• Journal of Animal Reproduction and Biotechnology
• /
• v.34 no.2
• /
• pp.100-105
• /
• 2019

The purpose of this study was to evaluate the effect of addition of ethylene glycol, glycerol and sucrose to TCG (Tris, Citric Acid, Glucose, Egg Yolk) and DMSO Frozen. The extender containing Egg yolk concentration (10%, 20%) affects viability and acrosome morphology of rabbit sperm. Sperm viability was then assessed for the freezing extenders TCGD (Tris + Citricacid + Glucose + DMSO), TCGED (Tris + Citricacid + Glucose + Egg yolk + DMSO), TCGGD (Tris + Citricacid + Glucose + Glycerol + DMSO) and TCGSD Tris + Citricacid + Glucose + Sucrose + DMSO) during thawing at 38℃. for 20 seconds, respectively. TCG + 10% egg yolk (viability: 77.0 ± 0.8, NAI: 73.3 ± 0.9) was significantly (sperm viability and normal acrosome interaction (NAI)) higher than TCG + 20% egg yolk (70.7 ± 1.1, 70.0 ± 0.9) in the sperm normalcy analysis according to the yolk concentration. TCGGD (53.4 ± 0.1, 62.3 ± 0.4), TCGSD (61.3 ± 0.0, 67.1 ± 0.1) sperm viability and normal acrosome interaction (NAI) in frozen spermatozoa are TCGD (46.4 ± 2.8 and 56.3 ± 1. 4) and TCGED (23.0 ± 1.1 and 54.6 ± 1.4) extenders was thawed at 38℃ for 20 seconds. According to the results from each frozen bulking agent, sperm membrane integrity by hypotonic swelling test (HOST) analysis in TCGGD (59.8 ± 0.7), TCGSD (59.3 ± 0.5) was significantly high compared to other experimental groups (p < 0.05). In conclusion, these results suggested that TCGGD and TCGSD extenders enhance survivability of rabbit sperm after frozen-thawing.

• Reproductive and Developmental Biology
• /
• v.34 no.1
• /
• pp.41-46
• /
• 2010

The objective of this study was to evaluated the efficiency of sperm cryosurvival using each extenders Triladyl and LEY containing egg yolk to the cryopreservation of separated sperm by percoll in miniature pig. The ejaculated semen from miniature pig was separated by 65% percoll and un-separated sperm as a control before freezing. The freezing of diluted semen added with Triladyl containing egg yolk and LEY solution (solution I: 11% Lactose or Triladyl + egg yolk; solution II: solution I + glycerol + OEP). Analysis of sperm ability was estimated by viability, capacitation acrosome reaction using chlortetracycline (CIC) the morphologic abnormality and hypoosmotic swelling test(HOST). The groups were designed that as separated sperm by Percoll with Triladyl(ST) or LEY(SL) for cryopreservation. And unseparated sperm with Triladyl(UT) or LEY(UL). As a results, the viability was higher significantly(p<0.05) in ST, SL, UT than UL extender. The morphologic abnormality was measured significantly (p<0.05) lower in ST than other extenders. The AR-patterned in CTC analysis was measured significantly(p<0.05) lower in SL and UL than other extenders. In conclusion, using Triladyl extender resulted in viability and morphology of separated sperm by percoll that were effective than using LEY extender, but it resulted in capacitation acrosome reaction was lower than using LEY extender.

• Reproductive and Developmental Biology
• /
• v.32 no.2
• /
• pp.127-133
• /
• 2008

The objective of this study was two folds: to investigate the relationship between paternal identification rate and sperm quality parameters such as motility and sperm chromatin structure assay after heterospermic insemination; to see if mutual complement between tests and development of useful technique to enhance the fertility in artificial insemination. In individual boar's fertilizing ability, 3 high fertility boars showed significantly high fertility (p<0.05) compared to 3 low fertility boars, but there was no difference in litter size between two groups. Sperm motility test in pooled and individual semen using computer assisted sperm analysis (CASA) revealed that no significant difference among boars. The high fertile boar showed tendency of low %Red (High red fluorescence/green+red fluorescence) in sperm chromatin structure assay (SCSA) but paternal identification rate from piglets did not differ after heterospermic insemination. The correlation coefficient between individual or pooled semen function test and farrowing rates were well correlated as follows: %Red with litter size (r= - 0.53, p=0.03); %Red with paternal identification rates (r=-0.51, p=0.03); paternal identification rates with litter size (r=0.57, p=0.02). These results indicate that sperm chromatin structure assay and sperm quality parameter test in pooled semen are useful method to predict and evaluate the fertilizing capacity after heterospermic insemination in boars.

• Reproductive and Developmental Biology
• /
• v.40 no.4
• /
• pp.33-37
• /
• 2016

This study was conducted to evaluate effect of ${\alpha}$-linolenic acid (ALA) and bovine serum albumin (BSA) on viability, acrosome reaction and mitochondrial intact in frozen-thawed boar sperm. The boar semen was collected by gloved-hand method and cryopreserved using freezing extender containing 3 ng/mL ALA and/or $20\;{\mu}g/mL$ BSA. Cryo-preserved boar sperms were thawed in $37^{\circ}C$ water-bath for 45 sec to analysis. Viability, acrosome reaction, and mitochondrial intact were analyzed using flow cytometry. In results, viability of frozen-thawed boar sperm was significantly higher in only ALA+BSA supplement group than control group (p<0.05), whereas there was no difference either in ALA or BSA supplement. However, acrosome reacted sperm in both of live and all sperm population were significantly decreased in all treatment groups than control (p<0.05). Interestingly, mitochondrial intact of boar sperm was enhanced in ALA and ALA+BSA groups compared with control (p<0.05). In this study, we showed that supplementation of ALA and BSA in freezing extender enhanced the sperm viability, mitochondrial intact and decrease acrosomal membrane damage. In conclusion, our findings suggest that quality of frozen-thawed sperm in mammalians could improve by using of ALA and BSA.

• The Korean Journal of Malacology
• /
• v.30 no.3
• /
• pp.211-218
• /
• 2014

Spermatid differentiations during spermiogenesis and sperm ultrastructures in male Mercenaria stimpsoni were investigated by transmission electron microscopic observations. In the early stage of the spermatid during spermiogenesis, a few granules and a proacrosomal granule, which is formed by the Golgi complex, become a proacrosomal vesicle. Consequently, it becomes an acrosome by way of the process of acrosome formation. The morphologies of the sperm nucleus type and the acrosome of this species have a curved cylindrical type and cap shape, respectively. The spermatozoon is approximately $48-51{\mu}m$ in length including a curved cylinderical sperm nucleus (about $4.18{\mu}m$ long), an acrosome (about $0.52{\mu}m$ in length) and tail flagellum ($42-45{\mu}m$ long). As some ultrastructural characteristics of the acrosomal vesicle, the peripheral parts of two basal rings show electron opaque part (region), while the apex part of the acrosome shows electron lucent part (region). These charateristics of the sperm belong to the family Veneridae in the subclass Heterodonta, unlike a characteristic of the subclass Pteriomorphia showing all part of the acrosome being composed of electron opaque part (region). Therefore, it is easy to distinguish the families or the subclasses by the acrosome structures. Exceptionally, In particular, a cylinder-like nucleus of the sperm is curved (the angle of the nucleus is about $80^{\circ}$), as seen in some species of Veneridae (range from $0^{\circ}$ to $80^{\circ}$). The number of mitochondria in the midpiece of the sperm of this species are four, as one of common characteristics appeared in most species except for a few species in Veneridae in the subclass Heterodonta. Cross-sectioned axoneme of the sperm tail flagellum shows a 9+2 structure.

• Journal of Aquaculture
• /
• v.11 no.1
• /
• pp.67-75
• /
• 1998

Experiments were performed to obtain cryopreservation techniques of black seabream (Acanthopagrus schlegeli) sperm. For sperm collection, brood stock reared in recirculating seawater system and fed with the commercial feed during experimental period. The results indicated that following cryopreservation method in block seabream sperm could be employed. Post-thaw survival rate of sperm revealed the highest value ($80{\pm}1.4$%) in 3% sodium citrate as a diluent for the cryopreservation. Cryopreserved sperm diluted with 5.4% glucose showed the highest fertilization rate to the ovulated eggs. Glycerol was a better cryoprotectant than dimethyl sulfoxide in sperm cryopreservation : survival rate and fertilizing capacity of cryopreserved sperm were decreased according to increase of glycerol concentration and varied in renges of 0.8~59.3% and 32.5~69.4% with 5~30% glycerol, respectively. A few of cryopreserved spermatozoa showed the enlarged head with granulated chromatin and ruptured plasma membrane by freezing and thawing injuries compared with unfrozen normal spermatozoa.

• The Journal of Korean Medicine
• /
• v.27 no.3 s.67
• /
• pp.63-76
• /
• 2006

Objective: These studies were undertaken to evaluate the effects of the different administration duration of Morindae Radix extract solution on spermatogenic abilities such as concentration, motility and morphological normality of sperm from the testes and the activities of sperm hyaluronidase. Materials and Method: We used 8-week-old ICR mice and administered 0.3mg/g extract solution of Morindae Radix once a day for 30, 60, 90 and 120 days. The control group was administered normal saline in the same way and duration. We examined the number of total, motile and normal sperm from the cauda epididymis. We also compared the testicular tissue, especially seminiferous tubules, between the control and treated groups by histochemical methods. Finally, we observed the difference of sperm hyaluronidase activities between the control and treated groups. Results: Significant administration duration-dependent differences were observed in the concentration of total sperm, motility and normality of spermatozoa of the Morindae Radix extract solution administered groups compared to the control group. In the histological analysis of the testicular tissues, the enlargement of testicular lobe diameter and apparent vasculogenesis between testicular lobes were observed in the Morindae Radix extract solution administered groups compared to the control group. Also, the activity of hyaluronidase was significantly increased in the Morindae Radix extract solution administered groups compared to the control group. Conclusions: This study shows that the beneficial effect of Morindae Radix extract solution on the concentration, motility and morphology of sperm, the testicular tissues and the activities of sperm hyaluronidase increased the greater the duration the mice were administered it. We suggest that Morindae Radix may be useful for the treatment of male sexual dysfunction and infertility.

• Development and Reproduction
• /
• v.18 no.3
• /
• pp.179-186
• /
• 2014

Characteristics of the developmental stages of spermatids during spermiogenesis and phylogenetic classicfication of the species using sperm ultrastructures in male Crassostrea ariakensis were investigated by transmission electron microscope observations. The morphology of the spermatozoon of this species has a primitive type and is similar to those of Ostreidae. Ultrastructures of mature sperms are composed of broad, modified cap-shaped acrosomal vesicle and an axial rod in subacrosomal materials on an oval nucleus, four spherical mitochondria in the sperm midpiece, and satellite fibres which appear near the distal centriole. The axoneme of the sperm tail shows a 9+2 structure. Accordingly, the ultrastructural characteristics of mature sperm of C. ariakensis resemble to those of other investigated ostreids in Ostreidae in the subclass Pteriomorphia. In this study, particularly, two transverse bands (stripes) appear at the anterior region of the acrosomal vesicle of this species, unlike two or three transverse bands (stripes) in C. gigas. It is assumed that differences in this acrosomal substructure are associated with the inability of fertilization between the genus Crassostrea and other genus species in Ostreidae. Therefore, we can use sperm ultrastructures and morphologies in the resolution of taxonomic relationships within the Ostreidae in the subclass Pteriomorphia. These spermatozoa, which contain several ultrastructures such as acrosomal vesicle, an axial rod in the sperm head part and four mitochondria and satellite fibres in the sperm midpiece, belong to the family Ostreidae in the subclass Pteriomorphia.

• Herbal Formula Science
• /
• v.14 no.1
• /
• pp.168-175
• /
• 2006

The purpose of this study was to investigate the effect of water extract of Dioscoreae Rhizome on the reproduction activity of Wistar male rat. The group treated with 1.0 g/kg/day oral administrations of water extract of Dioscoreae Rhizoma (SK0l6) during 8 weeks was compared with the normal group. Sperm count, sperm motility, body weight, and testis weight were investigated in two groups. Sperm count. body weight, and testis weight of two groups did not show the significant difference. But sperm motility of the treated group was reduced significantly (the normal group: 56.43 %, SK0l6 group: 53.47 %, p < 0.05). According to the results, SK0l6 have no effects on sperm count, body weight. and testis weight but reduced sperm motility significantly.

• Reproductive and Developmental Biology
• /
• v.30 no.1
• /
• pp.27-34
• /
• 2006

This study was conducted to compare the reproduction ability of the wild type boar and recombinant human erythropoietin (hEPO) transgenic boar semen. Ejaculated boar semen was analyzed by flow cytometry, Elisa and IVF methods. In experiment 1, flow cytometric analysis showed that the live sperm ratio of transgenic boar sperm significantly lower (P<0.05) than that of wild type boar after incubation at 20, 22, 24 and 26 hr. In experiment 2, the presence and levels of various cytokines (IL-6, IL-10 and $TNF-{\alpha}$) to related animal reproduction in the seminal and blood plasma were examined using specific enzyme immunoassay. There was no significant difference between both groups. In experiment 3, the fertilizing capacity and developmental ability of both boar sperm were compared. The transgenic boar sperm had a significantly low capacity of penetration, sperm-zona binding, embryo development, and blastocyst formation compared to wild type sperm (P<0.05). These results suggest that transgenic boar sperm harboring hEPO gene has low sperm viability than wild type boar, and it is a reason to decrease of fertility and litter size.