar spermatozoa, the head-to-head agglutinability changes in parallel with the development of the fertilizing ability. Namely, both abilities gradually increase in the distal caput and corpus epididymides, but are subsequently suppressed in the cauda epididymidis. It has been postulated that these changes of the agglutinability are controlled via sperm interaction with specific epididymal plasma factors including agglutination mediators (agglutinins) and inhibitors (anti-agglutinins). Expression of these abilities (sperm agglutination and capacitation) is hardly observed in spermatozoa immediately. after ejaculation, but it occurs during incubation in a capacitation medium. Recently, we have purified and characterized epididymal plasma anti-agglutinin for boar spermatozoa. Moreover, we have conducted a series of experiments to reveal biological significance and mechanism of the head-to-head agglutination and have accumulated data indicating that boar sperm agglutination is mediated by capacitation-supporting factors including calcium, bicarbonate and sterol acceptors. This review introduces our recent data and discusses a possible mechanism for suppression of the agglutinability in the distal epididymidis and relationship between agglutinability and fertilizing ability.

• Asian-Australasian Journal of Animal Sciences
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• v.10 no.6
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• pp.609-613
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• 1997

The reproductive performance of forty two male broilers divided into three similar groups and fed on isocaloric and isonitrogneous diets containing 0, 10 or 20% water washed neem seed kernel meal (WWNSKM) was investigated from 20 to 32-wks of age. Results on semen characteristics revealed that feeding of WWNSKM led to significant (p < 0.05) reduction in semen volume, sperm concentration associated with increased incidences of morphological abnormalities in the spermatozoa when compared to that of the control birds. A drastic reduction in the fertilizing ability of spermatozoa was observed, the adverse effects being more at higher inclusion level of the cake. Hatchability of eggs also declined in the WWNSKM fed group. Histological examination of testes revealed a higher number of degenerating cells and poor spermatogenesis along with multinucleated giant cells in the seminiferous tubules of the testes of birds receiving the high dose of WWNSKM in diet. It may be concluded that the feeding of WWNSKM by incorporating in isocaloric and isonitrogneous diets to cockerels is associated with adverse effect on their fertility.

• 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$^{\circ}C$.

• Journal of Embryo Transfer
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• v.25 no.4
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• pp.263-266
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• 2010

Prediction of semen's fertilizing ability used in artificial insemination (AI) is one of very important factors on pig reproductive performance. In vitro fertilization (IVF) has been used for indirect evaluation of sperm's fertilizing ability and it has been showed as highly correlated index. In swine industry, increasing interest in preservation of boar semen raises questions on the sperm motility from semen used in commercial AI centers. Mitochondria in sperm mid-piece generate the energy to support motility and could be an explanation of impaired fertility. Objective of this study was to suggest usable sperm motility to farms in measuring the effect of sperm motility and sperm abnormality on in vitro production of embryo in which sperm's fertilizing ability can be determined indirectly. Semen samples were provided from local AI center and used within 3 days after collection. Semen samples were divided by 4 different motile groups (>70%; 61~70%; 51~60%; <50%) using CASA (computer-assisted sperm analysis) on the days of IVF. Developmental rate to the blastocyst stage from over 61% motile sperm group showed significantly higher rate than below 60% motile sperm group ($16.5{\pm}0.7{\sim}18.4{\pm}0.8%$ vs $6.3{\pm}0.8{\sim}11.5{\pm}0.7%$, p<0.05). In experiment to determine the relationship between sperm motility and viability and abnormality, over 61% motile sperm groups showed significantly higher viability rate compared to below 60% motile sperm groups ($84.8{\pm}4.0{\sim}88.1{\pm}4.0%$ vs $69.1{\pm}4.0{\sim}74.2{\pm}4.0%$, p<0.05). On the other hand, morphological sperm abnormality showed significantly higher in over 70% motile sperm group ($10.2{\pm}2.2$ vs $16.0{\pm}2.2{\sim}21.0{\pm}2.2%$, p<0.05). In experiment to find the correlation between sperm motility of 4 different motile groups and amount of mitochondria, lower motility group also showed lower level of mitochondria (p<0.05). The mitochondria parameter used in this study showed another possibility to differentiate the sperm motility. Taken together, because below 60% motile semen used in AI reduce the fertility, AI centers should provide the over 60% motile sperm to the farms at the time of AI.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 1998.07a
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• pp.41-42
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• 1998

The changes in calcium binding protein(CBP) of mouse epididymal sperm during their post-testicular differentiation were analyzed by two-dimensional SDS-PAGE. According to dpididymal maturation, capacitation and acrosome reaction of spermatozoa, both quantitative and qualitative changes of CBPs in the epididymal sperm was detected. It suggested that the development of fertilizing ability of epididymal sperm was closely related to the changes in the CBPs profiles of sperm during epidiyaml transit.

• Clinical and Experimental Reproductive Medicine
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• v.23 no.1
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• pp.81-86
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• 1996

The assessment of acrosomal status is important in evaluating the ability of sperm to fertilize the egg. The acrosomal status of sperm from 47 normal volunteers with proven fertility and 167 subfertile men with not to achieve pregnancy for at least 1 year were evaluated with Acrobeads test(FUSO Pharmaceutical Industries, Ltd, Japan) using immunobeads coated with MH61 monoclonal antibody, which is specific for acrosome-reacted sperm. The mean${\pm}$SD of acrobeads score in 47 volunteer group was $2.8{\pm}0.7$, of which 46(97.9%)cases were ${\geq}$ 2. The mean${\pm}$SD of acrobeads score in 167 subfertile group was $1.7{\pm}0.8$, of which 73(79.3%)cases were ${\leq}$ 1. The aerobe ads score in subfertile group were significantly lower(r=0.294, p<0.05) than those in volunteer group. In subfertile group, acrobeads score were well correlated with the sperm density and motility(r=0.275, r=0.281, p<0.01), but not with semen volume(r=0.16) and serum hormone level(FSH r=0.084, LH r=0.036, testosterone r=0.058, prolactin r=0.006 and estradiol r=0.060)(p>0.05). Of 63 subfertile cases with normozoospermia, 22(34.9%)cases showed 0 or 1 of acrobeads score, which means to accompany with a functional defect in spite of normal morphology. As a results, Acrobeads test is not only a technically simple sensitive procedure with good reproducibility in evaluating the sperm fertilizing capacity but also an useful in the evaluation of effectiveness in the treatment of infertility and the separation of acrosome-reacted sperm in the assisted reproductive technique.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.6
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• pp.748-756
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• 2000

Effects of xanthine (X), xanthine oxidase (XO), and catalase (C), $H_2O_2$, and carbohydrates on sperm capacitation, acrosome reaction, and fertilizing ability in vitro were examined. Glucose alone, but not fructose, supported the maximum rate of sperm capacitation and acrosome reaction. However, in the combination of X, XO, and C (XXOC) or $H_2O_2$, fructose alone also supported maximum capacitation, acrosome reaction, and fertilization. Either insufficient or excessive amounts of $H_2O_2$ decreased sperm capacitation and the acrosome reaction. In order to understand how glucose generates $H_2O_2$ or other reactive oxygen species in sperm cells, 6-aminonicotinamide, an inhibitor of the pentose-phosphate pathway (PPP), and apocynin, an inhibitor of NADPH oxidase, were added to sperm suspensions in glucose-containing medium. Results appeared that sperm capacitation, acrosome reaction, and fertilization were consequently inhibited by either one of these compounds. These inhibitory effects were nullified by addition of XXOC. These results support the hypothesis that glucose, in addition to being a substrate for glycolysis, facilitates sperm capacitation and the acrosome reaction by generating reactive oxygen species through G-6-P dehydrogenase and NADPH oxidase.

• Korean Journal of Animal Reproduction
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• v.10 no.2
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• pp.211-217
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• 1986

This experiment was conducted to determine the effect of FSH on in vitro maturation and in vitro fertilizing ability of oocytes recovered from normal follicles of different sizes in superovulated rabbits. Follicular oocytes recovered were cultured in modified Ham's F12 medium containing 0, 0.1, 1.0 and 10$\mu\textrm{g}$ FSH/ml for 18 hours and investigated the degree of cumulus cells expansion and nuclear maturation, which were fertilized with in vivo capacitated rabbit sperm. 1. The number of normal follicles<1.5mm, 1.6 to 2.5mm and> 2.5mm in diameter at 16 to 18hrs after HCG administration was 4.8 (38.8%), 5.5(45.4%) and 3.3(15.8%), respectively. Average percent of oocytes recovered was 69.7% and larger follicles tended to have a higher percent, recovery rate than smaller follicles. 2. The degree of cumulus expansion in medium containing 0.1$\mu\textrm{g}$ FSH/ml was similar to that of control, but markedly decreased under the level of above 1$\mu\textrm{g}$ FSH/ml. The proportions of oocytes which reached the second meiotic metaphase were 57.1, 61.5, 43.8 and 45.0% in medium containing 0, 0.1, 1.0 and 10$\mu\textrm{g}$ FSH/ml, respectively. Oocytes from larger follicles showed a higher nuclear maturation than that from smaller follicles. 3. In vitro fertilization rate of oocytes matured under 1$\mu\textrm{g}$ FSH/ml was slightly, not significant, higher than that of others. 4. Progesterone level in follicular fluid was about 67 to 71ng/ml with no difference in follicular sizes and estradiol-17$\beta$ level was under 25pg/ml.

• Korean Journal of Animal Reproduction
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• v.10 no.1
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• pp.109-120
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• 1986

This study was conducted to define the effect of addition of lysolecithin (LC) and 20% v/v rabbit serum to sperm preincubation medium on the induction of acrosome reaction (AR) an fertilizing ability in vitro of LG-added sperm. Ejaculated rabbit sperm from New Zealand White buck was washed once by centrifugation, then preincubated for 2 or 4 hrs in a chemically defined medium (DM), DM plus 20% rabbit serum or BSA-free DM plus 20% rabbit serum at 37$^{\circ}C$ water bath or CO2 incubator. At the end of preincubation LC was added to the preincubated sperm, which was stained at 0.5 to 4 hr later and examined for AR and sperm motility. For in vitro fertilization, gametes were coincubated in DM up to 24 hrs and thereafter fertilized embryos were incubated in BSM -II up to 48 hrs. Addition of LC to 4-hr preincubated sperm was more effective for the AR and sperm motility than that to 2-hr preincubated sperm and optimal concentration of LC for AR was about 80${\mu}$g/ml. A significant increase in AR occured from 20 to 30 min. after addition of 80 to 100${\mu}$g/ml in 4-hr preincubated sperm. BSA-free DM plus 20% rabbit serum showed a higher AR and sperm motility than those of DM plus 20% rabbit serum in LC-added sperm after 4-hr preincubation. The incidence of AR after 4-hr preincubation and at 30 min after 60${\mu}$g/ml LC addition varied greatly among individual bucks. Sixty ${\mu}$g/ml LC-added sperm showed a slight high cleavage rate over control levels, but 100${\mu}$g/ml LC-added sperm showed lower cleavage rate rather than 60${\mu}$g/ml LC. It is concluded that optimal concentration of LC for high AR induction and sperm motility in 4-hr preincubated sperm was about 80${\mu}$g/ml, but 60${\mu}$g/ml level was more useful for in vitro fertilization.

• Korean Journal of Animal Reproduction
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• v.21 no.3
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• pp.239-246
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• 1997

The objective of this study was to evaluate the variation of fertilizing ability following the morphologically normal sperm ratio in porcine IVF using epididymal sperm The results obtained in this experiment were summarized as follows: 1. When the penetration rate (PR), polysper my rate (PSR), pronuclei formation (2PNF) and mean number of sperm (MNS) per oocyte were evaluated according to the percentage of morphologically normal epididyrnal sperm at insemination($\leq$lO%, 10~30% and $\geq$50%). the PR and PSR of $\leq$50% group (82.4, 87.4%) were significantly higher than those of other two groups ($\leq$lO%; 29.7%, 22.6% and 10~30%; 20.3, 37.0%) (p<0.01). Also, the 2PNF per examined oocytes was significantly high in $\geq$ 50% group (p<0.01). 2. When the $\geq$50% group in epididymal sperm was adjusted to 100% (5x1$^5$ cells/ml) , the PSR and 2PNF were not different between epididymal sperm (86.7, 35.1%) and frozen-thawed ejaculated sperm (86.0. 39.4%) although the PR in epididymal sperm (79.7%) was significantly lower than that in frozen-thawed ejaculated sperm (95.5%)(p<0.01). 3. Also. when the PR, PSR, 2PNF and MNS of epididymal sperm were evaluated according to the oocyte: sperm ratio (1:6000, 1:6650. 1:7700 and 1: 10000) at insemination. the PR, PSR and MNS were increased as the oocyte:sperm ratio increases. However, this result indicated that the 2PNF was high in the oocyte:sperm ratio (1:6000 and 1:6650). Therefore. these results suggested that when the percentage of morphologically normal epididymal sperm was more than 50. the fertilizing a ability was very similar to that of frozen-thawed ejaculated sperm and that the detailed evalu¬a ation of morphological normality in porcine IVF using epididymal sperm should be prerequisite to obtain the more effective fertilizing ability.