• Journal of Veterinary Clinics
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• v.28 no.6
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• pp.571-575
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• 2011

Sperm cryopreservation methods have been improved over the last few decades. However, an optimized thawing rate has not yet been established. Therefore, we investigated the effect of thawing rate on sperm function after cryopreservation. The ejaculates collected from beagle dogs were cryopreserved and then thawed at two different thawing rates ($37^{\circ}C$ for 1 min or $70^{\circ}C$ for 15 sec). The thawed sperm were evaluated for motility, viability, morphology, plasma membrane integrity, phosphatidylserine (PS) translocation, and intracellular $H_2O_2$ level. The sperm thawed rapidly at $70^{\circ}C$ showed improved motility, viability, normal morphology, plasma-membrane integrity and non-PS translocation compared to the sperm thawed slowly at $37^{\circ}C$ (P < 0.05). However, the intracellular $H_2O_2$ levels were not significantly different between the rapid- and slow-thawed sperm (P > 0.05). In conclusion, sperm rapid thawing at $70^{\circ}C$ could improve the function of cryopreserved canine sperm, and the appropriate thawing rate would enhance the quality of the cryopreserved sperm.

• Journal of Embryo Transfer
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• v.18 no.1
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• pp.51-59
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• 2003

This study was carried out to evaluate the effect of different freezing and thawing rates on the viability, motility and acrosomal changes of frozen canine spermatozoa. The ejaculated semen was extended with Tris-egg yolk buffer containing 8% glycerol and equilibrated for 60 min after cooled to 4$^{\circ}C$ for 58 min. The straws were cryopreserved gradually by slow-cooling at different distance(6, 10 and 17 cm, respectively) from the liquid nitrogen (L$N_2$) to achieve temperature rate of 3, 8.9 and 19$^{\circ}C$ /min. Thawing of the straws was performed in a water bath fur 2 min at 37$^{\circ}C$ and 55$^{\circ}C$ , respectively. The motility of frozen-thawed spermatozoa was assessed by phase-contrast microscopy. To assess their viability and acrosome content, spermatozoa were stained with a vital stain and Fluorescence conjugated lectin Pisum Savitum Agglutinin (FITC/PAS), respectively. Concentration of the ejaculated fresh semen was normal range of 3.44 $\times$ 10$^{8}$ /ml. Freezing temperature were reduced to -110, -70 and -35$^{\circ}C$, as higher distance from liquid nitrogen, 6, 10 and 17 cm, respectively. Freezing at 3$^{\circ}C$/min in distance of 17 cm from liquid nitrogen yielded better motility, viability and rate of intact acrosome than 8.9 or 19$^{\circ}C$/min and the optimal thawing was 37$^{\circ}C$ for 2 min.

• Korean Journal of Animal Reproduction
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• v.11 no.1
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• pp.63-72
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• 1987

Effects of sperm and seminal plasma isoant ibodies upon the rate of superovulation and ferlil ization were slud ied in both normal and immunized rabbits. The results obstained were summarized as follows: 1. On examin.ltion of the superovulation in the immunized animals, the average number of ovulation points was 22.1 and 25.3 for spermtreated animals and for seminal plasma-treated animals, respec tively. As compared to the con¬trol group of 41.0 in number, the immunized showed statisfical significance in ovulation (P<0.05). 2. In ovary weight and follicle's size there were no significant differences among the three groups, whereas sperm-and seminal plasma¬treated groups had an average rate of fertiliza¬tion of 62.X% and 5X .0%, respectively, in re¬markable contrast to the control hTfOUP of 91.4'1'0 (P<0.05). 3. When the animals were inseminated with a mixture of semen plus sperm or seminal plasma antisera, a sharp reduction of fertilization was observed with 5.6% and 16.0% as compared to the control group (P<0.05). Consequently, immunization with either sperm or seminal plasma had a substantial effect on fertilization.

• Parasites, Hosts and Diseases
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• v.31 no.3
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• pp.183-192
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• 1993

A transmission electron microscopic study was performed to observe the ultrastructures of the male germinal cells and spermatozoa of Fibricola seoulensis. Spermatogonia were found in the periphery of the testis and characterized by large nuclei and comparatively little cytoplasms. Spermatocytes contained an oval to spherical nucleus. Their nuclear volume was little larger in comparative to that of cytoplasm, and the chromatin was comparatively little. The early spermatids were characterized by a great amount of cytoplasm, and numerous mitochondria encircled the nucleus. In a more advanced spermatids the electron-dense strands of chromatin appeared in the nucleus, and a pair of rootlet of the axoneme and a microtubule-organizing center (MTOC) were observed near the nucleus. The sectioned spermatozoa were found in the testis and the seminal vesicle. Their cross sectional views were divided into 6 types when they were distinguished on the basis of the morphology and components. The spermatozoa of F. seoulensis showed two flagella of 9+1 type axoneme.

• Korean Journal of Ecology and Environment
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• v.35 no.3 s.99
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• pp.232-236
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• 2002

In an effort to uncover the reproduction of Korean brook perch Coreoperca herzi testis anatomy and sperm morphology were studied. Fish samples were collected in the Sooypcheon river from May to October 2001. White-colored testes have wedgeshaped external morphology, and developed symmetrically in the dorsal cavity of the trunk. Isogenetic germ cells developed in the cyst located in seminiferous lobule. Each lobule showed significant asynchrony in the spermatogenic stage of the cyst. Sperm was 43 ${\mu}$m in length. The round head was 2.2 ${\mu}$m long. The middle piece developed beneath the head was 0.5 ${\mu}$m long. Tail was 40 ${\mu}$m in length. Coomassie brilliant blue (CBB) gave rise the intense staining in the apex of sperm head and middle piece, suggesting the possible development of acrosome.

• Applied Microscopy
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• v.40 no.3
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• pp.169-176
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• 2010

Black molly (Poecilia sphenops) and sailfin molly (Poecilia latipinna) are a teleost belonging to Poeciliidae. The spermatogenesis between two species were investigated by light and electron microscope. The whitish testes of both black molly and sailfin molly were located between intestine and air bladder. The size of testis was major axis 7 mm, minor axis 2 mm. The testis contained numerous testicular cysts. In both black molly and sailfin, primary spermatocytes were comparatively large ellipsoidal, and mitochondria showed a marked development. The secondary spermatocyte was smaller than that of primary spermatocyte, highly condensed according to their development. The nucleus with electron-dense was round shape and flagella started to be formed. In spermiogenesis, chromatin was more condensed. The mitochondria were rearranged along the tail. The number of mitochondria was 2 to 4 in cross section and 8 to 10 in longitudinal section. The head of mature sperm was long cone shape and had not acrosome. The microtubules of flagella were arranged 9+2 structure. Also, the tail of sperm have lateral fins. In conclusion, spermatogenesis and sperm morphologies of these two species were same. These morphological similarity seems to be an indication of the Poeciliidae.

• Development and Reproduction
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• v.5 no.2
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• pp.145-150
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• 2001

The testis of Neoditrema ransonneti is testicular tubule type, each testicular tubule consists of numerous testicular cysts which contain numerous germ cells showing the same developmental stage. During spermatogenesis, well developed rough endoplasmic reticula and the Golgi complex are observed in the cyst cell. Secretory activity of cyst cell was the highest in the late spermiogenesis. Sperm binding materials of spermatozeugmata are secreted by testicular cyst cell. One spermatozeugmata is produced by a testicular cyst during spermatogenesis. The capsular structure was not found in the spermatozeugmata discharged from male. According to observations under transmission electron microscopy approximately 1,500 to 1,700 of sperm tails were observed in the cross sectioned spermatozeugmata.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.33 no.3
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• pp.171-175
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• 2000

The ultrastructures of germinal cells of male marsh clam, Corbicujar lena were studied. The mature sperm was primitive type, consisting of head, middle piece and tail. The mature sperm was whip-shaped and its head was divided into two parts; the acrosomal part shaped long hollow cone about $3{\mu}m$ in length and the sperm nuclear part shaped a long stick about $9\;{\mu}m$ in length. The posterior part of the sperm nuclear projected to centriole, The middle piece of the sperm-nuclear had four mitochondria and two centrioles. The sperm tail part had the 9+2 microtubular arrangement known as a typical pattern, During spermiogenesis, chromatin within sperm nuclear became fiberic materials by condensation.

• Journal of the korean veterinary medical association
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• v.37 no.10
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• pp.918-930
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• 2001

• Korean Journal of Ichthyology
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• v.19 no.4
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• pp.286-291
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• 2007

The bitterling, Acheilognathus koreensis spermatozoon has been examined by electron microscopy. The epididymal spermatozoa of A. koreensis are representing typical characteristic of cyprinid spermatozoa including the lateral insertion of flagellum, the organization of centriolar complex in shallow nuclear fossa and the asymmetrical arrangement of mitochondria. The sperm mid-piece contains a large mitochondrion characteristic enclosed by membranous vesicles. The mitochondria aspect is different from that of other cyprinid spermatozoa, which their mitochondria have a conventional aspect and never fuse to form a mitochondrial derivative. In term of sperm evolution, the fused mitochondria are regarded as the apomorphic character in comparison with the separate mitochondria. The single mitochondrion is not found in cyprinid spermatozoon except for Rodeus and Pungtungia.