• Clinical and Experimental Reproductive Medicine
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• v.25 no.1
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• pp.1-8
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• 1998

To investigate the influences on semen parameters and fertilizing capacity of immunoglobulin (Ig) isotypes and regional distribution of antisperm antibody (ASA) on the human sperm surface. Sixty-seven ASA-positive patients were compared with 96 ASA-negative donors. ASAs in semen showed significant negative effects on both semen parameters and fertilizing capacity; in those with ASAs in the sperm head and/or tail, the reductions were significant. In the head as well as the tail, there was close correlation between fertilizing capacity and both IgG and IgA. Both semen parameters and fertilizing capacity are significantly affected by the presence of ASA in semen. In particular, antibodies IgG to sperm head and/or tail, and antibodies IgA to sperm tail appeared to have a highly detrimental effect on fertilizing capacity.

• Clinical and Experimental Reproductive Medicine
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• v.30 no.1
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• pp.105-109
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• 2003

Objective s: To assess the fertilizing capacity using sperm penetration assay (SPA) to predict the outcome of the in vitro fertilization-embryo transfer (IVF-ET) outcome. Materials and Methods: Semen samples were provided by 129 patients undergoing IVF. We attempted to correlate the extent of sperm penetration under enhanced SPA protocol with the results of fertilization, cleavage, preimplantation embryo development, and pregnancy. Results: Univariate analysis demonstrated a statistically significant correlation between fertilizing capacity and motility, kinetics, fertilization, cleavage and embryo development, and pregnancy rate. By logistic regression analysis, fertilizing capacity was found to be the only variable that was statistically significant with respect to pregnancy rate. Fertilizing capacity, cleavage rate and pregnant rate were significantly higher in pregnant group. However, the fertilization rates was comparable with both group. Conclusions: Lower fertilizing capacity could denote a poorer prognosis for establishing a pregnancy, even after satisfactory fertilization rate is achieved.

• Journal of Embryo Transfer
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• v.7 no.2
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• pp.133-136
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• 1992

Glass wool filtration and swim-up method resulted in inreasing to 58.3% and 62.7% of the progressive motility in frozen-thawed boar sperm, compared to 34.2% in the untreated sperm. Glass wool filtration tended to be more successful than swim-up method for the survival sfter incubation of 38.5$^{\circ}C$ for 3h. Sperm recovered by both the swim-up method and the glass wool filtration method were tested in an in vitro fertilization to determine which of the two techniques would yield sperm with high fertilizing capacity. The results indicated that there was a significantly(p

• Journal of Veterinary Clinics
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• v.10 no.1
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• pp.1-10
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• 1993

To investigate the availability of hamster test in assaying the fertilizing capacity of dog sperm and the effect of canine sperm motility on sperm binding and penetration, semen were collected from four dogs(three dogs had been proven to be fertile and one dog to be subfertile during the past two years) and then preserved in BWW(Biggers, Whitten, Whittingham) medium for about 20 hours. The semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding) and ova penetrated by sperm(penetration), and also numbers of both bound and penetrated sperm per ovum. In comparison between fertile dogs and a subfertile dog, the rate of sperm binding was higher in fertile dogs than the subfertile dog(p<0.01, p<0.05). The number of bound sperm per ovum was considerably higher in a fertile dog than the subfertile dog((p<0.01), and also difference of number of the bound sperm was obtained among the fertile dogs(p<0.01, p<0.05). The rate of penetration as well as the number of penetrated sperm per ovum was higher in the fertile dogs than the subfertile dog(p<0.01, p<0.05). In fertile dogs. the canine semen preserved at 4$^{\circ}C$ for 18 to 22 hours showed from 30 to 80% motility at Insemination, however, no difference in hamster test was obtained according to different degree of sperm motility. These results indicated that hamster test would be of avail in assaying the fertilizing capacity of dog sperm.

• Fisheries and Aquatic Sciences
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• v.27 no.5
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• pp.306-313
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• 2024

The genus Chirostoma is endemic from the Mesa Central of Mexico. It is conformed of 18 species and six subspecies. Five species are in some category of risk, because of this, Chirostoma jordani is an excellent model species to implement biotechnologies like gametes cryopreservation. Aim of present study was to evaluate fertilizing capacity of cryopreserved C. jordani sperm, as alternative to conservation and assisted reproduction in this specie and genus. Males and females were collected from wild Atlangatepec dam stock, Tlaxcala State, Mexico. Seminal quality was evaluated in fresh and cryopreserved semen with three cryoprotective agents (CPAs): 10% dimethyl sulfoxide (DMSO), 10% methanol (MeOH), 14% ethylene glycol (EG) and it was determined its post-thaw fertilizing capacity. Sperm motility percentage decreased during cryopreservation process (p < 0.05). There were not significant differences in post-thaw motility percentage between EG (53.5 ± 1.9%) and MeOH (53.3 ± 1.3%), but DMSO (50.3 ± 0.5%) was significantly different (p < 0.05). Results showed that 0.2 μL fresh semen were enough to fertilize 100% oocytes (n = 60). 10 μL DMSO and 5 μL MeOH and EG cryopreserved semen were necessary to fertilize oocytes 100% (n = 60) (p < 0.05). Cryopreservation and fertilization protocol for C. jordani sperm was efficient and it could be used for its assisted reproduction.

• 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.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 Veterinary Research
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• v.32 no.3
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• pp.435-450
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• 1992

To evaluate the fertilizing capacity of domestic animal spermatozoa by hamster test, semen were collected from 15 boars(Duroc, Landrace, and Yorkshire) and 2 mixed dogs which had been proved to be fertile in the past then, the semen were preserved in BWW medium at $4^{\circ}C$ or $18^{\circ}C$ for about 20 hours and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of sperm binding to the ova, penetration and formation of a male pronucleus, and the numbers of both bound and penetrated sperm per ovum. Both the semen preserved at $18^{\circ}C$ for about 20 hours and that treated by swim up procedure showed considerably higher rates of sperm binding and penetration as well as higher number of penetrated sperm than that preserved at $4^{\circ}C$ for about 20 hours, respectively(p<0.01). Motility of boar sperm at insemination was from 40 to 90% and no difference in hamster test was obtained according to different degree of sperm motility. Abnormality in morphology of boar sperm at insemination was from 6 to 45% and no difference in hamster test was obtained according to different degree of sperm abnormality. The sperm concentrations of $7{\times}10^7$ and $7{\times}10^6$ showed considerably higher rates of sperm binding and penetration as well as higher number of bound sperm than that of $7{\times}10^4$ (p<0.01) along with the same higher results than that of $7{\times}10^5$(0<0.05), respectively. Boar sperm showed considerably higher rates of sperm binding and penetration as well as higher numbers of both bound and penetrated sperm than dog sperm, when both semen were treated by BWW＋heparin medium and swim up procedure, respectively. These results indicated that fertile boar sperm showed considerably lower rates in the results of hamster test, when preserved at $4^{\circ}C$ for about 20 hours and in lower concentration of sperm than when preserved at $18^{\circ}C$ for about 20 hours and in higher concentration of sperm, respectively, and at the same time considerably higher results than fertile dog sperm, consequently to prove that hamster test would be of great value in assaying the fertilizing capacity of boar sperm.

• Journal of Veterinary Clinics
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• v.9 no.2
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• pp.373-390
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• 1992

To assay the fertilizing capacity of domestic animal spermatozoa by hamster test, semen were collected from 13 boars(Duroc. Landrace and Yorkshire) which had been proved to be fertile in the past. then, were preserved in BWW medium or in raw state at 18$^{\circ}C$ or at room temperature. The preserved semen were given each different treatment according to the experimental design and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of ova bound with sperm(sperm binding). ova penetrated by sperm(penetration) and formation of a male pronucleus(pronucleus formation) and also numbers of both bound and penetrated sperm per ovum. Between BWW and TBM medium for boar sperm. no difference in the results of hamster test was obtained. The boar spermatozoa in BWW medium, BWW with caffeine, BWW with heparin, and BWW with both caffeine and heparin showed no difference in the results of hamster test. The boar spermatozoa in BWW medium containing both calcium and RSA showed considerably higher rates of sperm binding, penetration and pronucleus formation as well as higher numbers of both bound and penetrated sperm than those not containing calcium with or without BSA( p<0.01) and also the same results higher than that containing calcium without BSA( p< 0.05). The boar spermatozoa irradiated by X-ray(70 KVP, 20mA) for 3 seconds. then, maintained at 18$^{\circ}C$ for 18 hours showed considerably lower rate of sperm binding than all the other groups including the control and X-ray groups irradiated by smaller dose or maintained for shorter period(p<0.01), and also showed lower number of bound sperm than the other groups(p<0.01, p<0.05). All the control groups of both raw and diluted sperm in BWM medium showed higher rates of sperm binding, penetration and pronucleus formation as well as higher number of penetrated sperm than all the X-ray groups irradiated for 3 seconds(70KVP, 20mA) and maintained for either 3 or 18 hours (p<0.01, p<0.05). At the same time the control groups of diluted sperm showed considerably higher rates of sperm penetration and pronucleus formation than the control group of raw sperm( p<0.01). These results indicates that fertile boar sperm showed considerably lower rates In the results of hamster test, when incubated in the medium without calcium and irradiated by X-ray than when incubated in the medium with calcium and not irradiated by X-ray, respectively, to prove consequently that hamster test would be of great value in assaying the fertilizing capacity of boar spermatozoa.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.1
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• pp.57-66
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• 2002

Objective : This study was designed to investigate the interrelationship and clinical usefulness of sperm morphology by strict criteria (SM), acrosome reaction following ionophore challenge test (ARIC) and sperm penetration assay (SPA) using zona-free hamster ova as prognostic factors in in vitro fertilization. Materials and Methods: Semen samples were provided by 83 patients undergoing IVF. We first evaluated the differences between normal fertilization group and poor fertilization group on three andrologic tests. Secondly, we analyzed the relationship between the three andrologic tests and in vitro fertilization on IVF settings. Finally, we evaluated the effectiveness of the three andrologic tests as the prognostic indicators for fertilizing ability. Results: The fertilization rate of all men in the poor fertilization group was less than 30%; but there was no evidence that this poor fertilization was due to oocyte defects. The results of three andrologic tests were significatly higher in normal fertilization group. Fertilization rate (%) in vitro was highly correlated (p<0.001) with % normal sperm by SM, ARIC value (%), and SPA result. By using Receiver-Operator-Characteristic curve (ROC), we evaluated the effectiveness of these three tests. The sensitivity and specificity of SM, ARIC test and SPA in predicting fertilization potential in IVF setting were 76% and 75%, 84% and 90%, and 76% and 95%, respectively. Conclusion: Our data suggest that the three andrologic tests can be reliable tools as prognostic factors of sperm fertilizing ability. Among these test, ARIC test and SPA gave more accurate information on fertilizing capacity. ARIC test was shown to have a predictive value for fertilizing ability comparable to that of SPA that appears to be a simple and cost-effective addition to current andrology laboratory. Combined application of these three tests may give more information on predicting sperm fertilizing capacity.