• Proceedings of the KSAR Conference
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• 2004.06a
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• pp.277-277
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• 2004

The purpose of this study was to investigate the effect of different conditions (osmolity, solution, incubation times, comparison of fresh and frozen/thawed semen and storage times) on the swelling of canine spermatozoa. Employing the hypoosmotic swelling test (HOST), the membrane integrity of spermatozoa in different solutions (sucrose, fructose, latose, Na-citrate, Na-citrate plus sucrose, Na-citrate plus fructose and Na-citrate plus lactose were 61.4%, 66.2%, 62.5%, 68.1%, 62.0%, 68.5% and 60.2%, respectively. (omitted)

• Reproductive and Developmental Biology
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• v.30 no.1
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• pp.71-74
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• 2006

Hypoosmotic swelling test (HOST) is used for evaluating the plasma membrane function and fertilizing ability in mammal spermatozoa. However, HOS solutions and experimental conditions have not been determined clearly for assessing canine spermatozoa. This study was conducted to examine the HOS solutions and assay conditions, including incubation time (30 to 120 min), storage temperature (4, 17 and $20^{\circ}C$), semen status (fresh and frozen). Maximum spermatozoal plasma membrane swelling was obtained in an 150 mOsm Na-citrate/Fructose solutions with an incubation time for 45 min. The storage temperature and semen status affected the percentage of HOS positive spermatozoa. The HOS test adapted to canine spermatozoa in this study was simple and highly consistent assay with good repeatability. The optimal condition of HOST in canine spermatozoa is an 150 mOsm Na-citrate/Fructose solutions with an incubation time for 45 min regardless of semen storage temperature and semen status.

• Journal of Aquaculture
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• v.17 no.1
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• pp.8-11
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• 2004

Milt of the catfish was stripped into immobilizing solution containing 175 mM NaCl and 30 mM Tris at pH 7.8 and was successfully cryopreserved after a stepwise freezing procedure. After stepwise thawing, motility of spermatozoa was slightly lower than that of fresh sperm. Batches of 40-80 eggs were fertilized with cryopreserved spermatozoa, after thawing and activation in solution containing 50 mM NaCl, 20 mM Tris and HCl at pH 7.8; this resulted in 62.2% fertilization success, compared to 70.6 % with fresh sperm.

• Reproductive and Developmental Biology
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• v.35 no.3
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• pp.203-207
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• 2011

The objective of this study was to determine the effect of semen extenders on the motility, viability and fertility in vitro of spermatozoa during storage of fresh boar semen diluted in different commercial extenders used for pig artificial insemination (AI). In this experiment, semen were diluted in Androhep plus, Beltsville Thawing Solution (BTS), Modena, Seminark and Vitasem LD. Five ejaculates were collected from three Duroc boars and sub-samples were diluted ($30{\times}10^6$ spermatozoa/ml) in different extenders. Semen was stored at $170^{\circ}C$ for 10 days. Sperm motility and viability was assessed using Computer-Assisted Semen Analysis (CASA) and flow-cytometry on 1, 3, 5 and 10 day post collection The motility of spermatozoa stored in different extenders was gradually decreased by increasing the duration of storage of semen. However, there was not significant1y different in the sperm motility and viability among other extenders. On the other hand, the in vitro-matured oocytes were fertilized and cultured in vitro to assess the fertility of boar spermatozoa stored for 3 and 10 days in different extenders. The percentage of morula and blastocyst were taken as indicators of fertility in vitro of spermatozoa. Therefore, there were no differences in the rate of embryos developed to the molular and blastocyst stage. There were no differences in the motility and fertility in vitro among 5 kinds of commercial boar semen extenders.

• Development and Reproduction
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• v.5 no.2
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• pp.159-165
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• 2001

Follicular fluid has pivotal effects on attraction and motility of spermatozoa for successful fertilization. The effect of samples of human follicu1ar fluid(hFF) on attraction and motility of spermatozoa was investigated. Capillary tubes loaded with one of these samples, hFF sample A collected from patients with tubal factor, hFF sample B collected from patients with male factor, m-HTF and heated hFF sample were used for assessment of attraction and motility of spermatozoa following culture of 1, 2, and 4 hrs. Number and motile rate of spermatozoa in the tubes loaded with hFF sample A were significantly(P<0.05) higher than those of m-HTF, hFF sample B and heated hFF. Although the fresh hFF tended to increase the attraction of spermatozoa as compared to inactivated hFF, there was no significant difference between treatments.

• Journal of Aquaculture
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• v.10 no.3
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• pp.273-279
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• 1997

Condition for fresh storage of tiger puffer in the liquid state were investigated in several experiments. When marine fish ringer solution and 1% NaCl were used as the diluent for the short-term preservation method, spermatozoa activity index (SAI) and survival rate showed the best result among the various diluents tested. The dilution rate for the shortterm preservation of spermatozoa was suitable between 3 and 5 times with the 1% NaCi diluent. The appropriate range of temperature for the short-term preservation showed between 0 and $5^{\circ}C$. In order to keep high SAI and survival rate of spermatozoa, antibiotic addition (800 ppm neomycin) could be suggested. These results indicated that the short-term preservation method could be employed in tiger puffer spermatozoa.

• Asian-Australasian Journal of Animal Sciences
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• v.17 no.7
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• pp.885-891
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• 2004

To evaluate the feasibility of using sperm-mediated gene transfer (SMGT) for carrying foreign gene into chicken oocyte, a reporter gene, CX-EGFP, was used in this study. The reporter gene was first mixed with liposome or liposome-like compound and the mixtures were further combined with ejaculated cock spermatozoa. The spermatozoa treated with liposome and CX-EGFP mixture was subsequently coincubated with DNaseI to remove the extra DNA which insured the authenticity of positive signals. The treated sperms were then subjected to transgene (reporter gene) existence analysis and artificial insemination of laying hens. Obtained results indicated that the spermatozoa were able to take-in the foreign DNA; which was confirmed by polymerase chain reaction and Southern blot analysis. In the following experiment, fresh ejaculated sperms were mixed with CX-EGFP-liposome or CX-EGFP-liposome-like complex then used for artificial insemination of each of six laying hens. Eggs laid between day-3 and day-7 post insemination were collected. Newly hatched chicks, two out of 53 from CX-EGFP/liposome treated group and two out of 21 from CXEGFP/liposome-like treated group, were proven to be transgenic. This study suggests that SMGT is a powerful method for generating transgenic chickens.

• Fisheries and Aquatic Sciences
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• v.5 no.2
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• pp.87-96
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• 2002

The physico-chemical properties of fresh milt of marbled sole, Limanda yokohamae, brown sole, Limanda herzensteini, starry flounder, Platichthys stellatus and olive flounder, Paralichthys olivaceus among flatfishes, and the fine structure of their cryopresserved spermatozoa were investigated. The highest concentration of sperm among these four species was $3.60\pm1.35\times10^{10}/mL$ in marbled sole. Osmolality and pH of seminal plasma in four flatfish species were approximately 330 mOsm/kg and 7.6-8.1, respectively. Seminal plasma compositions showed interspecific differences. The sperm heads of marbled sole, brown sole and starry flounder were ellipsoidal and that of olive flounder was round. The numbers of mitochondria of these four species were eight in marbled sole, seven in brown sole and starry flounder, and six in olive flounder. Cross-sectional view of flagellum showed typical 9+ 2 structure in all species. Most of sperms cryopreserved with a proper method had no visible ultrastructural changes after freeze-thawing, compared with the fresh sperm, but in a few cases, swelling of their heads and midpiece regions were observed.

• Journal of Embryo Transfer
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• v.32 no.3
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• pp.235-241
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• 2017

To preserve genetic materials, cryopreservation of the semen from live animals is the main technique to establish cryo-banking system which could be used for artificial insemination and embryo transfer. However, the population of Korean black goat (KBG) becomes to dwindle in number and is now faced genetic erosion by crossbreeding with non-native breeds in small KBG farms. In this study, simple freezing method was used to preserve frozen semen from KBG using spermatozoa of cauda epididymis (CE) and electro-stimulated semen (ES). The negative effects of seminal plasma on fresh sperm was confirmed using precipitation test of Triladyl egg yolk diluent and sperm viability after thawing was compared between CE and ES spermatozoa. When seminal plasma of fresh ES semen was washed with semen washing media (SWM), the rates of live sperm shown no significant difference between CE and ES spermatozoa before freezing. However, the survival rate of frozen/thawed CE sperm was higher than ES ($74.6{\pm}10.6%$ vs $53.8{\pm}5.2%$) with significant difference (p < 0.05). The results of longevity test on frozen/thawed sperm from CE showed healthier sperm than ES. Therefore, spermatozoa from CE could be used for cryo-banking system in KBG lines. The more studies are needed to increase survival rate of ES semen.

• Reproductive and Developmental Biology
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• v.29 no.3
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• pp.149-154
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• 2005

The purpose of this study was the analysis of sperm ability in Specific Pathogen Free (SPE) miniature pig for production of bio-organ. The collected semen was diluted with extender and stored at $17^{\circ}C$t for up to 7 days. The semen samples were evaluated at 0, 1, 3, 5, and 7 days of storage for analysis of sperm ability. Sperm ability was evaluated by examining viability, progressive motility, sperm abnormality and intensity of the sperm membrane. Also, the semen was processed according to the convenient freezing method, and frozen-thawed sperm was evaluated by examining viability, capacitation and acrosome reaction using chlortetracycline (CTC) staining. Motility of spermatozoa of SPF miniature pig was significantly (P<0.05) lower on 3 days or later compared to the Duroc, Yorkshire and Landrace in domestic boar. The percentage of abnormal spermatozoa of Landrace were significantly (P<0.05) higher than in SPF miniature pig, Duroc and Yorkshire that had a similar percentage on 5 or 7 days of sperm storage. The percentage of spermatozoa with coiled tail decreased during the storage period but there were no significant difference. On the other hand, viability of frozen-thawed spermatozoa had a significantly (P<0.05) lower in SPF miniature pig than in other domestic boars. CTC patterns had no significant difference, but SPF miniature pig had higher percentage of capacitated spermatozoa and lower percentage of acrosome-reacted it than domestic boars. Therefore, this study suggest that it is necessary to develop the suitable extender and freezing methods methods for the high viable rate and fertilizing ability in vitro.