• Journal of Embryo Transfer
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• v.28 no.1
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• pp.31-39
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• 2013

Alpha-tocopherol as an antioxidant acts in preservation of chilled semen by preserving cell membrane damage from lipid peroxidation. Optimum concentrations of ${\alpha}$-tocopherol in egg yolk-citrate (EYC) extender need to be studied in crossbred bull's semen. Different concentrations of ${\alpha}$-tocopherol viz. 0, 1, 2, 4 and 6mg per ml of extender were used. Semen was collected once a week from four bulls used to regular collection, aged 4 to 7 years, weighing 320 to 450 kg, and with body condition score 4 to 4.5 and scrotal circumference 23 to 32 cm. Semen was evaluated routinely and sperm morphology was viewed under light microscope at ${\times}1,000$ magnification after fixing with buffered formal saline. Over 90% had normal head, acrosome, mid-piece and tail. Semen was diluted with egg-yolk-citrate extender to produce $15{\times}10^6$ spermatozoa/ml and 0, 1, 2, 4 and 6 mg/ml ${\alpha}$-tocopherol were added. The semens amples were kept at $8^{\circ}C$. Sperm motility and viability were examined daily up to 5 days under light microscopy at ${\times}200$ magnification. Sperm viability was acceptable (${\geq}40%$) up to the $4^{th}$ day with all concentrations of ${\alpha}$-tocopherol and up to the $5^{th}$ day with 2 mg/ml ${\alpha}$-tocopherol. Sperm motility was acceptable (${\geq}40%$) up to the $3^{rd}$ day irrespective of ${\alpha}$-tocopherol concentration, and up to the $4^{th}$ day with 2 mg/ml ${\alpha}$-tocopherol. It is suggested that the lifespan of chilled semen may be extended up to 4 days by adding 2mg/ml ${\alpha}$-tocopherol.

• Journal of Embryo Transfer
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• v.16 no.2
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• pp.127-131
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• 2001

The Jeju horse has been raised for centuries in Jeju island. Recently, as the number of this indigenous horse has been dropped dramatically, this breed became Natural Monument #347 to conserve and multiply this endangered breed. To provide the basic information for AI, sexual activity and semen characteristics in Jeju horse were investigated. Jeju horse semen was collected using Missouri style artificial vagina from fertile stallion.\\`she number of mount per ejaculation was 2..3$\pm$1.8, and the ejaculation time was 27.0$\pm$12.5 seconds. The total volume and gel-free volume of semen was 47.8$\pm$26.7 ml and 42.7$\pm$27.4ml, respectively, and the concentration of sperm and the total number of spermatozoa per ejaculation was 200.7$\pm$112.9$\times$10$^{6}$ ml and 7.6$\pm$3.9$\times$10$^{9}$ ml, respectively. The percentage of motile sperm and the number of live spermatozoa per ejaculation was 75.0$\pm$18.2% and 6.1$\pm$3.4$\times$10$^{9}$ ml, respectively, and the pH of gel-free semen was 7.3$\pm$0.2. The total percentage of abnormal sperm was 31.5%, and the percentage of sperm with abnormal head, midpiece and tail was 9.5$\pm$11.7%, 7.0$\pm$4.0% and 15.0$\pm$15.0%, respectively.

• Applied Microscopy
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• v.39 no.3
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• pp.219-226
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• 2009

The ultrastructure of spermatogenesis and sperm in Cichlasoma managuensis belonging to Cichlidae was investigated by light and electron microscopes. The testis of C. managuensis contained numerous testicular cysts, and spermatogenesis was synchronized in these testicular cysts. In the case of spermatogonia, the nucleus was comparatively large ellipsoidal, and mitochondria showed a marked development. The size of primary spermatocyte was smaller than that of spermatogonia, and that of secondary spermatocyte was smaller than that of primary spermatocyte. The chromatin of spermatocyte was highly condensed according to their development. The nucleus with electron-dense was round shape. In spermiogenesis, flagella started to be formed and chromatin was more condensed. The mitochondria were rearranged in a middle piece. The sperm was formed by loss of cytoplasm. The head of mature sperm was a spherical shape and had not acrosome. The microtubules of flagella were arranged 9+2 structure. Also, the tail of sperm have lateral fins.

• Applied Microscopy
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• v.31 no.2
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• pp.129-141
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• 2001

To investigate the spermiogenesis of the Saghalien Pygmy shrew (Sorex minutus gracillimus), the testis obtained from mature male shrew was studied by electron microscopy, and the following results obtained based on the morphological characteristics of cell differentiation of the seminiferous epithelium in the testis. According to the fine structural differentiation, spermiogenesis of S. minutus gracillimks was divided into Golgi, cap, acrosome, maturation and spermiation phases. Beside, the Golgi and cap phases were subdivided into three steps of early, middle and late phase respectively, and acrosome phase into two steps of early and late phase , and maturation and spermiation phases has only one step respectively. Thus, the spermiogenesis of S. minutus gracillimus was divided into a total of ten steps. The chromatin granules begin to be condensed in the acrosome phase, and a perfect nucleus of sperm was formed at the spermiation phase. Mancette were appeared from the late acrosome phase to the maturation phase. The formation of sperm tail began to develop in the late Golgi phase, and completed at the spermiation phase. Multivesicular bodies were appeared from the Golgi phase to the maturation phase, recognized with pale, pale and moderate, and dense at Golgi, cap and acrosomal and matulation phases respectively.

• The Korean Journal of Malacology
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• v.22 no.1 s.35
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• pp.39-49
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• 2006

Ultrastructure of germ cell differentiation during supermatogenesis and the reproductive cycle in male Patinopecten yessoensis was studied by histological and cytological observations. The gonadosomatic index (GSI) in males rapidly increased and reached a maximum in April when seawater temperature gradually increased. Then the GSI gradually decreased from May through July when spawning occurred. Accordingly, monthly changes in the GSI in males coincided with testicular maturation and spawning periods. The sperm morphology of P. yessoensis belongs to the primitive type and showed general characteristics of external fertilization species. The head of the spermatozoon is approximately $3.50{\mu}m$ in length: the sperm nucleus and acrosome are approximately $2.90{\mu}m\;and\;0.60{\mu}m$ in length, respectively. The nuclear type of the spermatozoon is vase in shape, and the acrosome is cone type. The axoneme of the tail flagellum consists of nine pairs of microtubules at the periphery and a pair of central microtubules in the center The satellite body (which is formed by the centriole) and four mitochondria appear in the middle piece of the spermatozoon. The spawning period was from April through July and the main spawning occurred from May to June when seawater temperatures gradually increased. The reproductive cycle of this species can be classified into five successive stages; early active stage (September to November), late active stage (October to March), ripe stage (February to August), spawning stage (April to July), and spent/inactive stage (July to November).

• Journal of Veterinary Clinics
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• v.21 no.4
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• pp.329-335
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• 2004

Dog spermatozoa were recovered from the caudae epididymides of 23 domestic dogs which were 11 pure breed and 12 mix-breed dogs ranging in age from 0.6 to 3 years. The experimental designs were as follows: 1) the effect of chilling to 0. 10 or 37$^{\circ}C$. 2) the kinetics of chilling injury at 0 or 4$^{\circ}C$, and 3) the effect of sugars at $0^{\circ}C$. Viable spermatozoa were recovered by percoll gradient separation and adjusted to 5${\times}$10$^{7}$ spermatozoa/ml. In experiment 1, spermatozoa were diluted with 0.33 M glucose supplemented with 3% BSA (G-BSA) at 1:2 dilution. Spermatozoa were loaded into straws and exposed at 0, 10 or 37$^{\circ}C$ for 30 min. In experiment 2, spermatozoa were prepared as the experiment 1 and exposed for 0.5, 5, 15, or 30 min at 0 or 4$^{\circ}C$. In experiment 3, spermatozoa were diluted with different sugars (0.33 M galactose, glucose, fructose, mannitol, lactose, sucrose, raffinose) and cooled to $0^{\circ}C$ for 30 min. Sperm membrane integrity, motility and acrosome integrity were assayed after rewarming at 37$^{\circ}C$ for 5 min. Sperm motility and membrane integrity abruptly decreased with decreasing temperature but acrosome integrity gradually decreased (P<0.05). Sperm motility was more sensitive to cold shock than membrane integrity and acrosome integrity. Spermatozoa cooled to $0^{\circ}C$ were more damaged than those at 4$^{\circ}C$. Sperm motility was not different among exposed times at both. 0 and 4$^{\circ}C$. However, membrane integrity of spermatozoa exposed for 30 min at both 0 and 4$^{\circ}C$ was significantly lower (P<0.05). Spermatozoa diluted in 0.33 M fructose or galactose showed lower motility and higher morphological abnormality with coiled tail at $0^{\circ}C$. These sperm characteristics were strongly related. These results indicate that dog epididymal spermatozoa are relatively sensitive to rapid cooling and higher morphological abnormality at $0^{\circ}C$ was shown in spermatozoa diluted in fructose and galactose.

• Asian-Australasian Journal of Animal Sciences
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• v.24 no.10
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• pp.1348-1357
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• 2011

The objective of this study was to compare the effect of Butylated Hydroxy Toluene (BHT), Pentoxifylline (PTX) and ${\alpha}$-tocopherol (Vit E) on semen quality parameters of Karan Fries bulls. The fortification of extender by various semen additives improves motility as well as fertility of spermatozoa. Split samples of 24 ejaculates of four Karan Fries bulls were extended in extender with or without various additives such as BHT, PTX and Vit E, and performance was evaluated at an interval of 0, 24, 48 and 72 h at refrigerated temperature (4-$7^{\circ}C$). Results of the present study revealed that addition of BHT, PTX and Vit E in extender improved sperm cell function, such as motility, viability, HOST, and acrosome integrity, as compared to the control during liquid storage up to 48 h of preservation at refrigerated temperature. There was no significant (p<0.05) difference between any of the additives up to 48 h of preservation. Overall, the results showed a significant (p<0.05) deterioration in motility after each storage interval. The results showed a significant deterioration in the acrosome integrity and plasma membrane integrity up to 48 h; subsequently, there was not much degradation of both the semen quality parameters. There was a significant increase in spermatozoal tail and total abnormality after each storage interval at refrigerator temperature (4 to $7^{\circ}C$); however, the head and mid-piece abnormalities were almost unaffected. Tail and total abnormality were least in extender fortified with BHT, PTX and Vit E at different hours of incubation as compared to the control. The addition of 1.5 mM BHT, 3.6 mM PTX and 1 mg/ml Vit E in the semen extender has more beneficial effect in terms of semen quality and preservability of spermatozoa.

• The Korean Journal of Malacology
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• v.21 no.2 s.34
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• pp.95-105
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• 2005

Gonadosomatic index, reproductive cycle, spermatogenesis and first sexual maturity of Chlamys farreri were investigated by cytological and histological observations, from January 1998 to December 1999. The gonadosomatic index (GSI) rapidly increased in April and reached a maximum in May when seawater temperature rapidly increase. Then the GSI gradually decreased from June to August when spawning occur. Accordingly, monthly changes in the GSI in males coincide with the reproductive cycle. The spermatozoon of Chlamys farreri is the primitive type found in external fertilization species. The head of the spermatozoon is approximately $2.75{\mu}m$ in length including the acrosome measuring about $0.50{\mu}m$ in length, and its tail was approximately $20{\mu}m$, the axoneme of the tail flagellum consists of nine pairs of microtubules at the periphery and a pair at the center. Five spherical mitochondria around the centriole (the satellite body) appear in the middle piece of the sperm. The spawning period was from June to August and the main spawning occurs from July to August when seawater temperatures are greater than $20^{\circ}C$ The reproductive cycle of this species can be categorized into five successive stages; early active stage (January to March), late active stage (March to April), ripe stage (April to August), partially spawned stage (June to August), and spent/inactive stage (August to January). Over 50% of male scallops attained first sexual maturity between 50.0 and 60.0 mm in shell height, and 100% of those over 60.0 mm in shell height achieved maturity. Accordingly, we assume that male individuals begin reproduction at three years of age.

• Development and Reproduction
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• v.20 no.1
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• pp.11-22
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• 2016

The ultrastructures of germ cells and the functions of Leydig cells and Sertoli cells during spermatogenesis in male Kareius bicoloratus (Pleuronectidae) were investigated by electron microscope observation. Each of the well-developed Leydig cells during active maturation division and before spermiation contained an ovoid vesicular nucleus, a number of smooth endoplasmic reticula, well-developed tubular or vesicular mitochondrial cristae, and several lipid droplets in the cytoplasm. It is assumed that Leydig cells are typical steroidogenic cells showing cytological characteristics associated with male steroidogenesis. No cyclic structural changes in the Leydig cells were observed through the year. However, although no clear evidence of steroidogenesis or of any transfer of nutrients from the Sertoli cells to spermatogenic cells was observed, cyclic structural changes in the Sertoli cells were observed over the year. During the period of undischarged germ cell degeneration after spermiation, the Sertoli cells evidenced a lysosomal system associated with phagocytic function in the seminiferous lobules. In this study, the Sertoli cells function in phagocytosis and the resorption of products originating from degenerating spermatids and spermatozoa after spermiation. The spermatozoon lacks an acrosome, as have been shown in all teleost fish spermatozoa. The flagellum or sperm tail of this species evidences the typical 9+2 array of microtubules.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.36 no.5
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• pp.480-485
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• 2003

Morphology of the spermatozoa from the testes of the catfish (Pseudobagrus brevicorpus) was studied by transmission and scanning electron microscopy. The spermatozoa of P. brevicorpus are approximately $82.25\pm0.06\;{\mu}m$ in length and relatively simple cells composed of a spherical head, a short midpiece and a tail as in most teleost fish, The nucleus measuring about $2.00\pm0.02\;{\mu}m$ in length is depressed with a deep nuclear fossa of about $1.05\pm0.03\;{\mu}m$ in length three fifths of the nuclear length. The nuclear fossa contains the proximal and distal centrioles. The two centrioles are oriented approximately $150^{\circ}$ to each other. The mitochondria are arranged in two layers and their number is 12 or more. They are separated from the axoneme by the cytoplasmic canal. The axoneme is the 9+2 microtubular pattern and has inner but no outer dynein arms as in other bagrids. The axonemal fins were the closed to axonemal doublet 3 and 8. The axonemal fins and lost outer dynein arm are shared in Bagridae and the deep nuclear fossa is shared in Siluriformes. The axonemal fins observed in Bagridae and Amblycipitidae of Siluriformes might be the apomorphic character in Ostariophysi.